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Annals 
of the 
Missouri 
Botanical 


Garden 
à m VG 


umber 


Annals of the 
Missouri Botanical Garden 


Volume 87, Number 1 
Winter 2000 


The Annals, published quarterly, contains papers, primarily in systematic botany, 
contributed from the Missouri Botanical Garden, St. Louis. Papers originating out- 
side the Garden will also be accepted. All manuscripts are reviewed by qualified, 
independent reviewers. Authors should write the Managing Editor for information 
concerning arrangements for publishing in the ANNALS. Instructions to Authors are 
printed in the back of the last issue of each volume and are also available online at 


www.mobot.org (through MBG Press). 


Editorial Committee 
Victoria C. Hollowell 

itor, 
Missouri Botanical Garden 
Amy Scheuler McPherson 
Managing Editor, | 
Missouri Botanical Garden 
Diana Gunter 
Associate Editor, 
Missouri Botanical Garden 
Vicki Couture 
Senior Secretary 
Barbara Mack 
Administrative Assistant 
_ Ihsan A. Al-Shehbaz 
Missouri Botanical Garden 


Gerrit Davidse 
Missouri Botanical Garden 


Roy E. Gereau 

Missouri Botanical Garden 
Peter Goldblatt 

Missouri Botanical Garden 
Gordon McPherson 
Missouri Botanical Garden 
P. Mick Richardson 


Missouri Botanical Garden 


Henk van der Werff 
Missouri Botanical Garden 


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http://www.mobot.org 


© Missouri Botanical Garden 2000 


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Volume 87 
Number 1 
2000 


Annals 
of the 
Missouri 
Botanical 


Garden 


NZ 


OUR UNKNOWN PLANET: 
RECENT DISCOVERIES AND 
THE FUTURE. 
INTRODUCTION! 


P. Mick Richardson? 


Recent years have seen many new discoveries in 
the plant, animal, and other kingdoms. Can we es- 
timate how many more organisms are out there to 
be discovered? An international group of experts has 
been invited to St. Louis to give their thoughts and 
predictions about this intriguing area of biology. The 
symposium will consist of an exciting set of presen- 
tations, ranging from flowering plants in the U.S., 
Australia, and the tropics, to freshwater fishes, mam- 
mals, and last, but not least in every sense, extre- 
mophiles and other bacteria.—F lyer advertising the 
45" Annual Systematics Symposium. 

In February 1998 I was in London, and purely 
by chance I was able to attend a talk at the Linnean 
Society on coelocanths, expertly delivered by Pete 
Forey. Later that year my wife and I were impressed 
by a coelocanth specimen in a delightful marine 
in Tulear, a specimen caught lo- 
cally off the southwest coast of Madagascar and 
thought to have been swept southwards from the 
populations around the Comoro Islands. However, 
coelocanths have recently been found in Indonesia, 


science museum 


and one wonders if there are more of them around 
the world to be discovered. The finding of this sec- 
ond species of coelocanth was headline news to bi- 
ologists, as was the discovery of the Wollemi Pine 
and several new large mammals, discussed in the 
following pages. However, the regular and contin- 
ued reporting of new species usually attracts less 
attention. For example, one of my former students 
recently published five new species in the brome- 
liad genus Cryptanthus (Ramirez, 1998). To my 
mind, such a publication may actually be more im- 
portant than the discoveries which make headline 
news because the new species of Cryptanthus are 
the result of full and detailed revisions of the genus 
based on field, herbarium, and laboratory studies. 
The readers can judge for themselves in the follow- 
ing papers that comprise the published proceedings 
of the symposium. 

1998 Annual Systematics Symposium dif- 
fered from the usual format in that there were ten 
speakers rather than the usual seven. This allowed 
for a greater diversity in composition of the program 


! This and the eight articles that follow it are the proceedings of the 45th Annual Systematics Symposium of the 
T 


Missouri Botanical Garden, Our Unknown Planet: 
October, 1998, at the Missouri Botanical Garden in St. Lo 


Recent оте and the Future. 


he symposium was held 9-10 
s, Missouri, U.S.A 


е symposium was supported in part by the National p d Foundation under grant number DEB-9420140. I 


thank Peter Raven for helping to select a fine diversity of speakers, Kathy Hurlbert and her e 


rt staff for wonderful 


help in organizing and administrating the symposium, Barbara Alongi for her fine illustration used for the cover of the 


sy mposium brochure, : 


? Missouri Botanical Garden, P.O. Box 299, St. 


and the symposium speakers for being such a pleasant. group of scientists. 
Louis, Missouri 63166, U.S.A. 


ANN. Missouni Bor. GARD. 87: 1-2. 2000. 


Annals of the 
Missouri Botanical Garden 


and allowed a wider variety of organisms to be cov- 
ered in some detail. The morning session began 
with Michael Madigan’s talk on prokaryotic organ- 
isms, a group that constitutes the bulk of evolu- 
tionary diversity on Earth and which is of increas- 
ing interest for use in biotechnology and related 
areas. Think about where systematics would be 
without the PCR methodology so necessary for cur- 
rent molecular systematics techniques. Literally, 
the book on molecular systematics of plants had to 
be rewritten within six years (Soltis et al., 1992, 
1998). Microbes were followed by Richard Brusca’s 
fascinating talk on arthropod diversification, and 
this made me think of eating deliciously tasty large 
crabs caught in the River Jurua in Acre, Brazil. 
Next was Ebbe Nielsen’s discourse on insects, un- 
fortunately not included in the published proceed- 
ings. Our current knowledge of freshwater fishes 
was the subject of John Lundberg and his col- 
leagues. If South American fishes are so incom- 
pletely known, I wonder if the diversity of fishes 
we ate alongside the aforementioned crabs may 
have been species new to science. Fortunately, they 
were photographed before they went into the frying 
pan, leaving some clues at least to their existence. 
Last in the animal line of talks was John 
Kinnon, who told us about new ungulates being dis- 
covered in Vietnam and his predictions about 
where future finds will likely be made. The morning 
session ended with some wonderful video footage 
and slides from Lynn Margulis, not published here, 
but see her books Five Kingdoms: An Illustrated 
Guide to the Phyla of Life on Earth (Margulis & 
Schwartz, 1998) and Symbiotic Planet: A New Look 
at Evolution (Margulis, 1998). While not everyone 
will agree with Margulis's concept of monophyly, 
there is no denying that she has a a very in- 
teresting viewpoint to overall ни of biodi- 
versity, and at the same time she makes an urgent 
appeal for all biologists and paleontologists to in- 


ac- 


tegrate their analyses and discuss criteria for es- 
tablishment of higher taxa 

he plant talks came after lunch. lain Prance 
informed the audience (and now the readers) that 
the number of angiosperms is currently underesti- 
mated, and he confidently predicted e there are 
in fact between 300,000 and 320, He used 
specific examples of the discovery of me in Mad- 
agascar and other areas, as well as detailed studies 
of all genera in areas in Brazil and Brunei, to de- 
velop his case for an intensification of the rate of 
collection to confirm his predictions before it is too 
late. Barbara Briggs's talk on botanical discoveries 
in Australia contrasted the media attention given to 
the discovery of a new genus of conifers compared 
to the uncharismatic discovery of 61 new species 
in the Restionaceae and allied families. Finally, 
Barbara Ertter made the surprising announcement 
that the rate of discovery of new plants in the Unit- 
ed States and Canada has been constant for the past 
century and shows no evidence of tapering off. 

Mike Donoghue gave a very entertaining and 
stimulating after-dinner talk, emphasizing that the 
current age of discovery may be different from ear- 
lier ones, but it is both richer and more illuminat- 
ing. It is the duty of all systematists to capture the 
imagination of other scientists and, even more im- 
portantly, the public at large. 


Literature Cited 
Margulis, L. 1998. паши |“ 
lution. rg Books, 
& V. Schwartz. " 1998, Fi ive Kingdoms: An Il- 
lustrated Guide to the Phyla of Life on Earth. Freeman, 

San Francisco. 
Ramirez, I. M. 1998. Five new species of wit scarce 
(Bromeliaceae) and some nomenclatural novelties. Har- 
2: 


vard T | 3: 215-224. 


A New Look at Evo- 


Soltis . E. Soltis & J. J. Doyle (Editors). s 
Noc n matics of Plants. Chapman & Hall, N 
Ene E., P. S. Soltis & J. J. 1998. 


Doyle (Editors). 
"dene Systematics of Plants П: DNA Sequencing. 
Kluwer, Boston 


EXTREMOPHILIC BACTERIA 
AND MICROBIAL DIVERSITY! 


Michael T. Madigan? 


ABSTRACT 


rokaryotic microorganisms inhabit *extreme environments"—habitats in which some chemical or physical 
variable(s) при significantly from that found in habitats that support plant and animal life. Great strides have been 


made in t years 


have "лије рај, metabolic properties and os evolutionary historie 
е ге gam s, as all c 
all known life forms. As our knowledge of bacterial diversity 


rs in the isolation and characterization of extremophilic pm and many of them turn out to 


Prokaryotes that grow at very high tem- 
ellular nic din need to be made heat M x and 


improves, primarily from the introduction шаша: tools for assessing bacterial phylogeny and diversity and from 
new nces in isolation and laboratory culture, it is becoming clear that the bulk of evolutionary diversity on Earth 
does not reside in plants and animals, but instead in the invisible prokaryotic world r great interest in 
mining the diverse genetic resourc h's smallest cells for use in biotechnology and related areas 


words: extremophilic bacteria, evolutionary history, microbial diversity, prokaryotes. 


Since the days almost 100 years ago when Robert 
Koch and his associates isolated the first pure cul- 
tures of bacteria, microbiologists worldwide have 
been isolating laboratory cultures of literally thou- 

ands of different bacteria. These include, of course, 
most of the causative agents of infectious diseases, 
but more important from the standpoint of the web 
of life on Earth, many of the bacteria that carry out 
critical chemical reactions that form the “life sup- 
port” system for plants and animals (Madigan et al., 
2000). Despite the diversity of organisms that are 
already known, it is now clear that microbiologists 
have only seen the tip of iceberg; most microorgan- 
isms that exist in nature, in particular the bacteria, 
have not yet been obtained in laboratory culture! 
Indeed, with the help of new molecular tools micro- 
biologists have explored a variety of microbial hab- 
itats and have detected not only new species of bac- 
teria, but new genera, families, orders, and even 
phyla (Barns et al., 1994; Hugenholtz, et al., 1998). 
Imagine finding a new phylum of plants or animals 
today! The challenge for microbiologists now is to 
isolate these organisms, learn about their basic bi- 
ology, and harness their vast genetic resources for 
the benefit of mankind. 


А NATURAL PICTURE OF THE BACTERIAL WORLD 


Great excitement has pervaded the field of mi- 
crobial diversity in recent years because of the 
new-found ability of microbiologists to experimen- 


tally determine the evolutionary relationships of 


bacteria. This giant leap forward emerged from the 
tools of molecular biology, especially as regards the 
development of rapid gene sequencing methods and 
powerful algorithms for the comparative analysis of 
nucleic acid sequences. But for these advances to 
impact microbial evolution, a gene or genes that 
reflected the evolutionary history of an organism 
had to be identified. Such an evolutionary “Rosetta 
Stone" had long been sought, but not until the ad- 
vent of comparative ribosomal RNA sequencing as 
a rapid and specific means for deducing bacterial 
phylogenies (Woese, 1987) did microbiologists have 
the tool they needed to classify bacteria in a nat- 
ural fashion—the way botanists and zoologists had 
classified their subjects for over a century using 
primarily phenotypic characteristics such as bones 
or leaf arrangements as evolutionary guideposts. 
Two key concepts have emerged from compara- 
tive molecular sequencing of ribosomal RNAs: (1) 
that cells evolved along three major lineages, the 
Bacteria, the Archaea, and the Eukarya, instead of 
just two, the prokaryotes and the eukaryotes (Fig. 
1); and (2) that the evolutionary difference between 
a mouse and an elephant (or between Chlorella and 
Trillium, for the more botanically oriented) pales by 
comparison to the evolutionary distance between 
virtually any two common soil bacteria you might 
want to mention, like Pseudomonas and Bacillus. 
The first of these conclusions, that prokaryotic 
life contains two major evolutionary lineages, is 
slowly but surely becoming mainstream thinking 
among microbiologists, and is even gaining support 


! The d of M. T. Madigan is supported by National Science Foundation grant OPP 980 


? Depa 


nt of Microbiology and Center for Systematic mus Mailcode 6508, Southern m University, Car- 
bondale, Illinois 62901-6508, U.S.A. madigan@micro.siu.e 


ANN. Missouni Bor. GARD. 87: 3-12. 2000. 


Annals of the 


Missouri Botanical Garden 


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adigan 
Extremophilic Bacteria 


from macrobiologists as evidenced by the inclusion 
of this concept in recent biology textbooks (Raven 
& Johnson, 1999; Raven et al., 1999). However, 
the second conclusion, that morphologically quite 
different plants or animals can be extremely closely 
related in a molecular evolutionary sense, has been 
for many a harder pill to swallow. If one steps back 
for a moment and considers that it is not the evo- 
lution of the mouse and the elephant, or the alga 
and the flowering plant, as intact entities, that mo- 
lecular sequencing speaks to, but instead, the evo- 
lutionary history of the cells that make them up, it 
is easier to understand why the bulk of evolutionary 
change has occurred in the prokaryotic world; pro- 
karyotes have existed for some 3.8 billion years, 
while the mouse and the elephant have only very 
recently evolved and diverged. 

Prokaryotes ruled the Earth for at least 2 billion 
years before the modern (organelle-containing) eu- 
karyotic cell appears in the fossil record. And 
metazoans (multi-celled plants and animals) have 
only existed for some half billion years or so. So by 
the time the stage was set for what botanists and 
zoologists consider the “evolutionary diversification 


29 


of plants and animals,” most of cellular evolution 
had already occurred. Diversification of the mouse 
and the elephant, for example, was simply a matter 
of arranging cells in different ways to yield what 
appears to the eye to be highly divergent organisms. 
But in terms of their evolutionary history, the mouse 
and the elephant are virtually identical organisms. 

In contrast to higher organisms, prokaryotes have 
had more evolutionary time to show great genetic 
divergence. However, unlike metazoans, evolution- 
ary change in prokaryotes is not manifest in mor- 
phological variation. For whatever reason(s), bac- 
teria maintained a very small size and changed 
relatively little (compared with metazoans) in mor- 
phology through billions of years of evolutionary 
history. But that is not to say they did not evolve. 
Molecular sequencing tells us that they have in- 
deed evolved but that the product of this evolution- 
ary change is invisible—instead of big changes in 
size or shape, evolutionary change in the prokary- 
otes focused on metabolic diversity and the genetic 
capacities to explore and eventually colonize every 
conceivable environment on Earth, including ex- 
treme environments. Thus we must go to the genes 
of the prokaryotes to see their true phylogenetic 
diversification, and with advances in nucleic acid 
sequencing, this world is now beginning to open up 
(Madigan et al., 2000) 

Using comparative ribosomal RNA sequencing 
microbiologists can now not only construct natural 


relationships of prokaryotes (Fig. 1), but can also 
use phylogenetic information to construct highly 
specific nucleic acid probes as a means of identi- 
fying and tracking specific microorganisms in the 
environment. А natural application of this technol- 
ogy has been to take these tools into various ex- 
treme environments and probe for the diversity of 
microbial life therein. The fallout from these stud- 
ies, which historically followed by many years more 
classical enrichment and isolation approaches, has 
been an awareness that extreme environments are 
not a place for “hangers on,” but instead are hab- 
itats that flourish with microbial life, especially pro- 


ave revealed for our understanding of the physi- 
ochemical limits to life. 


EXTREME ENVIRONMENTS AND EXTREMOPHILES 


Microbiological examination of extreme environ- 
ments has revealed many new prokaryotes. By “ex- 
here, it is meant an environ- 
at humans would consider extreme ог 
uninhabitable: extremes of heat or cold, pH, salin- 
ity, pressure, and even radiation. As previously 
mentioned, extreme environments are inhabited by 
diverse populations of microorganisms, most of 
which have evolved to live only in the presence of 


oe 


the extreme. These organisms are the “extremo- 


Oren, 1999). Several classes of extremophiles are 
recognized in microbiology, and Е odi cultures 
of representatives of each class are know 

1). Organisms in each class are denoted a de- 
scriptive term, usually a word with Greek or Latin 
roots followed by the combining form “phile,” 
Greek for “loving.” Thus there are thermophiles and 
hyperthermophiles (organisms growing at high or 
very high temperatures, respectively), psychrophiles 
(organisms that grow best at low temperatures), 
acidophiles and alkaliphiles (organisms optimally 
adapted to acidic or basic pH values, respectively), 
barophiles (organisms that grow best under pres- 
sure), and halophiles (organisms that require NaCl 
for growth) (Table 1). Instead of trying to be inclu- 
sive here, as literally hundreds of different species 
could be included, the organisms listed in Table 1 
in each of the ex- 
tremophile categories. The column of most interest 
in Table 1 is the one labeled “optimum,” for here 


are the current “record holders” 


6 Annals of the 
Missouri Botanical Garden 
Table 1. Classes and examples of extremophiles*. 
Descriptive 

Extreme term Genus/species Minimum Optimum Maximum 
Temperature 

High Hyperthermophile ^ Pyrolobus fumarii 90°C 106°C 113° 

Low Psychrophile Polaromonas vacuolata 0°C 4°C 12°C 
pH 

Low Acidophile Picrophilus oshimae —0.06 0.7 (60°C): 4 

High Alkaliphile Natronobacterium grego- 8.5 0 (20% NaCl)! 12 

ryt 

Pressure Barophile MT41 (Mariana Trench) 500 atm 700 atm (4?C) 21000 atm 
Salt (NaCl) Halophile Halobacterium salinarum 1596 2596 32% (saturation) 


* [n each category the organism listed is the current "record holder" for requiring a particular extreme condition for 
wth. 


» Strain MT41 does not yet have а formal genus and species name and is also a psychrophile. 
* P. oshimae is also а thermophile, growing optimally at 60°C. 
1 М. gregoryi is also an extreme halophile, growing optimally at 20% NaCl. 


it becomes clear that these organisms are not mere- 
ly tolerating their lot, but that they actually do best 
in their punishing habitats; indeed most actually 
require their extreme condition(s) in order to repro- 
duce at all. 

Extremophiles are of interest to both basic and 
applied biology. In a basic sense, these organisms 
hold many interesting biological secrets, such as 
the biochemical limits to macromolecular stability 
and the genetic instructions for constructing mac- 
romolecules stable to one or another extreme (Ma- 

Oren, 1999). But in an applied sense, 
these organisms have yielded an amazing array of 
enzymes capable of catalyzing specific biochemical 
reactions under extreme conditions (Adams & Kel- 
ly, 1995). Such enzymes have served as grist for 
industry in applications as diverse as laundry de- 
tergent additives aree ne and the ge- 
netic identification of с s (Taq DNA poly- 

ase and its use in [pm оа сһаіп 
reaction, РС 

Another important realization that has emerged 
from the study of extremophiles is that some of 
these organisms form the cradle of life itself. Many 
extremophiles, in particular the hyperthermophiles, 
lie close to the “universal ancestor” of all extant 
life on Earth (Fig. 1). Thus, an understanding of 
the basic biology of these organisms is an oppor- 
tunity for biologists to “look backward in time” so 
to speak, to a period of early life on Earth. This 
exciting realization has fueled much research on 
these organisms in order to understand the nature 
of primitive life forms, how the first cells “made a 
living” in Earth’s early days, and how early organ- 


isms set the stage for the evolution of modern life 
forms. 


LIFE AT HIGH TEMPERATURE 


Although thermophilic bacteria (organisms with 
growth temperature optima between 45°C and 80°C) 
have been known for over 80 years, hyperthermo- 
philic bacteria—organisms with optima above 
80°C—have only been recognized more recently 


in Brock, 1978), Karl Stetter and co-workers at 
gensburg (Germany) have proceeded to isolate over 
30 genera (> 70 species) of hyperthermophiles. 
Brock was the first to demonstrate, often using sim- 
ple but ingenious field experiments, that bacteria 
were present in boiling hot springs in Yellowstone 
National Park (Brock, 1978). By contrast, Stetter's 
group, whose focus has been on isolation and cul- 
ture, has isolated many of the hyperthermophiles 
known today, including Pyrolobus fumarii, a re- 
markable prokaryote that can grow up to 113°C (Ta- 
ble 1, Fig. 2) (Blóchl et al., 1997). 

ermophilic microorganisms can be isolated 
from virtually any environment that receives inter- 
mittent heat, such as soil, compost, and the like. 
But hyperthermophiles thrive only in very hot and 
constantly hot environments, including hot springs, 
both terrestrial and undersea (hydrothermal vents), 
and active sea mounts, where volcanic lava is emit- 
ted directly onto the sea floor (Stetter, 1999). It is 
also strongly suspected, and some supportive evi- 
dence exists, that hyperthermophiles reside deep 


Volume 87, Number 1 
2000 


Madigan 7 
Extremophilic Bacteria 


Figure 2. ‘Transmission electron micrograph of a cell 
of Pyrolobus Јитаги, the most thermophilic of all known 
TO 


ind can grow at up to 113°C. Even higher temper- 
atures are tolerated but do not support growth. Micrograph 


courtesy of Reinhard Rachel, Universität Regensburg. 


within the earth, living a buried existence and re- 
lying on geothermal heat for their metabolic activ- 
ities and reproduction (Stetter, 19¢ 
The most extreme of known hyperthermophiles, 
those with temperature optima above 100°C, have 
come from submarine hydrothermal vents (Stetter, 
1996, 1999), and examples include P таги 
(Blóchl et al., 1997, and Fig. 2) and the methano- 
gen Methanopyrus kandleri (Kurr et al., 1991). Both 
of these amazing prokaryotes are members of the 
Archaea (Fig. 1) and are chemolithotrophs (organ- 
isms that use inorganic compounds as energy 
sources), using molecular hydrogen, H,, as their 
electron donor (energy source), reducing either 
NO, (P fumarii) or CO, (M. kandleri) as electron 
acceptors to grow by anaerobic respiration (Madi- 
gan et al., 2000; Stetter, 1999). Besides requiring 
substantial heat for growth, these bacteria can sur- 
vive temperatures substantially above their upper 
growth temperature limits, making a conventional 
at 121°C) insufficient 
for sterilizing cultures of either species! 
P. fumarii and M. kandleri originated from 


autoclave regimen (15 min. 


hydrothermal vent chimneys (Blóchl et al., 5 
Stetter, 1999). These аге precipitated iron mineral 
deposits that form as extremely hot water (up to 
400°C) containing various minerals emerges from 
deep-sea hydrothermal vents (note that although 
this water is superheated, it does not boil because 
of the hydrostatic pressure of the water column, 
usually 2000-3000 m, that overlies these vents). 
Although the water that emerges is too hot for life, 


the chimneys, which are often only about 0.5 cm 
thick, show a temperature gradient from about 
300°C inside to 2°C outside. Because prokaryotes 
are so small, microenvironments differing in tem- 
perature exist across the chimney wall leading to 
ideal habitats for various species of heat-loving 
bacteria. 

Using nucleic acid probe technology several 
morphological types of bacteria have been detected 
in hydrothermal vent chimney walls (Harmsen et 
al., 1997), suggesting that these compact thermal 
gradients may contain many different microbial 
populations in addition to those already isolated. 
And for my botanical friends reading this paper, I 
would be remiss if I did not point out that P. fumarii 
and M. kandleri are good examples of primary pro- 
ducers totally divorced from sunlight, a capacity 
widespread in the microbial world. Besides growing 
at almost unbelievably high temperatures, P fu- 
marii and M. kandleri are also autotrophs, capable 
of growing in a simple anaerobic mineral salts me- 
dium supplied with СО, and H,; neither sunlight 
nor a key product of photosynthesis, O,, is required 
for either organism. Indeed, it has been hypothe- 
sized that long before the process of photosynthesis 
evolved, anaerobic H,-based chemolithotrophy was 
the major means by which new organic material 
was synthesized on Earth (Madigan et al., 2 

For an organism to grow at high temperatures, 
especially as high as those of the hyperthermo- 
philes discussed here, all cellular components, in- 
cluding proteins, nucleic acids, and lipids, must be 
heat stable (Adams & Kelly, 1995; Ladenstein & 
Antranikian, 1998; Wiegel & Adams, 1998; van de 
Vossenberg et al., 1998a). The thermostability of 
enzymes from various hyperthermophiles, referred 


een documented, and 
О, 


to as extremozymes, has 
some have been found to remain active up to 1 
(Adams & Kelly, 1995). The structural features that 
dictate thermal stability in proteins are not well 
understood, but a small number of noncovalent fea- 
tures seem characteristic of thermostable proteins. 
These include a highly apolar core, which appar- 
ently makes the inside of the protein “sticky” and 
thus more resistant to unfolding, a small surface- 


tends to remove options for flexibility and thus in- 
troduce rigidity to the molecule, and extensive ionic 
bonding across the protein’s surface that helps the 
compacted protein resist unfolding at high temper- 
ature (Ladenstein & Anthranikian, 1998). In ad- 
dition to these intrinsic stability factors, special 
proteins called chaperonins are synthesized by hy- 
perthermophiles. Chaperonins function to bind heat 


Annals of the 
Missouri Botanical Garden 


denatured proteins and refold them into their active 
form. The thermosome is a type of chaperonin that 
is widespread among hyperthermophiles capable of 
growth above 100°C, like P. fumarii and M. kan- 
dleri (Stetter, 1999). 

everal factors may combine to prevent DNA 
from melting in hyperthermophiles. However, the 
two most important features appear to be the en- 
zyme reverse DNA gyrase, which catalyzes the pos- 
itive supercoiling of closed circular DNA (by con- 
trast, nonhyperthermophiles contain DNA gyrase, 
an enzyme that supercoils DNA in a negative twist- 
ed fashion), and various types of DNA binding pro- 
teins, including histone-like proteins (Madigan & 
Oren, 1999; Pereira & Reeve, 1998). For various 
physicochemical reasons, positively supercoiled 
DNA is more resistant to thermal denaturation than 
is negatively supercoiled DNA. And the fact that 
reverse gyrase seems to be the only protein thus far 
found universally among hyperthermophiles (re- 
gardless of their metabolic pattern) (Madigan & 
Oren, 1999) points to an important role for it in the 
heat stability of DNA. 

Several hyperthermophiles contain DNA binding 
proteins that appear to play a role in maintaining 
DNA in a double-stranded form at high tempera- 
ture. Some of these proteins are structurally related 
to the core histones of eukaryotic cells and function 
to wind and compact the DNA into nucleosome-like 
structures (Pereira & Reeve, 1998). Others have no 
structural relationship to histones but when bound 
to DNA alter its structure in such a way as to sig- 
nificantly raise its melting temperature (Madigan & 
Oren, 1999). It is likely that the combination of 
positive supercoiling of DNA along with proteins 
that prevent DNA melting is a major solution to the 
maintenance and integrity of DNA in hyperther- 
mophiles. 

Heat can also affect membrane stability. As all 
biologists know, in organisms living at moderate 
temperatures cell membranes are constructed along 
the typical “lipid bilayer” model: hydrophobic res- 
idues (fatty acids) inside oppose each other and 
retain an affinity for one another while hydrophilic 
residues (such as glycerol phosphate) lie at the sur- 
face of the environment and the cytoplasm, respec- 
tively, maintaining contact with the aqueous phase. 
If one applies sufficient heat to such a membrane 
architecture the two leaflets of the membrane will 
pull apart, leading to membrane damage and cy- 
toplasmic leakage. To prevent this from occurring 
at very high temperatures, hyperthermophiles have 
evolved a novel membrane structure. Instead of 
forming a membrane as a lipid bilayer, as just dis- 
cussed, some hyperthermophiles chemically bond 


the opposing hydrophobic residues from each layer 
of the membrane together (van de Vossenberg et al., 
1998a). This forms a lipid monolayer instead of a 
bilayer, and prevents the membrane from melting 
at high temperature. Although the precise chemis- 
try of lipid monolayer membranes can vary some- 
what from species to species, they are common 
among hyperthermophiles and are likely an impor- 
tant evolutionary response to life at high tempera- 
ture. 


LIFE AT Low TEMPERATURES 


How about life at the other end of the thermom- 
eter? Cold environments on Earth are actually 
much more common than hot ones. For example, 
the oceans, which make up over one half the 
Earth's surface, maintain an average temperature of 
about 2?C. And vast land masses are intermittently 
cold and in some cases permanently cold, or even 
frozen. However, cold temperatures are no barrier 
to microbial life, as various microorganisms flourish 
in cold environments, even in ice (Horikoshi & 
Grant, 1998; Madigan & Marrs, 1997). Many mi- 
croorganisms have been isolated capable of growth 
at refrigerator temperatures (4—8°C). These are usu- 
ally psychrotolerant, meaning that although they are 
capable of growth in the cold, they grow better at 
warmer temperatures, usually 25-35°С. True psy- 
chrophiles, defined as microorganisms that grow best 

15°C or lower, are usually only present in per- 
manently cold environments like the Arctic, or in 
particular, the Antarctic (Horikoshi & Grant, 1998). 

А variety of microorganisms including algae and 
diatoms have been found in Antarctic sea ice— 
ocean water that remains frozen for much of the 
year. Sea ice is the habitat for one well-character- 
ized bacterium, Polaromonas vacuolata, the genus 
name indicating its affinity for cold temperatures 
). Polaromonas vacuolata grows 


— 


Irgens et al., 
optimally at 4°C and finds temperatures above 12° 
too warm for growth (Table 1). Other psychrophiles 
are known, but because some of them appear to be 
very sensitive to warming, great care must be taken 
in their isolation and culture to prevent killing them 
off at temperatures as low as room temperature. 
An understanding of the biochemistry and mo- 
lecular biology of psychrophilic bacteria is in a 
much earlier stage than that of the hyperthermo- 
philes. From what is known about the biochemistry 
of psychrophiles, it appears that their proteins func- 
tion optimally at low temperatures because they are 
constructed in such a way so as to maximize flex- 
ibility; this is essentially the opposite strategy from 
that of hyperthermophiles (see earlier). Moreover, 


Моште 87, Митбег 1 
2000 


Madigan 
Extremophilic Bacteria 


proteins from psychrophiles are typically more po- 
lar and less hydrophobic than proteins from hy- 
perthermophiles, a fact that undoubtedly also as- 
sists in their relative flexibility. 

Besides keeping their enzymes functional, psy- 
chrophiles have other biological problems to con- 
tend with, transport of nutrients across the mem- 
brane being chief among them. However, just as 
margarine, with its higher content of unsaturated 
fats, can stay softer than butter at cold tempera- 
tures, psychrophiles regulate the chemical compo- 
sition of their membranes, including in particular 
the length and degree of unsaturation of fatty acids, 
to keep them sufficiently fluid to allow for transport 
processes, even at temperatures below freezing 
(Horikoshi & Grant, 1998). Applications of en- 
zymes from psychrophiles include the cold food in- 
dustry, where enzymes that work at refrigerator tem- 
peratures are sometimes desirable, as well as 
producers of cold-water laundry detergents (see 
more on this below). 


LIFE IN BATTERY ACID OR SODA 


Many extremophiles have evolved to grow best 

t extremes of pH: these are the acidophiles and 
b. alkaliphiles (Horikoshi & Grant, 1998). Al- 
though extremely acidic or alkaline (below pH 3 or 
above pH 10) habitats are rare on earth, in such 
environments one can find a variety of microorgan- 
isms thriving in chemistry the equivalent of battery 
acid or soda-lime. Highly acidic environments can 
result naturally from geochemical activities, such 
as from the oxidation of SO, and H,S produced in 
hydrothermal vents and hot springs, and from the 
metabolic activities of certain acidophiles them- 
selves. For example, the iron sulfide-oxidizing bac- 
terium Thiobacillus ferrooxidans can generate acid 
by oxidizing Fe?* to Fe?*, the latter of which pre- 
cipitates out as Fe(OH), (Fe** + ЗЊО — Fe(OH), 
+ 3H*), or by oxidizing H8. to S0,- (НЗ + 20, 
= 502 + H^). Thiobacillus ferrooxidans is par- 
ticularly active in surface coal mining operations 
where exposure to oxygen of pyrite (FeS,) in the 
coal seam triggers acid production from the meta- 
bolic activities of this and related bacteria. Runoff 
from these habitats can often have a pH of less than 
2, fueling conditions for further acidophile activity. 

The most acidophilic of all bacteria known thus 
far is Picrophilus oshimae, whose pH optimum for 
growth is just 0.7 (Schleper et al., 1995) (Table 1). 
Picrophilus oshimae is also a thermophile (temper- 
ature optimum, 60°C) so this organism must be sta- 
ble to both hot and acidic conditions. Cultures of 
P. oshimae were isolated from an extremely acidic 


(< pH 1) solfatara in Italy, and the organism has 
clearly evolved to require these highly acidic con- 
ditions for its very existence. 

Interestingly, however, acid-loving extremo- 
philes, even those as extreme as P. oshimae, cannot 
tolerate great acidity inside their cells, where it 
would destroy such important molecules as DNA. 
They thus survive by keeping the acid out. The 
internal pH of P. oshimae is about pH 5, and it is 
the cytoplasmic membrane of this organism that 
keeps protons from passively entering the cell. 
However, studies of the P. oshimae membrane have 
shown that it can only retain its integrity in acidic 
solutions; above an external pH of about 4 the P. 
oshimae membrane spontaneously disintegrates. 
Major unanswered questions concerning the metab- 
olism of P. oshimae and other extreme acidophiles 
concern how they generate a proton motive force 
during respiration and related issues of bioenerget- 
ics involving membrane-mediated proton translo- 
cation (van de Vossenberg et al., 1998b). 

Various acid-tolerant enzymes from acidophiles, 
primarily ones located on the cell surface or ones 
excreted from the cell into the acidic milieu, have 
been studied and potential industrial applications 
identified. These are primarily as animal-feed sup- 
plements where the enzymes function to break 
down inexpensive grains to more nutritionally ben- 
eficial forms directly in the animal’s stomach. Such 
enzymes have been widely used in the poultry in- 
dustry and have been shown to reduce feed costs 
and the time necessary to get birds to market. 

Extreme alkaliphiles live in soils laden with soda 
(natron) or in soda lakes where the pH can rise to 
as high as 12. Natronobacterium gregoryi (Table 1), 
for example, was isolated from Lake Magadi, a soda 
lake located in the Rift Valley of Africa; N. gregoryi 
grows optimally at a pH of about 10 (Table 1) (Hor- 
ikoshi & Grant, 1998). In the opposite scenario 
from the acidophiles, alkaliphiles have to contend 
with the problems associated with high pH. Above 
a pH of 8 or so, certain biomolecules, notably RNA, 
break down. Consequently, like acidophiles, alka- 
liphiles must maintain their cytoplasm nearer to 
neutrality than their environment. Nevertheless, 
any proteins found in the cell wall or in the mem- 
brane that make contact with the environment must 
be stable to high pH. Indeed, many such enzymes 
have been studied and a number have found in- 
dustrial applications, especially in the laundry de- 
tergent industry. Detergents that are “enzyme en- 
riched” contain proteases and lipases (enzymes that 
degrade proteins or fats, respectively, in clothing 
stains) that function at the high pH of soapy solu- 
tions (Horikoshi & Grant, 1998). In addition, alkali- 


Annals of the 
Missouri Botanical Garden 


active enzymes from thermophiles and psychro- 
philes have been discovered and commercialized to 
better target detergent additives to hot water or cold 
water applications, respectively. 

Besides keeping their cytoplasm near neutrality, 
alkaliphiles have other biological problems to con- 
tend with. For example, consider the problem of 
membrane-mediated bioenergetics—protons ex- 
truded to the external surface of the membrane en- 
ter a sea of hydroxyl ions. Nevertheless, biochem- 
ical studies of this problem have shown that a 
proton motive force is indeed formed by extreme 
alkaliphiles and drives some of the energy-requir- 
ing reactions in the cell, such as motility and trans- 
port. Sometimes in ATP synthesis, an ion gradient 
of Ма“, rather than H+, drives this key bioenergetic 
process in extreme alkaliphiles шалк ры & Grant, 

1998). This is probably not surpri о 
considers that many (but not all) extreme alkali- 
philes are also extreme halophiles (see below), re- 
quiring high salt as well as high pH for metabolism 
and reproduction. 


ing when one 


LIFE IN A BRINE 


Another remarkable group of extremophiles are 
the halophiles—organisms adapted to grow best in 
salty solutions (Oren, 1999; Ventosa et al., 1998). 
And for extreme о like Halobacterium, a 

“salty solution” me e from 25% NaCl 
up to saturation (32% NaCl) (Table 1). Halophilic 
microorganisms abound in hypersaline lakes suc 

the Dead Sea, the Great Salt Lake, and solar salt 
evaporation ponds. Such lakes are often colored red 
by the dense microbial communities of pigmented 
halophiles such as Halobacterium (Javor, 1989). 


, and under- 
ground salt deposits. To date, a very large number 
of halophilic bacteria have been grown in culture 
including members of all domains of life, including 
the Eukarya (Kamekura, 1998). 
chaeal halophiles as exemplified by Halobacterium 
species remain the most halophilic organisms 


owever, the ar- 


own. 

Halophiles are able to live in salty conditions by 
preventing dehydration of their cytoplasm. They do 
this by either producing large amounts of an inter- 
nal organic solute or by concentrating an organic 
or inorganic solute from their environment (Hori- 
koshi & Grant, 1998; Oren, 1999). Th 
patible solutes” is often used to describe organic 
osmolytes, of which there are several types, but not 
all halophiles employ such solutes (Madigan & 
Oren, 1999; Oren, 1999). For example, as its os- 


e term “com- 


molyte, the archaeon Halobacterium (Table 1) con- 
centrates large amounts of potassium (K+, as KCl) 
from its environment. Dissolved KCl in the cyto- 
plasm of Halobacterium cells is present at a con- 
centration equal to or slightly above that of the dis- 
solved NaCl outside, and in this way cells maintain 
the tendency for water to enter and thereby prevent 
dehydration. As would be expected from such a 
salty cytoplasm, enzymes that function inside of 
cells of Halobacterium have evolved to require this 
large dose of K* for catalytic activity. By contrast, 
membrane or cell wall-positioned proteins in Hal- 
obacterium require Ма’ and are typically stable 
only in the presence of high Ма’ (Madigan & Oren, 
999) 


= 


Extreme halophiles are sources of а variety of 
biomolecules that can function under salty condi- 
tions. Applications of salt-active enzymes include 
those that can break down viscous materials pres- 
ent in oil wells (oil is often found in geographic 
strata that contain salt) as well as enzymes that can 
carry out desirable transformations in highly salted 
foods. In addition, some halophiles that produce 
organic compatible solutes have been commercial- 
ized for the production of these solutes as skin care 


supplements (Madigan & Oren, 1999). 


OTHER EXTREMOPHILES 


Extremophilic microorganisms adapted to high 
pressure or which show no deleterious effects from 
exposure to high levels of radiation are also known. 
Barophiles are microorganisms that grow best under 
pressure greater than 1 atmosphere. Extreme bar- 
ophiles are the most interesting in this regard as 
they actually require pressure, and in some cases, 
extreme pressure, for growth (Table 1). Strain 

T41, for example, a bacterium isolated from ma- 
rine sediments in the Mariana Trench near the Phil- 
ippines (a depth of greater than 10,000 m), requires 
at least 500 atmospheres of pressure in order to 
grow and grows optimally at 700 atmospheres (and 
at a temperature of 4°C because strain МТАЛ is also 
a psychrophile). Because laboratory culture of ex- 
treme barophiles is rather difficult, comparatively 
little is known about their important biomolecules. 
However, although probably all macromolecules in 
extreme barophiles need to be biochemically tai- 
lored to high pressure to some extent, experiments 
with moderately barophilic bacteria, some of which 
can be grown without pressure, have pointed to nu- 
trient transport proteins in the cytoplasmic mem- 
brane as key cell components requiring structural 
modifications in order to function at high pressure 


(Horikoshi & Grant, 1998). 


Volume 87, Number 1 
2000 


Маадап 
Extremophilic Bacteria 


The bacterium Deinococcus radiodurans is an 
amazingly radiation-resistant microorganism (Mur- 
ray, 1992). This remarkable organism can survive 
30,000 Grays of ionizing radiation, sufficient to lit- 
erally shatter its chromosome into hundreds of 
pieces (by contrast, a human can be killed by ex- 
posure to as little as 5 Grays). A powerful DNA 
repair machinery exists in cells of D. radiodurans 
that is able to piece the shattered chromosome back 
together and yield viable cells. Because of its re- 
markable radiation resistance, Deinococcus has 
been proposed as a cleanup agent for the biore- 
mediation of toxic materials in contaminated soils 
that are also radioactive from the leakage of radio- 
active materials; these conditions exist primarily at 
nuclear weapons production sites. 


EXTREMOPHILES IN THE EVOLUTION OF LIFE 


A focus of research on extremophiles has cen- 
tered on the hyperthermophiles. As discussed ear- 
lier, there is good reason to believe that at least 
some hyperthermophiles have evolved relatively lit- 
tle from their ancestors present on earth over 3.5 
billion years ago (Figs. 1, 2). If true, an understand- 
ing of the biology of hyperthermophiles may yield 
a glimpse of what life was like eons ago. In this 
connection the genomes of several hyperthermo- 
philes have been sequenced (Madigan & Oren, 
1999), and the large number of genes they contain 
that lack counterparts in other organisms suggests 
that their biological secrets have at this point only 
been partially revealed. As if living in boiling water 
isn’t enough, just imagine what other tricks hy- 
perthermophiles might be able to perform! 

As previously mentioned, the excitement in mi- 
crobial diversity these days comes from the fact that 
the evolutionary history of the prokaryotes can now 
be experimentally determined. Microbiologists no 
longer have to propose bacterial phylogenies based 
on speculation or “educated guesses” of what type 
of microbe likely preceded another; the phylogenies 
themselves are etched in the sequences of mole- 
cules, and all one has to do is read them. Moreover, 
the application of molecular phylogenetic methods 
to natural environments (Barns et al., ; Hu- 
genholtz et al., 1998) has given us the exciting 
news that the diversity of the microbial world is 
enormous—indeed it is beyond our wildest expec- 
tations. Thus, in the final analysis bacterial diver- 
sity will likely dwarf that of all of the rest of biology, 
perhaps by several orders of magnitude. But only 
continued and expanded research into the diversity 
of microbial life in all environments, extreme and 


otherwise, will yield the data needed to confirm 
this. 

It may indeed be humbling to many biologists to 
think that prokaryotes dominate living diversity. 
But within the rich genetic resources of the pro- 
karyotes undoubtedly lies more benefit for human- 
kind than we will extract from any other group of 
organisms. Antibiotics, fermentation, and biotech- 
nology are only the beginning. The best is yet to 
come. 


Literature Cited 


Adams, М. W. W. & E M. Kelly. prs Enzymes from 
Chemical and 


isms in 
Engineering News 73: нр 
5 R. Е. Fundyga, М. W. Jeffries & N. К. Расе 
1994. Веза сваеи diversity detected in а © 
lowstone National Park hot spring environment. Proc. 
Natl. Acad. Sci. U.S.A. 91: 1609-1613 
Blóchl, E., R. Rachel, S. Burggraf, D. Hafenbradl, H. W. 


tending the upper temperature "for life to 113?C. Extre- 
mophiles 1: 14—21. 

Brock, T. D. 1978. Thermophilic Microorganisms and Life 
at High Temperatures. Springer, New York. 

Harmsen, H. J. M., D. Prieur & C. Jeanthon. radit Dis- 
tribution of microorganisms in deep-sea hydro 
vent chimneys investigated by whole-cell hybridization 
н enrichment culture of I subpopulati 

pl. Environm. Microbiol. 63: 287 3. 

Ti eg . D. Grant. 1998. Т. NET 
crobial Life in Extreme oe Wiley, New his 

Hugenholtz, P., C. Pitulle, Hershberger & N. 

e. 1998. Novel Leda ed bacterial aes in 

a Yellowstone hot spring. J. Bacteriol. 180: 366—373. 

Irgens, R. L., J. J. Gosink & J. T. Staley. 1996. Polaro- 
monas sacuolata gen. nov. sp. nov., a psychrophilic, ma- 
rine, gas vacuolate pr from Antarctica. Int. J. 
Syst. Bacteriol. 46: 8 

Javor, B. J. 1989. E Environments—Microbi- 
ology and "PARI ie Springer-Verlag, Berlin 

Kamekura, M. 1998. Diversity yi = ае 
bacteria. sang odd UE 

< ана, Fricke, 

er. 1991. 


Ladenstein, R ntranikian. 1998. Proteins from hy- 
pert ermophiles Stability and enzymatic TM close 
to the ing pr of water. Advances Biochem. En- 
gin /Bitechnel. 6 

њу Ei Marrs. 1997. Extremophiles. Sci. 

6: oa 
. Oren. 1999. The rmophilic pos MES 
extremophiles Curr. O pin. оа 

————, J. M. Martinko & J. Park 2000. Brook po 
of Microorganisms 9th ed. ыш Hall, Upper Saddle 
River, New Jerse 

Murray, К. G. Е. 1992. The family Deinococcaceae. Pp. 
3732-3744 in A. Balows, H. G. Trüper, M. Dworkin, 
W. Harder & K-H. Sdhleifes (editors), The Рета, 
2nd ed., А Handbook on the Biology of Bacteria: Eco 


Annals of the 
Missouri Botanical Garden 


physiology, Isolation, Identification, 
Springer-Verlag, New Yor 

Oren, A. (editor). 1999. Mic robiology and Biogeochemistry 
of Hypersaline Environments. CRC Press, Boca Raton, 
Florida. 

Pereira, S. L. & J. N. Reeve. 1998. Histones and nucle- 
osomes in archaea and er A comparative analy- 
sis. Vou dud 141-148 

Raven, P. H B. Johnson. 1999. Biology, 5th ed. 
WCB/MeGraw- нї Dubuque. lowa. 

) rt & S. E. Eichhorn. 1999. Biology of 
Plants, 6th E W. H. СА New York. 

Schleper, С., , А. Сатђасопа, D. Ја- 

ekovic, U. aa H-P. Klenk & W. Zillig. 1995. 
Picrophilus gen. nov., fam. nov., a novel aerobic hetero- 
trophic, thermoacidophilic genus and family composing 
archaea capable of growth around pH 0. J. Bacteriol. 
177: 7050-7059. 

Stetter, K. O. 1996. Hyperthermophilic prokaryotes. 
FEMS Microbiol. Rev. 18: 149-158. 


Applications. 


. 1999. Volcanoes, e venting, and the 
origin of life. /n J. Marti & G. J. Ernest (editors), Vol- 
canoes and the sai Cambridge Univ. Press, 
ии. Сан (in press). 

Ventosa, A., J. J. Nieto & A. Oren. 1998. Biology of mou: 
erately halophilic porte bacteria. Microbiol. & Molec 
Biol. Rev. 62: ae 44. 

Vossenberg, J. L. c van de, A. J. M. Driessen & W. 
N. Konings. 1998а. pm essence of being extremophilic: 

The role of pe ое archael membrane lipids. Extre- 

mophiles 2: 70. 


. Zillig & W. N. Konings. 1998b. Bio- 
свете uni cytoplasmic membrane stability of the 
extremely acidophilic, thermophilic archaeon, Picrophi- 
lus oshimae. Extremophiles 2: 67—74. 

Wiegel, J. & M. W. W. Adams (Editors). 1998. Thermo- 
philes—T he Keys to Molecular Evolution and the Ori 
gin of Life? Taylor and Francis, London. 

Woese, C. R. 1987. Bacterial evolution. Microbiol. Rev. 
51: 221-271 


UNRAVELING THE HISTORY 
OF ARTHROPOD 
BIODIVERSIFICATION! 


Richard C. Brusca? 


ABSTRACT 


Current views of arthropod phylogeny are assessed in light of recent research in morphological and molecular 
phylogenetics, m biology. neurobiology, and po Recent fossil discoveries and molecular clock 


data inform us that 
were already 


pod diversification began in the Pre 
he аан а iose metazoan re im on earth. 


cambrian, and suggest that by the Cambrian the arthropods 
“The c ombination of metamerism and jointed appendages 


(with intrinsic musculature), and the evolutionary potential of homeotic genes, has profoundly affected arthropod evolution 


avors a monophyletic Arthropoda. Accumulating evidence 


and created many Wap aces ‘al homoplasies. | 
supports a hypothesis that in 


ects and modern crustaceans ато 


а phylogenetic sister group, and that they, and perhaps 


also trilobites, chelicerates, p myriapods, all could have evolved out of an ancient crustacean stem line. Two implic ations 


of this hypothesis are that Crus 
Key words: 


stacea comprise a paraphyletic taxon and insects may be viewed as “flying crustaceans.” 
Arthropoda, arthropod evolution, Crustacea, insects, Met 


PREFACE. THE CHALLENGE OF UNRAVELING 
METAZOAN PHYLOGENY 


Despite great progress made in zoology during 
the 20th century, there remain many fundamental, 
unanswered questions concerning metazoan evolu- 
nd relationships of 


e part, this stems from 
covering unambiguous йоген enc from 
ancient lineages. Recent molecular and paleonto- 
logical studies suggest that major splits among the 
Metazoa occurred in the Precambrian, some per- 
haps nearly a billion years ago (Wray et al., 1996; 
Ayala et al., 1998; Seilacher et al., 1998; Li et al., 
1998). In part, it may also be because the field of 
comparative morphology has lost popularity (and 
employment opportunities). And finally, the emerg- 
ing field of molecular phylogenetics is still so new 
that every year sees improvements in the data an- 
alyzed and the phylogenetic inference methods 
used. For example, prior to 1997 most molecular 
analyses were based on small numbers of taxa and 
short sequences of a single gene, usually the in- 
herently problematic 188 rDNA gene. Recently, 
however, new (and larger) molecular data sets have 
been developed based on other conserved nuclear 
genes, mitochondrial gene order, and gene dupli- 
cation data. Because it is unlikely that a single 
gene will recover the full phylogeny of Metazoa, the 
future will no doubt see analyses of multiple gene 
sets. 


Emerging molecular studies have corroborated 
many, and challenged some, paradigms of metazoan 
phylogeny. For example, whereas some molecular 
studies have supported the long-held close rela- 
tionship between annelids and arthropods (Wheeler 
et al., 1993), recent studies have not done so (Lake, 
1990; Halanych et al., 1995; Eernisse, 1997; Agui- 
naldo et al., 1997). Furthermore, the discovery of 
new animal phyla, and thus new fundamental body 
plans, continues to occur. The first edition of Lin- 
naeus's (1735) Systema Naturae listed 14 groups 
that we now recognize as distinct animal phyla. To- 
day, we recognize 34 animal phyla. Three former 
phyla have recently been sunk: Pentastomids are 
now placed within the Arthropoda (allied with the 
Maxillopoda), and vestimentiferans and pogonoph- 
orans are now regarded as annelids (probably high- 
ly modified polychaetes) (McHugh, 1997; Brusca & 
Brusca, in press). 

Most of the large-bodied animal groups were dis- 
covered by the end of the 19th century. We are now 
on a track of discovery of microscopic metazoa, and 
three new animal phyla have been discovered just 
since 1956: Gnathostomulida (1956), Loricifera 
1983), and Cycliophora (1995). There is a corre- 
lation between the discovery of new animal phyla 
and their body sizes: phyla described in the period 
of 1901-1920 have maximum body lengths of 3- 
10 mm; phyla described in the period of 1941— 
1960 have maximum body lengths of just 1 mm; 


P mm 


! This paper е from reviews by Wendy Moore, Lisa Nagy, and an anonymous reviewer. 
SF-PEET m ag B-9521649) to the author. Special thanks go to the inde СТА Peter Raven 


ported in part b by 
for encouraging "bs ы. of this 


This work was sup- 


? Columbia University, Biosphere 2 uda: P.O. Box 689, Oracle, Arizona 85623, U.S.A. 


ANN. MissounRi Bor. 


GARD. 87: 13-25. 2000. 


Annals of the 
Missouri Botanical Garden 


phyla described in the 1980s and 1990s have max- 


imum body lengths of less than 0.5 mm. Most of 


the small-bodied phyla are meiofaunal, although 
cycliophorans live as commensals on the mouth ap- 
pendages of various marine crustaceans (Funch & 
Kristensen, 1997). The discovery of these minute 
animals presents challenges to those of us interest- 
ed in animal phylogeny. They are so small that a 
great deal of their anatomy is reduced or otherwise 
altered. We know almost nothing about their de- 
velopmental biology, and they are so rare that mo- 
lecular biologists have not yet extracted gene se- 
quences from them. I predict that the discovery of 
new microscopic phyla will continue for another 
half-century. 

The challenges of unraveling animal phylogeny 
are not unique to molecular biology, small animals, 
or new phyla. Biology has a long history of skir- 
mishing over phylogenetic issues at all levels. The 
evolutionary history of the Arthropoda has been one 
of the most challenging issues biologists struggled 
with throughout the 20th century. What follows is 
an update (as of mid 1998) on what we know about 
arthropod evolutionary history. 


ARTHROPOD EVOLUTION: BACKGROUND 


There are five clearly distinguished groups of ar- 
thropods: trilobites (extinct since the end of the Pa- 
leozoic; ~ 4000 described species); Chelicerifor- 
mes (horseshoe crabs, eurypterids, arachnids, 
pycnogonids; ~ 75,000 described living species); 
crustaceans (crabs, shrimp, isopods, and their kin; 
~ 50,000 described living species); hexapods (in- 
sects and their kin; 878,000 to 1.5 million de- 
scribed living species); and myriapods (centipedes, 
millipedes, and their kin; ~ 14,000 described liv- 
ing species). And, there are two close allies of the 
arthropods, tardigrades (water bears) and опу- 
chophorans (Peripatus and their kin). The close re- 
lationship between the Tardigrada and the Arthrop- 
oda has never been seriously questioned (Brusca & 
Brusca, 1990), and recent molecular work contin- 
ues to support a sister-group relationship between 
these two phyla (Garey et al., 1996). There are now 
1.02 to 1.64 million described arthropods, known 
from virtually all environments on earth. Estimates 
of undescribed arthropod species range from 3 to 
100 million. The arthropods (Table 1) comprise 
about 85% of all described metazoan species. 

The arthropods also encompass an unparalleled 
range of structural and taxonomic diversity, have a 
rich fossil record, and have become favored ani- 
mals of evolutionary developmental biology. Arthro- 
pods were among the earliest animals to evolve. 


Table 1. 


Fossil record of major arthropod groups. 


Tardigrades: Middle Cambrian to present 
Onychophora: Middle Cambrian to present 
Trilobita: Early Cambrian to Permian 

Xiphosura: Early Ordovician/Silurian to present 
Eurypterida: Early Ordovician through mid-Permian 
Arachnida: Upper Silurian to present 
Pycnogonida: Devonian to present 

Crustacea: Early Cambrian (or Vendian) to present 
Hexapoda: Lower Devonian to present 

Myriapoda: Upper Silurian to present 


Recent work (Waggoner, 1996) suggests that even 
the Ediacaran (Vendian) fauna, of the latest Pre- 
cambrian, included early arthropod taxa, perhaps 
true Crustacea. 

Ever since Darwin, biologists have asked the 
question, “How has the incredibly successful di- 
versification of arthropods come about?” Why are 
there so many arthropods? Is there something “spe- 
cial” about these animals? What is the phyloge- 
netic history of the Arthropoda? Specifically, are 
the arthropods monophyletic and what are the re- 
lationships of the major arthropod groups to one 
another? There have been four great challenges to 
biologists in answering these questions. (1) Until 
the last decade of the 20th century, there had been 
a lack of hypotheses on arthropod evolution based 
on principles of explicit phylogenetic inference. (2) 
We have a very incomplete understanding of ar- 
thropod development, though this is improving 
quickly with the advent of molecular developmental 
biology. (3) There has been a paucity of compre- 
hensive studies based on fossils from the earliest 
ages of arthropod evolution (late Precambrian and 
early Paleozoic). (4) It is apparent that high levels 
of homoplasy exist among the arthropods. In just 
the past 10 years, major discoveries have begun to 
address each of these challenges, as discussed be- 
ow. 

Work by the great comparative biologist Robert 
Snodgrass in the 1930s established a benchmark 
in arthropod biodiversity research. Table 2 shows a 
classification of the arthropods at that time, and it 
is this classification that one still finds in most 
modern biology textbooks. The Snodgrass classifi- 
cation embraces three important hypotheses: 


(1) Arthropods comprise a monophyletic taxon. 

(2) Myriapods and hexapods form a sister group, a 
taxon called Atelocerata (= Tracheata, or Uni- 
ramia of some authors). The Atelocerata have 
been united by several seemingly powerful at- 
tributes: 


Volume 87, Number 1 
2000 


Brusca 
Arthropod Biodiversification 


Table 2 


Classification of the arthropods and their allies sensu Snodgrass (1938). 


Phylum Arthropoda 
Subphylum Trilobita 
Subphylum Chelicerata 
Class Merostomata 
Subclass Xiphosura. Horseshoe crabs 
Subclass | 
Class Arachnida. Land spiders, mites, etc. 
Class Pycnogonida. Sea spiders 
ge ibn Mandibulata 
Class Crustacea. Crabs, shrimps, isopods, etc. 
Class heata (= Atelocerata) 
Subclass He 
Superorder Protura. Proturans 


xapoda 


Superorder Insecta. Insects 
Subclass Myriapoda 
Chilopoda. Centipedes 
Diplopoda. Millipedes 
Symphyla. Symphylans 


Superorder 
Superorder 
Superorder 


Superorder Рапгорода. Pauropodans 


Eurypterida. Eurypterids; extinct Paleozoic arthropods 


(a) A tracheal риу system. 

(b) Uniramous legs. 

(c) Use of Malpighian tubules for excretion. 

(d) Loss of the second head appendages—the 
second antennae (as the name Atelocerata im- 
plies). Vestiges of the anlagen of this appendage 
can be seen during the embryogeny of some 
insects (e.g., Sharov, 1953; Brukmoser, 1965). 

(3) The Crustacea and the Tracheata form a sister 

the a name that Snod- 


grass himself coined. 


group, Mandibulata 


For a brief period of time in the mid-century the 
concept of a polyphyletic Arthropoda, championed 
mainly by S. Manton and D. Anderson, enjoyed 
some popularity (Manton, 1973, 1977; Manton & 
Anderson, 1979; Anderson, 1979), and Anderson 
(1996) still maintains this view. The Mantonian 
view of arthropods placed the myriapods, hexapods, 
and onychophorans in a separate lineage (Manton’s 
phylum “Uniramia”) with an origin apart from the 
rest of the arthropods. However, this idea, based on 
flawed phylogenetic argumentation and an inade- 
quate embryological foundation, did not long sur- 

ive the rigors of scientific testing and modern 
methods of phylogenetic inference (see below). In 
addition to phylogenetic analyses, studies of Perm- 
sae а insects (Kukalová-Peck, 
1991a, b, 1992; Kukalová-Peck & Brauckmann, 
1990) have hoan that early pterygotes probably 
possessed polyramous appendages, further under- 
mining the Manton-Anderson Uniramia hypothesis. 
Additional support for arthropod monophyly has 
come from studies of compound eyes using a mono- 


ian 


clonal antibody raised against a specific glycopro- 
tein (3G6), to crystalline cones, eucones, and other 
elements in a variety of insect and crustacean ret- 
inas (Edwards & Meyer, 1990). 

It was not until the i 1980s that Snodgrass’s 
long-standing view of arthropod relationships began 
to be seriously questioned with: (1) the appearance 
of explicit morphological and molecular phyloge- 
netic analyses, (2) the discovery of the amazing po- 
tential of homeobox genes in arthropod develop- 
ment and evolution, (3) the emergence of 
molecular-based evolutionary developmental biol- 
ogy, and (4) the discovery of exquisite new Cam- 
brian preservations from Sweden, China, and else- 


ere. 


MORPHOLOGICAL PHYLOGENETIC STUDIES OF 
ARTHROPODS 

Morphological phylogenetic studies of the arthro- 
pods are summarized in Table 3. Overall, these 
analyses suggest three important conclusions: 


(1) The arthropods are a monophyletic taxon. 

(2) The relationship of the Crustacea to the insects 
and myriapods is ambiguous; that is, Snod- 
grass's Mandibulata is a taxon of questionable 
validity. 

(3) The monophyly of the Atelocerata (insects + 
myriapods) is also questionable. 


Waggoner (1996) included in his analysis a num- 
ber of arthropod-like fossils belonging to the “Ven- 
dian fauna," from the latest Precambrian (= Edi- 
acara Period) that had generally been regarded as 


16 Annals of the 
Missouri Botanical Garden 
Table 3. Morphological views of monophyly within the arthropods. 
Arthropods Mandibulates Tracheata 

Year Author(s) monophyletic monophyletic monophyletic 
1990 Brusca & Brusca Yes Yes n.a. 
1991 k Yes No n.a. 
1992 Eernisse et al. Yes n.a. n.a. 
1993 Backeljau et al. Yes n.a. n.a. 
1993 Wheeler et al Yes Yes Yes 
1994 Wills et al Yes Yes Yes 
1995 Wills et al Yes No No 
1996 Nielsen et al. Yes Yes Yes 
1996 Waggoner Yes No No 
1997 Emerson & Schram Yes No No 
1998 Strausfelc Yes No No 


“problematica.” He also included 21 Cambrian ar- 
thropods, and various modern taxa. He concluded 
that: (a) the Arthropoda are monophyletic, (b) the 
Ediacaran arthropod-like fossils are, in fact, true 
arthropods, and (c) the anomalocarids (and their 
kin) fall out very close to the base, and are probably 
the most primitive known arthropods. Anomalocar- 
ids were giant predatory arthropods (arguably, true 
Crustacea) that reached a meter in length. They are 
known from both the Precambrian and the Cam- 
brian, and they were probably the m poe 
of that time (Briggs, 1994; Chen et al., 

he most recent phylogenetic us ji = 
pods was based on anatomical features of the сеп- 
tral nervous system (Strausfeld, 1998). Strausfeld 
used 100 conserved neural characters in the brains 
of a variety of segmented invertebrates to recon- 
struct phylogenetic relationships among the arthro- 
pods. His analysis suggested that insects and crus- 
taceans comprise a sister group, that the myriapods 
are a polyphyletic group (1.е., chilopods and dip- 
opods are not sister taxa), and that pycnogonids 
are true chelicerates. The most important neuronal 
synapomorphies of Crustacea—Insecta are elements 
of the optic lobes and mid-brain, particularly fea- 
tures of the midline neuropils and neuropils asso- 
ciated with the compound eyes. This analysis cor- 
roborated earlier neurological descriptive work by 
Strausfeld et al. (1995), which also concluded that 
insects are closer to crustaceans than to any other 
arthropod group. 

All arthropod central nervous systems use the 
same fundamental p plan of construction 
(Whitington et al., ; Thomas et al. 4; 
Strausfeld, 1998). ы. a fundamental distinc 
tion between the early embryonic development of 
the myriapod nervous system and that of insects + 
crustaceans was recognized some time ago. Whi- 
tington et al. (1991) found that in insects and crus- 


taceans longitudinal connectives are pioneered by 
segmental neurons, whereas in the centipede Eth- 
mostigmus rubipres longitudinal connectives are pi- 
oneered from neurons in the brain that send their 
axons posteriorly to set up the parallel connectives. 
This difference between centipede and insect-crus- 
tacean ventral nervous systems is compounded by 
the fact that the pattern of segmental neurons in 
centipedes is quite different from that found in in- 
sects and crustaceans; centipede ganglia receive 
contributions from more widely distributed neu- 
rons, and there are more neurons in the centipede 
ventral cord when segmental axons are laid down. 
Comparisons of early neuronal outgrowth during 
embryonic development of the brain and thoracic 
ganglia also suggest a close affinity Quar crus- 
taceans and insects (Harzsch et al., : 
anos et al., 1995; Whitington et al., T Paulus 
(1979) argued for arthropod monophyly on the basis 
of shared characters in the organization of photo- 
receptors and their satellite cells in compound and 
single-lens eyes. He further noted that insect and 
crustacean ommatidia, with their developmentally 
fixed numbers of cells, share more fine structural 
characters than either do with the chilopod om- 
matidia (which comprise an indeterminate number 
of elements). 


MOLECULAR PHYLOGENETIC STUDIES OF 
ARTHROPODS 


Molecular о studies of the Arthropo- 
e 4. Field et al. (1988) 
sequenced a short У вс of 185 rRNA but used 


da are summarized in 


representatives of just 10 phyla (only 4 of which 
were arthropods). Despite its limitations, the Fie 

et al. work was pioneering. It was the first molec- 
ular phylogenetic study to test the monophyly of the 
arthropods, which it supported, and the work ini- 


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2000 
Table 4. Molecular views of monophyly within the arthropods. 
Myriapoda + 
Arthropods Crustacea + Hexapoda 
Year Author(s) monophyletic Hexapoda — (Tracheata) Data 
1988 Field et al. Yes Yes No 18S rRNA 
1989 Patterson Yes Yes No 185 rRNA 
1990 Lake No No Yes 185 rRNA 
1990 Field et al. Yes Yes No 188 rRNA 
1991 Turbeville et al. Yes Yes No 185 rRNA 
1992 Ballard et al. Yes Yes No 125 rRNA 
(mitochondrial) 
1992 Winnepenninckx et al. Yes Yes n.a. 85 rDNA 
1993 Van « ге: Yes Yes f 185 rDNA 
1993 Wheeler et al. Yes Yes No 18S rDNA + ubiquitin 
1995 Winnepenninckx et al. Yes Yes n.a. 18S rDNA 
1995 Friedrich & Tautz Yes Yes No 18S + 28S rDNA 
1996 Garey Yes Yes n.a. 18S rDNA 
1997 Regier 8 Shultz Yes Yes No EF-la + POLII 
1997 Eernisse Yes Yes No 185 rDNA 
1997 Spears & Abele Yes Yes No 185 rDNA 


tiated a stream of follow-up studies, continuing to 

e 188 rRNA sequences, and later the 185 rDNA 
gene itself. Each subsequent study has tended to 
use more taxa and longer nucleotide sequences for 
its data base, but until very recently most also con- 
tinued to rely on the 18S gene. Problems associated 
with the 185 gene, use of short gene sequences, 
and single-gene phylogenetic inferences are well 
known and need not be repeated here. Further- 
more, although there are now over 300 metazoan 
185 sequences available, most published phyloge- 
nies have been based on fewer than 20 sequences 
(Eernisse, 1997). This is despite studies that sug- 
gest a minimum o taxa are needed to ac- 
curately identify the root node of a large clade (Le- 
cointre et al., 1993a, b; Sanderson, 1996; Hillis, 
1996). In spite of methodological and sampling 
problems, recent molecular studies are beginning 
to converge on some similar conclusions. However, 
as Spears and Abele (1997) pointed out, “. . . in the 
crusade for understanding relationships among 
crustacean and other arthropod lineages, the rDNA 
data represent but a relic, and not the Holy Grail 
itself.” 

The most recent 18S sequence data suggest that 
insects share fewer similarities with the myriapods 
than they do with the Crustacea. Spears and Abele 
(1997) analyzed 31 18S sequences, and their re- 
sults suggested that neither crustaceans nor insects 
were monophyletic. When they removed the “prob- 
lematic” long-branched crustacean taxa (Remipe- 
dia, Cephalocarida, Mystacocarida), a myriapod + 
chelicerate clade emerged first, with insects as the 


sister group to a paraphyletic Crustacea. The 
Spears and Abele analysis also strongly supported 
malacostracan monophyly. Eernisse (1997) ana- 
lyzed 103 sequences and concluded that (1) the 
Arthropoda are monophyletic, but only if the tar- 
digrades are included [probably another 185 arti- 
fact], and (2) hexapods are more closely related to 
crustaceans than they are to myriapods. Regier and 
Shultz (1997) made a complete and welcome break 
with the 18S gene, using sequences from two other 
nuclear genes, the elongation factor (EF-la) gene 
and the A polymerase II (POLII) gene. These 
trees were robust and mostly in agreement with the 
18S work, concluding that: (1) Arthropods are 
monophyletic, (2) Crustacea are paraphyletic, and 
3) insects are not the sister group of the myriapods, 
but arose from within the Crustacea. 

Recent work by Boore et al. (1995) examined not 
gene sequences, but the linear arrangement of mi- 
tochondrial genes. This new type of data corrobo- 
rates the gene sequence work and recognizes a mi- 
tochondrial gene arrangement that is unique to the 
crustaceans and insects alone. 

In summary, the majority opinion from the mo- 
lecular research, and the most recent opinions from 
both the morphological and molecular work, rec- 
ognize four key features in arthropod phylogeny: 


~ 


(1) Arthropods are monophyletic. 

(2) Neither the Mandibulata nor the Atelocerata 
are natural groups. 

3) Crustaceans and insects constitute а sister 
group, exclusive of the myriapods. 


~ 


Annals of the 
Missouri Botanical Garden 


(4) Crustacea are likely to constitute a paraphyletic 
taxon 


These last three conclusions are in conflict with 
150 years of morphology-based thinking. Thus, two 
profound implications of these new studies are that 
the morphological attributes linking insects to myr- 
iapods might all be convergences (e.g., uniramous 
legs, tracheal system, Malpighian tubules), and that 
insects are actu ying crustaceans” (in the 
same sense that birds are flying reptiles). 


EMERGING VIEWS FROM DEVELOPMENTAL STUDIES 


The unique combination of segmentation and 
jointed appendages has allowed arthropods to de- 
velop modes of locomotion and feeding, and body 
region specialization, unavailable to other metazoan 
phyla. We now know that the fates of these seg- 
mental units and their appendages are under the 
ultimate orchestration of homeotic genes. These 
genes select the critical developmental pathways to 
be followed by cells during morphogenesis. Ho- 
meobox genes determine such basic body architec- 
ture as the dorso-ventral and the anterior-posterior 
body axes, where body appendages form, and the 
general types of appendages that form (Averof & 
Patel, 1997; Panganiban et al., 1997; Shubin et al., 
1997). Homeobox genes can either suppress limb 
development, or modify it to create alternative ap- 
pendage morphologies. A growing body of evidence 
suggests that these unique genes have probably 
played major roles in the evolution of new body 
plans among arthropods and the Metazoa in general 
(Davidson et al., 1995; Williams & Nagy, 1995; 
Panganiban et al., 1995). 

The degree to which homeobox genes have been 
conserved is remarkable, and most of them proba- 
bly date back at least to the Cambrian. For exam- 
ple, homologues of the Pax-6 gene seem to dictate 
where eyes will develop in all animal phyla. Pax- 
6 is so similar in protostomes (insects) and deu- 
terostomes (mammals) that the genes can be ex- 
perimentally interchanged and still function 
correctly. Homeobox genes modulate the expression 
of dozens of interacting, downstream, developmen- 
tal genes whose products drive morphogenesis. The 
profound evolutionary potential of homeobox genes 
lies in this hierarchical nature. Variation in the out- 
put of these multigene networks can arise at many 
levels, simply by tinkering with the relative timing 
of gene expression—an evolutionary process we 
know as heterochrony. To understand the profound 
potential of homeobox genes to drive evolutionary 
change, consider that within the Drosophila genome 
85-170 different genes might be regulated by the 


product of a single homeobox gene, the Ultrabi- 
thorax (Ubx) gene (Carroll, 1995 
A good example of the evolutionary potential of 
homeobox genes is seen in the abdominal limbs of 
insects. Abdominal limbs (*prolegs") occur on lar- 
vae of various insects in several orders, and they 
are ubiquitous in the order Lepidoptera, i.e., cat- 
erpillars. Abdominal limbs were almost certainly 
present in adult insect ancestors. Hence prolegs 
may have reappeared in such groups as the Lepi- 
doptera through something as simple as the de-re- 
pression of an ancestral limb development program 
(1.е., they represent an atavism). We now know that 
proleg formation is initiated by a change in the reg- 
ulation and expression of the BX-C gene complex 
e., the Bithorax complex, which includes the Hox 
s Ubx, abdA, and abdB) during embryogenesis 
(Carroll, 1995 
Molecular and developmental biology also seem 
to have broken the deadlock on the arguments over 
origins of uniramous and biramous limbs (e.g., Po- 
padic et al., 1996; Panganiban et al., 1995, 1997; 
Shubin et al., 1997; Emerson & Schram, 1997). We 
now know that limb branching is a second-order 
phenomenon, probably orchestrated largely by the 
homeobox gene Distal-less (DII). This single gene 
initiates development of unbranched limbs in in- 
sects and branched limbs in crustaceans. Antibod- 
ies that recognize ОИ proteins show expression at 
the tips of insect limbs and also in biramous crus- 
1995). Branched 


limbs are formed when the gene is expressed ec- 


tacean limbs (Panganiban et al., 


topically in Drosophila (Diaz-Benjumea et al., 
1994). In fact, ОИ occurs in many animal phyla, 
where it is expressed at the tips of ectodermal body 
outgrowths in such different structures as the limbs 
of vertebrates, parapodia and antennae of poly- 
chaete worms, tube feet of echinoderms, siphons of 
tunicates, and appendages of arthropods. Further- 
more, recent work suggests that whether an arthro- 
pod mandible is *whole-limb" (i.e., built of many 
segments) or "gnathobasic" (i.e., built of only the 
basalmost segments) also depends on the expres- 
sion of the gene Distal-less. Thus, Dil is expressed 
in the whole limb (or multisegmented) jaws of myr- 
iapods, but not in the gnathobasic jaws of crusta- 
ceans and insects—still further testimony to the 
probable sister-group relationship of insects and 
Crustacea. 


THE PALEONTOLOGICAL DATA 


Recent work has shown the fossil record of ar- 
thropods dates back to the early Cambrian, or per- 
haps the late Precambrian. And, by the mid-Paleo- 


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Brusca 19 
Arthropod Biodiversification 


Table 5 


Some important Precambrian and Cambrian arthropod Lagerstdtten faunas. 


Age 


Name 


Principal location 


Orsten fauna Upper Cambrian (~ 
Burgess Shale fauna 
Chengjiang fauna 
Ediacaran fauna 


510 MYA) 
Middle Cambrian (~520 MYA) 
Lower Cambrian (~530 MYA) 
560-600 MYA) 


Latest Precambrian (~ 


Southern Sweden 
British Columbia 
Southern China 
Ediacara Hills, Aust. 


zoic, all five arthropod lineages were in existence 
and had already undergone substantial radiation. 
Arthropods are also the first land animals for which 
we have a geological record (Labandeira et al., 
1988; Kukalová-Peck, 1990), and by the Late 51- 
lurian the first terrestrial scorpions and myriapods 
were already present. In fact, both terrestrial and 
marine myriapods have been reported from this pe- 
riod (Almond, 1985; Hahn et al., 1986; Labandeira 
et al., 1988), although molecular data suggest that 
myriapods might have arisen as early as the Cam- 
brian (Friedrich & Tautz, 1995). The first centipede 
fossils occur in the Upper Silurian (— 414 MYA) 
and, along with trigonotarbid arachnids, constitute 
the earliest known land animals (Jeram et al., 
1990). The first millipede fossils occur in Devonian 
deposits (Almond, 1985; Robison, 1990); they are 
similar to the extant genus Craterstigmus (Shear et 
al., 1984) and are contemporaneous with the first 
terrestrial mites, pseudoscorpions, and scorpions 
(Størmer, 1969, 1977; Shear et al., 1987), as well 
as the first hexapods (Greenslade, 1988). The ear- 
liest known fossil hexapods are bristletails and col- 
lembolans from 390-million-year-old Gaspé mud- 
stone (Labandeira et al., 1988). Some good records 
of these early creatures now exist, and the presence 
of these predatory arthropods suggests that complex 
terrestrial ecosystems were in place at least as early 
as the late Silurian. Perhaps the most important 
ancient arthropod fossils are those in which even 
the soft parts of the animal were preserved—the 
so-called ancient Lagerstátten (Table 5). 

These ancient fossils have pushed the age of or- 
igin for the arthropods back to at least 600 MYA, 
and they provide us with critically important data 
on early arthropod anatomy and evolution. These 
extraordinary faunas are now telling us that Crus- 
tacea probably predate the appearance of trilobites 
in the fossil record, running counter to a long-held 
belief that trilobites were the most ancient arthro- 


pods. The recently exploited Chengjiang fauna of 


south China is Lower Cambrian, about 10 million 
years older than the Middle Cambrian Burgess 
Shale fauna (Chen et al., 1994). The Chengjiang 
fauna is very well preserved and includes at least 
100 species of animals, many without hard skele- 


tons, including the first known members of many 
modern groups. However, it is the arthropods that 
dominate this fauna, including trilobites and. bra- 
doriid “crustaceans” (and also tardigrades and on- 
ychophorans). The largest of the Chengjiang ani- 
mals is Anomalocaris, also known from Ediacaran 
and Middle Cambrian deposits (Briggs, 1994). The 
Chengjiang fauna is very similar to that of the Bur- 
gess Shale, and it demonstrates that the arthropods 
were already far advanced by this early date. 

The spectacular recent discovery by Klaus Müll- 
er and Dieter Walossek (Müller, 1983, 1992; Müller 
& Walossek, 1985; Müller et al., 1995; Walossek 
& Müller, 1992, 1997) of microscopic arthropods 
from the Upper Cambrian Orsten deposits of Swe- 
den, has brought to light a rich fauna of minute 
crustaceans, crustacean larvae, and various crus- 
tacean-like arthropods. Among them, for example, 
is Skara, a cephalocarid-, or mystacocarid-like 
crustacean for which both naupliar larvae and 
adults have been recovered (the nauplius larvae are 
only a couple hundred microns long; adults are 
about 1 mm in length) (Müller & Walossek, 1986). 
Skara, and many other Orsten Crustacea, were sure- 
ly meiofaunal animals not unlike modern marine 
meiofaunal crustaceans. 

Fossils from this Cambrian site in Sweden have 
been collected since the days of Linnaeus, who ac- 
tually described the first fossils from this area in 
1757 (trilobites and conodonts). However, a brand- 
new kind of collecting began with Müller and Wal- 
ossek's work in the 1980s. This new Orsten material 
is all microscopic, three-dimensional fossils. The 
Orsten arthropods show little or no signs of decom- 
position. They preserve details less than 1 micro- 
meter in size (e.g., cuticular pores, the bristles on 
setae). Dozens of Orsten microcrustacea have so far 
been described. The recovery of these three-dimen- 
sionally preserved animals and the developmental 
series that have been found (with successive larval, 
juvenile, and adult instars—in animals less than 1 
mm in length) have provided us with information 
on the detailed anatomy of body segments and ap- 
pendages of many ancient stem-arthropods. The Or- 
sten fauna shows that Cambrian Crustacea had all 
the attributes of modern crustaceans, such as com- 


Annals of the 
Missouri Botanical Garden 


pound eyes, a head shield, naupliar larvae (with 
locomotory first antennae), and biramous append- 
ages on the second and third head somites (the sec- 
ond antennae and mandibles). 

Taken together, this recent paleontological work 
corroborates Whitington’s observations long ago 
about the Burgess Shale fauna, that during Cam- 
brian times the non-trilobite arthropods were both 
morphologically more varied and more numerous 
than were the trilobites (despite popular belief). We 
also now know that arthropods have probably been 
the dominant animals in terms of species diversity 
since the Cambrian. Arthropods comprise over one- 
third of all species described from Lower Cambrian 
strata. 

Briggs and Fortey (1989) cladistically analyzed 
23 of the Cambrian arthropod taxa, plus 5 extant 
groups. Their tree placed the Crustacea at the very 
base, as a paraphyletic sequence of taxa, and 
placed the trilobites and chelicerates near the top 
of the tree. The most recent molecular work does 
not conflict with this tree, in viewing the Crustacea 
as a paraphyletic group from which the other major 
arthropod clades emerged. 

THE PENTASTOMIDA 

Pentastomids are obligatory parasites of verte- 
brate respiratory systems. There are about 100 de- 
scribed species, all of which infest various tetra- 
pods, including two cosmopolitan species that 
infest humans. The blood-sucking adults inhabit re- 
spiratory tracts of their hosts, where they anchor 
themselves by means of their hooklike head ар- 
pendages. For years it was believed that pentasto- 
mids were allied with the onychophorans as ver- 
miform, pre-arthropod creatures. However, several 
recent molecular studies (using 185 gene sequenc- 
es) have revealed the pentastomids to be highly 
modified crustaceans (Abele et al., 1989, 1992; 
1996). Corroborative independent 
work over the past few years has come from cla- 


Garey et al., 


distic analyses of sperm and larval morphology, 
nervous system anatomy, and cuticular fine struc- 
ture (Wingstrand, 1972, 1978; Storch, 1984; Storch 
& Jamieson, 1992). Furthermore, Miiller and Wal- 
ossek’s work on the Swedish Orsten fauna proves 
that the pentastomids (and also the tardigrades) had 
appeared at least by the Upper Cambrian, long be- 
fore the land vertebrates had even evolved (Miiller 
& Walossek, 1988; Walossek & Miiller, 1994). So, 
we must ask what the original hosts of these para- 
sitic crustaceans might have been. Walossek, Miill- 
er, and even Stephen Jay Gould have noted that 
Conodont fossils are common in all the Cambrian 


localities that have yielded pentastomids, and thus 
the conodonts (also long a mystery, but now widely 
fish-like vertebrates) 
might have been the original hosts of the pentasto- 


regarded as parts of early 
mids. 


THE ONYCHOPHORA 


As with pentastomids, onychophorans, too, were 
part of the amazing, early-Cambrian, explosive ma- 
rine diversification. They have been found in Bur- 
gess Shale-type faunas at several localities, in Cam- 
brian deposits from China and Siberia, and in the 
Swedish Orsten fauna (Xianguang & Weiguo, 1988; 
Xianguang & Junyuan, 1989; Ramskóld & Hou, 
199 nd, we now know that Conway Morris's 
original reconstruction of Hallucigenia (from the 
Burgess Shale) had the animal turned upside-down. 
Ramskóld and Hou (1991) recently turned Hallu- 
cigenia over and found a second pair of legs, con- 
cluding it was an onychophoran with long dorsal 
spines. And there is now an onychophoran known 
from the Chengjiang deposits of China with side 
plates and spines (Ramskóld & Hou, 1991). Ay- 
sheaia (also from the Burgess Shale) was originally 
described by Walcott as an annelid, but it, too, is 
now regarded as an early marine onychophoran. 


CONCLUSIONS 


Let us now return to our two fundamental ques- 
tions regarding arthropod evolution: Why are there 
so many arthropods, and what is the phylogenetic 
history of the arthropods? As to the first question, 
I propose six over-arching scenarios, each complex 
in its own right 


(1) The numerical superiority of arthropods is not 

a recent event. Recent fossil discoveries, and 
molecular clock data, inform us that arthropod 
diversification began very early in the history 
of the Metazoa, in the Precambrian, and by the 
Cambrian the arthropods were probably already 
the most speciose metazoan phylum on earth. 
Arthropods have been on a powerful phyloge- 
600 MY. They 
have had a great deal of time to PEN and 
with the exception of the trilobites and the eu- 
rypterids, all the major lineages have survived 
and continue to radiate. 


netic trajectory for well over 


S 


Their great size range, especially on the smaller 
end of the scale, adapts arthropods for a great 
variety of ecological niches. The Cambrian Or- 
sten. deposits tell us that a whole fauna of in- 
terstitial/meiofaunal arthropods already existed 
as early as the mid-Cambrian, and this habitat 


Volume 87, Number 1 
2000 


Brusca 21 
Arthropod Biodiversification 


has continued to be rich in adaptive radiation 

and specialized species ever since. Similar 
small-body-size niches are filled in a great 
many specialized environments today. We find 
high diversities of minute arthropods in habitats 
such as marine sediments, coral reefs, among 
the fronds of algae, on mosses and other prim- 
itive plants, and on the bodies of every kind of 
animal imaginable. There are even arthropod 
faunas that live strictly on the gills of other 
crustaceans (mites and small crustaceans). 
Small insects and mites have exploited virtually 
every terrestrial microhabitat available. 

(3) The close relationship and coevolution with 
flowering plants (on land) and algae (in aquatic 

een a powerful force in 


— 


environments) have 
the radiation of the arthropods. It is not just the 
insects that have been on a coevolutionary tra- 
jectory with plants—many crustaceans utilize 
algae as both a living substrate and a food 
source and show strong evidence of coevolu- 


tion. 

(4) The arthropods (insects) were the first flying an- 
imals, and the ability to fly led them into niches 
other invertebrates simply could not penetrate. 

(5) Metamerism (the serially repeated body seg- 
ments and appendages of arthropods) provides 
an enormous amount of easily manipulated 
body plan material upon which evolutionary 
processes can act. Given the great age, sheer 
diversity, and our emerging knowledge of reg- 
ulatory genes in these animals, a high level of 
homoplasy is no longer surprising. 

(6) The potential for major changes in body plans 
due to variations in homeobox genes, and the 
downstream genes they regulate, is just begin- 
ning to be realized, but this potential is clearly 
enormous. There seems little doubt that chang- 
es in homeotic genes over time have profoundly 
affected arthropod evolution. Considering the 
number and position of limbs in arthropods, 
and the flexibility of homeobox and regulatory 
switches, it is little wonder that arthropod an- 


atomical diversity seems so endless. 


As to the second question—what is the phylo- 
genetic history of the Arthropods—it seems the 
plasticity of the arthropod body and homeobox gene 
expression may have produced an even higher level 
of homoplasy than once thought. As a result, some 
traditional morphological classifications are in con- 
flict with molecular classifications. All the evidence 
suggests that the arthropods are monophyletic. 
However, fossil data, recent comparative neuroan- 
atomical research, and molecular data all suggest 


that Crustacea are a paraphyletic group, and that 
the Crustacea and Insecta are very closely related 
to one another, but not to the Myriapoda. In fact, 
the insects appear to have arisen from within a 
crustacean stem line. Further, recent molecular and 
fossil data are beginning to suggest that the trilo- 
bites, chelicerates, insects, myriapods, and recent 
crustaceans all might have emerged from crusta- 
cean stem-line ancestors. This view of a paraphy- 
letic Crustacea spinning off a series of other major 
arthropod lineages might explain why morpholo- 
gists have been unable to come to agreement on the 
sister-group relationships of the major arthropod 
lineages. Resolution of this conflict will come, | 
predict, within the next two decades, with further 
understanding of the genetic regulation of devel- 
opmental processes, examination of new nuclear 
and mitochondrial genes (and use of multiple gene 
data sets in phylogenetic analyses), and as more 
cladistic analyses include fossil species, particu- 
larly the growing series of Chengjiang, Orsten, and 
related arthropods. 


A SPECULATION 


The realization that insects might have arisen out 
of an ancestral crustacean stem line leads to many 
new implications concerning arthropod evolution. 
For example, given this scenario, one could search 
about among the Crustacea for a likely ancestor to 
the insects and in doing so recognize the presence 
of a “fixed” 19-segmented body plan in insects and 
certain crustaceans (or more likely a 20-segmented 
plan in each, Kukalová-Peck, 1991а, b; Scholtz et 
al., 1994; Scholtz, 1995). All insects are fixed on 
this body plan. Of all the crustacean higher taxa, 
this body plan consistently occurs only in the sub- 
class Eumalacostraca—the crabs, shrimps, isopods, 
and their kin. Thus, if the insects did evolve from 
a crustacean ancestor, one might spec ulate that 
they could have evolved from a 
Examining the Eumalacostraca for a piile i insect 
ancestry, there is only one group that is truly ter- 
restrial, has evolved gas-exchange tracheae (grant- 
ed, probably convergently to those of insects), has 
reduced/lost one pair of antennae (antennae one re- 
duced in oniscideans, antennae two reduced 
hexapods), and has strictly uniramous walking legs 
(as do the insects)—the terrestrial isopods (Isopo- 
da: Oniscidea). Could it be that insects are not only 
flying crustaceans, but flying isopods? 

The concept of a Eumalacostraca—Insecta sister- 
group relationship finds strong support in the com- 
parative anatomy of arthropod central nervous sys- 
tems. Development of the compound eye follows 


„пала 


Annals of the 
Missouri Botanical Garden 


similar morphogenetic events in insects and eu- 
malacostracans (Hafner & Tokarski, in press). In 
addition, the optic lobes of pterygote insects and 
eumalacostracans are distinguished by nested reti- 
notopic neuropils, each of which represents the 
whole eye. In these two taxa, these neuropils com- 
prise an anatomically distinct lamina, medulla, and 
lobula complex (Strausfeld, 1996). The presence of 
these structures in pterygote insects and eumala- 
costracans was viewed as a homology indicating a 
sister-group relationship between these taxa 
Osorio and Bacon (1994) and Nilsson and Osorio 
(1997). Further, eumalacostracans that have so far 
been examined also possess a distinctive form of 
neuron, called a bushy T-cell, which was first rec- 
ognized in insects on the basis of its characteristic 
dendritic “tree” situated near the inner face of the 
medulla (Strausfeld, 1976). Bushy T-cells in insects 
and eumalacostracan crustaceans send their axons 
to large tangential dendrites that extend across sub- 
stantial areas of the retinotopic mosaic. In those 
pterygote orders investigated, bushy T-cells com- 
prise part of an evolutionarily conserved subset of 
retinotopic elements that contribute to elementary 
motion detector circuits (Strausfeld & Lee, 1991; 
Douglass & Strausfeld, 1995, 1996). The presence 
of these cell types in eumalacostracan crustaceans 
and pterygote insects implies that either identical 
circuits have independently in the two 
groups, or the circuit for motion detection evolved 
in a common ancestor to insects and crustaceans 
has been maintained basically unchanged through- 
out the history of both groups. That the latter is 
more likely is suggested by the presence of small 
field retinotopic neurons that arise from the inner 
layer of the medulla of the apterygote Thermobia 
and extend into the lateral lobe of the protocere- 
brum (Strausfeld, 1998). 

All crustacean nervous systems so far examined 
possess the architectonic and positional equivalent 
of a fan-shaped body, the neurons of which extend 
laterally into the рое с) lobes, as they do іп 
insects (Strausfeld, 1998). However, except in iso- 


insects. Further, in pterygote insects and isopods 
(but not in decapods or apterygotes) the fan-shaped 
body is supplied by a bridge of neuropil that lies 
posteriorly in the brain and connects the left and 
right protocerebral hemispheres. Strausfeld (1998) 
concluded that, while fan-shaped bodies are syna- 

morphic to insects and crustaceans, the proto- 
cerebral bridge may have evolved independently in 
insects and isopods. 

Many morphological features are in conflict with 


a close malacostracan-insect relationship, includ- 
ing differences in tagmata arrangement and loca- 
tions of the gonopores. The fossil record also does 
not support an isopod + insect sister-group rela- 
tionship. The oldest known isopod fossils are only 
300 million years in age (Phreatoicidea: Hesslerella, 
Carboniferous) (Brusca & Wilson, 1991). However, 
a recent analysis of phreatoicidean phylogeny sug- 
gests the isopods might have had their origin con- 
siderably earlier than this (Wilson & Keable, in 
press), and further examination of this unconven- 
tional idea may be warranted. 


Literature Cited 
Abele, L., W. Kim & B. E. Felgenhauer. 1989. Molecular 


evidence for inclusion of the phylum ооа іп 
the Crustacea. Molec. Biol. Evol. 6: 685—69 
Spears Kim & M. Applegate. ж РҺу- 
т of selected одан and other crustacean 
taxa based on 185 ribosomal ПРИ sequences: А 
РАИ are Acta Zool. 73: : х 
Aguinaldo, А. М. А., J. М. Turbeville, бе 5. intend, M. 
C. Rivera, J. | Garey, R. А. Raff & J. А. Lake. 1997. 
Evidence for a clade of nematodes, arthropods and other 
moulting dep Nature 387: 489—493. 
Almond, J. E. 1985. The Silurian-Devonian fossil record 
of the s Philos. Trans., Ser. B 309: eder 
Anderson, D. T. 1979 Ae fate же and the 
logeny of anhropads: Pp. 59-106 in A. P. Gupta (еф. 
tor), РА Phylogeny. Van Nostrand, New York. 
Atlas of Invert ce Anatomy. Univ. New 
South ы Press, NSW, Aus 
ч М. & М. Н. Patel. 1907. Cee appendage 
volution associated with changes in Hox gene expres- 
Kn ME 388: 682—686. 
A. Rzhetsky & F. J. Ayala. 1998. Origin of 
rm paleon- 
1 


. Winnepenninckx & L. de Bruyn. 1993. 
Cladistic analysis of metazoan relationships: А геар- 
praisal. a 9: 167-181. 

а J. W. O., С. J. Olsen, D. P. Faith, W. A. Odgers, 

. Ro i & P. W. Atkinson. 1992. Evidence from 
Y boron] RNA sequences that onychophorans are 
48. 


phylogeny from mitochondrial DNA rearrangements. 
Nature 376: —165. 

Briggs, D. Е. i н Pes о from the Cambrian 
of China. aues 264: 284. 

A. Fortey. Dd The early radiation and 
relationships of the major arthropod groups. Science 
246: 241-243. 

Brukmoser, P. 1965. Untersuchungen über den Kopfbau 
der Collembole Orchesella villosa. L. Zool. Jahrb. Anat. 
Ontog. 82: 299-364. 

Brusca, R. C. & G. J. Brusca. 1990. Invertebrates. Sinauer 
Associates, Sunderland, Massachusetts. 

& . Invertebrates, 2nd ed. Sinauer Asso- 
ciates, Sundedand, Massachusetts (in press). 

— а G. D. Е Wilson. 1991. A phylogenetic е 
of the Isopoda with some classificatory гесоттепда- 
tions. Mem. Queensland Mus. 31: 143-204. 


Моште 87, Митбег 1 
2000 


Brusca 23 
Arthropod Biodiversification 


Carroll, 5. В. 1995. Homeotic genes and the evolution of 
е and chordates. Na ture 376: 479-485. 

Chen, Ramskéld & G.-Q. Zhou. 1994. Evider 
for d and anhropod affinity of Cambrian wn 
predators. Science 264: 1304—13( sa 

Davidson, К. K. J. Peterson & R. A. Cameron. 1995. 
Origin of lave body plans: p of develop- 
mental regulatory mechanisms. Science 270: 1319— 
1325. 

Diaz-Be Kroa a, F. J., 
‘ell interaction between 
alaa distal axis of Drosophila legs. 
175-179 


ES 


. Cohen & 5. M. Cohen. 1994. 
compartments established the 
Nature 372 


1995. Visual motion 


notion computa- 


Douglass, J. & N. J. Strausfeld. 
detection circ dh in flies: Репрћега 
tion. by identified small. field Sirm neurons. J. 
Neurosci. 15: 5596-5611. 

& . Strausfeld. 1996. Visual motion detection 

'arallel direction- and non-direction- 

edulla and lobula 


circuits in к | 
sensilive pathways between the т 
4562. 


1990. Conservation of an- 
a cie cone conslituent in the com- 
мон” еуе of arthropods. J. Neurobiol. 21: 441—452 
Eernisse, D. S. 1997. Море апа = relationships 

re-examine з Рр. 43—56 т В. А. Fort . Thom- 
as (editors), Arthropod Кылыр: ране & Нап, 
1 


~ 


J. S. Albert & F. E. Anderson. 1992. Annelida 
and Arthropoda are not sister taxa: A phylogenetic anal- 
ysis | oe metazoan morphology. Syst. Biol. 41: 
305— 

Emerson, er J. & F. R. Schram. 1997. Theories, patterns 
and es Game ue for arthropod phylogeny. Pp. 67— 


86 in R. A. Forte R. H. Thomas rue rs), Arthropod 

Ke laii vo & Hall, Гот 
T^ 2 из ‚ D. J. rw 5. ]. oe 
Pace & R. Raff. 


P G Ps lin, 
uae uw: c на hg ny ii ^s pee erus Sci- 
ence 23¢ 
bat мм К. А. Raff & В. А. Best. 1990. 

иуи relationships of phylum Cnidaria inferred 
from 185 rRNA sequence data. Fourth Int. Congr. Syst. 
Evol. E | Abstract]. 

Friedrich, M. & D. Tautz. 1995. Ribosomal DNA phylog- 
eny of ^ major extant А classes and the evo 
lution of Б Машге 376: 165—166. 

R. от Cycliophora. Pp. 

. Ки T A speek 

3, Lopho- 


Funch, Kristensen. 
ren т F. W. Harrison & E. 
Microscopic Anatomy of и вечни Vol. 
phorates, Entoprocta, and Cycliophora. A oe li Ass, New 

or 

Garey, 1. H., М. Krotec, D. К. Nelson & J. Brooks. 1996. 
Molecular analysis supports a tardigrade-arthropod as- 
sociation. Invertebrate Biol. 11501): 79-88 

Greenslade, Р. 88. Reply to К. А. Crowson's ‘Com- 
ments on sie of the Rhynie Chert’ [1985 Entomol. 
Gener. 11: 09 Piee Entomol. Gener. 13: 115-117 

Hafner, G. S. & í R. Tokarski. Morphogencsis and patte 
formation in the retina of the crayfish Ка АУ 
clarkii. p 2 Res, (in pres 

Hahn, G., R. С. Brauc E 1986. Zur Kennt- 
nis von fe (Myriapoda; Ober-Karbon). Geol. 

Palaeontol. 7 125-1: 
~ th, К. M., J. D. Вас deter A. M. A. Aguinaldo, S. 
Liva, D. E Hillis & J. A. Lake. 1995. Evidence 


from 185 ribosomal DNA that the Ais ei are 
protostome animals. Science 267: 1641-1643. 
Harzsch, S., K. Anger & R. R. Dewers. pe Immun 
cytoc hend detection of к бк. ие alpha-tubulin and 
ius ups synapsis in the onic crustacean ner- 
ous system. Int. J. Deca: Biol 41: 411-494. 
Hillis, D. M. 1996. Inferring complex phylogenies. Nature 
383: 130-131. 
P. A. Selden & D. Edwards. 1990. Land 
Silurian: Arachnids and myriapods from 
Shropshire, England Science 250: 658—661. 
Kukalová-Peck, J. 1990. Fossil history and the evolution 
of hexapod structures. /n T. D. Naumann (editor), ne 
Insects of Australia, 2nd ed. CSIRO and Melbourn 
Univ. poen Melbourn 
la. The * ‘Uniramia” do not exist: The кама 
р!ап of i Pterygota as revealed by Permian Diaphan 
opteroidea from Russia ee сане a} 
Canad. Lt Zool. 70: 236- 
991b. Fossil hsion and the evolution of hexa- 
jda structures. Pp. 141—179 in I. D. Naumann (editor), 

The Insects of jd т Vol. 1. Cornell Univ. Press, 
York. 

. New Carboniferous Diplura, Monura, and 
nu the hexapod ground plan, and the role of 
thoracic side lobes in the origin of wings (Insecta). Can- 
ad. J. Zool. 65: 2327—2345. 
2. Brauckmann. 1990. Wing folding in ptery- 

gote insects, and the oldest Diaphanopteroidea from the 
early E jm Md of West Germany. Canad. J. 
Zool. 68: 111 
Labandeira, т В. Beall & F. Hueber. 1988. Early insect 
dE Uo: Evidence from a lower Devonian bristle- 
tail from Quebec. Science 242: 913-916. 
к: J. А. 1990. Origin the Metazoa. Proc. Natl. Acad. 
. U.S.A. 87: 763—766 
Locos G., H. Philippe, Н. L. Van Le & Н. Le Guyader. 
a. Species sampling has a major impact on phy- 
Кока inference. Molec. Phylogenetics & Evol. 2 
24. 


o0- 


Ithaca, pes 


3b. How many 
АЯ аге зг to айе, а phylogenetic prob- 
lem? The use of a new statistical method applicable to 
available sequences. Molec. Phylogenetics & Evol. 3: 
292-309 


6 ЖАЗ Chen & T.-E. Hua. 1998. Precambrian 
TENA S with cellular structures. Science 279: 879-882. 

58. Systema Naturae. Regnum Animale, 
10th ed., uo Salvii, Hominae. 

Manton, S. M. 1973. Psi i ови modern 
D cag pd J. 22 171: 111—130 

——. 7. The Arthropoda. Oxford Univ. Press, Ox- 


Linnaeus, C. 


ford. 

& D. T. Anderson. 1979. Polyphyly and the evo- 
lution of arthropods. Pp. 269-321 in M. R. House (ed- 
itor), The Origin of Major pipra Groups. System- 

atics Assoc. Special Vol. 12. Academic Press, 
London. 

McHugh, D. 1997. Molecular evidence that echiurans and 
ври are derived annelids. Proc. Natl. Acad. 
Sci. 0.5 4: 8006—8009. 

Miiller, K. 7 m Crustacea with preserved soft parts 
from the Upper Cambrian of Sweden. Lethaia 16: 93— 


on я Cambrian “Orsten.” Рр. 274—2 
гоміћег ймы. Palaeo- 


ЈЕ. 
inda ^v ura Blackwell Sci. Publ., London 


24 


Annals of the 
Missouri Botanical Garden 


Walossek. 1985. A remarkable arthropod 
fauna ‘ies the Upper Cambrian “Orsten” of Sweden. 
Trans. Roy. Soc. е Earth Sci. 76: 161-172. 

——— & ————. Martinssonia be rud gen. et 


= 


sp. n., a crustacean- “like e d fi inc: Cam- 
brian "Orsten" of pn Zool. S els 15: —92. 
——— & ——. 1988. Arthropod larvae from the Up- 


per Cambrian of Sweden. Trans. Roy. Soc. Edinburgh, 


Earth Sci. 77: 157— rae 


. Zakharov. 1995. “Orsten” type 
phosphatized soft- enn preservation and a new 
record from the Middle Cambrian Kuonamka depu 
in Siberia. Neues Jahrb. Geol. Palüontol., Abh. 

-118 

Nielsen, C., N. Scharff & D. Eibye-Jacobsen. 1996. Cla- 
distic analyses of the animal kingdom. Biol. J. Linn. 
Soc. 57: 385—410 

Nilsson, D.-E. & p. Osorio. 1997. Homology hus paral- 
lelism in arthropod pn processing. Pp. 333-347 in 

. Forte Thomas eh i od Re- 
ка а Pad & Hall, Londo: 

Osorio, D. & J. P. Bacon. 19€ т А т eye for arthropod 
раке BioEssays 16: 4 сн 

Panganiban, G., A. Sebring, L. у & S. Carroll. 1995. 

e development of crustacean Де ла and the evolution 
of arthro pods. Science 270: 136 Pas 

‚ S. M. Irvine, C. Lowe, H. Roehl, L. S. Corley, B. 

ra J. K. Grenier, J. F. Fallon, J. Kile M. Walk- 

Wray, B. J. Swalla, M. Q. Martindale & S. B. 
Carroll. 1997. The origin and own of animal ap- 
poe Proc. Natl. Acad. Sci. U.S.A. 94: 5162- 


Patterson C. 1989. Phylogenetic ОГ of major 
о Conclusions and prospects. Pp. 471—488 in B. 

Tial (editors), The 
Hierarchy of Life. Elsevier, Amsterdam. 

Paulus, H. F. 1979. Eye structure and the monophyly of 
the Arthropoda. Pp. 299-383 in A. P. Gupta (editor), 
о = МР ч Arthropod Phylog- 

y. Van Nostrand Reinhold, ork. 
TER Y J. van der, M. Neefs, Р. ^ m & R. De Wachter. 
constructing evolution from eukaryotic small- 
ти е RNA sequences: peas tion of the 
molecular clock. J. Molec. Evol. 37: -232. 
Popadic, А D. Rusch, M. Peterson, B. A Rogers & T. ( 
<aufma - 199%. Origin of the arthropod mandible. Na- 


T 


80: : 

Ramskóld, А E X. Hou. 1991. New early Cambrian an- 
imal a ay hophoran affinities for enigmatic metazoa. 
Natur 28. 

Regier, ]. © & J. W. Shultz. 1997. Molecular phylogeny 

of the major maod groups indicates polyphyly of 
crustaceans ane а new Vh pes of d origin of hexa- 

. Biol. Evol. -9]: 

990. bd i uniramous arthro- 
pod. Nature 343: 163 

Sanderson, M. J. 1996. How many taxa must be sampled 
to identify the root node of a large clade? Syst. Biol. 

at 


Scholtz, G Patel & W. Dohle. 1994. Serially ho- 
mologous Ede stripes are alin via difiere nt 
cell lineages in the germ band o 
Int. J. ner Biol. 38: 471—478 

995. Head segmentation in Crustacea—an im- 

uno  cytoc о al study. Zoology 98: 104—114 

Se ram. F. R. 1. Cladistic analysis of metazoan » phyla 

nd the Pen nt of fossil problematica. Pp. 35—46 in 


Li 


Turbeville. J. M., 


A. M. Simonetta & S. Conway Morris (editors), The Ear- 
ly Evolution of the Metazoa and the Significance of 


the tg of insect phylogeny]. Trudy Inst. Morf. Zhi- 
201. 8: 63-127. 

Shear, W. n P. M. Bonamo, J. D. Grierson, W. D. I. Rolfe, 
E. : Smith & R. A. Norton. 1984. Early land animals 
i th America. Evidence from Devonian age arthro- 
Bi from Gilboa, New York. Science 224: о 

A. Selden, W. D. I. Rolfe, P. M. Bona 

1987. New terrestrial arachnids from the 

Devonian of Gilboa, New York (Arachnida, Trigonotar- 

bida). Amer. Mus. Novit. 2901: 1–74. 

. 1997. Fossils, pp 

and ihe: evolution of animal limbs. Nature 388: 


D. г. 


Snodgrass, R. E. 1938. Evolution of the Annelida, жа 
SNET кү [оке Smithsonian Mise. Collec 
-15 


TNR T. E Abele. 1997. ae phylogeny 
ur э. ү rDNA. Рр. 169-187 ir orte 
& В. Thomas sari Arthropod в. 
NE a Hall, London. 

Storch, V. 1984. 37. enatis Pp. 709-713 in J. Ber- 
ndn p С. Matols . 5. Richards (editors), 
Biology of the Integument. 1, im Springer, 

erlin. 

& B. С. M. Jamieson. 1992. Further spermatolog- 
ical evidence for including the Pentastomida бирме 
worms) in the Crustacea. Int. J. Parasitology 22: 95— 
108. 

Stormer, L. 1969. Oldest known terrestrial arachnids. Sci- 
ence 164: 1276-1277. 

1977. Arthropod invasion during late Silurian 
and Devonian end Science 197 4. 

Strausfeld, N. J. 

Heidelberg. 


— 


. Atlas of an Insect Brain. Springer, 


. 1996. Oculomotor control in flies: From muscles 
to elementary motion detectors. Pp. 277-284 in P. S. 
G. Stein & D. Stuart (editors), Neurons, Networks, and 
Motor Behavior. Oxford Univ. Press, Oxforc 

1998. Crustacean-insect iciaionship s: The use 

of бай sharacters to derive phylogeny А вер- 
mented invertebrates. Brain, Behavior, Evol. 52: 186 
206 


Jushbeck & R. S. Gomez. 1995. The аг- 
TOR puris body: Its roles, evolutionary enigmas 
nd mistaken identities. Pp. спева in О. Breidbach 
us Systems of Inver- 
volutionary and дана Арргоасћ. 
Birkhiuser ыш. Basel. 
& J.-K. Lee. 1991. Neuronal basis for parallel 
visual processing in the fly. Visual Neurosci. 7: 13-33. 
Therianos, 5., 5. a euzinger, F. Hirth, C. S. Goodman & 
5. Embryonic development of the Dro- 
sophila к на of commissural = descend- 
ing pathways. Development 121: 3849-3861 
Thomas, J. В., М. J. Bastiani S. и 1984. 
From grasshopper to Drosophila: A common plan for 
euronal ок ment. Nature 310: 203—207. 
р. М. Pfeifer, К. С. Field & R. А. Raff. 


Volume 87, Number 1 
2000 


Brusca 25 
Arthropod Biodiversification 


1991. The phylogenetic status of arthropods, as inferred 
1 185 rRNA sequences. Molec. Biol. Evol. 8: 669— 


Waggoi oner, B. M. 1996. Phylogenetic hypotheses of the 
i PER of arthropods to Precambrian and Cam- 
brian рки fossil taxa. Syst. Biol. 45: 190—222. 
Walossek, D. K Müller. 1992. The “Alum Shale 
Wi nao "—Contribution of “Orsten” о to the 
phylogeny of Crustacea. Acta Zool. 73: : 312 
——& 1994. Pentastomid parasites 1 from the 
. Edin- 


Lower асои “ы Sweden. Trans. Roy. 
burgh, Earth Sci. 85: 1-37 

& . Cambrian “Orsten”-type arthro- 
pods and the E of Crustacea. Pp. 139-153 in 
R. A. Fortey & omas (editors), Arp Re- 


lationships. Systematics Assoc. Special Vol. 55. Chap- 
man y а n. 
Wheeler, W. P. Cartwright & C. Y. Hayashi. 1993. 


лаје Аш а А nd Eu Cladistics 


«uix P. M., T. Meier & P. King. 1991. m 

tion, neurogenesis and formation of cud» axonal path- 
s in the centipede ie nien rubripes (Brandt). 
Roux's Arch. Developmental Biol. d 3. 

, D. Leach & R. Sa pipe Меки 
Tu in neural от „а is arthro 
Axonogensis in the e two crustaceans "Deve - 
opment 118: 449-46 |. 

Williams, Т. А. & L. М. Nagy. 1995. Brine shrimp add 
salt to the stew. Gun. Biol. 5: 1330-1333. 

Wills, M. A., D. E. Briggs & R. A. Fortey. 1994. Disparity 
as an evolutionary index comparison of une 
and Recent arthropods. Paleobiology 20: 93-13( 

———, — —, — —— & M. Wilkinson. 1995. a sig- 


nificance of fossils in war dup arthropod evolu- 
tion. Verh. Deutsch. Zool. Ges. 88: 203-215 
Wilson, 6. D. F. & 5. Keable. A phylogenetic analysis of 
the n suborder Phreatoicidea. /n: R. C. Brusca & 
3. Kensley, Proc ne d ч the Second Int. Isopod Con- 
. Crustacean Issues, Bal 


trand, K. G. Cor 

pentastomid, Raillietiella cunei à and a ie пс 

ап crustacean | 

pentastomid po Kongel. 

Selsk. Biol.-Skr. 19: 1-72. 

. 1978. Comparative spermatology of the Crustacea 

Entomostraca I. Subc gg "pu Kongel. Dan- 
ske Vidensk. E Biol. -Skr. : 1-66 

Winnepenninc 
Brooks, a de W te & J. В. 
18s rRNA data а. that Aschelminthes аге poly- 
nhi in ann and consist of at least three distinct 

clades. Molec. Biol. & Evol. k 1132-1137. 

; R. De Wachter. 1992. 
Sienie of the rs dox subunit RNA of the 
үн snail Limicolaria kambeul, and phylogenetic 

nalysis of the Metazoa. Federation of European Bio- 
ie mic al Societies 309: 123-126. 

Wray, G. A., J. S. Levinton & L. H. Shapiro. 1996. Mo- 
licer poi for deep Preca Pod oe 
among metazoan phyla. Science 274: 56 

Xianguang. ‹ S. Weiguo. 1988. ^ a covery of Che- 
ngjiang зай at Meichucun, в Yunnan. Acta Ра- 
laeonto Du 27: 1—1 

2 Junyuan. 1989. xd в ag arthropod- 

ace intermediate sea animal, gen. nov. from ea 

jiang, Yunnan. Acta Кш. Sin. 28: 207-213 


Danske Vidensk. 


E 


SO MANY FISHES, SO LITTLE Ай Kottelat,” 
Ger ith,’ Melanie 

TIME: AN OVERVIEW OF Stiassny,? and Anthony C. Gille 

RECENT ICHTHYOLOGICAL 

DISCOVERY IN 

CONTINENTAL WATERS’ 


ABSTRACT 


Although freshwaters contribute only about 0.01% to Earth’s water supply, their fishes now number more than 10,000 
species and thus account for at least 40% of all fish species. The continental fish faunas differ greatly in taxonomic 
composition and species richness, our state of knowledge of them, and the rate of discovery of unknown kinds. The 
ichthyofaunas of North America (about 1050 species), Europe (about 360), and Australia-New Guinea (about 500), are 
the most thoroughly documented, but new species continue to be described based on discovery of previously unseen 
orms and species-level taxonomic splits of known species. The ichthyofaunas of tropical Asia (perhaps > 3000), Africa 
(perhaps > 3000 species), and South and Central America (perhaps >> 5000 species), are species-rich yet inc omple tely 
known. Tropical freshwaters are the hot spots of recent and likely future ichthyological discoveries. Especially in the 
species that signal new generic-level taxa are common, and new family-level groups are found 


occasionally. Everywhere ongoing phylogenetic studies oft gg r | unsuspected relationships. These аге 
imes of exciting disco advancement of knowledge in freshwater ichthyology. New discoveries beckon us to 
seek the many remaining unknowns in the diversity of life on our planet. These are also times of rapid and destructive 


change i in freshwater habitats around the pe se threats alert us to the increasing potential for permanent loss 
and i icd of ve of our planet's rich aquatic biota. 

Key words: Actinopterygii, Characiformes, Craniata, Cypriniformes, freshwater, Gymnotiformes, ichthyofaunas, ich- 
ыл. Otophysi, з finned fish, Siluriformes 


The species of craniate animals (hagfishes + ver- 
tebrates) are probably the most thoroughly docu- 
mented of all the large clades in the tree of life. 
Yet new species of living craniates are described 


i ichthyologists. Characterized in this way, the total 
mber of living fish species is about 25,000 and 
accounts for roughly 50% of the extant species rich- 
ness of the Craniata, while the other half are tetra- 
pods (Nelson, 1994). Thus delimited, fishes are not 
a сна не group because their subgroups аге 
along a phylogenetic "ladder" below the 
asd. (Fig. 1). The vast majority of fish species, 
owever, belong to one clade: the Actinopterygii or 
ray-finned fishes. There are at least 23,700 living 


frequently, and most of these are fishes. In a recent 
tally, Eschmeyer (1998) found that about 200 fish 
species are described annually 

Fishes are water-dwelling craniates with perma- 
nent gills borne on the walls of pharyngeal arches 


or pouches. Fishes have median fins supported by 
cartilaginous or bony rays, and most have paired 


actinopterygian species (Nelson, 1994). The species 
richness of the ray-finned fishes strongly contrasts 


fins, but never limbs bearing digits (Bond, 1996; with the e present species poverty of the other higher 
Helfman et al., 1997). Uniquely, fishes are studied fish clades (Fig. 1). 


! We applaud and and acknowledge Peter Raven and P. Mick Richardson for creating the 45th Annual System- 
atics Symposium at the Missouri Botanical Garden, and for inviting us to participate. We ч оиг тапу usse s 
who provided helpful Sc and suggestions on drafts of this manuscript: Alberto Akama, na C rnandes, 
Carl Ferraris, John Friel, lan Pos ui Hopkins, Michel Jegá, Dave Johnson, Luiz Миљан, Lucinda ме Dade, 
Steven Norris, Lynn Parenti, Мапо C. C. de Pinna, Lácia Rapp Py-Daniel, јен = sis, Ole Seehausen, Monica 
Toledo-Piza, Guy Teugels, P. J. Unmeck, Richard Vari, Stan Weitzman, and anonymous reviewers. Editors Amy Mc- 
Pherson and Victoria Hollowell at the MBG Press offered us patient and wise assistance өн чаша ће MER: process. 
Permission roduce ursi ade or photographs was granted by the authors or creators noted in figure captions, 
and the publishers of: American Museum Novitates; Copeia; Ichthyological Е xploration of Freshwaters; Occasional Papers 
of the Museum of Zoology, ay of Michigan; Proceedings of the Academy of Natural Sciences, Philadelphia; and 
Revue Suisse de Zoologie. 

? Department a enis and Evolutionary о The University of ta Tucson, Arizona 85721, U.S.A. 
осће 12, Саве > postale 57, < 19-2952 Cornol, Switzer lan 


S. 
Museum of Natural History, New Yo rk, New York 10024, U.S 
• Department of Zoology, The Natural History Museum, Cromwell Road, London SW7 5BD, United ETE 


ANN. Missouni Bor. GARD. 87: 26—62. 2000 


Моште 87, Митбег 1 


Lundberg et al. 27 
Ichthyological Discovery 


| Hagfishes 25 | 
| | 
Lampreys 
| 99 F 
ISharks, rays 1,200 ! ! 
|S 
IRay-finned fishes «423,700! Н 
E 
| Coelacanths 1(2?) | S 
| | 
|Lungfishes — — — 7 | 
Tetrapods 24,000 


Figure l. The phylogenetic tree of the major groups 
of living craniate animals and estimated number of extant 
species in each (based on Nelson, 1994, and Compagno, 
pers. comm.). 


The large-scale ecological distribution of fishes 
is strongly bimodal: 5846 of fish species are marine, 
41% (about 10,000 species) live principally in 
freshwater, and only about 160 (~ 1%) regularly 
migrate between salt and fresh water (Cohen, 1970; 
McDowell, 1988). This diversity of freshwater fishes 
causes some pause in light of the impressive fact 
that freshwater makes up a tiny amount, only about 
0.01%, of Earth's water supply. Thus, Horn (1972 
calculated that freshwaters hold a far greater “den- 
sity" of fish species than the oceans—greater by 
7500 times! But, of course, most marine species 
live in a relatively small volume of seawater in the 
productive photic zone, especially around coral 


МУ 


reefs. 

In this paper we focus on the diversity of fresh- 
water fishes, those living in continental lakes, riv- 
ers, streams, and swamps. In addition to their great 
species numbers, freshwater fishes are interesting 
to us because of their tremendous variety in form, 
function, and habit. Freshwater fishes provide evo- 
lutionary biologists with some . the best — 
of natural selection and adaptation, e.g., gupp 
(Endler, 1983) and а (Bell & Foster, 
1994), divergence and speciation, e.g., Laurentian 
Great Lakes whitefishes (Smith & Todd, 1984) and 
North American Great Basin fishes (Hubbs & Mill- 
er, 1948; G. R. Smith et al., in press), reticulate 
evolution, e.g., Catostomidae (Uyeno & Smith, 

1972), species flocks (African cichlids, Stiassny, 
this paper), and Lake Titicaca Orestias (Parenti, 
1984a; Costa, 1997), and the historical develop- 
ment of biotas on scales from regional, e.g., post- 
glacial age North America (Bailey & Smith, 1981) 
to continental, e.g., North America (Patterson, 
1981; Grande, 1994) and Africa-South America 
(Lundberg, 1993). 

Here we are particularly interested in the ex- 
panding knowledge of freshwater fishes: the 


amounts, natures, and sources of recent ichthyolog- 
ical discoveries and prospects for the future. Our 
first concern is with the new species of freshwater 
fishes that are being found and described. In ad- 
dition, we call attention to discoveries of other im- 
portant aspects of fish diversity: new phylogenetic 
lines and unexpected relationships, newly foun 
extraordinary characteristics of phenotypes and 
natural histories, and new understanding of the pro- 
cesses that control rates of speciation and extinc- 
tion. The major sources of previously unseen fishes 
are explorations in biotically uncharted waters, es- 
pecially in remote tropical areas, and in difficult- 
to-sample aquatic habitats such as deep river chan- 
nels, cataracts, and leaf litter. Yet survey work at 
long-visited sites, such as the African Great Lakes 
and Amazon margins, continues to yield many new 
species. Unrecognized and even formerly unseen 
species are routinely found in the course of thor- 
ough taxonomic studies, especially in revisions of 
species-rich, freshwater groups of ostariophysans, 
atherinomorphs, and percomorphs. Another source 
of recent species descriptions involves taxonomic 
concepts and practice. Application of species cri- 
teria that emphasize diagnosability or distinctness 
of populations over traditional geographically wide- 
spread, apes Auges results in an increase of 
recognized speci 

ollowing some : additional background informa- 
tion aimed mainly at the non-specialist, we take a 
regional approach to showcasing freshwater fish di- 
versity and recent discoveries. Based on our indi- 
vidual expertise, authorship responsibility follows 
geography: Smith—North America, Lundb 
South and Central America, 
tropical Asia, Stiassny—Africa, Gill—Australia 
and New Guinea. 


erg— 


FRESHWATER FISH FAUNAS: TAXONOMIC 
COMPOSITION 


Most higher fish clades include species that 
sometimes or permanently live in freshwater. All 
lamprey species reproduce in freshwater; some re- 
main there whereas others migrate to the sea to 
feed. Modern lungfishes are restricted to freshwater. 
Several species of sharks, sawfishes, and rays reg- 
ularly enter freshwater, and the Potamotrygonidae 
are a moderately diverse clade of sting rays con- 
fined to South American rivers and lakes. Scores of 
actinopterygians live in freshwater. Familiar ex- 
amples of these are sturgeons, gars, carps, pira- 
nhas, tetras, electric eels, catfishes, pikes, trouts, 
guppies, sticklebacks, black basses, sunfishes, 
darters, cichlids, bettas, and gouramies. If biotic 


Annals of the 
Missouri Botanical Garden 


success is measured by species numbers and 
breadth of overall distribution, the dominant grou 
of freshwater fishes is the Otophysi (Ostariophysi in 


(carps, minnows, barbs, suckers, loaches) of North 
America, Europe, Asia and Africa, presently with 
roughly 2700 species; (2) Characiformes (tetras, pi- 
ranhas), now with at least 1300 species, distributed 
today in the Neotropics and Africa, and formerly 
extending to Europe and the Arabian Peninsula; (3) 
Gymnotiformes (electric eel and knifefishes) of 
South America with over 90 described species; (4) 
Siluriformes (catfishes) currently with more than 

0 species and an overall distribution including 
all continents, even Antarctica as recorded by fos- 
sils from Seymour Island (Grande & Eastman, 


BIOGEOGRAPHY AND PHYSIOLOGICAL ECOLOGY OF 
FISHES 


Freshwater fishes play important roles in a va- 
riety of biogeographic studies. Some are endemic 
markers that delimit regional biotas. Fish species 
are censused in determinations of regional and 
global patterns of species richness or biodiversity. 
Each clade of fishes has its singular biogeographic 
history. Many freshwater fish clades are compo- 
nents of biotas that share with others common vi- 
cariant or dispersal-based biogeographic histories. 
Many fish species and higher taxa serve as biotic 
indicators of former connections among regions 
from watersheds to continents. 

Monophyletic groups of strictly freshwater fishes 
have special significance in continental biogeogra- 
phy because they require freshwater for dispersal, 
and thus their distributions are correlated with the 
evolution of topography and watersheds. The degree 
to which fishes are physiologically and behaviorally 
restricted to freshwater varies widely. Species and 
higher taxa of fishes are commonly grouped ac- 
cording to their observed present and presumed 
historical habitat and physiological salinity toler- 
ances (Myers, 1938; Darlington, 1957). So-called 
“primary” freshwater fish groups, such as most oto- 
physans, spend their entire lives in freshwater hab- 
itats and are physiologically incapable of coping 
with seawater. It is important to know the рћујо- 
genetic level at which subgroups within a clade 
show strict primary freshwater distributions (Pat- 
terson, 1975; Lundberg, 1993) 

"Secondary" freshwater fish taxa, such as cypri- 
nodontiforms and cichlids, are usually limited to 
freshwater, especially for reproduction, and have 


North America Europe 360 
1,050 9 | ЧЩ Tropical 
EMD = Asia 
> 3,000 


SA ING 


Australia & 
New Guinea 
500 


South & Central 
America »5,000 


Africa 
«3,000 
Figure 2. Estimated number of living fish species in 
(ће major freshwater faunas. Base map created by Xerox 
Corporation's Palo Alto Research Center, Map Viewer. 


their greatest diversity and abundance there, but 
occasional individuals or member species may be 
found regularly in coastal saltwater habitats. Sec- 
ondary freshwater fishes might, therefore, disperse 
through the sea. "Peripheral" fishes are marine 
groups that include species with individuals that 
may move sporadically into freshwater or species 
that are permanent freshwater residents; common 
examples include herrings, anchovies, needlefishes, 
puffers, pipefishes, drums, gobioids, and soles. All 
continental ichthyofaunas include peripheral 
groups with persistent freshwater populations, and 
the freshwater faunas of many islands comprise en- 
tirely peripheral fishes. These are generally regard- 
ed as "marine dispersants" but, unless or even if 
their closest intragroup relatives are known to be 
resident marine taxa, peripheral fishes should not 
automatically be ignored in comparing interconti- 
nental faunas. 


DISTINCTNESS OF FRESHWATER СНТНУ‹ 


ГА ОМА 


At the scale of continents (and Alfred Wallace’s 
Zoogeographic Realms) ichthyofaunas differ greatly 
in their taxonomic composition and species rich- 
ness (Fig. 2). Australia and Europe each have hun- 
dreds of fish species, North America has somewhat 
more than 1000, tropical Asia and Africa each have 
about 3000, and there may be more than 5000 fish 
species in the American tropics. Especially at the 
species level, there are also major differences in 
thoroughness of our knowledge of continental fish 
faunas. Higher-level taxonomic differences are 
mostly caused by the long and distant isolation of 
the freshwaters of most continents. Differences in 
species diversity probably have more to do with 
factors that control both terms of biological diver- 
sification: speciation and extinction. Differences in 
the extent of knowledge of species result from dif- 
ferent histories of exploration and study, as well as 
species richness. It is not surprising that we know 


Volume 87, Number 1 Lundberg et al. 29 
2000 Ichthyological Discovery 

Figure Fish icons of North America. —A. Petromyzon marinus, sea lamprey. —B. Polyodon ie ли p 
—C. Lei 25165 05565, _ gar. —D. Amia NO bowfin E. Lavinia exilicauda, hitch. nus pla- 
tyrhynchus, mountain suc —G. Noturus furiosus, Carolina шайган, —Н. Aphredoderus nm pirate perch, —1. 
Percopsis omiscomaycus, trout-perch. . Spe leti poulsoni, Alabama cavefish. —K. Umbra limi, central mud- 
minnow. —L. Esox темин, musk БШ ge. —M. randria bimaculata, spottail кае —N. кези notatus, 
blackstripe topminnow. —O. Goodea atripinnis, blac kfin goodea. —P. Percina caprodes, logperch. —Q. Archoplites 
interruptus, Sacramento perch. —R. Elassoma evergladei, Everglades pygmy sunfish. Drawn n Tum Шу, 


more about the less diverse European, North Amer- 
ican, and Australian faunas, than the rich African, 
tropical Asian, and South American assemblages. 


NORTH AMERICA 


The North American freshwater fish fauna oc- 
cupies the Nearctic Realm: from Canada and Alas- 
ka, south to the Transvolcanic Axis south of the 
Mexican Plateau (Fig. 2). Over 1050 species (e.g., 
Fig. 3) in about 175 genera and 32 primarily fresh- 
water families (plus a few freshwater species be- 


longing to 35 genera in 24 mostly marine families) 
occupy rivers and lakes currently or recently drain- 
Caribbean, and 


ing to the Arctic, eastern Pacific, 


western Atlantic. The numerically dominant fami- 
lies are: Cyprinidae (305 species), Percidae (172), 
Poeciliidae (75), Catostomidae (68), Ictaluridae 
(48), Goodeidae (40), Fundulidae (37), Centrarchi- 
dae (32), Atherinidae (35), Cottidae (27), and Cich- 
lidae (21). These 11 families make up about 80% 
of the species in the fauna. 

Historical biogeography of the fauna was sum- 


30 


Annals of the 
Missouri Botanical Garden 


marized in the comprehensive treatise Systematics, 
Historical Ecology, and North American Freshwater 
Fishes edited by Mayden (1992). Regional bioge- 
ography was thoroughly analyzed in Zoogeography 
of North American Freshwater Fishes edited by Ho- 
cutt and Wiley (1986). Systematic accounts and 
distributional records have been documented in 
about four dozen regional, provincial, or state fish 
monographs and the Atlas of North American Fresh- 
water Fishes (Lee et al., 1980). Nine families and 
128 genera are now endemic to North America, but 
some (e.g., Amiidae, Hiodontidae) were more wide- 
spread in the past. The principal intercontinental 
relationships of most of the fauna (e.g., Cyprinidae, 
Catostomidae, Salmonidae, Esocidae, Percidae) are 
with Eurasia, whereas the Characidae, Pimelodi- 
dae, Cyprinodontidae, Fundulidae, Profundulidae, 
Poeciliidae, and Cichlidae have neotropical rela- 
tionships (Patterson, 1981; Grande, 1994). 

The combined fauna of Atlantic and Gulf of Mex- 
ico drainages is several times as diverse as the Pa- 
cific drainage fauna because its land area is larger 
and has been geologically and climatically more 
stable during the Cenozoic. The Mississippi Basin 
is the center of North American freshwater fish di- 
versity; its fauna is the largest with 375 species. 
The Eastern Highlands, mostly in the Mississippi 
drainage, have the most distinctive fishes with 57 
species (Mayden, 1985, 1987). Diversity decreases 
with distance away from the Mississippi Basin 
heartland. The vast northern glaciated areas are de- 
pauperate because of long winters and slow post- 
glacial recolonization (Fig. 4; see also C. L. Smith, 

5). Western and southern faunas are depauper- 
ate because they are mountainous, arid, and iso- 
lated by barriers. The Great Basin (Hubbs & Miller, 
1948; Hubbs et al., 1974) and Colorado River fau- 
nas, in western North America, have the lowest di- 
versity, but highest endemism (Miller, 1959), over 
50%, indicating a high extinction rate (G. R. Smith 
et al., in press). 

Freshwaters of the United States were thoroughly 
explored in the 19th century and the first half of 
the 20th century; North America now has few un- 
discovered species except in Mexico and Central 
America. Recently described additions to the North 
American fauna are mostly allopatric, divergent lo- 
cal populations of darters, minnows, and suckers 
that are elevated to species status based on the 
phylogenetic and evolutionary species concepts 
(Mayden & Wood, 1995). An unusual recent dis- 
covery was Scaphirhynchus suttkusi Williams & 
Clemmer, a new species of sturgeon from the Mo- 
bile Basin. Molecular data have also revealed di- 
verse populations of salmons, trouts, and whitefish- 


50 
150°, 
a 
40 
440 
э}. 
Е з0 
20 
420 


ZA во-1 
49-79 
25-48 


LJ 10-24 
СО 1-9 
Figure 4. Fish species diversity gradients displayed 
as nu 


cies richness from 80 to 105; 49 to 79; 25 to 48; 10 to 
24; 9 


es (Bernatchez & Wilson, 1998), some of which are 
sympatric but ecologically different. Some of these 
forms fit species concepts based on inferred genetic 
and ecological differences between populations that 
spawn in different times and places. With emphasis 
on historical processes of origin, however, G. R. 
Smith et al. (1995) defined species as lineages sep- 
arated from each other by genetically based mor- 
phological, reproductive, ecological, or behavioral 
barriers sufficient to confer long-term historical in- 
dependence. 

Monographs on groups of the North American 
freshwater fish fauna have been produced recently. 
The most magnificent are The Evolutionary Biology 
of the Threespine Stickleback, edited by Bell and 
Foster (1994), and Native Trout of Western North 
America, by Behnke (1992). Two of North America’s 
famous relictual fishes and their fossil relatives, 
paddlefishes (Polyodontidae) and bowfins (Ami- 
idae), are RT treated by Grande and 
Bemis (1991, so notable as popular as 
well as мен а valuable treatments аге The 
Handbook of Darters by Page (1983) and The Amer- 
ican Darters by Kuehne and Barbour (1983). The 
fauna has also benefited from outstanding ecologi- 
cal and evolutionary treatises, such as that by Mat- 
thews and Heins (1987). The field eagerly awaits 


Моште 87, Митбег 1 
2000 


Lundberg et al. 31 
Ichthyological Discovery 


the appearance of the Freshwater Fishes of Mexico, 
in preparation by Robert R. Miller 

Because North American fish species are well 
described, the most exciting recent discoveries are 
of evolutionary relationships and patterns. New in- 
sights on relationships are the result of research in 
molecular and morphological phylogeny, an area of 
systematics substantively pioneered by North 
American ichthyologists (Wiley, 1981; Burr & May- 
den, 1992). Systematic summaries of most of the 
important groups can be found in Mayden (1992). 
The higher-level systematics of representative fish 
taxa is treated in Stiassny et al. ineteen 
of the 21 richest North American families have 
been the object of substantial phylogenetic work 
(Burr & Mayden, 1992: 32). More than 250 phy- 
logenetic papers on North American fishes have 
been published, but much more cladistic work is 
needed (Burr & Mayden, 1992: 59). Unexpected 
recent discoveries include the close relationship of 
the live-bearing Goodeidae (Fig. 30) of the Mexi- 
can Plateau to the egg-laying Crenichthys and Em- 
petrichthys of the Great Basin (Parenti, 1981; Webb, 
1998) and the relationship of the pygmy sunfishes, 
Elassoma (Fig. 3R), to the sticklebacks (Johnson & 
Springer, 1997). Other relationships, based on DNA 
sequences, include spinedace (Hubbs & Miller, 
1960) and creek chubs among cyprinids (Simons & 
Mayden, 1997), and Pacific trouts and salmons 
among salmonids (Stearley & Smith, 1993; Philips 
& Oakley, 1997). In addition, studies of phylogeny 
have revealed the frequent influence of introgres- 
sion on evolutionary patterns of North American 
freshwater fishes (G. R. Smith, 1992; Dowling & 
Secor, 1997). 

New evolutionary analyses relate patterns in fish 
diversity through time to the geological control of 
rates of evolution and extinction. In tectonically 
fragmented and volcanically disturbed areas, high 
extinction rates control diversity. Comparison of 
fossil and Recent North American fishes suggests 
that they have not evolved substantially in response 
to Pleistocene environmental changes. These re- 
sults are consistent with those based on discovery 
of early to middle Cenozoic fishes closely related 
and similar to extant species in North America and 
South America (Cavender, 1986; Wilson & Wil- 
liams, 1992; Lundberg, 1998). Molecular and mor- 
phological data in their geological context indicate 
that anagenic change and speciation are much 
slower than geological and climatic changes (G. R. 
Smith et al., in press). These results suggest that 
the current (latest of the past two dozen or so cy- 
cles) post-glacial species assemblages in glaciated 


regions, such as the Great Lakes basin, are not 


Б 5 N Q 
л © о о 
1 1 1 J 


E 
o 


T T T T T Џ 


MEAN NUMBER OF DORSAL SPINES 


Џ ct i. 
70 80 90 100 110 
TIME ( х 1000 YEARS) 
ure 5. Short-term fluctuations within a 100,000-yr. 
trend in stickleback spines (from Bell et al., 1985) den 
onstrating limited significance of ка Баев over the 
long course of phenotypic evolution 


likely to be ecological communities with fine-tuned 
interspecific interactions. 

Studies of rates of fish evolution were pioneered 
by Hubbs and Miller (1948) in the Great Basin. 
They documented rapid evolution based on ob- 
served changes attributed to post-pluvial isolation 
(in the past 10,000 years) of populations assumed 
to have been uniform when waters were connected 
in pluvial times (e.g., Kocher & Stepien, 1997). 
Current studies negate the assumption of geneti- 
cally uniform species (G. R. Smith et al., in press). 
Studies of Pliocene and Pleistocene morphological 
changes in the Great Basin show that the early, 
rapid responses to environmental change such as 
those documented by Hubbs and Miller (1948, 
1974) do not usually lead to new species (Bell et 
al., 1985; G. R. Smith et al., in press). The isolation 
of small populations in rapidly changing environ- 
ments promotes rapid changes, i over the lon 
term these appear to be short-term fluctuations 
within slow trends (Fig. 5; Bell & "Haglund, 1982; 
Bell et al., 

Fossil TM suggest that the modern North 
American families probably date back more than 
65 million years to the Cretaceous, most genera to 
the Miocene, and species mostly to the Pliocene or 
early Pleistocene. Species formation averages slow- 
er than one branch per million years per clade in 
rivers (depending on the family) but may be more 
rapid in lakes (Echelle & Kornfield, 1984). Molec- 
ular evolution (Kocher & Stepien, 1997) may be 
stochastically constant within 10-25% error; for 
certain mitochondrial genes, it varies from 0.596 
sequence divergence per million years in salmonids 
to about 196/m.y. in cyprinids and cyprinodontoids 
Fig. 6; G. R. Smith et al., in press). The a 
estimates suggest that adaptive evolutionary re- 
sponses to the current global ecological crisis are 
unlikely. 


— 


Annals of the 
Missouri Botanical Garden 


32 
8 ‚25 
c Cyprinidae 
5 .20- 
o 
$ 15 Cyprinodontoidei 
8 ло 
5 Salmonid 
5 .05- опіаае 
© 
Р) Ч Т Т Т 
0 


5 10 
Million years before present 


sure 6. Estimated rates of mitochondrial gene se- 
quence divergence vary from 0.5% to 1% per million 
years among salmonids, cyprinids, and id E dE 
Fossil discoveries enable eed. of these rates when 
synapomorphies of earliest 1 fossils can be used to 
identify the internode segment or pi of the lineage in 


which the fossil fish w ember. The age of the fossil 
can then be e to е а minimum sana of bis 
age in millions of years—the deno the 
equation; The оме sequence eae eg eke 


xa for which an age estimate is available Же becomes 
ifa num cud in the rate equation. The rates are then 

sed to e e the ages of other vicariance barriers and 
ages of 


Prospects for future ichthyological discoveries in 
North America include some new species in Mexico 
and important advances in our knowledge of pat- 
terns and processes of fish diversification. 


SOUTH AND CENTRAL AMERICA 


The vast Neotropical ichthyofauna, estimated to 
contain between 5000 and 8000 species (Schaefer, 
1998; Vari & Malabarba, 1998), inhabits the fresh- 
waters of South and Central America (Fig. 2), with 
a handful of cichlids, pimelodid catfishes, and 
characids extending north into Mexico or the south- 
ernmost U.S. The great majority of Neotropical fishes 
belong to one of five dominant groups: characi- 
forms, siluriforms, gymnotiforms, cyprinodontiforms, 
or cichlids. 

Within the Neotropics the Amazon Basin con- 
tains Earth's most diverse riverine fish fauna that 
certainly far exceeds 1000 species. The Orinoco, 
Paraná, and other large, tropical rivers flowing to 
the Atlantic are also species-rich. Fish diversity 
drops sharply in the watersheds emptying into the 
Caribbean and Pacific, and southward into temper- 
ate South America where the taxonomic composi- 
tion also changes markedly. Central American 
idees contain roughly 300 species (Bussing, 

. In this region the San Juan Basin of Nica- 
ragua and Costa Rica has the most diverse fauna 
with about 54 species (Bussing, 1985). 

Neotropical cichlids and gymnotiforms are en- 
demic, monophyletic clades. On the other hand, 
Neotropical characiforms (Vari & Malabarba, 
1998), siluriforms (de Pinna, 1998), and cyprino- 


dontiforms (Costa, 1998) each contain several sep- 
arately monophyletic subgroups with incompletely 
known extralimital relationships. Some Neotropical 
fish clades have their closest relatives of today in 
African freshwaters (Lundberg, 1993; Vari & Mal- 
abarba, 1998; de Pinna, 1998): lungfish, arapaima, 
ctenoluciid + erythrinid and some characid char- 
aciforms, doradoid catfishes, aplocheiloid and poe- 
ciliid cyprinodontiforms, cichlids, and nandids. A 
few of the ed cyprinodontiforms are most 
closely related to American taxa (Parenti, 
1981). Many ibd fishes, including individ- 
ual species and some small clades, have their prox- 
imate relatives and presumed ancestry in coastal 
marine waters, e.g., river sting rays, various her- 
rings and anchovies, drums, soles, needlefishes, 
toadfishes, and a puffer. 

It is scarcely surprising that no comprehensive 
treatise yet exists for the Neotropical fish fauna. 
However, two recent publications have immensely 
advanced access to information about the fauna. 
The massive Catalog of Fishes (Eschmeyer, 1998) 
provides the most thorough listing ever retrieved of 
the binomials applied to all fishes and also contains 
an extensive bibliography of descriptive ichthyolo- 
gy. The electronic version of the Catalog has great- 
ly facilitated an estimate of the historical account- 
ing of published description of Neotropical fishes. 
The symposium volume Phylogeny and. Classifica- 
tion of Neotropical Fishes (Malabarba et al., 1998) 
contains 28 papers that summarize much up-to- 
date systematic knowledge of higher Neotropical 
axa. 


c 


a 
The discovery of Neotropical fish species became 
an active enterprise by about 1825 and it continues 
at a high rate. The latest ca. 50-year trends in spe- 
cies description for the Neotropical siluriforms, 
gymnotiforms, characiforms, cyprinodontoids, and 
cichlids are shown in Figure 7 (data from Eschmey- 
er, 1998). Overall about 1400 species were de- 
scribed during this period, and the vast majority of 
these are considered valid. Using the recent esti- 
mates for total Neotropical fish species richness, 
this amount of discovery and description could rep- 
resent about 25% of the whole ichthyofauna. 

The levels of recent discovery and estimates of 
total species richness for the five major groups are 
truly i impressive. The siluriforms are the richest of 


species (Nelson, 1994). 
491 Neotropical catfishes were described, and more 
than half of these were published in the last 20 
years. Several known catfish species await descrip- 
tion, and certainly other unrecognized and unseen 
species will turn up in natural history collections 


Volume 87, Number 1 
2000 


Lundberg et al. 
Ichthyological Discovery 


<< Characiformes 
m æ< Silurifor ^. 
noti < 
Сутпо{ Ног = 4 
165 Se >= = = 
я Cic < ч a 
чч o аф 
<< << <a 
150 < < << 
< < < 
< << << 
- = = 
ок. + 
135 «ec a < 
o «v 4 < 
c «< E ~ 
о а сас «< 
~ а җы ~ 
120 а ee o 
- о и a 
а. И 
= «ьс «асс абс аА ё 
a ee чада: с «йч 
© 105 < «і сас сс ойыс аф 
[7] << «< ae «адаг седи чв 
Ф << ас сана с ен А 
аф --— анас «адас андаг фр 
б << aime age т a 
90 я << to tee Em o n 
<< < а €—X abe ес А 
o = pres > LEM eee або te — 
Ф 75 с a$ = 2 m 
—_ <_< << L сан abe abe адаг адас 
[5] > dn БЕ б ьс «ee abe «бе M ан 
Ф << << << << « d с або «бо ес и 
а. ы LJ << 4 ~ I ee tee або ве лес tee 
<< «v | а сас афс чо с << ate — = — 
<< = сана ee аа ed — «ча о 4 cee mee 
~ ас сас саас афс са афс эы о a бе бе әдә со | ос 
45 | << ЧЕ чс ада аф ол ас о о ве бе йэ 4 до ә 
саас dM NE o b ete eee << бе ве Фи abe «she 
ee ee а eee ee << бе ве 4 -— ado 
p de Lm orn 
A^ | ———— «abe a —- ад 4 бе n 4 4 = & o ate 
= a =й abe abe eo ——— «ae «ш ae а & Ф ate ate 
15 | бе a аа --— abo = ee —— чё —— = = == & «ae ate 
aio an чш -— «e ди а - «до де dde 4 XR & 4 
-— <= «alte m ебә o © m 4 --—- ле с 4 = = 
4 =e ate @ 4 ate ate ote 46 4 + а & = = 
= 4 4 = а 4 = & 46 T 46 T TE & & 
1952 1955 1958 1961 1964 1967 1970 1973 1976 1979 1982 1985 1988 1991 1994 1997 
3 - Year periods ending in date shown 
Figure 7. Neotropical fish species "d "sc ds dry 1950-1997 for characiforms, siluriforms, gymnotiforms, cyprino- 


dontiforms, and cichlids based on data 


and new field samples. Thus, any estimate of the 
total species richness of Neotropical catfishes is 
now imprecise, but close to or more than 2000 spe- 
cies should be expected 
Mago-Leccia (1994) NE 94. valid species 
of electric fishes; Campos da Paz and Albert (1998) 
estimated the total to exceed 100 species. In just 
the last 25 years, descriptions of 31 new gymnoti- 
form species and 13 new genera were published, 
and we are aware of an additional 32 undescribed 
species. As with catfishes, it is difficult to estimate 
the total species richness of Neotropical characi- 
forms because of their taxonomic complexity and 
the high rate of discovery of undescribed species. 
Nelson (1994) estimated an overall total of 1300 
characiform species, and Vari (1998) predicted that 
far greater than 2 exist overall. Of these only 
about 225 are African. Between 1950 and 1997, 
over 450 characiform species were described in the 
Neotropics, and although annual rates of descrip- 
tion vary, there is no indication of reduction. Ex- 
perts working with characiform taxonomy are aware 
of many dozens of undescribed forms. 
South and Central American cyprinodontiforms 
are estimated at about 375 species (Huber, 1998; 


n Eschmeyer (1998). Each whole fish i 


partial fish icons equal one or two desc dcin each column covers a three-year peri 


icon represents three descriptions, 
iod. 


Nelson, 1994). From the Neotropics 203 cyprino- 
dont species have been described since 1950, and 
of these over 80% (166) were published after 1974. 
These numbers would be higher if new species in 
shared and closely related genera from Mexico, the 
U.S., and the West Indies were included. Neotrop- 
ical cichlids are not as species diverse as African 
cichlids, but their numbers are high and increasing. 
Kullander's current (1998) estimate is that the total 
will be about 450 Neotropical cichlids, of which 
more than 100 are undescribed. About 30% (134) 
of the estimated total have been described since 
1974. 

The continuing high level of discovery of fish 
species in the Neotropics may be explained by the 
combination of an increased amount of field and 
taxonomic work applied to a very large, taxonomi- 
cally complex, and undersampled fauna (see also 
Vari & Malabarba, 1998). We could cite dozens of 
recent examples of ichthyological discovery coming 
out of small and incidental to large, focused field 
exploration throughout the Neotropics. However, 
the recent history of ichthyological exploration in 
Venezuela illustrates the great magnitude of dis- 
covery, on one large, hydrographically complex re- 


Annals of the 
Missouri Botanical Garden 


gional scale, that is still possible in many areas of 
South America. About 30 years ago Mago-Leccia 
(1970) published the first comprehensive list of Ve- 
nezuelan freshwater fishes, which included 474 
species. In 1997 Taphorn et al. were able to revise 
the Venezuelan list upward to 1065 species (a 
125% increase) based largely on the combined re- 
sults of much survey work carried out during the 
intervening approximately 25 years. 

The number of taxonomists involved in describ- 
ing Neotropical fishes has increased over the years, 
although we may have reached a plateau. Using 
characiform taxonomy as an example, for the 10 
quarter-century periods between 1758 and 1990 
the numbers of authors of species descriptions were 
4, 5, 9, 18, 12, 31, 28, 38, 64, 57, and in the 7 
years since 1990 already 54 authors have been in- 
volved in describing characiforms. Between 1970 
and 1997, 228 authors contributed to papers de- 
scribing approximately 960 species of ee 

characiforms, siluriforms, gymnotiforms, cyprino- 
dontiforms, and cichlids. As expected, рана 
productivity of species descriptions varies tremen- 

ously: the range of descriptions across author is 
1-102, with a strong mode of 1 description per au- 
thor, and a median of 2. Seven highly productive 
individuals were sole or co-authors of at least 40 
species descriptions and, remarkably, the total con- 
tribution of these 7 persons accounts for over half 
(524) of the 960 recent descriptions. 

A general pattern of biodiversity is that species 
richness increases with sample size, e.g., with area, 
time, specimens, taxonomic scope, etc. The effect 
of taxonomic scope on Neotropical fish diversity is 
evident at different ranks. For example, differences 
in the amounts of species discovery among catfish 
families are correlated with the species richness of 
those families (Fig. 8). The same relationship ap- 
pears to hold at the genus level with the very large 
genera Corydoras, Aspidoras, Trichomycterus, and 
Hypostomus accounting for over 3096 of all Neo- 
tropical siluriform species described during the last 
ca. 25 years. Among characiforms, too, the large 
genera Leporinus, Hyphessobrycon, Cyphocharax, 
Curimata, Astyanax, and Creagrutus account for 
about 3096 of the species described in the same 


riod. 

Exemplifying the great yield of new species that 
can be found by in-depth study of relatively small- 
bodied characiforms, Vari and Harold (1998) have 
raised the species number of Creagrutus (east of 
the Andes) from 15 to 56, a 37396 increase. Re- 
visionary work at the species level seems more of- 
ten than not to produce a net increase in the 


ber of Neotropical fish species. Kullander (1980). 


150- 
m 
~ 
ш г = .96 

Lo 
ш 
о 
2100 
2 
a 
ul 
a 
Ca 
* 504 
d Pi 
та Тг 
Ф sc Au 
t Do 
Di 100 200 300 400 500 600 
CURRENT TOTAL SPECIES 
Figure 8. Relationship between species richness of 


Neotropical catfish families and numbers of new species 
described during the last ca. 25 yr. based on data from 
Eschmeyer ipu Ар = Азрг єй йе, At = Astroble- 
pidae, Au = Auchenipteridae (inc luding Ageneiosidae 
and Centromoc Ме), Са = Callic 


5 
& 
d. 
E 
Ф 

| 


Trichomycteridae. (One fossil but no extant species of Ме- 
matogenyidae were described in this period.) 


for example, reported that by 1977, 31 nominal 
species of the cichlid genus Apistogramma had 
been described, of which he judged only 20 to be 
valid. In his 1980 revision, Kullander added 18 
new Apistogramma; he recognized 36 overall and 
estimated that the genus contains > 40 species. By 
1997 an additional 17 species of Apistogramma had 
been described, about a 50% increase in the 17 
years following the initial revision. On the other 
hand, revisionary work may sometimes suggest con- 
specificity that might yield a net reduction in the 
number of recognized forms. In a revision of the 
widely distributed catfish genus Rhamdia, with 
about 100 nominal species, Silfvergrip (1996) pro- 
posed that only 11 species are valid (the author 
described 3 of these in the wor 

Whereas the numbers of new species are high 
and the common taxa are becoming more so, the 
qualities of recent ichthyological discoveries in the 
Neotropics are perhaps more impressive than quan- 
tity. Neotropical fishes are famous for their diversity 
of diet and trophic apparati. New species with strik- 
ingly distinct feeding specializations are common. 
Among characiforms the most unusual recently 
found dietary specialist is the serrasalmine Ossub- 
tus xinguense (Fig. 9A; Jégu, 1992), a species from 
turbulent cataracts that feeds on aquatic macro- 
phytes, especially Potemogeton. During its devel- 


Volume 87, Number 1 
0 


Lundberg et al. 35 
Ichthyological Discovery 


New dietary specialists from the Neotropics. 


Figure 9. 
—A. Ossubtus xinguense, left, 
mouth; right, an adult specimen about 153 mm long (re- 
produced with permission from Jegu, 1992). —B. Magos- 


drawings of teeth and 


ternarchus duccis, left, anterior third of adult specimen 
skeleton of teeth and jaws (re- 
1996). — 

. Sternarc AI sp. (Copy right John С. Lundbe TE and 
hn P. Sullivar 


about 235 mm long; right, 
produc ‘ed with permission from Lundberg et al., 


opment the snout, jaws, and gill chamber of Ossub- 
tus undergo a metamorphosis, twisting ventrally so 
that the mouth and blade-like teeth are directed at 
the plants attached to the rocky substrate. An in- 
teresting twist was added to this case with the dis- 
covery of a parasitic isopod living in the gill cham- 
ber of the fish that also develops a contorted 
morphology, presumably in response to its host’s 
cursive ontogeny (Thatcher, 1995). 


Most gymnotiforms have generalized diets of 


small invertebrates or fishes. However, the recently 
described genus Magosternarchus (Fig. 9B; Lund- 
berg et al., 1996) includes two species of Amazon 
Basin apteronotids that, with greatly enlarged jaws 
and teeth, feed on the tails of other electric fishes. 
Another “new” specialist among electric knife fish- 
es is Rhabdolichops zareti (Lundberg & Mago-Lec- 
cia, 1986), from deep channels of the Orinoco, that 
feeds mostly on allochthonous zooplankton washed 
into the rivers mainstem channels from the pro- 


1987). 


Providing an example of an odd newly discovered 


ductive marginal savannas (Lundberg et al., 


structure possibly related to electrolocation of prey, 
an undescribed species of the apteronotid Sternar- 
chogiton bears a probe-like, somewhat protrusible 
organ on its lower jaw (Fig. 9C) 

The evolution of small to miniature-size species 
is commonplace among Neotropical fishes (Weitz- 
man & Vari, 1988). It is no surprise that very small 
fishes are discovered relatively late in survey work; 
40 of the 85 little species listed by Weitzman and 
Vari were described in the latest 25-year period, 
more than twice that of the previous quarter cen- 
tury, and several more have been discovered re- 
cently that fall below or approach their size crite- 
rion of 26 mm. In the Neotropics the evolution of 
reduced size is most common among characiforms, 
siluriforms, and cyprinodontiforms, and there are a 

andful of remarkably small engraulids (Amazon- 
sprattus scintilla, Roberts, 1984) and gobioids (Mi- 
crophylipnus species, Myers, 1927). Weitzman and 
Vari (1988) pointed out that miniature species of 
most taxa live in slow-flowing or still waters. The 
most strikingly modified small and miniature cat- 
fishes, however, are found in swiftly flowing and 
often deep, river channels. Examples of these in- 
clude the aspredinid Micromyzon akamai (Fig. 
10A; Friel & Lundberg, 1996), a blind, nearly pig- 
mentless, heavily armored species that matures at 
about 12 mm, making it one of the smallest known 
catfish species. Bathycetopsis oliveirai (Fig. 10B; 
Lundberg & Rapp Py-Daniel, 1994) is a deep-water 
ca. 35-mm cetopsid catfish that has completely lost 
its eyes and pigment but has enormously hypertro- 
phied olfactory organs. There are two new groups 
of very small, microphthalmic, channel-dwelling pi- 
melodids: Horiomyzon retropinnatus (Fig. 10C; 
Stewart, 1986), with wing-like pectoral fins, and an 
undescribed clade with at least four species bearing 
thickened bones and skin studded with sensory or- 
gans. 

At the other end of the size spectrum, there are 
occasional discoveries of large to very large fish 
species. Toledo-Piza (1997; Toledo-Piza et al., in 
press) has determined that the well-known fang- 
toothed cynodontine Hydrolycus, long considered 
monotypic, contains at least four species of which 
two are undescribed. Lundberg and Akama (1999, 
and in prep.) have identified an unrecognized spe- 
cies of “goliath” catfish, Brachyplatystoma (Fig. 
10D), in the central and lower Amazon. This im- 
portant species is now known to reach close to a 
meter in length, and it is among the most common 
in the commercial and artesanal fisheries where it 
is confused with B. filamentosum. 


Annals of the 
MESE Botanical Garden 


кк 10. 
fishes 


long ^ rode ed wi ith pe 
996 


Recent discoveries of very small and large 
the Amazon. —A myzon akamai, 
rmission n from 


14 mm 
Friel & Lundberg, 


~ 
X 


S 
N 
N 
© 
= 
т 
® 
LI 
= 
S 
— 
5 
= 
= 
5 
& 
= 
= 
z 
~ 
= 
= 
ge 
= 
% 
О 


produced with permission from Stewart, . —D. Bra- 
chyplatystoma sp., specimen about 500 mm long. (Сору- 
right John С. Lundberg.) 


- 


Novel aspects of reproductive biology are being 
discovered in Neotropical fishes. In addition to the 
continuing discovery of new genera and species 
& Menezes, 1998; Malabarba, 
1998), recent investigations of the glandulocaudine 
and cheirodontine Characidae have turned up a 


see Weitzman 


new mode of reproduction for characiforms. These 
with 
males developing a variety of specialized, court- 
ship-related features of the caudal fins such as bent 
fin rays with hooks and fleshy tissues that appear 
to be glandular (Fig. 11A; Burns et al., 1995, 1997; 
Malabarba & Weitzman, 1999). Almost all male 
glandulocaudines develop a cutaneous caudal 
gland thought to produce a pheromone, and the 
gland is surrounded by hypertrophied scales and in 
some species putative *pumping" 


fishes are convergently sexually dimorphic, 


muscles. Oddly, 
although the males lack any obvious intromittent 
organ, many of these species are internal fertilizers. 

The known limits of sexually dimorphic charac- 
teristics are being stretched by some Neotropical 
fishes. Recent collections made during the high- 
water season near Manaus have turned up breeding 
associations of electric fishes and morphologically 
intermediate individuals showing two cases of ex- 
treme sex dimorphism that have misled taxonomic 
decisions. The nominal species Aperonotus anas is 


‘igure 11. 


E p sexual dimorphism found in 
| X 


fishes. —A. Left, Xenurobrycon 


Fig 
Neotropical 
male о characin 
dal gland; right, 1 
in male Mimagoniates T (from Weitzman & Fink, 
1985; 


mac ropus, 
with a cau- 


odified scales surrounding caudal gland 


‚ 15.4 mm long, 


ermissio n to 


офисе images granted by Smith- 
The 


NMNH. Division: of Fishes). —B. 


sonian Institution, 


а ия exodon. (Copyright John 
undberg and John P. Sullivan). —C. Neblinichthys pilo- 


sus, male about 50 mm long (reproduced with permission 
from Ferraris et al., 1986). 


based on a large, hypermorphically long-snouted 
male of A. hasemani (Cox Fernandes, 1998; Cox 
Fernandes & Lundberg, 1999). The nominal, mono- 
typic Oedemognathus exodon is based on large 
males of Sternarchogiton nattereri that develop 
strong externalized teeth, perhaps for use in combat 
over mates (Fig. 11B; Cox Fernandes & Lundberg, 
in prep.). Ferraris et al. (1986) described a bizarre 
catfish, Neblinichthys pilosus 
(Fig. 11C), in which males develop on their snouts 
a cartoon-like bush of elongate odontodes, i.e., cu- 


loricariid armored 


taneous teeth. 

Among discoveries during the last 25 years, the 
greatest scientific interest attaches to previously 
unseen fishes that represent new phylogenetic lines 


Volume 87, Number 1 
2000 


Lundberg et al. 37 
Ichthyological Discovery 


at or above the family level. These discoveries have 
been most common among catfishes and often result 
in broad changes in higher classification. The lor- 
ic — Scoloplax dicra (Fig. 12A, B; Bailey & Bas- 
kin, 1976) was first described as a new subfamily 
of es 'ariidae, but was soon elevated to family 
rank by Isbrücker (1980). Three more Scoloplax 
species were described by Schaefer et al. (1989), 
and Schaefer (1990) determined that this clade is 
the likely sister taxon of the vast assemblage of 
Loricariidae + Astroblepidae. 

е Trichomycteridae presently include about 
200 species (perhaps over 100 in the large genus 
Trichomycterus) arranged in eight or nine subfami- 
lies (de Pinna, 1998). The most widely known tri- 
chomycterids are the *candirás? or “parasitic cat- 
fishes” that feed within the gill chambers of larger 
fishes on blood and epithelia. (Candirás finally 
earned their notoriety for supposed inadvertent en- 
try into the urethrae of humans with the first med- 
ically verified case of such behavior in. Brazil in 
1998.) Of more scientific interest, however, is the 
recent explosion of systematic knowledge of the 
perplexing diversity of trichomycterids. Although 
16 of the 27 trichomycterid species described be- 
tween 1975 and 1997 belong to Trichomycterus, the 
remaining 11 (4096) are placed in new or previ- 
ously rare, poorly known genera and subfamilies. 
The highly derived Trichogenys (Fig. 12C, D; Brit- 
ski & Ortega, 1983) has been placed in its own 
subfamily by Isbrücker (1986). Two 
scribed genera, Copionodon (Fig. 12E, F) and Gla- 


newly de- 


phyropoma, form a small, plesiomorphic clade of 
three species that de Pinna (1992) placed as the 
sister taxon to all other trichomycterids. The sub- 
families Sarcoglanidinae and Glanapteryginae, 
known from very few specimens, contain perhaps 
the most extraordinarily spec alized and diminutive 
catfishes in the world (Fig. 12G, n addition to 
major range extensions, in just the last decade the 
ranks of the sarcoglanidines were increased by four 
new monotypic genera (e.g., Fig. 12K, L; de Pinna, 
1989; de Pinna & Starnes, 1990; Costa & Bock- 
mann, 1994b; Costa, 1994), and two species and 
one genus were added to the glanapterygines (de 
Pinna, 1988, 1989; Costa & Bockmann, 1994a). It 
is no surprise that this remarkable amplification of 
trichomycterid diversity at the genus and family- 
group levels has caused a significant revision of our 
phylogenetic understanding (de Pinna, 1992, 1998; 
Stiassny & de Pinna, 1994). This case emphasizes 
the major impact that taxon sampling can have in 
Neotropical fish systematics (see also Schaefer, 


Discoveries of new species and new character 


evidence in known clades often lead to altered phy- 
logenetic arrangements and classifications. Exam- 
ples include the catfishes Helogenes, discovered to 
be closely related to the Cetopsidae (de Pinna & 
Vari, 1995), and Hypophthalmus (Fig. 13A), which 
is now confidently placed within Pimelodinae in- 
stead of in isolation in its own family (Howes, 1983; 
Lundberg et al., 1991). The first examinations (only 
within the last three years) of skeletal specimens of 
the monotypic, по Зао Francisco endemic Cono- 
rhynchos conirostris (Fig. 13B) have revealed that 
this supposed pimelodid, described in 1840, does 
not belong to any diagnosed family group of silur- 
iforms (Bockman, de Ferraris, 
comm., pers. obs.). There are noteworthy recent 
higher-level discoveries in other Neotropical groups 
as well. Stiassny (1991) has found synapomorphies 
that unite New World cichlids as the monophyletic 
sister taxon of some African cichlids. On the other 
hand, there is increasing evidence of multiple sis- 


inna pers. 


ter-taxon relationships between African and South 
American characiform and cyprinodontiform clades 
(Parenti, 1981; Vari, 1995; Buckup, 1998; Costa, 
1998) 

Thus, at every level knowledge of Neotropical 
fishes continues to grow at remarkable rates: myr- 
iad species, their intra- and intercontinental rela- 
tionships, and their richness and novelty of char- 
acteristics and natural history. Prospects for 
ichthyological discovery in South and Central 
America continue to be as high as ever. 


EUROPE 


Europe west of the Ural Mountains comprises 
roughly one-third of the Palearctic Realm (Fig. 2). 
The major groups accounting for most (ca. 8096) of 
the fish species are Cyprinidae (129), Salmonidae 
54), Coregonidae (43), Gobiidae (31), Cobitidae 
21), Petromyzontidae (11), Clupeidae (11), and 
Percidae (11). The “classical” checklists of Euro- 
pean freshwater fishes are outdated and reflect a 
state of knowledge and species concepts of 30—40 
years ago. Maitland (1976) listed 215 species for 
Europe west of the Urals and 170 species in Europe 


йл адар 


exclusive of the former U.S.S.R. In а recent review 
Kottelat (1997) recognized 358 species in Europe 
(excluding the former U.S.S.R.), an increase of 

Northern Europe, much of it glaciated during the 
Pleistocene, has low fish diversity, usually with only 
2 or 3 native species in any watershed. The number 
of species increases to the south. North of the Alps 
from France to Russia, the fauna of about 30 spe- 
cies is relatively uniform. The Danube basin, with 


38 Annals of the 
Missouri Botanical Garden 


(251 
iu 


А 
NS АА 


Figure 12. Recent discoveries of phylogenetic importance among South American sc oloplac ‘id and trichomycterid 
à , B. The sc oloplac ча Scoloplax dicra, scale bars | mm (reproduced with permission from Bailey & Baskin, 
1976). —C, p. The trichogenine Trichogenys longipinnis (from Britski & Ortega, 1983). —E, F. The горот 
Copionodon pecten, about 60 mm long (reproduced with permission from de Pinna, 1992). —G, H. The glanapterygine 
E an киша, about 55 mm long (reproduced with permission from de Pinna, 1989). — 1, J. The ТИ ine 
амтигта camposi, about 38 mm long (reproduced with permission from de Pinna, 1988). —К, L. The sarcoglanidine 
Stauroglanis gouldingi, about 23 mm long (reproduced with permission from de Pinna, 1989). 


el 
T 


Volume 87, Number 1 
2000 


Lundberg et al. 39 
Ichthyological Discovery 


Figure 13. 
among long-known South 


а fimbriatus. — В. Соп 


Recent discoveries of phylogenetic impor- 

бн ‘an catfishes. —A. 

orhynchos conirostris. 
2 


(Copyrights John G. Lundberg and jue. P. Sullivan 


about 90 species, has the most diverse fauna on the 
continent. The fish faunas of the southern European 
peninsulas contain relatively high numbers of en- 
demic species but are otherwise species-poor as- 
semblages: 87 species in the Balkan Peninsula, 29 
in the river basins of the Italian Peninsula, and 26 
in the Iberian Peninsula. The only endemic Euro- 
pean fish family is the cyprinodontiform Valenci- 
idae, with two species. This fauna has its closest 
relationships with Asia, North America, and north- 
ernmost Africa. 

The cornerstones of European fish systematics 
were provided in the 16th century in Rondelet's 
Libri de piscibus (1554) and Gesner’s Nomenclator 
aquatilium (1560) and Fischbuch (1563). Modern 
European ichthyology started with Peter Artedi's 
Ichthyologia (1738), whose system and nomencla- 
ture were adopted by Carl von Linné in 1758. 

Having thus enjoyed 450 years of study (about 
double that of any other continental fish fauna), one 
would expect that our modern systematic knowl- 
edge of European fishes is definitive. This is not 
the case, and even the most basic question, “How 
many species?," has no easy answer. That approx- 
imately 2000 names have been applied to Euro- 
pean fish species suggests that their taxonomy is 
not simple (Kottelat, 1997). As with North America, 
the discovery of previously unknown fish species in 
European waters is now uncommon. How then do 
we account for great increase in the number of valid 
European fish species proposed by Kottelat (1997)? 

First, there have been some recent discoveries 
of a few previously unknown species. Twenty spe- 
cies were discovered in European waters between 


gure 14. Valid European cyprinid species previous- 


ly dnd by копу us . Scardinius acarnanicus, 152 
m long. —B. Scardinius graecus, 140 mm long. (Copy- 
rights Maurice Какы t.) 


1978 and 1998, and some 10 known species are 
still unnamed. These are cyprinids, cobitids, and 
gobiids of small to very small size (3 to 10 cm). 
Most were found in southern Europe on the Iberian, 
Italian, and Balkan peninsulas. In 1998, two new 
cyprinids and one new cobitid were described from 
the Iberian Peninsula. 

Second, several species described by earlier au- 
thors and considered invalid by later authors, have 
since been demonstrated to be distinct. This also 
mainly concerns species from southern Europe. For 
example, the species of the cyprinid genus Scar- 
dinius have all been treated as synonyms of S. er- 
ythrophthalmus. It appears that at least five species 
should be recognized, some of which are conspic- 
uously distinct from 5. ү (Fig. 14) 
(Iliadou et al., 1996; Kottelat, 1 ) 

Another interesting case is the E Carassius 
gibelio, originally described from Germany in 1782. 
This species is traditionally considered as either 
the wild form of the goldfish, a feral goldfish, or a 
hybrid. These theories ignore the Japanese litera- 
ture on morphology and karyotypes of goldfishes 
that documents the existence of at least five species 
of Carassius in east Asia, all of them distinct from 
C. gibelio (see Hosoya, in Nakabo, 1994, for a sum- 

ary). They ignore the fact that C. gibelio was ap- 
pret recorded in European waters as early as 
9 (Kentmann’s Codex; Hertel, 1978), long be- 
E goldfish were first imported to Europe in 1611 
or 1691, and maybe even before it was imported 
from China to Japan (between 1502 and 1748). The 
hybrid theory does not seem to have support. So 
given our current knowledge, there is no alternative 
but to recognize Carassius gibelio as a distinct spe- 


cies. 
Third, application of the Phylogenetic Species 


Annals of the 
Missouri Botanical Garden 


Concept (PSC) has played a role in increasing the 
number of recognized ee (Kottelat, 1997). In 
this view, a species is “an irreducible (basal) clus- 
ter of organisms diagnosably distinct from other 
such clusters, and within which there is a parental 
pattern of ancestry and descent” (Cracraft, 1989: 
34-35). Most earlier work on European fishes ei- 
ther used some sort of implicit Biological Species 
Concept or, more frequently, was not explicit re- 
garding concepts, definitions, and criteria. Incon- 
sistencies in application of criteria are common. 
Detailed studies of geographic variation and sys- 
tematics reveal that fish diversity often has been 
underestimated, and there is no theoretical or prac- 
tical advantage in underestimating biodiversity. 
Subspecies of earlier authors that satisfy species 
criteria under the PSC must be treated as species. 
Further, some traditionally recognized subspecies 
have been found to be nothing more than arbitrarily 
delimited sections of clines, and these entities must 
be abandoned because there is no sense in recog- 
nizing arbitrarily defined units. 

e systematics of salmonids and coregonids has 
been notoriously difficult because of real or per- 
ceived high degrees of character plasticity and fail- 
ure to find consistent morphological differences 
among populations. Most earlier treatments avoided 
dealing critically with the systematics of salmonids 
and coregonids, and this resulted in a much un- 
derestimated number of species. Some “classical” 
lists merely include a single coregonid taxon, *Cor- 
egonus sp." The species-level systematics of these 
families is still far from resolved, but a pragmatic 
handling of their taxonomy is possible. Using a few 
simple concepts and definitions, Kottelat (1997) 
tentatively recognized 44 monophyletic evolution- 
ary units within the genus Coregonus in Europe that 
can be recognized as species (cf. 7 species recog- 
nized by Blanc et al., 1971), and 27 within the 
genus Salmo (cf. 5 recognized by Stearley & Smith, 

). Careful comparisons often show constant 
differences in morphology and in color patterns, 
and this is now supported by molecular data (e.g., 
Bernatchez & Dodson, 1994; Bernatchez et al., 
1992). The distinctive endemic species of Salmo 
provide examples (Fig. 15). Among these, S. trutta 
(Rhine basin) and S. rhodanensis (Rhóne basin) oc- 
cur in creeks only a few kilometers apart along the 
divide between the two basins in western Switzer- 
land. These species have always been empirically 
treated as different on the basis of coloration and 
morphology, a fact corroborated by molecular data. 

Some may object that it will be impossible to 
handle the large number of names that application 


of the PSC will produce. This is, however, not the 


Fig e 15. 


c of three endemic and one wide- 


pean Salmo. —A. Salmo carpio, 
33 mm long, endemic to Lake Garda, Italy. — 
. Salmo peristerus, 236 mm long, endemic to the Lake 
B евра basin, Greece-Albania-FYROM. —С. Salmo rho- 
danensis, 305 mm long, endemic to the Rhóne basin. — 
D. Salmo trutta, 235 mm long, spec imen from Rhine ba- 
sin, Switzerland, caught about 5 km from C. (Copyrights 
Maurice ud ) 


point; if they are different species we have to han- 

le them as species, give them names as species, 
and manage them as species. The problem is not 
the names, but with our perceived limitations. Ar- 
tificially squeezing all trout species into the same 
pigeonhole risks confusing critical information 
about species distinctions. If details of ecology of 
one species are mixed with the migration data of 
another and the reproduction of a third one under 
a single name, it is not surprising that patterns of 
species diversity are blurred. 

Salmonoids are notorious also for the existence 
in many lakes of several sympatric stocks that have 
been о considered as different “forms” of 
a single species. In many cases, the “forms” are 
not only MADE M different, but have differ- 
ent habitats, prey on different organisms, have dif- 
ferent spawning seasons and spawning grounds, 
and are genetically distinct (but few have been in- 
vestigated on this last aspect). Under any species 
concept, they are different species, and several of 
i thus are groups of 


these “polymorphic species” 


Volume 87, Number 1 


Lundberg et al. 41 
PAM D Discovery 


species or possibly species flocks. Presently, the 
best example is probably the three or four species 
of Salvelinus of Lake Thingvalla in Iceland. Euro- 
pean examples are the Salmo of Lough Melvin (Ire- 
land) and Lake Ohrid (Albania and the former Yu- 
goslavian Republic of Macedonia) and the 
Coregonus of Lake Konstanz and Lake Thun. 


AFRICA 


According to current estimates, Africa (Fig. 2) 
harbors some 2850 species arrayed, depending 
upon taxonomic convention, within 40—50 families. 
While perhaps not as rich in species as other trop- 
ical regions, what Africa lacks in terms of raw num- 
bers it makes up for in an ichthyofauna that in- 
cludes some striking examples of evolutionary 
phenomena ranging from relictual “living fossils” 
and strange morphological isolates, to stunningly 
diverse species flocks exhibiting unparalleled spe- 
ciation rates and adaptive radiations. It is notewor- 
thy that African fish diversity is marked by an in- 
teresting preponderance of archaic, phylogenetically 
isolated taxa. Fishes such as bichirs (Polypteridae), 
lungfishes (Protopteridae), the butterflyfish (Panto- 
dontidae), weakly electric elephantfishes (Mormy- 
ridae), the denticle herring (Denticipitidae), the 
snake mudhead (Phractolaemidae), and knerids 
(Kneriidae) are among the many examples found 
today only in African freshwaters. Perhaps more fa- 
miliar, and certainly more numerous, are members 
of the two major freshwater fish radiations: the oto- 
characiforms, and siluriforms 


— 


physan cyprinids, 
and the percomorph cichlids. In Africa an inter- 
esting taxonomic dichotomy between riverine and 
lacustrine communities exists. The former are dom- 
inated by otophysans: cyprinids, characiforms, and 
a few dominant catfish families (Clariidae, Mochok- 
idae, and Claroteidae), along with mormyrids and 
cyprinodontiforms (particularly in rivers of west and 
central Africa). On the other hand, Africa's many 
lacustrine ecosystems, particularly those of the 
Great Rift Valley, are almost completely dominated 
by remarkable adaptive radiations of cichlid spe- 
cles. 

ven the enormous size of the African conti- 
nent, an ichthyological tally of 2850 species, al- 
though unquestionably an underestimate of actual 
species richness, is remarkably low. By way of par- 
tial explanation, the common vision of Africa’s in- 
land waters as characterized by the great Nile, Ni- 
ger, Congo (Zaire), and Zambezi Rivers is 
misleading. In fact, over 90% of Africa’s rivers are 
less than 9 km in length and many flow only sea- 
sonally. Surprisingly, Africa has more arid and 


semi-arid area than any other continent on the 
planet, and as a result the great majority of Africa’s 
sh biodiversity is concentrated in the tropical, 
moister (and historically forested) regions. 

Roberts (1975), building on foundations laid by 
Boulenger (1905) and Pellegrin (1911, 1921), di- 

vided Africa into 10 so-called ichthyofaunal prov- 
inces reflecting broad areas of endemicity. Green- 
wood (1983) has likened the state-of-the-art to 
bioaccountancy rather than biogeographic analysis, 
and Leveque (1997) noted recently that the bioge- 
ography of African freshwater fishes remains poorly 

nown. The problems are manifold: new taxa are 
constantly being discovered, distribution patterns 
for most taxa are poorly known, and range exten- 
sions often encompassing thousands of kilometers 
are far from uncommon, but perhaps most impor- 
tantly there is a profound lack of adequate hypoth- 
eses for the phylogenetic relationships of the spe- 
cies involved. Despite these shortcomings, the 
ichthyofaunal provinces as currently recognized 
(see Leveque, 1997, for an up-to-date summary) are 
a useful first-pass approach to subdividing the ich- 
thyofauna into meaningful biogeographic units. Un- 
questionably, the fauna is phylogenetically richest 
and most varied in the rivers of tropical west and 
central Africa, with numbers attenuating in the 
provinces to the north and south of the tropical 
band. However, in terms of raw species numbers, 
it is the cichlids of the lakes of east Africa that 
dominate the continent's inland waters. 

Overall African fishes exhibit a very high level 
of endemism; almost 100% at the species level and 
upward of 40% at the familial, no doubt a reflection 
of the long-term stability and isolation of the Afri- 
can continent. To the extent that extracontinental 
affinities are known, it seems that the dominant re- 
lationship is with Asia (e.g., Notopteridae, Cypri- 
nidae, Bagridae, Schilbeidae, Clariidae, Channidae, 
Anabantidae, Mastacembelidae, Scatophagidae, 
pellonuline clupeids, and etropline cichlids), al- 
though links with South America (e.g., Lepidosiren- 
idae, arapaimid osteoglossomorphs, some characi- 
forms, siluriforms, cyprinodontiforms, and cichlids) 
and Europe (some cyprinid genera) are also evident 
in a few clades. Unfortunately, our current under- 
standing of the phylogenetic relationships of most 
African clades is insufficient to enable historical 
interpretation of many distributional data. 

While the first African fishes to reach European 
collections were assembled by the French naturalist 
Michel Adanson during his stay in Senegal (1749— 
1753), much early exploration in Africa was cen- 
tered on the Nile, a river that has played a major 


42 Annals of the 
Missouri Botanical Garden 


Number of species described 


0 тиз т == Т LI Li т Тт 
1758- 1779- 1799- 1819- 1839-  1859- 1879- 1899-  1919- 1939- 1959.  1979- 
1768 1788 1808 1828 1848 1868 1888 1908 1928 1948 1968 1988 


Species discovery plotted for three African fish clades (Citharinidae, Mormyridae, and Cichlidae). Black 
bars in ndic rate the number of species described in 10-year increments from 1748 until 1998. Data compiled mainly from 
Eschmeyer (1998). 


role in the history of Mediterranean civilization (see 1992) for most of west Africa and Skelton (1993) 
Beadle, 1974, for an excellent discussion of early for southern Africa (including the Zambezi prov- 
exploration of Africa's inland waters). Паре! (1994) ince), have comprehensive coverage. Additionally, 
recounted an amusing anecdote in which it is much up-to-date regional information is to be found 
claimed that on Geoffroy Saint-Hilaire's return from in Teugels et al. (1994), and references to major 
the Napoleonic Campaigns in Egypt (1799) his fish — revisional studies are to be found in Leveque 
collections were deposited at the Paris Museum. (1997). As was noted earlier, the great majority of 
hen Cuvier examined for the first time the ex- African species are members of just two groups: the 
traordinary bichir (Polypterus), he is reputed to — otophysan cyprinids (ca. 475 species in 23 genera), 
have declared, “This alone is justification for the — characiforms (characids and citharinids with ca. 
Egyptian Campaign." Whatever the importance of 208 species in 39 genera), and clariid (ca. 74 spe- 
nilotic fishes, the real beginnings of modern African cies in 12 genera), clarioteid (ca. 98 species in 18 
ichthyology are perhaps best marked by the pub- genera), and mochokid (ca. 167 species in 10 gen- 
lication of Cuvier and Valenciennes’ multi-volume era) catfishes; and the percomorph cichlids (ca. 870 
Histoire naturelle des poissons (1828-1849), in species in 143 genera). Together these seven fam- 
which about 140 African fishes were described. ilies comprise more than two-thirds of the conti- 
Some 50 years later when Boulenger published his — nent's ichthyofauna. 
four-volume Catalog (1909-1916) he was able to Despite almost 200 years of scientific study, 
record over 1400 species. And by 1991, with the there remains a remarkable amount to learn about 
publication of the last in an extremely useful series — African fishes, and many surprises no doubt re- 
of checklists (Базе! et al., 1984, 1986, 1991), the main. At the most basic level, monitoring species 
tally had risen to about 2850. Although Boulengers ^ numbers (Fig. 16) illustrates the cumulative dis- 
Catalog remains the only pan-African faunal study, covery rates for three typical African fish clades 
numerous regional accounts have since been pub- and, as elsewhere in tropical freshwaters, species 
lished. Two in particular, Leveque et al. (1990, discovery continues, sometimes at an extraordinary 


Volume 87, Number 1 
2000 


Lundberg et al. 43 
а Discovery 


Ole Seehausen 


Figure 17. 
with permission; иын Ole Seehausen. 


rate. For example, recent sampling in east Africa’s 
Lake Victoria has revealed the existence of a pre- 
viously unknown assemblage of over 120 stenotopic 
rock-dwelling cichlid species (Fig. 17) (Seehausen, 
1996; Seehausen et al., 1998). The quite unex- 
pected discovery of this extraordinary radiation not 
only has major implications for our understanding 
of the evolutionary dynamics of Africa's cichlid 
flocks (Kaufman et al., 1997; Turner, 1997) and 
their conservation (Kaufman, 1992; Kaufman & 
Ochumba, 1993; Seehausen et al., 1997), but it also 
illustrates that even in supposedly well-sampled lo- 
cales there still remain many unknowns. 

Equally illustrative is a recent study of the Cross 
River drainage of Cameroon and Nigeria. Although 
the Cross was considered reasonably well sampled 
when Teugels et al. (1992) thoroughly reviewed re- 
cently collected and historical material in muse- 
ums, they found that previous figures underesti- 


^cent disc оуегу of an unknown assemblage 


Lake Victoria. (Reproduced 


of rock-dwelling cichlids in 


mated true diversity by more than 70% (Stiassny, 
1996). Even after that comprehensive study new 
taxa are turning up. For example, Stiassny, Schliew- 
en, and Freihof (in prep.) are describing an enig- 
matic new genus of cichlid fish recently collected 
in the Cross (Fig. 18). This taxon, which is highly 
distinctive in appearance, displays a fascinating 
combination of character states that render its phy- 
logenetic placement problematical and may result 
in significant changes to current views of relation- 
ships among tilapiine cichlids (a major clade of Af- 
cichlids of considerable economic 


rican impor- 


ance). Another recent discovery altering our view 


= 


of the evolutionary dynamics of tilapiine cichlids is 
that of the Lake Bermin species flock (Fig. 19). 
ake Bermin, a small Cameroonian crater lake of 
little more than 0.5 km? has recently been deter- 
mined to be home to the continent’s first species 


— 


flock of substrate-spawning tilapiines. The flock in- 


Annals of the 
Missouri Botanical Garden 


Teugels « et al. 


ЖЕ 


85 


Estimated Number of species 
8 


ANN 
bs zie 


00: 


m 


Before 1992 study 


After 1992 study 


~ 


е 18. о estimates of fish diversity in the Cross River basin of West 
992). 


cludes the smallest known tilapia, Tilapia snyderae, 
a species that attains sexual maturity at only about 
25 mm. The discovery of endemic cichlid flocks in 
small crater lakes in the Cameroonian highlands 
provides a marvelous series of models for the study 
of evolutionary diversification and speciation mech- 
anisms (Trewavas et al., 
1992). For example, Schliewen et al. (1994) rec- 
ognized the Lake Bermin and Lake Barombi Mbo 
cichlid flocks as providing the most compelling ex- 


1972; Stiassny et al., 


Africa. 


Data compiled from 


amples of probable sympatric speciation known 


among vertebrates 


The cichlid radiations of the 
“living laboratories of evolution 


likened to 


east African lakes, 
es pro- 


vide some of the most extraordinary examples of 
vertebrate speciation, adaptive radiation, and eco- 
morphological diversification known on the planet 
and have intrigued biologists since their initial dis- 
covery at the end of the last century (e.g., Moore, 
1903; Fryer & Iles, 1972; Fryer, 1976; Greenwood, 


Range of interspecific variation 
in LPJ morphology among 


Lake Bermin tilapias 


T.snyderae 
7 


А > 
DEL 


T T thysi 


2 117 у ^, ^ / т hi { у 
tte S T bemim 
nel 


Figure 19. 


The Lake Bermin (Cameroon) Tilapia species flock. Data and illustrations after Stiassny et al. (1992). 


Volume 87, Number 1 
2000 


Lundberg et al. 45 
Ichthyological Discovery 


а Whee ли ы 
В. brevicephalus 


CLERK 
B. peer 


Volume-percentage 


т 
УУУ 


Analysis of gut contents of Lake Тапа 
Barbus species 


Figure 20. 


1981; Coulter et al., 1986; Galis, 1998; Stiassny & 
Meyer, 1 e recent explosion in molecular 
sequencing techniques and analytical methods has 
resulted in a plethora of studies aimed at unravel- 
ling phylogenetic, biogeographic, and speciation 
mechanisms among these fishes (e.g., Sturmbauer 
& Meyer, 1992; Meyer, 1993; Kocher et al., 1993; 
1994; Kornfield & Parker, 1997; 


1996, and references therein). The 


Schliewen et al., 
Verheyen et al., 
results have been stimulating to the field generally, 
have added significantly to our understanding of 
evolutionary modes and speciation mechanisms of 
the lake cichlids, and have served to highlight the 
important roles for both morphological and molec- 
ular analyses. 

Another recent discovery of considerable interest 
and evolutionary implication is found in Ethiopia’s 
Lake Tana, the highland source of the Blue Nile. 
In an elegant series of taxonomic and ecomorphol- 
ogical studies of trophic diversification, Nagelkerke 
and his colleagues have established the existence 
of an ecologically and reproductively segregated ra- 
diation of 14 species of the cyprinid genus Barbus 
(Fig. 20; Nagelkerke et al., 1994; 
1997). Since the demise of the Lake Lanao cyprinid 
radiation in the Philippines, the Lake Tana flock 
becomes the planet’s only remaining lacustrine cyp- 
rinid radiation. 

Lest the impression remains that new discovery 
is limited to Africa’s lakes, a few riverine examples 
serve to redress the balance. Recent work that com- 


agelkerke, 


The Lake Tana (Ethiopia) Barbus species floc 


k. Data and illustrations after Nagelkerke (1997). 


bines neurobiology, behavior, and morphology has 
revealed remarkably rich communities of sympatric 
mormyrid taxa living in small rainforest rivers of 
west and central Africa (Hopkins, 1981; Moller, 
1995). This is especially well illustrated for species 
in the genus Brienomyrus from Gabon (Fig. 21), 
where 11 new species have been discovered by re- 
cording distinctive electric organ discharges (Al- 
ves-Gomes & Hopkins, 1997) and detailed mor- 
phological study (Teugels & Hopkins, 

Teugels & Hopkins, in prep.). Mormyrids ереси 
electric pulses many times per second for purposes 
of active electrolocation, and as a consequence spe- 
cies-typical and sex-specific signals are broadcast 
nearly continuously. The diversity of wave patterns 
among sympatric species suggests that individuals 
exploit differences in waveform, polarity, and du- 
ration to identify the species and sex of signalers. 
Playback experiments using computer-synthesized 
EODs have confirmed their importance in repro- 
ductive isolation and sex recognition (Hopkins & 
Bass, 1981). There are even individual differences 
in EODs that are sufficient to permit the tracking 
their in small 
streams monitored daily (Friedman & Hopkins, 
1996), thus enabling non-intrusive ecological mon- 


of individuals and movements 


itoring of populations. 

Electric catfishes (family Malapteruridae) are en- 
demic to Africa and are one of the continent's ich- 
thyological icons. They owe their common name to 
their peculiar ability to produce stunning discharg- 


46 


Annals of the 
Missouri Botanical Garden 


Stomatorhinus — 


| ¢ 


)  Boulengeromyrus кповртел 

l Mormyrops zancltrostris 
NĚ 

} Isichthys henryi 

үү ——$% 


Г ... Polimyrus marche: 


. Marcusentus тооги 


Ivindomyrus opdenboschi 
Г б 

ү 
Paramormyrops gabonensis 
а 


| Petrocephalus simus 


| Petrocephalus stuhimani 


Marcusenius сопсерћа 5 


i Brienomyrus kingsieyae 


> Bnenomyrus sp. 7 


а. 
„Л Brienomyrus longicaudatus тб 
V 47 ~ 
—— M S. 
ү SE 


А Brienomyrus sp 1 
лава 
| 


) ~ Brienomyrus sp 2 
E v 


р А Brienomyrus sp. 3 


Bnenomyrus sp. 6 


0 5 10 


e 2l. 


Mormyrid diversity in the Ivindo River (Gabon). Species-specific 


Electric Organ Discharges (EODs) 


Mei in middle columns. (With permission and modified after Hopkins, 1981.) 


es, up to 350V in large specimens. The first species 
was formally described 200 years ago as Silurus 
(now Malapterurus) electricus, and for much of the 
time since then a single species was recognized. 
However, electric catfishes are widely distributed 
across Africa, and careful revisionary work (Norris, 
in prep.) has recovered a complex of upward of 20 
species previously unrecognized in museum collec- 
tions and cataloged simply under the name M. elec- 
tricus. Interestingly, the diversity of electric catfish 
species matches that of other groups conforming to 
the basic division of ichthyofaunal provinces of the 
continent. Beyond taxonomic diversity, Norris has 
uncovered a large amount of ecological and ana- 
tomical variety. For example, some taxa are highly 
depressed bottom-dwellers, others are fusiform and 
presumably active swimmers. By far the most as- 
tonishing discovery in this family is a clade of 
dwarf forms in which females can be gravid at sizes 
as small as 6 cm, a size range in which specimens 
of other species show little gonadal differentiation. 
Most of the dwarf catfishes have reduced cephalic 


and lateral line systems, elongate adipose fins, and 
a highly unusual three-chambered swim bladder. 
The function of the three-chambered bladder is as 
yet unclear, but Norris has suggested that it may 
play a role in the acoustic biology of these fishes, 
either in sound reception or production, or both. 

There is little doubt that a similar taxonomic 
complexity will be revealed when other supposedly 
widespread "species," such as the enigmatic char- 
acoid Hepsetus odoe, are subject to the same sort of 
critical review. New species, and even whole new 
communities, abound in Africa, and their discovery 
and documentation will continue to require effort 
and render important results. But without doubt the 
greater scientific challenge will be the unraveling 
of the phylogenetic relationships of the African ich- 
thyofe auna. Such information is critical to our un- 
derstanding of the evolutionary history of the fauna, 
the geologic history and relationships of the conti- 
nent, and increasingly, as an aid in informing con- 
servation work. In all these aspects, this is a task 
that has hardly begun. 


Volume 87, Number 1 
2000 


Lundberg et al. 47 
Ichthyological Discovery 


The Malagasy region has long been recognized 
as a biotic entity distinct from that of mainlan 
Africa. The region includes the Grande Ile of Mad- 
agascar and its offshore islands, the volcanic Mas- 
carenes and Comores, and the granitic Seychelles. 
As noted by Roberts (1975) the region, which is 
something of an enigma to biogeographers, harbors 
a highly endemic ichthyofauna. More recently 
Stiassny and Raminosoa (1994) provided a review 
of the fauna and, in addition to recognizing many 
as occupying phyloge- 
netically basal positions in their respective clades, 
they noted the enigmatic absence of a great many 
of the families currently represented in Africa and 
India. Because of its singular nature, the Malagasy 
ichthyofauna is not treated in detail in this essay. 
For more information on the Malagasy ichthyofauna 
see Teugels et al. (1985), Stiassny and Raminosoa 
(1994), and Benstead et al. (2000) 


of the Malagasy endemics 


TROPICAL ASIA 


As treated here Tropical Asia approximately 
equals the Oriental Realm, extending from the In- 
dus basin eastward to South China and to the Mo- 
luccas (ca. *Wallace's Line") in Indonesia (Fig. 2). 
The dominant groups among primary and secondary 
freshwater families are Cyprinidae (about 1000 
species), Balitoridae (about 300), Cobitidae (about 
100), Bagridae (about 100), Osphronemidae (85): 
and among peripheral families, Gobiidae (about 
300). This pattern applies principally to the main- 
land areas and in the insular parts to the major 
river basins draining onto the continental shelf. On 
oceanic islands and on the mainland in streams not 
draining to the continental shelf, the proportion of 
primary freshwater fishes is lower, and there is an 
increase in representation of peripheral families. In 
the Moluccas, Sulawesi, and most Philippine is- 
lands, there are no primary freshwater fishes, and 
gobies constitute about half of the inland fish fauna. 

While recognizing an element of arbitrary taxo- 
nomic convention in doing so, fish families are of- 
ten used as a rough guide to higher-level taxonomic 
diversity of faunas. A distinguishing feature of the 
Tropical Asian inland fish fauna is its high number 
of families. One-hundred-twenty-one families have 
been recorded from inland waters, either as per- 
manent, temporary, or occasional residents (cf. 45— 
50 in Africa and about 55 in South America). Thir- 
ty-four of these families are primary or secondary 
division freshwater fishes, and 18 are endemic to 
Southeast or tropical Asia. The remaining 87 fam- 
ilies are peripheral freshwater fishes, most repre- 
sented by few species. The high number of periph- 


eral families in Asian freshwater is explained by 
the existence in adjacent seas of the highest fish 
family diversity in the world. The northern tropical 
Asian ichthyofauna has some affinities with that of 
northern temperate Asia, with which it shares some 
cyprinid and cobitoid lineages, but on the whole, 
tropical Asia shares several lineages with Africa 


— 


g., Notopteridae, several cyprinid lineages, Bag- 
ridae, Schilbeidae, Aplocheilidae, Nandidae, Ana- 
bantidae, Channidae, Mastacembelidae). 

An estimated total number of species for tropical 
Asian freshwater fishes is 3000. More precise fig- 
ures have been compiled for some parts of South- 
east Asia (Irrawaddy, Salween, Mekong, Red River 
basins and intermediate areas, Hainan, Malay Pen- 
insula, the Philippines and Indonesia eastward to 
the Moluccas), where 2100 valid species are pres- 
ently known (Kottelat, in prep.). However, consid- 
erable differences exist in our ichthyological knowl- 
edge among regions and countries because of 
incomplete surveying and variable ichthyological 
practices among countries. 

The Indian ichthyofauna remains in need of in- 
depth systematic study. Recent work on Sri Lankan 
and southern Indian fishes indicates that many spe- 
cies thought to be widely distributed and conspe- 
cific with species originally described from north- 
eastern India are in fact assemblages of allopatric 
species (Pethiyagoda, 1991; Pethiyagoda, pers. 
comm.; MK, pers. obs.). In Kerala (South India) 
10-20% of the fishes in any basin of reasonable 
size are likely to be undescribed (Pethiyagoda & 
Kottelat, 1994). Habitat deterioration in this region 
poses a serious threat to most species, including 
those that are poorly known or unknown. 

The development of ichthyology in China has 
been largely independent of that in the rest of Asia, 
but many scientific names applied to China's fresh- 
water fishes are taken from adjacent faunas without 
adequate comparison. Survey work is very incom- 
plete, especially in the hilly areas in the south. 
Species diversity is underestimated especially for 
small-sized fishes. For example, the loach identified 
as Schistura fasciolata is probably an assemblage 
of at least half a dozen species. Unfortunately, eco- 
nomic changes in recent years have made illegal 
electric fishing widely available everywhere. This 
poses a serious threat to the survival of most en- 
demic hill stream fish communities. In 1996 one of 
us (MK) observed in western Sichuan that most 
small streams surveyed were without fish. 

Knowledge of the ichthyofauna of tropical Asia 
is still in an exploration and discovery phase. The 
fish fauna of Laos jumped from 216 species re- 
corded in the scientific literature as of early 199 


48 


Annals of the 
Missouri Botanical Garden 


to some 370 known by mid-1997 (Kottelat, 1998). 
This represents an increase of about 80% in about 
11 weeks of fieldwork. Some 85 of these species 
were unnamed, though 


of the family Balitoridae that inhabit hill streams 
and have a relatively small distribution area, often 
restricted to part of a single river basin. Similar 
figures can be recorded for most Asian countries 
(Kottelat & Whitten, 1996b). The situation is likely 
similar (but much more acute) in Vietnam, where, 
as in China, ichthyology has been largely isolated. 

Survey efforts in little-known areas and poorly 
sampled habitats continue to yield new discoveries. 
Kottelat et al. (1993) listed 964 species known in 
1 rom western Indonesian inland waters. Ап 
addendum published in 1996 added 79 species de- 
scribed in the interval, and others have been dis- 
covered but not yet reported. Changes in systematic 
or nomenclatural status affected 56 other species 
(Kottelat & Whitten, 1996a). Fieldwork in reason- 
ably remote areas easily results in the discovery of 
1 or 2 new species per actual day of fieldwork, but 
also in the discovery of an almost equal number of 
identification problems involving earlier known 


cies. 
“Sundaland” (Java, Borneo, Sumatra, Malay Pen- 
insula) has a rich fauna (total ca. 1200 species). 
This is due to its historical fragmentation into 
smaller isolated basins and very diverse habitats 
ranging from coastal mangroves to high altitudes. 
One unusual habitat type has been discovered to 
harbor a very specialized and diverse fauna. This 
is the peat swamp forest, characterized by a peat- 
like soil (not made of sphagnum, but of tree roots) 
with a morphology somewhat similar to northern 
peat bogs, culminating into peat domes with a cen- 
tral “lake.” The black water of these oligotrophic 
lakes is darkly tinted by tannin and has a very low 
pH (3 to 5). Once thought to be depauperate, there 
are now more than 100 species known exclusively 
from these habitats. Most are small fishes: fighting 
fishes (Betta), licorice gouramies (Parosphromenus), 
and numerous diminutive cyprinids. A particularly 
interesting discovery in the peat swamp is Bihun- 
ichthys monopteroides (Fig. 22), а chaudhuriid 
earthworm eel (Kottelat & Lim, 1994). This family 
was considered endemic to Inle Lake in Myanmar, 
ut it is now known from about a dozen species 
with an overall wide range in Asia. Bihunichthys 
monopteroides is the sais chaudhuriid (maxi- 
mum known size of 36 mm SL) with extreme re- 
duction or loss of scales, ег line, fins, and a 
number of cranial bones. It has fossorial habits in 
soils around the swamps. 


A diminutive, fossorial earthworm eel, Bi- 
blackwater 
a Ng; drawing by Kelvin Lim). 


Figure 22. 
hunichthys топоріе ur 5, from an 
swamp (photo by Peter 


Asian 


In tropical Asia, as in Africa and South America, 
discovery of miniature je is fairly 
3B). Danionella pelluci- 


common. 
Many are cyprinids (Fig. 2 
da from Myanmar and Boraras micros from north- 
east Thailand are adult at about 12 mm SL, and 
females of the loach Kottelatlimia katik are ripe at 
about 13 mm SL. In mangroves the smallest known 
species is an undescribed goby, Gobiopterus from 
Singapore, that reaches about 10 mm Minia- 
tures provide one of the most striking examples of 
discovery of a new fish clade, the family Sundasa- 
Wesen (Fig. 23C), described based on two species 

1981 by Roberts. These are tiny (25 mm SL) and 
cae fishes with many reduced structures, 
and, in places, they are extremely abundant. Four 
additional species have been found on Borneo since 
their first discovery, and Siebert (1997) provided 
evidence that the group is related to herrings (Clu- 
peidae). 

At the other end of the size scale, occasional 
large-sized species are still being described. Some 
of these were reported by early explorers, but it is 
only in recent years that they have been carefully 
diagnosed and described. In Asia an impressive ex- 
ample is Himantura chaophraya (Fig. 23A), the 
Mekong sting ray known to science since 1880 but 
just named in 1990 (Monkolprasit & Roberts, 

his immense fish can attain a disc width 
of 2 m, an estimated total length of 4 m, and a 
weight of 600 kg 

Probably the most interesting communities of 
Southeast Asian fishes discovered in recent years 
are the species flocks of the lakes of central Sula- 
wesi, Indonesia. A group of five tectonic lakes (Mal- 
ili lakes) and the connecting streams host a fauna 


Volume 87, Number 1 
2000 


Lundberg et al. 49 
sil eres Discovery 


Figure 23. Recent discoveries of very large and small 
бі in hs Asia. —A. Freshwater sting гау, Himan- 
tura chaophraya. —B. A tiny, undescribed cyprinid, 9.9 
mm long, from Bo . Sumatra, Malay Peninsula. — 
One of the Mig puer и EUN Sundasalanx аек 
hynchus, 23.8 mm long. (Copyrights Maurice Kottelat.) 


consisting of 26 known native species, all but 4 
endemic to this system. Within this lake system, 
two units can be distinguished, Lake Matano and 
lakes Towuti, Mahalona, and Wawontoa; only 2 en- 
demic fish species are shared by these two units. 

e fish species include 3 endemic Hemiramphi- 
dae (halfbeaks), 6 endemic Gobiidae (gobies 
demic Oryziidae (ricefishes), 


‚ 3 en- 


м 


4 endemic Telmath- 
erinidae (sailfin silversides); also present are two 
endemic genera and 4 non-endemic species (one 
each in the families Telmatherinidae, Gobiidae, 
Synbranchidae, Anabantidae). The lakes also con- 
tain at least 4 endemic crabs, a dozen endemic 
shrimps, about 60 endemic molluscs, and endemic 
ostracods, sponges, other invertebrates, and mac- 
po (Kottelat, 1990b, с, 1991; Larson & Kot- 
telat, 1992; Bouchet, 1995). 

The family Telmatherinidae includes five genera, 
two endemic to the lakes, one with eight species 
endemic to the lakes plus one riverine species oc- 
curring also in an adjacent basin, one monotypic 
genus endemic to southwestern Sulawesi, and an- 


Endemic lake s of Sulawesi. 


Figure 2 : 
courting pair ой Telma reni атотае. —B. 
scale-eating Telmatherina prognatha. (Copyrights Maurice 


Kottelat.) 


Fin-ray and 


other monotypic genus from баби | an island off 
Irian Jaya (the Indonesian рам 

(Aarn et al., 19 

matherinids are 


New Guinea) 
Ше the bis of all tel- 
“pelagic,” the adults live close to 
the bottom and are substrate spawners. They are 
strongly sexually dimorphic (Fig. 24A), and males 
of several species exhibit color polymorphism. One 
species (Telmatherina prognatha, Fig. 24B) appar- 
ently feeds on fin-rays and scales of other species, 
and another (T. sarasinorum) preys on eggs of other 
spe 
The и lakes of central Sulawesi, lakes Poso 
and Lindu, have less diverse fish faunas. Lake Lin- 
du apparently hosts a single native species, Xeno- 
poecilus sarasinorum while 
Lake 


three endemic 


(Adrianichthyidae), 
Poso is inhabited by two endemic Gobiidae, 
Adrianichthyidae, and two endemic 


Oryziidae (Kottelat, 1990a). The Adrianichthyidae 


are noteworthy for their breeding behavior. AI- 
though originally recorded as live-bearers, it now 


appears (Fig. 25A) that the female carries a clutch 
of eggs with her pelvic fins, in a ventral depression 
of the belly, for about 10 days until they hatch. 
Adrianichthyidae are closely related to Oryzii- 
dae, which are known from India to Japan and Ti- 


Annals of the 
Missouri Botanical Garden 


Figure 25. Recent a 'overies of oddly specialized 
. The 


fishes i in n tropical ris e adrianichthyid Xenopoe- 
cilius оорћоги: fertil n ized clutch of eggs. —B. Th 
cave-dwelling balitorid loach Schistura Oedipus from Thai- 
land. welling cyprinid Puntius sp. from 
Java. (Copyrights Maurice Kottelat.) 


~ 
у 


mor, in fresh and brackish waters. Interestingly, 
Adrianichthyidae are known on Sulawesi from that 
part of the island (the western half) that plate tec- 
tonics suggest was earlier connected to elements 
now constituting part of Southeast Asia. On the oth- 
er hand, the distribution of telmatherinid species 
(except one) on the eastern half of Sulawesi corre- 
sponds to a plate earlier connected with the New 
Guinean one, on which Misool is located, where the 
family is also represented by an endemic genus 
(Kottelat, 1991 


Tropical Asia contains several of the most exten- 


Мм 


sive karst areas of the world, and it is not surprising 
that at least 31 species (some undescribed) of cave 
fishes are known from this region and many more 
should be expected. Some of the largest karsts 
(Laos, Vietnam) have barely been explored speleo- 
logically. Sixteen species of cave fishes are known 
m China, seven from Thailand, four from India 
(three actually from wells), two from Indonesia, and 
one each from Malaysia and Laos. Most of these 
belong to genera known from surface waters, but 
five cannot be linked with known surface species 
and are treated as distinct genera. Noteworthy is 
the cyprinid genus Sinocyclocheilus, which has 
evolved independently at least seven hypogean spe- 
cies in China (Guizhou, Yunnan, and Guangxi prov- 
inces). The epigean species of Sinocyclocheilus are 
usually found associated with springs or in streams 
under overhanging rocky shores, which make them 
“pre-adapted” to colonize cave habitats. The known 
cave fishes in Asia belong to the families Balitori- 


dae (13 species, e.g., Fig. 25B), Cyprinidae (12, 
e.g., Fig. 25C), Cobitidae (2), Synbranchidae (2), 
Gobiidae (1), and Clariidae (1). 


AUSTRALIA AND NEW GUINEA 


Australia and New Guinea are perhaps better 
known for their diverse marine fish faunas than for 
their freshwater ones. Nevertheless, freshwater fish- 
es of Australia and New Guinea are distinctive, in- 
teresting, and have been the subject of significant 
ichthyological discovery over the past 20 or so 
years. 

With an area roughly equal to the contiguous 
‚ Australia (Fig. 2) is generally 
characterized by lower topographical relief, lower 


continental U.S 


rainfall, and a much less extensive system of river 
drainages. However, largely as a consequence of 
the latitudinal spread of the country (from about 
10°40’S to about 43?40'S), and of the influence of 
the Great Dividing Range on climatic conditions, 
Australia possesses a wide variety of freshwater 
habitats. These include artesian springs and 
ephemeral desert lakes and streams (Great Austra- 
lian Bight, Lake Eyre, and other internal drainag- 
es); tropical streams that undergo extensive flooding 
during summer monsoonal rains (northwestern Aus- 
rainforest 


= 


tralia and Gulf of Carpentaria drainages); 
streams (northeast coast); coastal streams and sand- 
dune lakes (east coast); and alpine lakes (Tasman- 
ia). The Murray-Darling River System is by far the 
largest in Australia, extending southwestward about 
1900 km from the interior of southern Queensland 
to the Southern Ocean. 

Stretching around 1600 km, New Guinea is the 
world’s second-largest island (Fig. 2). Biogeograph- 
ically and geologically, New Guinea is divided into 
more-or-less northern and southern provinces, sep- 
arated by an extensive mountain chain along the 
long axis of the island. This mountainous topogra- 
phy, in combination with high rainfall, results in 
numerous drainage systems, and a large array of 
freshwater habitats that include: short coastal 
streams, large lowland rivers, coastal swamps and 
floodplain lakes, alpine streams and lakes, and 
large highland rivers. 

Australia and New Guinea are much more than 
just adjacent land masses, as they have been con- 
nected throughout most of their history. The Sahul 
Shelf, beneath the shallow Arafura Sea and Torres 
Strait that now separate the two areas, was emer- 
gent until as recently as about 6000 years ago dur- 
ing the latest glacial lowering of sea level, and 
southern New Guinean streams were confluent with 
those of the adjacent Australian coast. 


Volume 87, Number 1 
2000 


undberg et al. 51 
Ichthyological Discovery 


Australian freshwater fishes were reviewed by 
Allen (1989), who recorded 187 species and sub- 
species; subsequent collecting and research have 
taken this count to around 215. The dominant 
freshwater taxa in the country are m gobioid fam- 
ilies Eleotrididae and Gobiidae (about 50 species), 
the superfamily Galaxioidea, an austral group of 
salmoniform fishes related to the northern smelts 
(26 species), Terapontidae, an Indo-West Pacific 
group of perch-like fishes (22 species), Percichthyi- 
dae, a family of southern Australian and southern 
South American perch-like fishes (21 species), Me- 
lanotaeniidae, an Australian-New Guinean endem- 
ic family (but, see below) of atherinomorph fishes 
(16 species and subspecies), Plotosidae, an Indo- 
West Pacific catfish family (about 15 species), and 
Atherinidae, a worldwide atherinomorph family (15 
species and subspecies). Other distinctive compo- 
nents include the Queensland lungfish, Neocerato- 
dus forsteri (Neoceratodontidae), and the bony- 
tongue (osteoglossid) species, Scleropages jardinii 
and 5. leichardti. 

Freshwater fishes of New Guinea were reviewed 
by Allen (1991), who listed 320 species, including 
some estuarine forms; subsequent collecting and 
research have taken this count to about 350. The 
most diverse freshwater taxa are eleotridids and go- 
biids (about 115 species), Melanotaeniidae (53 spe- 
cies), Ariidae, a circumtropical catfish family (21 
species), Terapontidae (16 species), Chandidae (— 
Ambassidae), an Indo-West Pacific family of small, 
perch-like fishes (15 species), Pseudomugilidae, a 
family of atherinomorph fishes endemic to Australia 
and New Guinea (13 species), 
(about 13 species). Most of these taxa are also spe- 
cies-rich in northern Australia, emphasizing the 
historical link between the areas; indeed, about 50 
species from southern New Guinea also occur in 


and Plotosidae 


northern Australia, and many of these are restricted 
to the two areas. 

The amount of recent ichthyological activity in 
Australia and New Guinea can be gauged conser- 
vatively from the number of recently described spe- 
cies and subspecies. Since 1970 about 70 Austra- 
lian (33% of the total) and about 125 New Guinean 
(36% of the total) freshwater fish species and sub- 
species were described or are awaiting description. 

Increased understanding of freshwater fish sys- 
tematics and distributions in the region has largely 
stemmed from the application of modern collecting 
and systematic techniques by professional ichthy- 
ologists. However, no less significant has been the 
input from amateur ichthyologists and aquarists. In 
particular, during the last three decades a profound 
increase in interest in keeping freshwater fishes has 


Some of the taxonomically challenging and 
fishes of Australia and New Сите 
cairns”) form lacking 
—B. Melanotaenia 
Guinea” form with distal stripe on anal 


Figu re 26. 
diverse rainbow 
ия maccullochi, typical (< 
distal stripe on anal fin, 53.0 mm long. 
maccullochi, “New 
fin. 33.0 mm long. 


ea. —А 


resulted in the formation of various specialty soci- 
eties (e.g., Australia New Guinea Fish Association, 
"ANGFA") and in numerous amateur collecting ex- 
peditions. Aquarist interest has particularly con- 
centrated on the endemic rainbowfishes (Melano- 
taeniidae, Fig. 26), and this, in combination with 
taxonomic and field studies by G. R. Allen, has 
resulted in a dramatic increase in the number of 
species recognized in this family: of the 67 species 
and subspecies currently recognized, 38 (57%) are 
either undescribed or were described since 1970. 
Other significant new discoveries in recent years 
have included: Oxyeleotris caeca (Fig. 27A), a blind 
butidid gobioid from sink holes and caves in the 
Upper Kikori New Guinea (Allen, 
1996); two new giant percichthyids of the genus 
Maccullochella from coastal drainages of southern 
Queensland and northern New South Wales (Row- 
land, 1993); Scaturiginichthys vermeilipinnis, a new 


iver, 


Papua 


genus and species of pseudomugilid from an arte- 
sian spring in central western Queensland (Invant- 
soff et al., 1991); four new gobiids of the genus 
Chlamydogobius (Fig. 278) from artesian spring 
systems in central Australia (Larson, 1995); and 
numerous lacustrine atherinid, eleotridid, and me- 
latotaeniid spec ies endemic to various lake systems 


n & Hoese, 1986; Crowley 


1996). 

There also have been significant advances in our 
understanding of the phylogenetic relationships of 
Australian and New Guinean freshwater fishes. For 
example, Johnson (1984) diagnosed a monophylet- 
ic, freshwater Percichthyidae that included the 
southern South American genera Percichthys and 


52 


Annals of the 
Missouri Botanical Garden 


Figure 27. 

coveries in the Australia-New Guinea region. —A. The 
cave-dwelling gobioid, Oxyeleotris caeca, 107 mm id 
from Papua New Guinea (based on Allen, 1996). —B. One 
of the artesian spring-dwelling n oe 
japala, 35.5 mm long, Northern Territory, Australia. —C. 
Bostockia porosa (based on Merrick & Se lit 1984). — 
D. Gadopsis marmoratus (based on Mer Schmida, 
1984). —E. The burrowing лык E Lepidogalax- 
ias ее from Western Australia (copyright by 
Tim ra). 


A potpourri of recent ic | dis- 


Percilia, and the southern Australian genera Bos- 
tockia (Fig. 27C), Gadopsis (Fig. 27D), Edelia, Nan- 
noperca, Nannatherina, Maccullochella, Percalates, 
Plectroplites, and Macquaria (the latter three com- 
bined into a probably paraphyletic Macquaria by 
MacDonald (1978) and most recent authors). Pre- 
viously (e.g., Gosline, 1966), the Percichthyidae 
had been a catchall for basal perciform taxa that 
included, among others, marine genera now as- 
signed to the Acropomatidae, Serranidae, Moroni- 
dae, and Polyprionidae. Johnson further hypothe- 
sized that the southeastern Australian Gadopsis is 
sister to the southwestern Australian Bostockia, and 
that these genera form a clade with the diminutive 
Edelia, Nannatherina, and Nannoperca (previously 


classified in the Kuhliidae). Prior to this, the rela- 
tionships of Gadopsis were enigmatic, and it had 
been variously ‘ed in its own order (Scott, 
1962), allied to ophidioids (Gosline, 1968), 
cated to a monotypic percoid superfamily (Nelson, 
1976), or suggested as a possible relative of the 
perciform suborders Trachinoidei or Blennioidei 
(Rosen & Patterson, 1969) 

No Australian freshwater fish has attracted as 
much attention from systematists as the southwest- 
ern Australian salamanderfish, Lepidogalaxias sal- 
amandroides (Fig. 27E). Originally described as a 
galaxiid (Mees, 1961), subsequent authors have ei- 
ther retained it within the Galaxiidae (or at least 
within the austral taxon Galaxioidei), or suggested 
radically different positions for the taxon, such as 
the sister of a clade consisting of the Northern 
Hemisphere freshwater families Esocidae and Um- 
bridae (Rosen, 1974), or in an unresolved trichot- 
omy with the Salmonidae and Neoteleostei (Fink, 
1984). However, recent studies have nested the ge- 
nus within the galaxioids. Johnson and Patterson 
(1996) concluded that Lepidogalaxias is the sister 
of the Tasmanian genus Lovettia, these together 
forming the sister of a clade consisting of the typ- 
ical Galaxiidae plus the southern South American 
genus Aplochiton. Williams (1997) concluded that 
Lepidogalaxias is the sister of a clade consisting of 
the typical Galaxiidae and the Aplochitonidae (the 
latter including Aplochiton + Lovettia). 

No less controversial in recent years has been 
the position and composition of the family Melan- 
otaeniidae. This has particularly centered around 
its relationships to other atheriniforms, notably to 
the Australian—New Guinean family Pseudomugil- 
idae, the western Indonesian and New Guinean 
family Telmatherinidae, the Madagascan family Be- 
dotiidae, the southeast Asian Phallostethidae, and 
the West Pacific Dentatherinidae (e.g., Allen, 1980; 
Parenti, 1984b; Stiassny, 1990; Saeed et al., 1994; 
Dyer & Chernoff, 1996; Aarn & Ivantsoff, 1997). 

There has been a substantial increase in our un- 
derstanding of the biology of Australian and New 
Guinean freshwater fishes in recent years. Partic- 
ularly noteworthy have been several studies on the 
biology of the aestivating galaxioid Lepidogalaxias 
salamandroides (reviewed by Berra & Pusey, 

Jnfortunately, two unwarranted assumptions 
about the nature of Australian-New Guinean fresh- 
water fishes prevail in recent literature (e.g., Allen, 
1989; Pollard et al., 1990; Berra, 1998): (1) the fau- 
na is highly impoverished; and (ii) it is dominated 
by species that have recently evolved from marine 
ancestors. Both serve to devalue the significance of 
the fauna, and both are unquestionably premature. 


Volume 87, Number 1 
2000 


Lundberg et al. 53 
Ichthyological Discovery 


For example, Allen (1989: 8) noted that Australia 
has remarkably fewer freshwater species than does 
the continental U.S.A. However, no less remarkable 
is the difference in effort devoted to the taxonomy 
of the two faunas. 

Clearly, there is need for much more survey work 
to be done in Australia and New Guinea, as some 
areas remain poorly collected (particularly in Irian 
Jaya). However, perhaps more important, there is a 
need for more careful study of the many widespread 
species, as it is highly likely that such study will 
lead to a significant increase in the number of rec- 
ognized species. For example, geographic variation 
has been reported in various Australian freshwater 
fishes, including representatives of the Atherinidae, 
Eleotrididae, Galaxiidae, Latidae, Plotosidae, Me- 
lanotaenidae, Terapontidae, Percichthyidae, Retro- 
pinnidae, and Pseudomugilidae. Indeed, one me- 
lanotaeniid species, Melanotaenia trifasciata, has 
been divided into as many as 35 geographic forms, 
each with highly restricted, allopatric distributions 
(Hieronimus, 1 

ile it is clear that the taxonomic reevaluation 
of widespread species will benefit from recently de- 
veloped molecular techniques (e.g., Rowland, 
1993; Musyl & Keenan, 1996; Jerry & Woodland, 
1997), it is likely that the most significant advances 
will not come directly from technology, but rather 
from a sounder philosophical view of species and 
associated systematic method. Much could be 
gained from careful analysis of many morphological 
characters already at hand, such as coloration char- 
acters noted for many of the melanotaeniid “vari- 
eties." (Coloration characters, when not supported 
by other characters, have generally been dismissed 
by ichthyologists working on Australian and New 
Guinean freshwater fishes, although such prema- 
ture judgment is unjustified; a similar problem ex- 
ists for Indo-Pacific shore fishes; see Gill, 1999. 
There is urgent need for such studies in order that 


м2 


species Бе properly conserved and managed. 

One could argue about the biogeographic rele- 
vance of whether the freshwater fishes of Australia 
and New Guinea are of marine ancestry or not, as 
it is incorrect to equate a marine habit with mobil- 
ity and a random biogeography. Indeed, there are 
many endemic Australian marine fish taxa. How- 
ever, even if marine ancestry was an issue, the as- 
sumption that Australian and New Guinean fresh- 
water fishes are recent invaders from the sea is 
simply not supported by available evidence. While 
it is true that many of the dominant freshwater taxa 
also include marine species (e.g., Plotosidae, Ter- 
apontidae, and Gobioidei), it does not automatically 
follow that a marine habit is ancestral. Consider, 


for example, Vari’s (1978: fig. 9) phylogeny of the 
Terapontidae. Most of the genera are exclusively 
freshwater and restricted to Australia, New Guinea, 
and several other Gondwanaland fragments in the 
Indo-Australian area. Three of the four genera with 
marine species (Mesopristes, Terapon, and Pelates) 
are relatively widely distributed in the Indo-Pacific, 
but occupy relatively derived positions in the p 

logeny. The phylogenetic position of the fourth “ma- 


” 


rine” genus (Amniataba, which includes two spe- 


cies, an Australian freshwater species, and 
southern New Guinean and Australian euryhaline 
species) is unresolved, but it could be near basal. 
Leipotherapon, the basal-most genus in the phylog- 
eny, is exclusively freshwater. Therefore, one inter- 
pretation of available evidence, following a center 
of origin concept, suggests that terapontids are 
freshwater fishes, with several relatively recently 
evolved marine species 

hereas many previous authors have acknowl- 
edged that a small component of the Australian— 
New Guinean freshwater fish fauna probably has a 
relationship with Gondwanaland (e.g., galaxioids, 
and the osteoglossid genus Scleropages), there is no 
justification for a priori rejection of such a rela- 
tionship for the other taxa. For example, although 
the Australian Percichthyidae are often cited as be- 
ing recently evolved marine invaders, this is largely 
of the previously loose definition of 
the family, and Johnson’s (1984) cladistic definition 
of the family (see above) suggests that percich- 
thyids, too, predate fragmentation of Gondwana- 


a consequence 


land. 

Unfortunately, phylogenetic information is lack- 
ing for most Australian and New Guinean fresh- 
water fish taxa, but the need for such studies is 
obvious. 

In summary, the past few decades have seen sig- 
nificant advances in our understanding of the sys- 
tematics, biology, and distribution of Australian and 

ew Guinean freshwater fishes. However, there is 
need for continued work, particularly in the re- 
evaluation of purported widespread species and in 
phylogenetic studies. There is every reason to pre- 
dict that such studies will lead to a significant in- 
crease in recognized diversity, and a conclusion 
that the freshwater fishes are just as distinctive and 
historically/biogeographically informative as mar- 
suplals, birds, and the other, more famous compo- 
nents of Australia and New Guinea's biota. 


DISCUSSION 


hese are exciting times of discovery in fresh- 
water ichthyology. In tropical freshwaters, new fish 


54 


Annals of the 
Missouri Botanical Garden 


species are being found and described at rates as 
high as any in the history of ichthyological explo- 
ration. Many recently discovered fishes exhibit 
highly unusual characteristics, and some represent 
previously unknown lineages (described as new 
genera and family-group taxa). New fish species are 
still found occasionally in Europe and North Amer- 
ica, where the ichthyofaunas are already well doc- 
umented. Phylogenetic studies of fishes from all 
over the world commonly reveal unsuspected high- 
er-level evolutionary and biogeographical relation- 
ships. Investigations of even long-known fish spe- 
cies continue to discover novel characteristics and 
strange life histories. In addition, as the extent and 
patterns of fish diversity and relationships are more 
thoroughly documented, we are better able to de- 
termine the correlates and causes of fish diversifi- 
cation in evolutionary time. 

iese are also times of serious concern for the 
present and future health of populations, species, 
and communities of freshwater fishes and other 
aquatic organisms (e.g., Stiassny, 1999). А recent 
analysis of the North American situation by Ric- 
ciardi and Rasmussen (1999) estimates the extinc- 
tion rate for freshwater animals at five times the 
terrestrial fauna and similar to that for tropical rain- 
forests. Many human activities are increasingly dis- 
turbing and, in cases, destroying freshwater biotas 
(McAllister et al., 1985; Minckley & Deacon, 1991; 
Warren & Burr, 1994; Harrison & Stiassny, 1999). 
Wherever human populations are large or expand- 
ing, so increase dams and impoundments, dredged 
and straightened channels, erosional runoff and tur- 
bation, and the harmful waste products of mining, 
industry, agriculture, and urban growth. These im- 
pacts have unintended but negative and sometimes 
devastating effects on aquatic habitats and life. 
Consider the vast extent of interruption of natural 
river flows and fish movement, and altered benthic 
habitat and community structure that has already 
attended the proliferation of dams. According to the 
World Commission on Dams (1998), in 1997 there 
were an estimated 800,000 in the world, and “more 
than 45,000 of these dams have been categorized 
as large (dam height more than 15 meters above 
the natural river bed)." Among other horrific as- 
pects of an ecological catastrophe centered on 
freshwater, witness, in the years following diversion 
of the A 
collapse of the Aral Sea biota (including a fishery 


mu Darya and Syr Darya rivers, the total 


that once produced 45,000 metric tons per annum). 

Freshwater fishes are important and valued re- 
sources for food, sport, ornament, and biological 
control. Like so many other resources, populations 
and some species of freshwater fishes are already 


overexploited. Well-documented cases of overfish- 
ing include several species in the Laurentian Great 
Lakes (S. Smith, 1968) and the large pimelodid riv- 
er catfishes of the Amazon (Barthem & Goulding, 
1998). Freshwater fish communities and species 
also have been placed in harm's way by introduc- 
tions of non-native species. Here, too, the problem 
is current and global in extent. Some of the best 
known examples are the sea lamprey in the Lau- 
rentian Great Lakes, Nile perch in Lake Victoria, 
African tilapia cichlids and American largemouth 
bass in many parts of the world, and trouts in An- 
dean rivers. The world's most spectacular species 
flock of cyprinid fishes in Lake Lanao in the Phil- 
ippines went to extinction following introduction of 
several exotics. Lake Dianchi in Yunnan, China, 
formerly held about 25 native species (12 endemic 
in the lake). About 40 species live there now, but 
most are introduced. All the lacustrine endemics 
are gone, including the monotypic TR 
dianchiensis, which was described about 25 year 

after the last living specimen was documented. 

Human dependencies on and benefits from 
aquatic species and the communities and biotas 
they form are many, and some are critical, includ- 
ing food and indicators of water quality. Daunting 
and increasing as the threats are to freshwater bi- 
otas around the world, efforts must be made to re- 
duce and mitigate their impacts by reducing habitat 
destruction, over-fishing, and introduction of alien 
species. 

To meet the immense challenges confronting 
aquatic conservation biology, an accurate and thor- 
ough baseline knowledge of species-level diversity 
is essential. Such knowledge is also fundamental 
for research in systematics, ecology, and other sci- 
ences. The freshwater fish faunas reviewed herein 
exemplify how much we are still discovering about 
species and higher-level diversity, and suggest that 
much more remains to be discovered. Fishes are, 
of course, among the better known components of 
freshwater biotas. Even so the pace of habitat 
change is so rapid that it is likely that some, pos- 
sibly many, freshwater fish species will disappear 
before they are known to exist. What else will dis- 
appear along with those unknown casualties? How 
much do we not yet know of the species of aquatic 
invertebrates, green plants, fungi, protists, and pro- 
karyotes? We do not know, and possibly will never 
know, quite a lot of these cniin and the roles 
they play(ed) in their communitie 

Fishes, because they are bue ен and rel- 
atively easy to monitor, serve as indicator organisms 
to monitor the health of aquatic ecosystems. But to 
achieve the needed baseline knowledge of Earth’s 


Volume 87, Number 1 
2000 


Lundberg et al. 55 
Ichthyological Discovery 


freshwater fish species will require much work. 
There are praiseworthy and productive programs 
that support biotic surveys and discovery, e.g., the 
US NSF Biotic Survey and Inventories Program. 
Some natural history museums, governments, and 
universities promote the enterprise, but the task re- 
maining will depend on the continuing dedicated 
efforts of many individual ichthyologists in explo- 
ration, discovery, and description. The task remain- 
ing to make the world’s fishes known is a big one. 
Casting, as we must, the larger taxonomic net to 
capture knowledge of Earth’s entire biota is an im- 
mense and urgently needed job, but one that will 
yield a uniquely wonderful result. 


Literature Cited 


Aarn, A initial] & W. Ivanstoff. 1997. Descriptive anat- 
omy of Cairnsichthys rhombosomoides and Iriatherina 
werneri (Teleostei: Atheriniformes), and a phylogenetic 
analysis of Melanotaeniidae. Ichthyological Explor. 
Freshwaters ы 107—150. 

W. Ivantsoff & M. Kottelat. 1998. n A 
analysis of Telmatherinidae (Teleostei: Ather or- 
pha), with description of Marosatherina, a new genus 
from Sulawesi. Ichthyological Explor. Freshwaters 9: 


311-32: 

Allen, G. R. 
А (family Melanotaeniidae). Кес. 
8(3): 449—490. 

. 1989. Freshwater Fishes of Australia. T. F. H. 
Publication, Neptune City. 

——— 1991. Field Guide to the Freshwater Fishes of 
New Gui inea. Christensen Ps 'search. Institute (Madang, 
Papua s w Guinea), Publ. 9: 1—26 

—— ———. 1995. Rainbowfishes in Кае and іп the Aquar- 


1980. A generic classification of the rain- 
Austral. Mus. 


ium. k oa Verlag, Melle. 
——. 1996. Oxyeleotris caeca, a new species of blind 
cave dish (Eleotridae) from Papua New Guinea. Rev 
‘rang. па 23(1–2): 43-46. 
298. А new genus and species of rainbowfish 
mos "s from fresh waters of E t In- 
donesia. Rev. Franç. Aquariol. 25(1–2): 1 
i Е Ноезе. 1986. The eleotrid el of Lake 
Kutubu, Papua New Guinea with descriptions of four 
new species s. Кес. W. Austral. Mus. 13(1): 79-100. 
. J. Renyaan. rae Kleotrid fishes of the Tri- 
ton T" 's, Irian Jaya, with descriptions of four new spe- 
cies. Rev. "m Non 23(1–2 > —56. 
Alves-Gomes, J. ). Hopkins. 
sights into the e of е fishes and the 
organs. Brain Behaviour & 


Molecular in- 


ears | their electric 

Evol. 49: 324—351. 
Artedi, P. T 38. [с hthyologia sive opera omnia S Bec 'ibus 

scilicet: Bibliotheca ichthyologica. Philos 


E 


tiora quam antea ulla Me cm vindicavit, recognovit, 
coaptavit et edidit Carolus Linnaeus. Wishoff, Lugduni 
Batavorum. 

Bailey, R. M. & Baskin. 1976. Sc Ip dicra, a 
new armored A from the si ig Amazon. Occas. 
Pap. pur - Zool. Univ. Michigan No. 674: "m 

— & С. К. Smith. 1981. Origin en geography of the 


fish fauna of the | aurentian Great Lakes basin. Canad. 

1539-1561. 

М. јато 1998. The Catfish Connec- 
ken and Conservation of Amazon 
Predators. Columbia Univ. Press, New York. 

Beadle, L. C 4. The Inland Waters of Tropical Africa. 
An Bou to Tropical Limnology. Longman, Lon- 
dor 

Be Imke R. J. 1992. Native Trout of Western ds Amer- 

"и See e Avg 6: 1-27: 

Be ll. 994. The a Bi- 
xe a с {ы оаа i kleback. Oxford Univ. 
Press, Oxford. 

& T. R. Haglund. 1982. Fine scale temporal var- 
iation of the Miocene Du Gasterosteus doryssus. 

Paleobiology 8: 282— 

. Baumgartner à E. C. Olson. 1985. Patterns 

: temporal change in single hological characters 

a Miocene vocc а fish. ы hid 11: 


271. 
Benstead, J. Р., M. L. J. Stiassny, P. М. Loiselle, К. 1. 
ha g & М. А 2000. Ri i 
Tuba ar. In P. J. Boon 
(editors), Global Perspectives on R 
Science, Policy and Practice. John Wiley & Sons, 


г. 
Bernatchez, L. & J. J. Dodson. 1994. Phylogenetic rela- 
tionships among palearctic and nearctic whitefish (Cor- 
egonus sp.) populations as revealed by game ка 
DNA А Canad. J. Fish. Aquat. Sci. 51: (Su 
1): 240— 
———— & d С Wilson. 1998. Comparative phylogeog- 
raphy of Nearctic Rn Palearctic fishes. Molec. Ecol. 
—452. 


7(4): 431 


Guyomard & Е. Bonhomme. 1992. DNA se- 
ps variation of the mitochondrial control region 
mong ean and morphologically remote Eu- 
ropean brown trout Salmo trutta populations. Molec. 
161— 


13. 
8. А quum History of Australia. Aca- 
demic pee San 

А иѕеу. 1997. Threatened fishes p = 
orld: Le idog и salamandroides Mees, 


(Lepidogalaxiidac) Environm. Biol. Fishes 50(2): ae 


D P. Ban u, J.-L. Gaudet & J.-C. Hun 
1977. Ба ү Water =a A Multilingual Cat. 
alogue. Lr News, London 

996. Biology of IEEE 2nd ed. W. B. Saun- 


Bond, С 
ders, ord 
Bouchet, P. 1995. A major new mollusc radiation discov- 


ered in the ancient pis of Sulawesi. rig n rnational 
Malacological Euren Vigo. Abstract 
iei G. A. 5. The Puch of о fresh- 
wa "н ЫШ 72: 413—4 
909-1916. Catalog of Ed freshwater fishes of 
Africa in the British Museum (Natural History. I-IV). 
ae of the British Museum (Natural History), Lon- 


Britski, H. A. & H. ks у 1983. Trichogenes longipinnis, 
novo género e de Trichomycterinae do sudeste 
do Brasil (Pisces, Siluriformes). Rev. Bras. Zool. 1(3): 
211-216 

Buckup, Р. А. 1998. Relationships of the Characidiinae 
and Bo of characiform fishes | gee Ostar- 
iophysi). P —144 in L. К. Malabarba, R. Е. Reis, 
R. P. Vari, С. T `$. Lucena & 2. M. S. Dicen (editors), 


56 


Annals of the 
Missouri Botanical Garden 


Phylogeny and Classification of Neotropical Fishes. 
Mus. Ci. PUCRS, Porto Alegre. 

Burns, J. R . Weitzman, & L. R. Malabarba. 1997. 
Insemination in eight species of cheirodontine fishes 
(Tele ostei: P idae:  Cheirodontinae). Copeia 


1997(2): 43 


Tecnol. 
s. 


А , E J. Grier & N. A. Menezes. 1995. 
Internal. fertilization, and sperm morphology in 
glandulocaudine fishes (Te 2. era idae: Glan- 
duloc А 4 Morphol. 224: 


testis 


Burr, B. M. é „ Mayden. i го netics and 
North Nene freshwater fishes. Pp. 18-75 in » 
Mayden (editor), Systematics, е al Ecolog 8 
North American Freshwater Fishes. Stanford Univ. 


Press, Stanford, Californi 

Bussing, 276. о distribution of the San 
Juan ic ашин к South Americ | with remarks on 
its origin & ecology. Pp. 157-175 in T. Thorson (editor), 
Inve | of the Ichthyofauna of Үз araguan Lakes. 


~ 


~ 


Univ. о . Lincoln. 
985. patas of distribution of o i 
S rican ichthyofauna. Рр. 453-473 in F. С. Stehli 


). Webb (editors). o Great American a жы 
А Plenum Press, 

Campos da Paz, В. & J. S. Albert. "1998. The gymnotiform 

eels of tropical America: A history of classification and 
phylogeny of the South American electric knifefishes 
(Teleostei: Ostariophysi: Siluriphysi). Pp. see in L. 

Malabarba, R. E. Reis, R. P. Vari, C. A. S. Lucena 
& 7. M Luc ‘ena (editors), Phyloge ny ки Classifi- 
cation of Neotropical Fishes. Mus. Ci. Tecnol. PUCRS, 
Porto Alegre. 

Cavender, T. M. 1986. Review of a fossil Vene. of North 
American freshwater fishes. Pp. 699-724 . H. Ho- 
cutt & E. O. Wiley (editors), The а ue ‘of North 
oo ап Freshwater Fishes. Wiley-Interscience, New 


"ios `D. M. 1970. How many recent fishes are there? 
Proc. Calif. Ac va Sci. 4th ser. 38: 341-346. 

Costa, W. J. Е. 1994. A new genus and species of 
Sarc E isturiforaé s: Trichomycte -ridae >) from 
the Araguaia basin, central Brazil, with notes on subfa- 
milial phylogeny. Ichthyological Explor. Freshwaters 5: 
207-2 a 


7. Phylogeny and classification of the Cypri- 

о revisted (Teleostei: Cyprinodontiformes): 

Andean and Anatolian killifishes sister taxa? 
Comp. = 2(1): 1-17. 

— ———. 1998. Phylogeny and classification of the Cypri- 
m (Euteleostei: АРА А геар- 
prais ee 537-560 in L. R. Mala iba rba, R. E. Reis, 
R. P. Var S. Lucena (анын, 
Pide ns Classification ч шол Fishes 

s. Ci. Tecnol. PUCRS, Porto Ale p 
с К. А. Boc kinan 1994a. 
mycterus, a new blind Meroe fish (Siluriformes 

Trichomye pes from the Rio Tocantins, Brazil. Trop. 


Zool. n 


= 
Е 


phlobelus macro- 


Sarcoglanidinae (Siluriformes: Trichomycteridae) from 
re-examination | subfami- 
lial phylogeny. J. Nat. Hist. (London) 28: 715-730. 

V. B. R. Күү, М. М. Bruton, E H. Green- 
wood, R. C. Hart, P. B. N. Jackson & A. J. Ribbink. 
1986. е qualities aal special problems of the Af- 

an Great Lakes. Im Biol. Fishes 17: 161—183. 
Cox ae. С. 1998. Sex-related morphological vari- 


19 )Ab. А new genus and species of 


ation in two species of apteronotid fishes (Gymnotifor- 
mes) from the Amazon River basin. Copeia 1998(3): 
730-735. 
J. G. Lundberg. 1999. The largest of all elec- 
tric-fish snouts: Hypermorphic facial growth in male Ap- 
teronotus hasemani, and comments on the nominal spe- 
cies A. anas (Gymnotiformes: Apteronotidae). Amer. 
. le о" & Herpetology, и & Abstracts 
for "199 
Cracraft, x HA Speciation and its ontology: The empir- 
species concepts 


T 


ical consequences of alternative r 
understanding pum. апа processes of differentiation. 
Pp. 28-59 in A. Endler (editors), Specia- 
tion and Its Cac: Sinauer, Sunderland, Mas- 
sachuse a 

‘rowley, L. . M.. W. Ivantsoff & С. В. Allen. 1995. 
Desc xe а a new species of hardyhead, Crateroce- 
phalus а eg меш from Irian Jaya. 


~ 


ес. W. stral. 17(3): 325-3 
Daget, J. 1984. CA 1991. Check List i the Freshwater 
Fishes y Үш а. ORSTOM, Paris. 


Aus EE historique sur l'ichtyologie afri- 

сате. qe —149 J. Е. Guégan & 

bs (editors), Bisa Diversity of African 

Fresh- and Brackish W Fishes. Ann. Sci. Zool., Mu- 

sée ` Royal de L'Afrique C eine Tervuren, Belgique Vol. 
20 


'eugels, 


Duc инв; P. J. 1957. Zoogeography: The Geographical 
Jistribution of Animal | ‘iley & Sons, New York. 

owling, T. E. & C. I . 1997. The role of hybrid- 

izalion and ена in the diversification of апі- 

a Annual Rev. Ecol. Syst. 28: 571—593. 

Dyer, B. S. & B 1996. о relation- 


— 
= 


Chernoff. 


ships among егы) fishes (Teleostei: Atherino- 
oo u^ J. Linn. Soc. 117(1): ) 

schelle, A. A. & I. Kornfield (editors). 1984. Evolution of 
En Spas ies Flocks. Univ. Maine at Orono Press, 
Orono, ae 


Endler, J. 983. Natural and sexual selection on color 
atoms | in poeciliid fishes. Environm. Biol. Fishes 9: 
173-19 

Esc Wi yer, X. N. 1998. Catalog of Fishes. Calif. Acad. 

San Lie isco. 

быы С. J.. Jr. . Isbrücker & H. Bern. 1986. 

Neblinichth ys pilos sus, a new genus and species of 

mailed catfish from the Río Baria inm southern Ven- 

ezuela (Pisces, Siluriformes, Loricariidae). Rev. Franç. 
Aquariol. 13(3): 69-72. 
i V 1984. 


Fink, W. L Basal euteleosts: Relationships. Pp. 
202—206 in H. С. Moser, W. J. Richards, D. M. Cc e. 
P. Fa V. Kendall, Jr. & S. L. Richardson 


(editor) Cale ny dnd Systematics of ae Amer. 
. lc hthyologists & d airs en As ла! Publ. 
Friedman, А. & C. D. Hopkins. 1996. Tracking indi- 
vidual mormyrid ге fish in the field using electric 
organ discharge waveforms. Animal b bone 51: 391- 
401. 


Friel, J. P. & J. G. Lundberg. 1996. Micromyzon akamai, 
gen. et sp. nov. a small and eyeless Banjo Catfish (Silur- 
iformes: Aspredinidae) from the river channels of the 
lower Amazon Basin. Copeia 1996(3): 641-048. 

Fryer, G. 1976. Evolution of species flocks of cichlid fish- 
es in African lakes. Evol. Syst. Evolut.-forsch. 15: 141— 
165 


-& T. D. Цез. 1972. The Cichlid Fishes of the 
Great Lakes of Africa: Their Biology and Evolution. Ol- 
iver & Boyd, Edinburgh. 


Volume 87, Number 1 
2000 


Lundberg et al. 
Ichthyological Discovery 


Galis, F. 1998. Why are there so many cichlid species? 

Tree 13: 
Gesner, C. 1560. Nomenclator aquatilium animantium. 
Icones animalium pend їп таг and. dulcibus 
aquis degentium, plus qua ec 1 nomenclaturis 
singulorum Latinis, Grecis, "taliei 18, - Hispaniois, Galli- 
cis, Germanis, аа alijsq; interdum per certos or- 
dines digestae. C. Froschoverus, Tiguri [Zürich]. 

1563. Fia 'hbuch, das ist ein kurtze doch voll- 
аде Beschreybung aller Fischen so in dem Meer und 
süssen Wasseren, Seen, Flüssen pai е Васһеп 
ir Wohnung ћађепа, sampt irer м 1 Conterfactur 
L RA Froschower, Zürych [also 1 575. 1 508 & 1670 edi- 


. 1999, Subspecies, geographic forms and wide- 


(editors), Proceedings of e 5th Indo-Pacific 
Fish Conference, Nouméa, 1997. Soc. Franç. Ichtyol., 
aris. 
Gosline, W. A. 19 
jd 


. The limits of the fish family = rran- 

рон notes оп а в percoids. Proc. Calif. 

ad. S 4, 33(6): 91-112. 

68, “The т "rS E pe reform fishes. Proc. 

. Mus. 124(364 

1994. Studies of un. nts and his- 
toric il vie ography in the Fossil Butte and Laney 
Members of the wer run Formation. Univ. Wyoming 
Contr. Geol. 300): 

— & ;. Bemis. gr Osteology and phylogenetic 

уне of | en and recent paddlefishes (Poly- 

omments on the interrelationships of 

УИ, 

. (1). 

‚ 1986. A review of Antarctic ich- 
thy ox in light of new fossil disc ‘overies. Paleontol- 
ову 0 13-117. 

— а —— ———. 1998. A comprehensive phylogenetic 

study ч amiid files (Аана based on comparative 


skeletal anatomy. n УТС а! зеагс ћ (ог interconnect- 
ed patterns d natural history. Soc. Vert. Paleontol. 
Mem. 4: i-x, 1—690. supplement to J. Vert. Paleontol. 


1). 

Greenwood, P. H. 1981. The Haplochromine Fishes of the 
East African Lakes. Collected Papers on their Тахопо- 
my, Biology and Evolution (with an introduction and 
species index). Kraus International Publications, Mu- 
nich 

. 1983. The zoogeography of African freshwater 

fishes: Bioaccountancy or жүүт" Рр. 179-199 

т В. W. Sims, J. Н. Price Walley (editors), 

Evolution, Time and Space: The БР ye of the Bio- 

. Syst. Assoc. Special Vol. No. 23, Academic 

у 


k. 
Hirion I. J. & М. L. т Stiassny. 1999. The quiet crisis: 
A preliminary liuius of freshwater fishes of the worl 
that are either extinct or “missing in action." Pp. 271- 


[om 


331 in R. D. E. MacPhee (editor), Extinctions in Near 
Time SNAM Contexts, and Consequences. Plenum 
Press, New ork & Lond 

Helfman, G B. Collette . E. Facey. 1997. The 


pivcrsity | af Fishes. Blackwell E ience, Malden. 

sachuse 

‚ В. 1978. Uber die “Ic ‘hthyographie der Elbe” des 

Neun Kentmann—Eine Studie 

süchsischen Fischfaunen (Pisces). 
1-13. 


den 35: 75-100. pls. 


Mas- 


über die ältesten 


Zool. Abhandl. Dres- 


Hieronimus, H. 1998. Eine variable Art: der Regenbogen- 
fisch Melanotaenia trifasciata. TI Magazin 142: 47-51. 
Hocutt, C. Н. & Е. O. Wiley. 1986. The Zoogeography of 
North Americ ‘an Fre E NW Wiley-Interscience, 

ew 
Hopkins, C. D. 1981. On the diversity of electric signals 
inac RS г m electric fish in West Africa. 
Amer. Zool. 11-222. 
& A E ле 1981. Тетрога! m of species 
recognition page in electric fish. Science 212: 85—87. 
Horn, M. H. 1972. The amount of space elite for ma- 
rine and еве fishes. Fish. Bull. U.S. 70: 1295- 

1297 


l. 

Howes, G. H. 1983. Problems in fish anatomy and phy- 
logeny exemplified by the Neotropical Hypophthalmi 
dae (Teleostei: Siluroidei). Bull. Brit. Mus. (Nat. Hist.), 

-39. 

Hubbs, C. L. & R. R. Miller. 1948. The Zoological Evi- 

rrelation between fish distribution and hydro- 
graphic i in the de i Wi 

Great Basin, with Е 


Tw 


States the ( 


1 

and Jis ‘ial Times. Bull. "Wish 38(20): 17— 

166 

—— & —————. 1960. The Spiny-Rayed Cyprinid 
Fishes (Plagüpterinir of the Colorado River System. 
Univ. Michigan Mus. Zool. Misc. Publ. 

———, ——— & L. С. Hubbs. 1974. Hydrographic h his- 
tory and relict er of the North-Central Great Bas 

Mem. Calif. Acad. Sci. 7: 1-257. 


Huber, J. 1998. „БЕД Data 1996. Vs тш 
Е.Д /l 


К.. J. Sourdis, P. S. авар. T A. Wheeler. 

990. куен differentiation among species of 

the genus Scardinius (Pisces: Cyprinidae) in Greece. J. 
Nat. ime aue 30: 459-473. 

Isbriicker, 80. Classification and catalog of the 
mailed B гіс ЕТА (Pisces, Siluriformes). Verslagen en 
Technische Gegevens, Instituut voor Taxonomische 
Zoölogie, Zoólogische Museum, Universiteit van Am- 
sterdam, No. К 1–181. 

——. 198 


Iliadou, 
1 


ж. 


Tichomycteridae, mysterieuze meervallen. 
let Vari. биа: | 4–279. 
Р, ласк, В. Saeed & L. Е. L. M. Crowley. 
1991. A Mise blue-eye, a new species and genus 
of the eL Pseudomugilidae from central western 
АЧ ueensland. Fishes of Sahul 6(4): 2 
. M. 1992. RE xinguense, nouveaux genre et 
+ се du Rio Xingu, Amazonie, Brésil (Teleostei: $i 
rasalmidae). Ichthyological Explor. Freshwaters 3: 235- 
252. 


| 
Ivantsoff, W. 


Jerry, D. К. & D. J. Woodland. nity Electrophoretic ev- 
idence for the presence of = = cribed ‘Belinger’ 
Plotosidae) in four 
onthem oastal rivers. Marine 
nd F b ice Res 48(3): 2 5-240 
Johnson, G. D. 1984. Фа и and геја- 
tionships Pp. idi 38 in H. G. Moser, W. J. Ric ha 
D. M. Cohe ‚ А. W. Kendall, Jr. & S 
ee. x editor) e. and Systematics of E 
es. Amer. Soc. Ichthyologists & Herpetologists Special 
Publ. 1 
& C. Patterson. 1996. а а of lower eu- 
teleostean fishes. Pp. 251—332 in M. L. J. Stiassny, L. 
R. Parenti & G. D. [nu tr Interrelationships 
of Fishes. үүт mic Press, San Diego. 
& 7. Springer. 1997. oen] Another look. 


3d 


58 


Annals of the 
Missouri Botanical Garden 


Amer. Soc. е & Herpetology, Program & Ab- 
stracts for 199 

Kaufman, L. S. s y change in species-rich 
freshwater ecosystems, the lessons of Lake Victoria. 
Bioscience 42: 846—858. 

‚ В. О. Ochumba. 1993. Evolutionary and con- 

servation biology of cichlid fishes as revealed by faunal 

remnants in northern Lake Victoria. Conservation Biol. 

11 лут 

L. J. Chapman & С. А. Chapman. 1997. Evolu- 

tion in fast forward: Haplochromine fishes of the Lake 
Victoria Mr Endeavour 21: 23-30. 

Kocher, T. С. А. Stepien (editors). 1997. Molecular 
о. of Fishes. Academic Press, New York. 

‚ А. Сопгоу, К. К. McKaye & J. R. Stauffer. 
1993. < Similar 's of cichlid fish in Lakes Tan- 
ganyika and Malawi er due to convergence. Molec. 
ее Evol. 2: 165. 

Kornfield, 1. & A. Parker. pes Molecular systematics of 
a rapidly araling species flock: The Mbuna of Lake 
Malawi and the search Зи phylogenetic signal. Pp. 25- 
38 in T. D. Kocher & С Stepien (editors), Molecular 
и. of Fishes. ак Press, San Diego. 

Kottelat, M. а. Synopsis of the endangered Buntingi 
(Osteichthyes т and Oryziidae) of Lake 

, Central Sulawesi, Indonesia, with a new герго- 
бы. guild and description of three new species. Ich- 
thyological Explor. Freshwaters 1: 49—67. 

. 1990b. The ricefishes (Oryziidae) of the Malili 
„akes, Sulawesi, Indonesia, with description of a new 

species. Ichthyological Explor. Freshwaters I: 151-166. 

1990c. Sailfin silversides (Pisces: Telmatherini- 

dae) of Lakes Towuti, Mahalona and Wawontoa (Sula- 
wesi, Indonesia) with descriptions o ew genera 
and (мо new pe Ichthyological Explor, Freshwa- 

ters l: i 


Sail silversides (Pisces: Telmatherini- 
dae) of Lake Matano, Sulawesi, Indonesia, with descrip- 
tions of six new species. Ichthyological Explor. Fresh- 
waters S Eid 

7. European freshwater fishes. Ап heuristic 
еш, of the freshwater fishes of Europe (exclusive 


of former Mini ж an introduction for non-system- 
atists and c nomenclature and conservation. 
Ku (Bratislava), pre don 52 (Suppl. 5): 1-271. 
of the Nam Theun and Xe Bangfai 
s p wi sh ignes of twenty-two new species 
(Teleostei: Cypri , Balito ridas, Cobitidae, Coiidae 


de ШОП) ae ththyological Explor. Freshwaters 
9: 


г K. K P. Lim. 1994. Diagnoses of two new gen- 
de mr ын new species of earthworm eels from the 
ae and Borneo (Teleostei: ee 
се казые Explor. Freshwaters 5: 181–1% 

& А. Whitten. 1996a. 


F мен | E 
estern [ndonesia p Sulawesi: Additions and corre 
tions. Taa Jaka 

— ё ——. 19 ББ. Freshwater Biodiversity in 

Asia with Special дерин to Fish. World Bank 

Techn. e . 343: i-ix + 1–59. 

Я А] Whitten, 5. М. Kartikasari & 5. Wirjoat- 
тодјо. 1993. Freshwater Fishes of n Indonesia 
and Sulawesi. Periplus Editions, Hong Kor 

Kuehne, R. A. & R. W. Barbour. 1983. The American 
Darters. Univ. Kentucky Press, Lexington 

Kullander, S. O. 1980. A taxonomical stud col the genus 
Apistogramma Regan, with a revision of Brazilian and 


Peruvian species (Teleostei: Percoidei: Cichlidae). Bon- 
ner ae acd No. 14: 1-152. 
A phylogeny and c ions 'ation of the South 
"e 'an ү Schlidae: Pp. "с + L. R. Malabarba, 
R. Reis, R. P. sa ase &Z.M.S 
Lucena oo. oe at Classific ‘ation of Neo- 
tropical Fishes Тес по]. | 
Larson, Н. К. 
e genus Chlamydogobius with the dese о of 
five new species. The Begle Records of the Museums 
& es Galleries of the жене Те "rritory 12: 19-51 
Kottelat. 1992. A d species of Mugilo- 
gobius (Pisces: Gobiidae) from Lake Maiano, с pin Su- 
lawesi, Indonesia. Ic usd Explor. Freshwaters 3: 
25-234. 


> D. S., С. В. Gilbert, C. H. Hocutt, R. E. Jenkins, D. 

. Mc Allister d]. R. Stau ARE ue conse 1980. Atlas 
" North American Freshwater Fishes. North Carolina 
State Museum of Natural History, Raleigh, North Car- 


olina. 

Leveque, С. 1997. Biodiversity Dynamics and Conserva- 
tion of Freshwater Fishes of Tropical Africa. Cambridge 
Univ. Press, bare ridge. 

y & G. G. Teugels. 1990. Faune des pois- 
sons d’eau en ге et saumátre d'Afrique de l'Ouest, Vol. 


. MRAC-ORSTOM, Tervuren & aed 
1992 


e des poissons 
d'eau ‘done ге et saumâtre d Afrique de l'Ouest, Vol. 2. 
MRAC-ORSTOM, Tervuren 


& P aris. 


s 1. Editio decima, reformata. Salvius, Hol- 
24. 

Lundberg, J. G. 1993. African-South American freshwater 
fish clades and continental drift: Problems with a par- 
афет. Pp. 156-199 in P. Goldblatt (editor), Biological 
Relationships between Africa and South America. Yale 
Univ. Press, New Haven. 

. 1998. The temporal context for diversification of 
Neotropical dishes: Chapter 2. d 49—68 in L. В. Mal- 
ab: E. Reis, В. P. Var 9. пека & 7. 
М. 5. Lucena (е dios. Ple ny and Classification of 
Neotropical Fishes. Mus Tecnol. PUCRS, Porto 

Alegre. 

. Akama. 1999. Identification of an unrecog- 

nized yet diet tinctive and commercially important spe- 

cies of pimelodid catfish (Pimelodinae: Siluriformes) 
from the Amazon basin. Amer. Soc. lc па & Нег- 
53. 


petology, di dn am & Abstracts for 19° 
—— & Е 


хо-Гесста. 1986. А review Rhabdoli- 
chops Карашса Sternopygidae), a genus of South 
American freshwater fishes, with descriptions of four 
new species. Proc. Philade Johi Acad. Sci. 138(1): 58— 


„ Rapp Py-Daniel. 1994. Bathycetopsis olivei- 
rai gen. et sp. nov., a blind and depigmented cetopsid 
catfish from the Brazilian Amazon. Copeia 1994(2): 
381-389. 

. H. Bornbusch & F. Mago-Leccia. 1991. Gla- 
PON CHEM conquistador n. sp. from Ecuador, with diag- 
noses of the subfamily Rhamdiinae Bleeker (1862) and 


Pseudopimelodinae n. subf. (Siluriformes: Pimelodi- 
dae). Copeia 1991(1): 190-2 8. 
, С. Fernandes, J. 5. Albert & М. Garcia. 


1996. Magosternarchus, a new genus with two new spe- 
cies of electric fishes (Gymnotiformes: Apteronotidae) 


Volume 87, Number 1 
0 


Lundberg et al. 59 
Ichthyological Discovery 


from the Amazon River Basin, South America. Copeia 
aan 657-670. 
. M. Lewis, Jr., J. Е Saunders, Ш & F. Mago- 
Leccia. 1987. F ;vidence for a major food web component 
in the main c Бына of the Orinoco ee based on spe- 
cialized electric fishes. Science 237: 81-83. 
MacDonald, C. M. 1978. Morphological and biochemical 
d of Australian freshwater and estuarine per- 
ichthyid fishes. Austral. J. Marine & Freshwater Res. 
29(5) 067—698. 
Mago-Leccia, F. 1970. Lista de los Peces de Venezuela. 
Ministerio de Agricultura y Cria, Oficina de Pesca, Ca- 
racas 


ET. 94. Electric fishes of the continental waters of 

America Biblioteca de la Academia de Ciencias Físi- 

as, Matemáticas y Naturales, v. XXIX, Fundación para 

el Desarrollo de las Ciencias Físicas, Matemáticas y 
Naturales, Caracas. 

Maitland, P. S. 1976. Hamlyn guide to freshwater fishes. 

Hamlyn, London [Data based on 1977 French trans- 

5 et ace d'Europe en 


lation, Les poissons des lacs 
couleurs. = sevier-Séquoia, Par 
Malabarba, L. R. 1998. Characters Pod major clades of 
the Che н (Teleostei: Сһагас a Pp. 19 ya 
234 in L. R. dep гђа, R. E. Reis, R. , C. 
5. Lucena & Z S ena (editors), Pyle mi 
Classification w Te Fishes. Mus Tecr 
PUCRS, se rto Alegre 
& S Mets п. 1999. A new genus and зре- 
cies of Sub th American fishes (Teleostei: Characidae: 
derived caudal fin, including 


a 


Cheirodontinae) with a 


comments about кш cheirodontines. Proc. 
Biol. Soc. Wash. 112(2): 410-432. 
Reis, R. P. Vari, С. a & Z. 


. Lucena. 1998. Phylogeny and abun. cation of = 
Mus. Ci. Tecnol. PUCRS, Porto Alegre. 
D. C. Heins (editors). 1987. oo 
and Evolutionary Ecology of North American Stre 
Fishes. Univ. Oklahoma Press, Norman, Oklahoma 
Mayden, R. L. 1985. Biogeography of the Ouse hita High- 
land Fishes. Southw. Naturalist 30: 195-21 
. 1987. Historical ecology and na Кабай 
highland fishes: A research program in community ecol- 
ogy. Pp. 210-222 in W. J. Matthews & D. C. Heins 
(e ditors), C аши and кайни, Ecology of North 
Stream Fishes. Univ. Oklahoma Press, Nor- 


эйе Fishes. 


Matthews, W. J. & 


ла. 
1992. Systematics, Historical Ecology, 
and North American Freshwater Fishes. Stanford Univ. 

Press, Stanford, 1 
& В. food, 1995. Systematics, spec ies con- 


cepts, and the e Mice ein significant unit in biodiver- 
чы and conservation biology. Amer. Fisheries Soc. 

: 58-115. 
= D. E., B. J. Parker & P. M. McKee. 1985. 


Rare, Endangered a Extinct E in € AT Natl. 


Mus. Nat. Sci., Natl. s. Canada, Otta 

Mc Dowell, R. M. prem Yee in Fishes. c room Helm, 
London 

Mees, G. F. . Description of a new fish of the family 


Cai a Western Australia. J. Roy. Soc. W. Aus- 
tralia 4 38. 

Merrick, | n & G. Е. Schmida. 
water Fishes. Biology & Management. J. R. 


1984. Australian Fresh- 

Merrick, 
North Ryde. 

Meyer, A. 1993. Phylogenetic relationships and evolution- 


(rocesses т e African. cichlid fishes. Trends 
Ecol Evol. 8: 279-28 

Miller, R. R. 1959. ons and affinities of the freshwater 

~ fauna America. Pp. 187-222 in 

. Hubbs (editor), Zoogeography. A. A. A. S. Publ. 


Minckley, W. L. & J. Deacon. 1991. Battle against Ex- 
tinction: Native Fish Management in the American 
Univ. Arizona Press, Tucson 
ectric Fishes. Hisidiy and Behavior. 
Санап & Най, ee. 
Monkolprasit, S. & T. R. Roberts. 1990. Himantura chao- 
phraya, a new giant о stingray from Thailand. 
Japan. J. Ic ie 37: 203-208. 


Moore, J. E. S . The Tanganyika Problem. Hurst & 
Blackett, open 
s Ka . P. Keenan. 1996. Evidence for cryptic 


ius ralian freshwater eel- a catfish, 
нле ad il Copeia 1996(3): 526— 
5. escriptions of new ee а ап 


Bull. 


ers, G. S. 1927. De 

fresh-water fishes collected by Dr. Carl Ternetz. 
Mus. Comp. T. 68(3): 107-135. 

= resh-water fishes and West ш 200- 
тын Soin Кер. 1937: 339-36 

Nagelkerke, I. A. J 7. The Barbs of y in ia, Ethi- 
opia: Morphologic x a and its Implications for 
begr Trophic Resource Partitioning, and Fisher- 

schrift, Landbouwuniversiteit, Wageningen, 

The Netherlands 

Е Sibbing, J. а 
R. R. Res г & J. » Osse. The 
(Barbus spp.) of Lake d A е species flock? 
Environm. Biol. Fishes 39: 1—22. 

Nakabo, T. decus 1994. [Fishes of Japan with Pictorial 
|" (ће Species]. Tokai Univ. Press, Tokyo. |In Jap- 

e.| 

Nelson, m S. 1976. Fishes of the World, 1st ed. John Wiley 

& Tn ог 
4. Pishes of the World, 

& seda [o York. 

Page, L. M. 1983. Handbook ш Darters. T. 
cations, Neptune City, New 

Parenti, L. R. 1981. A ph i. and bioge rini 

is of cyprinodontiform fishes (Teleostei, ign 

nomorpha). Bull. Amer. Mus. Nat. Hist. 168(4): 3 
551 


. van ~ ече Е. Н. 
barbs 


3rd ed. John Wiley 


Publi- 


Е H. 


. A taxonomic revision of the Andean kil- 
lish genus reto оаа C ой 
tidae). v . Amer. Mus. Ма 178(2): 214. 
— wis On је ado i ken dn hid fish- 

Ses оный ма РИ with. notes on the anatomy of 
Pha је dunckeri Regan, 1913. Amer. Mus. Моуп. 
2779: 1-12 


Patterson, C. 1975 ae ое of Mesozoic fresh- 
water fishes. Pp. 15 n Biogéographie et liaisons 
intercontinentales au cours un Mésozoique. ap *m. ae 


Paris, sér. А, Zool. € 
North fia ‘rican 


nouv. sér. 
Тће de кее of the 
fish банын рай of historical biogeography. Рр. 
265-281 in P. L. Forey (editor), The Evolving Bio- 
e. Cambridge Univ. Press, Cambridge, England. 
Pellegrin. J. 1911. La distribution des poissons d'eau 
douce en Afrique. Compt. Rend. Hebd. Séances Acad. 
Sci., Paris 153: 287—299. 
. 1921. Les poissons des eaux douce de l'Afrique 
du Nord francaise et leur distribution géographique. 


Natl. His . Nat., 
p 


60 Annals of the 
Missouri Botanical Garden 
Compt. Rend. Assoc. ge, Avancem. Sci., Congrès де Rowland, S. J. 1993. ik o nt ikei, an endangered 
rasbourg, pp. 269-2 species of freshwater cod (Pisces: Percichthyidae) from 
Pethiyagoda 199 еа Fishes of Sri Lanka. the Clarence River System, NSW. and M. peeli marien- 


Wildlife Heritage Tus of Sri Lanka, Colombo. 

ottelat. 1994. Three new species of fishes 
of the genera reo: kitte hthys (Cyprinidae), Travancoria 
(Balitoridae) and Horabagrus (Bagridae) from the ~ ћа- 
ws dy River, Kerala, India. J. South Asian Nat. Hist. 


: 97-1106. 
Bios R. B. & T. H. Oakley. 1997. оона о rela- 


мени eps the Salmoninae based o 


mitochon ial DNA sequences эр 145. 162 in T. D 
Kocher A. Stepien (editors), Molecular Systemat- 

ics of Fishes Ac ademic Press, San Diego, California 
Pinna, M. С. € 1 w genus of trichomycterid 


catfish (Siluroidei, Gla s к жч with comments on 
its ee nee relationships. Rev. Suisse Zool. 95(1): 
3-128. 

1989. A new Sarcoglanidine catfish, phylogeny 
of its subfamily, and an appraisal of the phyletic status 
of the Trichomycterinae (Teleostei, Trichomycteridae). 
Mus. Novit. 2950: 1—39 
1992. A new subfamily of Trichomycteridae (Te- 

(світ. Siluriformes), lower loricarioid relationships and 

a discussion on the impact of additional taxa for phy 

logenetic analysis. Zool. J. Linn. Soc. 106(3): 175-229. 
—— ———. 1998. Phylogenetic relationships of neotropical 
: Hist torical overview and synthesis of ћу- 

279-330 in L. R. мања, R. E. Reis, 

. A. S. Lucena & 7. M. S. Lucena oo 
Гоу and Classification e Neotropical Fishes 
Mus. Ci. Tecnol. its RS, Porto Alegre. 

Ч E C. Starnes. 1990. A new genus and species 
of Sarc o ee from the Río Mamoré, Amazon Ba- 
sin, with comments on subfamilial Боору (Teleostei, 
Tric ‘homyc id J. Zool. (I ina 222(1): 75-88. 
= а В.Р. . 1995. Monophyly and Phylogenetic 

Diagnosis of ic ‘Family Cetops idac. with Synonymiza- 
tion of the Helogenidae (Teleostei: Siluriformes). Contr. 


Amer. 


ао 
phere Pp. 
R. P. Var 


Zool., 571. Smithsonian Institution Press, Washington, 
D.C. 

Pollard, D. A., B. A. Ingram, J. H. Harris & L. F. Reyn- 
olds. 1990. Ап over- 


и fishes in Australia 
ee ae 37(Suppl. A): 6 3. 
J. 


. Rasmussen. 


view. J. –7 
Ricciardi, 1999. Extinction rates 
of North Е ап ое fauna. Conservation Biol. 
13: 1220-1222. 
Roberts, T. R. 1975. Ge Mois a 'al d of African 
freshwater fishes. Zool. J. Linn. Soc. 57: 249-319 
1981. Sundasalangidae, a new far 
southeast 


nily of minute 
Гере EESO Asia. 
Proc. ps Acad. Sci. 42: 2 
984. Пао scintilla, new genus and 

egro, Brazil, the smallest known 

Ac Sci. (Ser. 4 


и в 
302. 


species pon the Rio 
clupeomorph fish. Proc. Calif. Acad. 
43(20): 317-321. 
ч G. 1554. Libri de ан marinis, in quibus 
erae piscium effigies e expressae sunt. Quae in tota a pis- 
cium historia contineantur, indic at elenchus pagina 
nona et decima. Bonhomme, Lugdunum. 

Rosen, E. 1974. Phylogeny and zoogeography of sal- 
moniform fishes and relationships of Lepidogalaxias sal- 
amandroides. Bull. Amer. Mus. Nat. Hist. 153(2): 265- 
326 


1969. The structure and relation- 
Amer. 


›. 
& С. Patterson. 
of the Appels ad rygian ue ш 


shipe 
Mus. Nat. Hist. 141(3): 357-474, pls. 5 


sis, a new subspecies from the Mary River System, Qld. 
Rec. Austral. Mus. 45(2 © 2 eae 

Saeed, B., W. Ivantsoff & L. a M. Crowley. 1994. Sys- 
tematic о of US families within Di- 
vision I of the series Atherinomorpha (Ac ERN соки 
with relevant Шоп al perspectives. J. Ichthyol. 34(9): 
27-72. [Originally published in Russian in Voprosy 
ikhtiologii, Ney 5), 1994, 579-610. ] 

Schaefer, 5. А. 1990. Anatomy and relationships of the 
sc ae ла cathahiess Proc. Acad. Nat. Sci. Philadelphia 

42: 167-210 
1998. Conflict and resolution: Impact of new taxa 


on phylogenetic studies of the neotropical Casc iren 


(Siluroidei: Loricariidae). i 375-418 in L. R. Mala- 
barba, R. E. Reis, R. P. Vari, C. A. S. Lucena & Z. M. 


Lucena (editors), Phylogeny and Classification of 
Neotropical Fishes. Mus Tecnol. PUCRS, Porto 
Alegre. 

. Weitzman & H. A. Britski. 1989. Review 
of the и ‘al catfish genus Scoloplax (Pisces: Lor- 
icarioidea: Scoloplacidae) with comments on reductive 
characters in phylogenetic analysis. Proc. Acad. Nat. 
Sci. Philadelphia 141: 181-211. 

Schliewen, U. K., D. Tauz & Pääbo. 1994. Sympatric spe- 
ciation о Д А of crater lake cichlids. 
Nature bg) 62 


Scott, T. rd "e Маг пе & Fresh Water Fishes of 

South ы кашы Printer (South Australia), 
Adelaide. 

Е ама 1996. Lake Victoria rock cichlids, taxon- 


omy, ec a and oe Verduijn Cichlids, Zev- 
as lar 
—— ап Apei & F. Witte. 1997. Cichlid 
Sa diversity Паје ned by | that curbs 
sexual selec ion. Sci ee e 277: 8-1811. 
jtsch, N. Boutor s H. Zwennes. 1998. 
M iei rock- dle cic lids of Lake Victoria: De- 
scription of ге nera and fifteen new Мт 
(Teleostei). le hthyological Explor. Freshwaters 9: 129— 


enhuizen, 
a 


Siebert, D. 1997. Notes on the anatomy and relationships 
of ла а Roberts (Teleostei, Clupeidae), sae à 
scriptions of four new species av m Bull. 

Hist. Mus. London (Zool.) бит 

Silfvergrip, А. М. С. 1996. А ѕуѕіе Roe revision of the 
Neotropical catfish genus Rhamdia (Teleostei, Pimelod- 
idae). Swedish ape of Natural History, Stockholm. 

Simons, A. M. & R. L. Mayden. 1997. Phylogenetic re- 
lationships of the creek chubs and the spinefins: An 

American cyprinid fishes (Ac- 

tinopterygii: Cyprinidae). Cladistics 5 205. 


ee Inland Fishes of New York State. 
e New a State Department of Environmental Con- 
servation. Albany, New York. 

Smith, G. R. 1992. Introgression in fishes: Significance for 

ТЕ cladistics, and evolutionary rates. Syst. 

Jiol. 4 ed 

"e ‚ Todd. i. о rs (d ies flocks 

in north yis rate lakes. ›8 т А. А. Echelle 

& 1. Kornfield (editors), pues on of Fish Species 
Flocks. Univ. Maine at Orono Press, Orono 


dber 61 


Volume 87, Number 1 Lun g et al. 
2000 Ichthyological Discovery 


‚ J. Rosenfield & J. Porterfield. 1995. Processes of Teugels, С. С. & C. D. Hopkins. 1998. Morphological 2^ 


origin and criteria for Pu ation of fish species. Amer. 
E 4—51. 
owling, к. E Gobalet, T. Lugaski, S. K. 
теза г js ) n press. Pai iu and 
rates е : “olution among Great Basin fishes. /n R. Her 
shler . Curry (editors), Great Basin Aguai г stem 
History a Institution Press, Washington, 
D. 


F isheries E 


shith, S. H. 1968. Species succession and fishe 
ploitation in the Great Lakes. J. Fish. Rev. Brad a 
ada 25(4) ) 


а > В. К. & С. В. Smith. 1993. Phylogeny of the 
Pacific trouts and salmons (Oncorhynchus) and genera 
of the family Salmonidae. Trans. Amer. Fish. Soc. 122: 


Sevan, D. J. 1986. А new pimelodid catfish from the 
deep Pub of the Río Napo, eastern Ecuador (Pisces: 
Pimelodidae). Proc. Acad. Nat. Sci. Philadelphia 138: 
46—52 

Sliassny, 
tionships of the bedotiid fishes of 
taxonomic revision of the genus Rheocles die rinomor- 
pha: pue Amer. Mus. Novit. 2993: 1—14. 

—— ———. 1991. Ph ylogenetic intrarelationships " the fam- 
ily Cic s An overview. Pp. 1—35 in M. H. A. Keen- 
leyside (editor), Cichlid Fishes: Behaviour, Ec aka and 
Evolution. Chapman Hall, London. 

———. 1996. An overview of freshwater biodiversity: 
With. some lessons from African fishes. Fisheries 21: 7— 


990. Notes on the anatomy and rela- 
Madagascar, with a 


. The on is the message: Freshwater 
E in peril. I 3—71 in J. Сгасгаћ & F. Grif- 
fo (editors), The b Planet in Crisis: Biodiversity 
Science апо Policy. Columbia Univ. Press, New York. 


Meyer. 1999. Cichlids of the east African 
lakes. Sci. Amer., February 1999: 64-66 
— —— & M. C. C. de Pinna. 1994. Basal taxa and the 


role of с Лаф вис patterns in the evaluation of conserva- 
w frc Pp. 235—240 in 
Forey, C. J. Humphries & R. I. Мапе Wrigh (ed- 
itors), Systematics and Conservation Evaluation. 5 
Assoc. Special Vol. 50: 235-249. Clarendon Press, Ox- 
ford. 
& N. Raminosoa. 1994. The fishes а the inland 
waters of Madagascar. Pp. 133-149 in G. Teugels, 
J. Е. Guégan & J. J. Albaret (editors), Бор 'al Diver- 
sity of African Fresh- and Brackish Water Fishes. Ann. 
Sci. Zool., Musée Royal de L'Afrique Ce 'entrale Tervuren, 
pone Vol. 275. 
К. Parenti & G. D. Johnson (Editors). 1996. 
mous of Fishes. Academic Press, San Die- 


go, Califorr 
sU 


en & W. J. Dominey. 1992. A n 
species flock of cichlid fishes from Lake Bermin, Cae 
eroon with a description о new species of Tilapia 
(Labroidei: ее [е оде ‘al Explor. Freshwa- 
ters 3: 311— 

Sturmbaur, С. 9 | Meyer. 1992. G 

ciation and morphological stasis in a MAE of Afican 
cichlid fishes. Nature 359: 578—581. 
Taphorn, D., R. A. Mac 


Royero 


dulce de Venezuela. Pp. 55-100 гл E. La 


tor), Vertebrados Actuales y Fósiles de Venezuela. Ser. 
"l Mérida, 


Cat. Zool. Venezuela, Vol. 1. Mus. Ci. Tecnol., 


Venezuela. 


Belgique, Vol. 2 


Thatcher, V. E. 199 


Toledo-Piza, M 


Р 'hado-Alison & Е. Mago 
Léccia. 1997. Lista ас шавд de los ресеѕ de agua 
у Marca (edi- 


osteological evidea for the generic position of M 
Myrus Pare шй in n the g enus oo Аш (Tele ostei: 
Mormyric dae). ia 1998: 199-204. 

іёрап & J.-J. Albaret. 1994. по ^" 
ume st Afric ‘an fresh- and brackish wa 
Geographic 'al overviews presented at the PARADI cm m- 
posium, Senegal, 15-20 November, 1993. Ann. Sci. 
Zool., Musée = de VAfrique Centrale Tervuren, 


, С. McG. ne & В. P. King. 1992. Fishes of the 


Cross River basin (Cameroon-Nigeria) taxonomy, zoo- 


geography, ecology and conservation. Ann. Mus. r. Afr. 
Centr. Zool. ru 4—132. 
L. . Janssens, t & M. mei 


1985. Sud une i T in de E s riviere de ( 
mores. Cybium 9: 41-56. 

5. Anphira xinguensis sp. nov. (Isopo- 
da, Cymothoidae) a gill chamber parasite of an Ama- 
zonian serrasalmid fish, ip em xinguense Jégu, 1992. 
Amazoniana XIII(3/4): 293— 

Е Revision of the Супо- 
dontinae (Teleostei: Ub eub: Characiformes). Ph.D 
Dissertation, The City University of New York, New 
fork 

A. Menezes & G. ndes dos Santos. In 
press. Re vision of the neotropical “fish genus Hydrolycus 
(Ostariophysi: Characiformes: Cynodontinae) with the 
dese SS of two new species. Icthyological Explor. 


а rs. 
.. J. Green & S. A. J. Corbet. 1972. о ? 
stuc fa on crater lakes in west = раб о of B 


ombi Mbo. Zool. J. Linn. Soc., London, : 41 ‘95. 
Turner, С. F. T. 1997. Small E go ми time. New Sci. 
2093: 36—40. 
Џуепо, Т. & С. В. Зтић. 1076 Tetraploid origin of the 


karyotype of catostomid fishes. Science 175: 644—646. 

Vari, R. Р. 1978. The terapon perches (Percoidei, Tera- 
ponidae). A didus Tap and taxonomic revision. 

t. 159(5): 175—340. 

. The Re тр al fish family Ctenoluciidae 

Supra and in- 


y. е. Contr. Zool. 564: 1— 
8. Higher level phylogenetic cone cepts within 

RS ae (Ostariophysi), a historical review : 
111—122 in L. R. Ma R. E. Reis, R. P. Vari, C 

S. Lucena & Z. M ena ш, pee 

ind : lassification of с AVENA Fishes. . Tec 
nal b Porto Alegre. 
. Harold. 1998. The genus Creagrutus (Te- 
leostei: au iformes: Characidae): Monophyly, rela- 
tionships, and ree d diversity. Pp. 245-260 in L. 
R. Malabarba, R. E. Reis, R. P. Vari, C. A. S. Lucena 
& Z. M. S. Lucena (Жїн), Phylogeny and Classifi- 
cation of dE a Fishes. Mus. Ci. Tecnol. PUCRS, 
Porto E 


IR. Malabarba. ис Neotropical ichthyolo- 
E E ug JE 


gy: An overview. Pp. 
. Lucena 


‚ Vari, C. A. Bt & Z 
{юг Phylogeny ki Саса; of мот 
Fishes. Mus. 

Verheyen, E., L. 
tochondrial урша! of rock deiliog cic -hlid 
fishes reveals evolut influence of historic lake lev- 
el fluctuations т lale a Africa. Philos. 
Trans., Ser. B. 351: 797—805 


Annals of the 
Missouri Botanical Garden 


Warren, M. L. M. Burr. 1994. Status of freshwat 
fishes of the Unite States: Overview of an nd 
fauna. Fisheries 19(1): 6-18. 

Webb, A. 1998. Phylogenetic Studies of the Family 
SP oa (Teleostei, Cyprinodontiformes). University 

f Michigan doctoral Sem pe Ann Arbor. 
мя 5. & 5. У. Fir 


uem onin phy- 
loge M ая ршапуе ка eromone ерй s in glandulocau- 
dine mee 


ystei: Characidae). Smithsonian Contr. 
Zool. ye 
& 


{ snezes. 1998. Relationships of the 
tribes and genera of the Glandulocaudinae (Ostariophy- 
si: Characiformes: Characidae) with 


a description of a 
new p Chrysobryeon. Pp. 171-192 in L. I 


in t. Ma за 


barba, R. Е. Reis, Е. P. Мап, С. А. 5. Lucena & 7. 

А de ena (editors), ую ny and Classification P 
Neotropical Fishes. . Ci. Tecnol. PUCRS, Porto 
Aleg 


R. P. Vari. 1988. Miniaturization in South 
American fishes: ч overview and discussion. Proc 
‚. Wash. 101: 

1981. ани The Theory апа Ргас- 
псе of Phylogenetic | Wiley, New York. 
Wilson, M Н. & R. G. Williams. 1992. Phyloge- 
netic, bio s oa ec ological significance of early 
fossil records y North American freshwater fishes. Pp. 
224— .. Mayden (editor), Systematics, Histor- 
ical Ecology, d North American oe Fishes. 
Stanford Univ. es пе ен Californ 
i G . Bones and adc t of the sus- 
xioids Lepidogalaxias salaman- 

droides бине Osmeriformes) and wn кашан 

significance. Rec. Austral. Mus. 49(2): 139-166 
World ерд оп Dams. 

http://www.dams.org/ 


pensorium in the Ha 


1998. рар site address: 


NEW MAMMALS IN THE 21st John MacKinnon' 
CENTURY? 


ABSTRACT 


50 years in which only one new large mammal had been found worldwide, three new ungulates were found in 
the same region of Vietnam within 4 years. The context of the finds is discussed in relation to the continuing finds of 
other mammals and birds. This paper draws conclusions about the types of places that may still conceal undiscovered 
mammals and predicts where future finds may be made into the next centu 

Key words: Bovidae, Cervidae, Laos, mammals, Megamuntiacus, Muntiaclis muntjac, Pseudoryx, Saola, Vietnam. 


NEW MAMMAL FINDS AMONG ScARS OF WAR The Chaco Peccary, Catagonus wagneri, already 
known and named from fossils, was found to be still 

Out of the several million species of animals eytant (Wetzel et al., 1975). A few new species were 
known to inhabit this planet, only a small number: „dded by taxonomists splitting known forms: Afri- 
are mammals (just over 4600) and birds (11,000); can colobine monkeys, Sulawesi macaques. Some 
yet these are the creatures the general public best zoologists believed the large mammals were, by and 
know, like, and show most concern for. That there Би EE CER MN 

ge, а useum and cataloged. 

Discoveries of new birds have been equall 
scarce. Ninety-eight percent of all 1135 Palearctic 


den = make Me ans. ды ико gey monn birds were described between 1758 and 1900. Onl 
mists see the job of describing the varied life forms 


of our planet as incomplete. The general public see 
it as a job nearly finished with just a few last hid- 


remain over 10 million microscopic insects to dis- 
cover and name may fascinate biologists, but this 


seven new Palearctic birds have been described 
since 1920, with five of these being in the less- 
explored regions of China (Roselaar, 1994). More- 
over, three of the seven finds lay for years unrec- 
ognized in museum collections before being 
recognized as new: pink-rumped rosefinch, Carpo- 
dacus eos, Vaurie’s nightjar, Caprimulgus centralas- 
icus, and Sillem's mountain finch, Leucoslicte sille- 
mi. The last had been collected 62 years before 
being recognized as new in the Museum of Am- 
sterdam (Roselaar, 1994) 

n 1992, a completely new, large mammal was 


den creatures still to be found. 
ver since Linnaeus began describing and 
counting the species with which Man shares the 
Earth, the number of known species has continued 
to grow. For some groups, the species discovery 
curve continues to rise ever steeper, but for the 
warm-blooded vertebrates, especially the large 
ammals, the curve is leveling off (Medellín & 
Soberón, 1999), and we can hazard some guesses 
as to how many more species there are still to find. 
Medellín and Soberón (1998) estimated another found in the North Annam mountains in the Vu 
247 mammals, mostly small, will be discovered be- Quang Nature Reserve of Vietnam: the Saola or Vu 
tween 1992 and 2032. Quang ox. Assigned to its own genus, Pseudoryx, 
DNA showed the ox was a primitive member of the 
cow and goat family Bovidae (Dung et al., 1993). 
Two sets of the unique horns were initially found 


During the first three decades of this century, 
only a handful of new large mammals were discov- 
red. The finding of okapi, Okapia johnstoni, in 
1901 in the forests of Congo created enormous pop- hanging as a hunter's trophies in a Vietnamese vil- 
ular interest and speculation that this was the last lage. Subsequent morphological and genetic studies 
great new mammal. But other new African finds have shown the animal is so unlike anything else 
were quickly made: giant forest hog, Hylochoerus that it should be regarded as a new subfamily. A 
meinertzhageni, in 1904, and Mountain Nyala, Tra- great amount of media interest was devoted to the 
gelaphus buxtoni, in 1910, before the African vein find, but it took two years before anyone actually 
started to dry up. In 1937, Urbain described the saw the animal alive, when two young animals were 
Kouprey, Novibos sauveli, a large ox of Cambodia, caught by farmers and brought to Hanoi. 
actually found in a zoo in Paris. During the next The find was exciting for several reasons. It 
50 years, only one new large mammal was found. caused a major revision of the taxonomy of the fam- 


! University of Kent, Canterbury CT2 7NX, United Kingdom. 
ANN. Missouni Bor. GARD. 87: 63—66. 2000. 


Annals of the 
Missouri Botanical Garden 


ily Bovidae. It helped highlight the conservation 
status of the neglected Annam Mountains Region. 
But most of all, it made people realize we really do 
not know all our large mammals, and it is still worth 
looking for more. 

Subsequent surveys in Vu Quang turned up a 
second new ungulate, the giant muntjac, Mega- 
muntiacus vuquangensis (Tuoc et al., . The 
finds were made in the same village as was found 
the type specimen of the Saola. This muntjac spe- 
cies was initially given a new genus name because 
it did not match the existing genus description for 
Muntiacus (Groves & Grubb, 1990). Further DNA 
studies have shown the new genus may be insuffi- 
ciently distinct to warrant its own genus (Слао et 
al., . There is some argument (Schaller & 
Vrba, 1996; Groves & Dawson, in press) as to 
whether Muntiacus should be redefined to accom- 
modate the giant muntjac. This cervid mammal dif- 
fers from the other seven known muntjacs by its 
larger size, short pedicles, and much larger, more 
spreading antlers. 

Both Saola and giant muntjac were also found in 
Laos, with wider searches in the Annamite Moun- 
tains revealing a new pygmy muntjac, Muntiacus 
truongsonensis (Giao et al., 1998). In Laos, the Roo- 
sevelt’s muntjac (previously known from only one 
specimen) was rediscovered together with a beard- 
ed pig, Sus bucculentus (previously known from 


— 


only one lost specimen in Shanghai Museum 
(Schaller & Vbra, 1996). A new striped rabbit was 
also found in Laos (formerly thought to be а Su- 
matran endemic genus) and has now also been 
found in Vu Quang and Pumat Reserves in Viet- 
nam, and still awaits scientific description. Another 
small muntjac has also been found in the central 
Annam Mountains of Vietnam, awaiting analysis 
and description (Hulse, pers. comm.). 

Vu Quang, itself in a remote evergreen part of 
the North Annam Mountains on the border between 
Vietnam and Laos, continues to reveal novelties. 
Two new species of fish have just been described 
there (WWF, 1996). 

orth Annam was already recognized as a small 
pocket of local endemism with such local special- 
ities as Owston’s palm civet, Chrotogale owstoni, 
Hatinh leaf monkey, Ls (francoisi) ha- 
tinhensis, Vietnamese pheasant, Lophura hatinhen- 
sis, and sooty babbler, Stachyris herberti. These new 
finds of undescribed mammals add significantly to 
the biological and conservation importance of this 
region. 

These exciting new discoveries in Vietnam and 
Laos seemed to stimulate a new wave of search and 
discovery around the world. A new antelope, Pseu- 


donovibos spiralis, was described from Cambodia on 
the basis of several sets of unique spiraling horns 
Peter & Feiler, 1994). But the failure of efforts to 
find the animal alive suggest we may be too late to 
see or save this species. A new tree kangaroo (the 
bondegezou) was discovered in the Jayawijaya 
Mountains of Indonesian New Guinea. No less than 
seven new marmosets (the latest being Callithrix 
mauesi and C. nigriceps) were added to the mam- 
mals list in Brazil. Two new bushbabies, Galaoides 
rondoensis and G. udzungwensis (Kingdon, 1997), 
have been described from Tanzania. A new horse 
was reported in the popular press from Xinjiang, 
China. 


Are we on a new wave of discovery? In fact, the 


— 


discovery of new mammals has been rather steady 
throughout the century. More than a hundred new 
mammals have been described without attracting 
much public attention (Wilson & Reeder, 1994) 
These have been largely bats, rodents, and insec- 
tivores, or cases of splitting up previously recog- 
nized taxa such as the Sulawesi macaques and Su- 
lawesi tarsiers. 


WHERE WILL FUTURE FINDS BE MADE? 


The Annam Mountains of Vietnam and Laos do 
seem to be a rich, and still not fully explored, 
source of diversity. It is one of the world’s over- 
looked “biodiversity hotspots.” The spectacular 
mammalian finds are largely due to a time warp in 
an area where zoological exploration had been held 
up for 50 years due to constant warfare and polit- 
ical trouble. But these finds do abi us clues about 
where to find yet more new species 

Despite being in one of the most E populéted re- 
gions of the earth and a region heavily devastated 
by both chemical and physical bombardment dur- 
ing the Vietnam War, the North Annam Mountains 
are rugged, difficult to access, unattractive for ag- 
riculture, and ecologically isolated from much drier 
surrounding lowland forests. Highland peaks are 
small and separate, resembling a small archipelago 
of evergreen montane islands. The region is part of 
an evergreen tropical continental system that has 
enjoyed climatic stability for thousands of years 
and where climatic oscillations of the Pleistocene 
could be easily accommodated by species by mak- 
ing minor vertical movement in the steep terrain 
(Giao et al., 1998). These are conditions ideal for 
the creation of local endemic species as well as for 
the survival of primitive and relict forms. The re- 
gion is both a classic Pleistocene refuge and a 
source of new vertebrate radiation. 

The following key characters can be identified as 


Volume 87, Number 1 
2000 


MacKinnon 65 
New Mammals 


indicators of the likelihood of a given geographic 
area still hiding undiscovered forms: 


(1) area of long geological stability; 

(2) area of tropical richness; 

(3) region of long-term humid conditions 
(Pleistocene refugia); 

(4) remote and poorly explored; 

(5) semi-isolated and archipelago-like habitat 
islands; 

(6) relatively small size of habitat islands; 

(7) high levels of endemism in other groups. 


North Annam fulfills all these criteria admirably. 
Most of the localized endemic vertebrates of Asia 
Western 


mountains, Mt. i (Sumatra), Mt. 
(Borneo), Taiwan, Mt. Victoria (Burma), West Javan 
mountains, Sulawesi, the Philippines, and the Mol- 
lucan islands. Contrast these with the generalized 
Asian large mammal fauna of elephant (Elephas 
maximus), tiger (Panthera tigris), leopard (Panthera 
pardus), gaur (Bos gaurus), wild boar (Sus scrofa), 
sambar deer (Cervus unicolor), and red muntjac 
(Muntiacus muntjak), which occur broadly from 
northwest India to Borneo, and through a huge 
range of altitudinal and rainfall differences. These 
latter large mammals probably constitute a fauna 
that has followed Man south and east through Asia, 
benefiting from human opening and burning of the 
forests and displacing the original, more evergreen- 
forest fauna. Glimpses of this richer fauna can be 
seen in the fossil record of the Siwaliks of north 
India. 

In Africa, one also finds that endemism and spe- 
cies richness are concentrated around relict ever- 
green mountains and Pleistocene refugia: west Af- 
rican rainforest, Mt. Cameroon, eastern Rift forests, 
and east Tanzanian forests and mountains. In con- 
trast, the huge forests of the Congo basin and the 

uge savanna plains of east and southern Africa 
have little endemism. 

e new mammals of the 21st century will be 
found in the still unexplored regions that meet the 
criteria mentioned above of isolation, tending to be 
tropical and evergreen systems, and lying within 
the regions of high species diversity or endemism. 
Such unexplored areas remain in northeast India, 
Burma, Laos, southeast Tibet, northwest Yunnan, 
south Philippines, New Guinea, peripheral moun- 
tains of the Amazon Basin, isolated mountains of 
Central America, as well as smaller neotropical 
drainage systems. 

In addition, there are many new descriptions to 
be made among lesser explored taxa and the less 


spectacular and more difficult small mammals. 

irds are much better known than mammals be- 
cause they are mostly diurnal, can mostly be rec- 
ognized in the wild at long range or by vocalization, 
and because the world is swarming with rather pro- 
fessional amateur birdwatchers. In contrast, most 
mammals are nocturnal, live in concealed 
spots, and are very difficult to identify. And some- 
times they smell and bite! 

However, the scientist now has several important 


sma 


new tools to help predict where species are likely 
to be and also to record, catch, and distinguish spe- 
cies. Satellite imagery allows habitats to be more 
easily recognized and mapped, which enables po- 
tential species distributions to be determined. Au- 
tomatic cameras allow shy secretive animals to be 
recorded. Tape recorders and sonographs allow vo- 
calizations to be used for identification and to dis- 
tinguish between forms. GPS (geographical posi- 
tioning systems) allow much greater accuracy in 
locality information, which enables a tighter defi- 
nition of species habitat requirements. IT (infor- 
mation technology) allows much faster comparison 
of material by scientists. Air travel and better com- 
munications allow biologists access to areas previ- 
ously only accessible by major expedition. DNA 
analysis provides a whole new high-resolution tech- 
nique for discriminating relationships between pop- 
ulations and species. 

For instance, the common Grant’s gazelle (Ga- 
zella granti) of East Africa resolves into at least 
three different species on the basis of DNA differ- 
ences (P. Arctander, pers. comm.). The whole con- 
cept of what is a species is raised once again. Some 
species seem morphologically very distinct, but 
DNA reveals they are not. Other species are mor- 
phologically inseparable but found to be very dif- 
ferent genetically. Efforts to define objective mor- 
phological criteria for defining species start to 
break down. Whether one adopts a biological spe- 
cies concept or a phylogenetic species concept, it 
is expected that good species in nature maintain 
discrete breeding. However, more and more evi- 
dence of cross-breeding between wild species is 
discovered. Most organisms accept alternatives if 
the perfect mate is not available, and breeding 
among closely related species does result in hy- 
bridization. All members of the family Cervidae can 

e made to hybridize with all other members in 
captivity (Arctander, pers. comm.). However, even 
wide, stable hybrid zones can be accepted without 
invalidating species status if hybrids remain at a 
disadvantage in the mating game. With birds, for 
instance, it can usually be demonstrated by play- 
back experiments that there is a stronger response 


66 


Annals of the 
Missouri Botanical Garden 


to the true species call than to either the call of the 
second species or a hybrid. As more of these spe- 
cies come under DNA scrutiny we will be able to 
move closer to a phylogenetic species concept, and 
many forms now placed conservatively within one 
species will split. 

e question of where species begin and end 
becomes more complicated when legal aspects be- 
come involved for protection, control of trade, and 
ownership of genetically modified organisms. The 
least recognized unit is ultimately the individual. 
Species evolve from one into another and split 
gradually from one to two through isolation. There 
is no objective cut-off point. We must accept that 
our criteria are rather subjective and not consistent 
across the board. We must redefine satisfactory new 
ways to label the individual. The question of “How 
many species are there?” returns again to the age- 
old taxonomists’ puzzle of “What is a species?” 

The world loses species and genetic variety at 
an unprecedented pace, but thanks to taxonomists, 
our lists grow longer and longer and we should be 
on guard lest a sense of loss is dulled by new dis- 
covery. It is important that taxonomists become 
aligned to the conservation movement, where their 
skills are sorely needed. Monitoring biodiversity is 
as important as describing biodiversity. 


Literature Cited 


99; А new neris e living 
bovid бош da Nature 363: 443—445. 
Giao, P. M., D. Tuoc, E. D. Wikramanayake, G. Amato, P. 


Actander & J. = бено ны Desc ription of Mun- 
tiacus truongsonensis cies of muntjac (Artio- 
dactyla: Манаа sidae) from ees Vietnam, and impli- 
cations for conservation. Animal Conservation 1: 61– 
68 


Groves, C. P. & S. Dawson. The phyletic position of Mega- 
muntiacus vuquangensis: Resolution of a conflict. Mam- 
malia (in press). 

& P. Grubb. 1990. Muntiacidae. Pp. 134—164 in 
G. A. Buberick & A. B. Buberick (editors), Horns, 
Pronghorns and Antlers. Springer-Verlag, New York 

Kingdon, J. 1997. The Kingdon Fieldguide to African 

mals. Academic Press, New York. 

Medellín, R. A. & J. Soberón. 1999. Predictions of mam- 
mal diversity on four land masses. Conservation Biol. 
13: 143-149. 

Peter, W. P. & A. Feiler. 1994. Horner von einer unbek- 
ug Bovidenart aus Vietnam (Mammalia: Ruminan- 

па). Faunistische по oo Museum 
fur rae Dresden 19(30): 2 

Roselaar, C. S. . Notes on јеле Mountain- finch, a 
recently desc ЕЕ species from western Tibet. Dutch 
Birding 16: 20-26. 

Schaller, С. B. & Е. 5. Vrba. 1996. Description of the 
giant muntjac (Megamuntiacus vuquangensis) in Laos. 
J. Mammal. 77: 675—683. 


wson, P. Arctander & J. 


est Inventory and Planning Institute (Hanoi), March 
1994: 4—13. P Vietnames -€— 

Wetzel, В. M., . Dubos, . Martin & P. Myers. 
1975. слаби ап пре а рессагу alive in Paraguay. 
Science 189: 379-381. 

Wilson, D. Е. ^i B M. Wird це), 1994. Mammal 
Species of the World: A Taxonomic and Geographic 
Reference, 2nd ed. а Institution, Washing- 
ton, D.C 
WF. 996. Vietnam nature reserve reveals another 
"new" species. WWF News Release 27 Sep. 1996. 


THE TROPICAL FLORA 
REMAINS 
UNDERCOLLECTED' 


Ghillean T. Prance,? Henk Beentje,? John 


Dransfield,? and Robert Johns? 


ABSTRACT 

Recent fieldwork of the Royal Botanic Gardens, 
Darus 
Reserve near 


areas such as the Ducke Forest 


Manaus, Brazil, and in Brunei where detailed studies of sm 


Kew, in many parts of the tropics reveals the extent to which they 
are still undercollected and poorly studied. Recent studies of palms in Madagascar, Cameroon, 

salam have produced many novelties, for example, in Madagascar, 3 new genera and 85 
юрса from Atlantic coastal Brazil, central Amazonia, and New 


., and Brunei 
ew species. Recent 
n. Even in a ен well collected 
ла areas are 


Guinea are given 


made, many novelties are found. It is recommended that more such intensive studies of restricted areas are 


Ё 
fact between 300, 000 and 320,000 к In order to develop conservation and sus 
ensify the rate of collection before it is too 


it is essential that we continue to in 


new species that are being described, an average of 2350 over the past nine years, and the 


һе 
rate of ит овим to 


ainable use of tropical ecosystems, 


ate 
Key words: Amazonia, е кы Brunei Darussalam, Cameroon, conse т field inventory, Lao P.D.R., 


Madagascar, New Guinea, palms, tropic 


The purpose of this paper is to show, with ex- 
amples from a few places, that the tropical flora 
remains severely undercollected and that many 
new species of angiosperms continue to be dis- 
covered and described every year. The destruc- 
tion of tropical habitats both in areas of rainforest 
and in savannas continues at an alarming rate 
and this before we have completed the inventory 
of what exists. As a result of the incomplete in- 
ventory, we seem to be underestimating the total 
number of seed plants in the world. We have 
identified a few areas of the world where further 
collecting is urgent and destruction of vegetation 
is continuing at alarming rates from a variety of 
causes. It is our hope that this will encourage 
greater efforts in both collection of material and 
habitat conservation before it is too late. The ar- 
eas described are places in which the Royal Bo- 
tanic Gardens, Kew, is involved, and they serve 
as examples of what is also occurring in many 
other places and in many different ecosystems, 
both tropical and temperate. 


THE MATA ATLANTICA OF BRAZIL 


We commence with this area because its de- 
struction is now so well documented and only 
about 6% of the Atlantic coastal rainforest of 
Brazil remains 


intact. It has been devastated 


mainly by the establishment of cacao plantations 
and sugar cane fields as well as by other types 
of farming. Many studies have shown the hig 

level of endemism in this area. For example, Mori 
et al. (1981) showed from a sample of 127 trees 
described in Flora Neotropica monographs that 
53.5% were endemic to the Mata Atlantica. 
There is no doubt that some of these locally de- 
scribed species are now extinct. However, recent 
collecting in the remaining fragments of this eco- 
new species 


system continues to turn up many 
and also interesting disjunct distributions. Two 


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(Cowan, 1979, 1981). An interesting range ex- 
tension is the recent first collection of Anthodis- 
cus (Caryocaraceae) in the Mata Atlantica, a ge- 
nus that was previously only known from the 
Guianas, western Amazonia, and the Chocó in 
Colombia. Anthodiscus amazonicus Gleason was 
found in the forests of Bahia (Prance & Mori, 
1980) and added yet another Amazonian disjunct 
to the many cited by Lima (1953). If the remnants 
of the Mata Atlantica are yielding so many nov- 
elties, what have we lost in the 9446 of the region 
that has been deforested? The conservation of the 
remnants, now declared a World Heritage site, 1s 
vital for the future reconstruction of this habitat. 


! We thank the many local institutions and collaborators who have assisted in the fieldwork in the areas discussed 


here. GTP thanks the Instituto Naci 
ов ч help with the Brent from which data ed. 
oyal Botanic Gardens 


nal de deos da Amazona, Michael Hopkins, and all the Reserva Ducke team 


, Kew, Richmond, Surrey TW9 3AB, United Kingdom. 


ANN. Missouni Bor. GARD. 87: 67—71. 2000. 


68 


Annals of the 
Missouri Botanical Garden 


ew species found in the 100 km? Reserva 
Florestal Adolpho Ducke, Manaus, Brazil in the course of 


preparing a flora. 


No. of 

Family species 
Lauraceae 11 
Sapotaceae 10 
Annonaceae 6 
Araceae 6 
Arecaceae 3 
Lecythidaceae 3 
Passifloraceae 3 
Chrysobalanaceae 2 
Rubiaceae 2 
Other families 9 
Total 55 


THE RESERVA FLORESTAL ADOLFO DUCKE, 
AMAZONAS, BRAZIL 


This 100-km? forest reserve near Manaus in cen- 
tral Amazonia was believed to be one of the best- 
collected areas in Amazonia. In 1993, we set out 
to prepare a Flora and a field guide to the reserve 
and initiated intensive collecting. The preliminary 
list from the INPA herbarium database contained 
825 species (Prance, 1990), and so we assumed we 
were working with a flora of about 1000 species. 
Five years later we now have 2175 species to be 
included in the Flora (M. Hopkins, pers. comm. 
This includes over 50 new species (see Table 1). 


This work shows the value of the intensive study of 


a small area of tropical forest and the value of flo- 
rulas. The recently published florula of the forest 
reserves around Iquitos, Peru, published by the 
Missouri Botanical Garden, is another good exam- 
ple of a local flora that has helped to improve the 
stocks of the botanical inventory of an area (Vás- 
quez Martinez, 1997). In order to complete the in- 
ventory of the tropics, we need many more of these 
detailed floristic studies of small areas. 


MADAGASCAR 


Madagascar is another place that is notable both 
for its high level of endemism of plants and animals 
and for the rapid destruction of the natural vege- 
tation. It is also badly undercollected despite the 
recent efforts of both the Missouri and Kew gar- 
dens. The latter has made a recent study of the 
palms, which serve as an example of what remains 
to be done and what can be found when specialists 
concentrate on a particular group of plants. 


Like many tropical countries with a half-finished 
flora, the families that have been treated in such 
floras are incomplete. The Palmae were written in 
1945 for the Flore de Madagascar et des Comores 
by Jumelle and Perrier de la Bathie, acknowledged 
experts on palm taxonomy. The volume described 
and keyed 115 species, but descriptions were often 
based on scrappy specimens and therefore the keys 
failed to work for most taxa. Consequently, palm 
information for Madagascar was confused, and this 
for a family in which many ethnobotanical uses 
were recorded: palms are used for food, construc- 
tion, basketry, medicine, and many other items in 

adagascar. 

When Uhl and Dransfield (1987) published their 
monumental Genera Palmarum, their main prob- 
lems with generic delimitation and tribal relation- 
ships were in Madagascar. Therefore Dransfield 
embarked on a series of field trips to the island, 
but soon started discovering so many new species 
and additional mysteries that he decided a special 
Madagascar project was needed. With funding from 
McDonald's Restaurants (UK) the project was set 
up, and Henk Beentje started working for it in 
1991. He lived in Madagascar for a year and a half, 
traveling all over the east coast in search of palms. 

The results of this fieldwork plus further visits 
by Dransfield showed how much we did not know: 
3 new genera and 85 new species were discovered 
(Dransfield & Beentje, 1995). These included the 
world's smallest palm (Dypsis tenuissima Beentje, 
less than a foot high) but also 80-feet-high canopy 
palms such as Voanioala, Lemurophoenix, and Or- 
ania ravaka Beentje, and an amazing aquatic palm. 
The latter, Ravenea илы у, Beentje, has seeds 
that float on the water. A small bump splits the 
fleshy fruit wall, and releases the seed, which then 
sinks. The seed has already developed a half-inch- 
long hooked seed leaf within the fruit (a strategy 
analogous to several mangroves), and this hook 
catches on projections on the streambed, enabling 
the seedling to start rooting. The worrying thing is 
that this palm only seems to grow in a single river, 
in which 450 trees have been counted. Many of the 
newly discovered palms have very restricted distri- 
bution, and coupled to the threats to the vegetation 
where they occur, this emphasizes two things: the 
need to conserve, and the need to inventory such 
dwindling habitats. 

'et another new genus, Satranala, was known 
from a few tens of trees in a single reserve; on a 
recent trip, Dransfield visited a second newly dis- 
covered site, bringing the known numbers to the 
low hundreds. Why are such palms so rare? In the 


Volume 87, Number 1 
2000 


Prance et al. 69 
Tropical Flora 


case of Satranala, there is an intriguing hypothesis. 
The fruit endocarp is hard and shows strong flang- 
es, unlike any other palm endocarp—apart from a 
few taxa from New Guinea, distributed by casso- 
waries. The hypothesis is that Satranala was dis- 
tributed by the giant elephant bird, the roc of the 
Thousand and One Nights, the Aepyornis of science: 
a bird now extinct, though its subfossil eggs are still 
found on Madagascar. Therefore, because Aepyornis 
is extinct, long-distance dispersal of Satranala does 
not take place any more, and its area of distribution 
dwindles. 

If the palm project had started 10 years later, 
several of the new species would not have been 
discovered, as they would have become extinct in 
the meantime. With information from projects such 
as these, focused conservation can take place next 
to general conservation, and save species. Without 
the inventory being completed, these rare species 
will just go extinct. 

Lao P.D.R., 


THE PALMS OF CAMEROON, AND 


BRUNEI DARUSSALAM 


Palms are one of the groups of plants that one 
might expect to be well known and well collected 
because they are often both large conspicuous 
plants and almost all are used in some way by local 
peoples. On the other hand, non-specialist collec- 
tors have tended either to avoid collecting palms or 
to make poor specimens because of the special dif- 
ficulties of collecting such large-leaved and often 
spiny plants. A brief summary of three recent field 
projects involving John Dransfield shows what still 
remains to be done in the field to get even a basic 
catalogue of palms. 


CAMEROON 


With relatively few species (about 20 in all), the 
continent of Africa has a poor rattan flora when 
compared with Southeast Asia (Uhl & Dransfield, 
1987). The rattans that do occur are certainly con- 
spicuous and are sometimes visible in great thick- 
ets along roads in perhumid areas of equatorial Af- 
rica. largely 
because they have rarely been adequately collect- 
ed. For some time it has been assumed that in the 
genus Laccosperma (ca. 5 species) there was one 
large-diameter species, Laccosperma secundiflorum 
(P. Beauv.) Kuntze. Beccari had described a second 
large species, L. acutiflorum (Becc.) J. Dransfield 
(as Ancistrophyllum acutiflorum Becc.), based оп 
very poor material. Subsequent workers have been 
content to include the latter as a synonym with L 
secundiflorum. During fieldwork in December 1997 


hey remain poorly understood, 


near Kribi in Cameroon, Terry Sunderland and 
John Dransfield found extensive populations along 
the main road of two clearly distinct, easily sepa- 
rable large-diameter rattans that match the two 
named taxa. They found it astonishing that two taxa 
so clearly distinct, could have been confused; no 
doubt the difficulty of collecting these spiny plants 
was responsible for their poor representation in her- 
baria. Terry Sunderland is now conducting a critical 
survey of all African rattans to put their taxonomy 
and economic development onto a firm basis. 


LAO PEOPLES’ DEMOCRATIC REPUBLIC 


Lao represents one of the largest gaps in our 
knowledge of palms. Until two years ago, only three 
palms were recorded in the literature as occurring 
in Lao P.D.R. As a result of a preliminary survey 
of rattans by the Lao Forestry Department, funded 
by the International Development Research Centre 
and the International Network for Bamboo and Rat- 
tan, we know that there are at least 30 species of 
rattan in Lao P.D.R., but these have yet to be crit- 
ined named. In order to provide a firm base for 
e rattan development in the country, the Lao 
a 'st Department, together with Oxford Forestry 


= 
= 


Institute and Royal Botanic Gardens, Kew, has 
started a new critical survey of Lao rattans and eco- 
logical work aimed at understanding the demogra- 
phy of Lao rattans. This is funded by the UK Dar- 
win Initiative. One of the first and most surprising 
results of the survey is that the premier large-di- 
ameter cane that is being harvested in huge quan- 
tities in Lao, much of it being shipped over the 
border to Vietnam, appears to be an undescribed 
taxon. 


BRUNEI DARUSSALAM 


Until the start in 1988 of Kew’s project to pre- 
pare a checklist of the plants of Brunei, there were 
a mere 17 palms recorded for the country. Yet this 
small nation has one of the highest collecting in- 
dices for the whole of the Flora Malesiana region— 
just the sort of statistic that added to the difficulty 
in justifying further intensive work in the country. 

owever, as in the case of the Reserva Ducke, in- 
tensive fieldwork in a relatively well collected area 
by specialists always yields results. By the end of 
the project in 1995 the palm list had risen from 17 
to 140 different species, showing that almost half 
the Bornean palm flora occurs within the well pro- 
tected forests of Brunei. During fieldwork several 
unusual new taxa were unearthed, including the 
smallest species of Livistona, L. exigua J. Drans- 
field, the handsome Pinanga yassinii J. Dransfield, 


70 Annals of the 
Missouri Botanical Garden 
Table 2. Data from /ndex Kewensis on number of taxa. 
Subspecific taxa 
Genera Species subspecies and variety 
ll incl. new All incl. new All incl. new 
New taxa combinations New taxa combinations New taxa combinations 
Year only new names onl ne es only & new names 
1989 2752 
1990 2653 
1991 2220 
1992 1705 
1993 77 88 2049 3436 508 1663 
1994. 92 102 2126 3557 517 1398 
1995 107 118 2413 4053 487 1400 
1996 95 106 2409 4168 467 1493 
1997 79 94. 2770 4342 543 1586 
5- Year total 450 508 11,767 19,556 2522 7540 
9-Year total 21,097 


and, among the non-palms, the extraordinary Or- 
chidantha holttumii K. Larsen (Lowiaceae). 


NEW GUINEA 


Of all the places in the tropics, New Guinea 
probably remains the least known. This is certainly 
evident from the results of recent fieldwork orga- 
nized by Kew in Irian Jaya, where a large number 
of the collections turn out to be new species. In 
Irian Jaya, collecting density estimates show that 
less than 25 specimens have been collected per 
100 km?, and there are only slightly under 140,000 
collections from a territory with an estimated flora 

‚000 species. The neighboring Papua New 
Guinea has a collection index of well under 50 
specimens per 100 km? and is also poorly known. 

ew Guinea offers a particularly large range of 
habitats because of its rugged topography, where 
you can go from coastal mangrove to the glacier on 
Mount Jaya in a distance of only 110 km. The flora 
varies from tropical rainforest to high alpine. New 
Guinea is a center of diversity for many important 
tropical groups such as tree ferns (Cyathea and 
Dicksonia), Pandanaceae (a recently collected new 
species has thick fleshy, fruity smelling bracts that 
are eaten by bats, which are probably its pollina- 
tors), and Myristicaceae (for example, the recently 
каке кш species Myristica inaequalis У. 
J. de W 

Ped in Irian Jaya is also turning up many 
interesting new records such as the recent collec- 
tion of Ternstroemia magnifica Stapf ex Ridley, 
which was thought to be distributed from the Malay 


Peninsula to Sulawesi. However, it has now been 
shown to occur in [лап 

The importance of Ени these basic data 
about the plants of New Guinea has been empha- 
sized by the fact that Robert Johns has been able 
to prepare maps of proposed conservation areas for 
both Irian Jaya and Papua New Guinea. These 
maps are based largely on plant and vegetation data 
assembled as the result of Johns’s collecting activ- 
ities. This application to systematic data is a vital 
activity in an area where it is not too late to con- 
serve large tracts of this highly endemic flora, 
which is now under severe pressure from both min- 
ing and timber extraction. 


How MAny SPECIES TO Go? 


The most recent estimate of the number of spe- 
cies of angiosperms in the world is 250,000 and 
was originally based on a calculation by Stebbins 
(1974) and confirmed or cited by many other au- 
thors, including myself (Prance, 1977; Groom- 
bridge, 1992). These calculations were based on a 
family-by-family listing of species and provided a 
good estimate based on data of the 1970s. However, 
I am sure that this estimate must now be raised 
considerably. Data in Table 2 show the descriptive 
activity of botanists between 1989 and 1997, dur- 
ing which period 21,097 new species were de- 
scribed. Furthermore, the annual rate has not de- 
clined over the nine years involved. This is backed 
up by data from other sources: for example, 29% 
of the species treated in Flora Neotropica mono- 
graphs are new (Wm. Wayte Thomas, pers. comm.). 


Volume 87, Number 1 
2000 


Prance et al. 71 
Tropical Flora 


These data show two interesting points. Firstly, 
there is obviously still a lot more collecting to be 
done, if an average of about 2350 species are being 
added each year; and secondly, despite the many 
claims that descriptive taxonomy is on the decline, 
we are still describing species at the same rate. 

The fact that we have added 21,097 species over 
nine years, and also the comparison between 1970 
and 1997 data for a few selected plant families, 
leads us to conclude that there are actually between 


300,000 and 320,000 species of angiosperms. 


CONCLUSION 


Data from both the undercollected areas de- 
scribed here and the study of the rate of description 
of new taxa clearly demonstrate that the field in- 
ventory of the angiosperms is far from complete. It 
is necessary to continue to invest considerable re- 
sources into fieldwork and descriptive taxonomy 
and not be tempted to divert them all to the equally 
important and exciting new techniques of molecular 
systematics. More detailed studies of selected small 
areas in the tropics are likely to yield many more 
new species as well as most useful demographic 
data about those that are already described. The 
more complete the inventory is the better the data 
we will have to provide the rationale for conserva- 
tion and for the sustainable use of ecosystems. 


Literature Cited 


Cowan, R. S. 1979. Harleyodendron, a new D of Le- 
guminosae о Brittonia 31: 72-7 


. 1981. New taxa of harry к» 
e from Bahia, Brazil. ee 33: 9-1 
Dransfield, J. & H. Beentje. 1995. The E of Mada- 
gascar. Royal Botanic Carers Kew and International 
Palm Society, HMSO, Norw 
А B. (editor). 1992. G lobal Biodiversity: Sta- 
n ;arth's Living Resources. Chapman & Hall, 


Mou H. & H. Perrier м la Ваше. 1945. 30° Famille, 
Palmiers. Pp. 1-186 in H. Humbert (editor), Flore de 
Madagascar et pie Comores Imprimerie Officielle, Ta- 
nanarive, Madaga 
Lima, D. de A. 1953. p sôbre a dispersão de algumas 
espécies vegetais no Brasil. Anais Soc. Biol. Pernam- 
buco 111): 25-49. 
Mori, s. A., m & G. T. Prance. 1981. Оа 


bution patterns snl conservat 


7 of the tropics: 
ere do we stand? Ann. Missouri Bot. Gard. 64: 659— 


990. Floristic composition of the forests of cen- 
tral Amazonian Brazi 112-140 in A. Gentry (ed- 
itor), Four Neotropical Forests. Yale Univ. Press, New 
Haven. 

& 5. A. Mori. 
ocaraceae), um géne 


1980. Review. Anthodiscus (Cary- 
1 а Amazônia e o 
_65. 


88. Tropical floristics tomorrow. Taxon 37: 
549-56 
Stebbins, с L. 1974. Flowering Plants: Evolution above 
the Species Level. Edward Arnold, London. 

. W. 


Classification. of ie sed on the 
oore Jr. L. H. Bailey Ho onum zd (Bises ы 
Palm Society, Lawrence, Kansas. 

Vásquez Martínez, R. 1997. Flórula de las Reservas Bio- 
lógicas de Iquitos, Perá. Allpahuayo-Mishana Explor- 
napo Camp, рона Lodge. Monogr. Syst. Вог. Mis- 
souri Bot. Gard. 6 


WHAT IS SIGNIFICANT—THE Barbara G. Briggs? 
WOLLEMI PINE OR THE 
SOUTHERN RUSHES?! 


ABSTRACT 


The discovery in 1994 of Wollemia nobilis, a new conifer genus and species of Araucariac бас, attracted media and 
ара attention that was probably phe 'edented for a botanical discovery. If a px ре ay | 

s this one: tall, handsome, rare, of a lineage dating from the Jurassic, surviving undisc a mountain gor ge. 
| was м news around the dy and soon became one of Australia's most е a species. Scientists shared 
the enthusiasm, offering research collaboration to investigate its many aspects. Wollemia has contributed to understand- 
ing of structures in fossil Araucariaceae and conifer-mycorrhizal associations; its survival has added to the picture of 
long-term regional floristic change. After an extended period of small population size it shows no detectable inter-plant 


genetic diversity—releva 


the community the value and nature of biological research and the need for habitat conservation. At the o 


nt to the management of rare plant species. Its discovery helped us explain and emphasize to 


posite end 


of the charisma scale are the southern rushes, Australia's relatively inconspicuous Restionaceae and their allies (Cen- 


trolepidaceae, Ecdeiocoleaceae, Anarthriaceae). D 


espite their links with othe 


т southern continents and close relationship 


to the Poaceae, these had been greatly neglected for over 100 years and were largely misclassified generically. Study 


and fieldwork in recent decades have revealed 


61 formerly undescribed species, nearly 4096 of the total now distin- 


guished in these families for Australia. DNA sequencing of plastid genes gave surprising results, with evidence that 


needs. Many specie 


other groups. Wollemia and fe southern аа exemplify the significance of these new finds а 
e the oe of the 
rom research in many fields. 

Araucariaceae, Australia, conservation, Hoes Southern нео floras, Wollemia. 


understanding of relationships. In each с 
knowledge of organisms and their pac a ge cor 
Key words: 


findings are E to better understanding of the ancestry of 
related еа е о Hemisphere floras. Distinguishing the new rare species allows focus on t 
still being recognized in Australia’s e. among flowering plants = ш as well as 


eir conservation 


y discovered 
dise 'overies is realized на је in Fel ME of the 


WoOLLEMIA—A CHARISMATIC SUBJECT FOR 
RESEARCH 


In 1994 my colleagues and I at the National Her- 
barium of New South Wales in Sydney realized that 
we had been given a remarkable opportunity—the 
discovery of a new plant species that would catch 
public and scientific attention in a truly outstanding 


id Noble of the New South Wales National 
Parks and Wildlife Service had found about 40 
trees of a previously unknown species, soon to be 
named Wollemia nobilis W. G. Jones, K. D. Hill & 
J. M. Allen, a new member of the conifer family 
Araucariaceae (Jones et al., 1995). This rare and 
highly restricted species had been discovered about 

m northwest of Sydney, in a deep gorge 
bounded by sandstone cliffs (McGhee, 1995; Duffy, 
1997). Such a site would differ from most of the 
surrounding area in the constancy of water supply, 
more equable climate, and especially in being 


largely protected from wildfire, although one tree 
showed evidence of fire, followed by resprouting. 
The trees are mostly emergents overtopping both a 
dense fern layer and a canopy of closed forest 
(warm temperate rainforest) of Doryphora sassafras 
Endl. and Ceratopetalum apetalum D. Don. Seed- 
lings are present (about 200 juveniles were record- 
ed; Nash, 1997; Offord et al., 1999), but most only 
produce a few leaves, failing to grow to maturity 
unless a break occurs in the canopy 

The fossil record of the Araucariaceae has been 
studied extensively. The family appeared in the late 
Triassic, with a peak of diversity in the Jurassic and 
a continued decline since the end of the Cretaceous 
(Miller, 1977). Some of the earliest Araucariaceae 
are reported from the Northern Hemisphere, and 
fossil pollen with Araucaria-like features is wide- 
spread in both hemispheres in the Jurassic and 
Cretaceous. Its present survival in the south is thus 
relictual, rather than implying a Gondwanic origin 


I thank my colleagues John Benson, Sue B 
Carolyn Porter, and B 


ullock, Ken Hill, Adam Marchant, Patricia Meagher, Cathy Offord, 
rett Summerell for discussions and for making available unpublished findings. Deborah McGerty 


assisted with ie lle, while Neville Marchant, Director of the PERTH herbarium, and Bruce Fuhrer gave 


permission to us 


? Royal Botanic сюн Mrs Macquaries Road, Sydney 2000, Australia. bgb@rbgsyd.gov.au. 


ANN. Missourt Bor. GARD. 87: 72—80. 2000. 


Volume 87, Number 1 
2000 


Briggs 
Wollemi Pine or Southern Rushes? 


73 


igure l. Wollemia nobilis in its natural habitat. Photo 
Jamie Plaza. 


(Gilmore & Hill, 1997; Setoguchi et al., 1998). Be- 
ore the discovery of Wollemia, the family was 
known from South America, New Zealand, north- 
eastern Australia, New Guinea, and, in especially 
rich diversity, in New Caledonia. Two extant genera 
had been recognized, Araucaria and Agathis. 

The newly discovered Wollemia created a sen- 
sation. It was handsome, and so large—up to 40 m 
tall—that it was almost unbelievable that it had 
been unknown to science until now (Fig. 1). It 
linked with fossils that connected to ancient groups, 
back to the Jurassic (Fig. 2), and this in an age 
when dinosaurs have an unrivalled fascination for 
adults as well as children. Its habitat, in deep 
mountain ravines, held an almost sinister appeal. 

What followed would not apply to the average 
newly found species. 

A media conference was called to announce the 
discovery. This brought a response that exceeded 
our expectations: it was briefly front-page news in 
the press around the world, with journalists and 
science writers seeking further information. Tele- 
vision programs and tapes were prepared, featuring 
its discovery and the subsequent research. 


—A (top). Foliage of Wo "iu deos SN 


Figure 2. 
ONT (also known as A 


g t 
Jurassic fossil member of the Araucariaceae. Photos 
Plaza 


Ап interagency government committee was set 
up to develop a conservation plan (Nash, 1997) and 
to monitor threats and actions affecting its survival. 
Strict protocols for visits to the habitat were estab- 
lished, especially changing shoes at entry to the 
site, to avoid bringing in pathogens. Approvals to 
visit were highly restricted. 

Milton Silverman, who had gone from San Fran- 
cisco with Ralph Chaney in 1948 to collect the 
dawn redwood (Metasequoia glyptostroboides Hu & 

heng) in western China, wrote to congrat- 
ulate us on our efforts and the discovery, which was 
almost ironic considering how much more acces- 
sible our find had been than theirs. The name 
*Wollemi pine" was coined so that we would not 
seem too lacking in words in reporting the discov- 
ery of a plant that had not yet been botanically 
named. The intrepid travelers to China had coined 
“dawn redwood” partly because Metasequoia glyp- 
tostroboides would not fit across a newspaper col- 
umn in reporting their collections of that species, 


Annals of the 
Missouri Botanical Garden 


which, shortly before, had been discovered and 
compared with fossil species. 

Horticultural research and propagation started 
immediately, using cuttings and seeds (Fensom & 
Offord, 1998; Offord et al., 1999). The aims were 
to learn the propagation requirements, establish a 
conservation population in cultivation, and even- 
tually to safeguard the species by widespread use 
in horticulture. As in other Araucariaceae, gro 
was found to be plagiotropic, with plants raised 
from cuttings of lateral branches mostly continuing 
to grow horizontally, whereas those from erect stems 
continue erect growth. Advertisements for commer- 
cial partners in raising large numbers of plants 
brought many proposals. When a young tree was 
planted in the Royal Botanic Gardens in Sydney, 
this was done with ceremony by a senior govern- 
ment кааш and the plant was enclosed in a very 
stout c 

Offers t to collaborate in a diversity of research 
approaches flowed in, 30 within two weeks of an- 
nouncing the discovery on the TAXACOM listserv- 
er (Brooks, 1997). Studies of genetics, chemical 
constituents, embryology, and anatomy, as well as 
associated fungi and insects were soon focused on 

оЏетла. пао of the plastid gene rbcL (Gil- 
more & 997) confirmed the distinctness of 
Wollemia, Шо different analyses using differ- 
ent ranges of other taxa gave sharply contrasting 
phylogenies for the Araucariaceae. Gilmore and 
Hill (1997) and Stefanovic et al. (1998) found Wol- 
lemia to be sister to Agathis, with those two genera 
forming a clade that is sister to Araucaria. By con- 
trast, Setoguchi et al. (1998), using the same se- 
quence data for Wollemia but combined with a dif- 
ferent range of other conifer taxa, concluded that 
Wollemia diverged before the separation of Arau- 
caria and Agathis. It is hoped that the study of 
other genes will resolve this discrepancy. 

Comparison of Wollemia’s adult and juvenile fo- 
liage, stomates, pollen, and cone scales with other 
living and fossil Araucariaceae (Chambers et al., 
1998) helped in the interpretation of fossil Arau- 
cariaceae, especially in the structure of the cone 
scales and seed. Its tree architecture was described 
as unique (Hill, 1997), differing from previously 
described structural models and other Araucari- 
aceae. Male and female cones are each terminal on 
a first-order, short-lived lateral branch, and coppic- 
ing is a consistent feature. Leaf anatomy has been 
studied (Burrows & Bullock, 1999) and so has re- 
production (Offord et al., 1999). The pollen was 
found to be indistinguishable from the fossil pollen 
form-genus Dillwinites, which is recorded in Aus- 
tralia and New Zealand extending back to the late 


rele (Macphail et al., 1995; Chambers et al., 
e most recent fossil records of this pollen 
— from Bass Strait, are about 2 million years 


© 
у 


An endophytic fungus, Pestalotiopsis guepinii, 
was isolated from Wollemia (Strobel et al., 1997) 
and found to produce taxol, which has anti-cancer 
properties and is effective in controlling oomyce- 
tous fungi. The mycorrhizal associates and suscep- 
tibility to common pathogens were studied (B. Sum- 
merell, pers. comm.). More than 50 taxa of fungi 
have been recovered from the trees and their im- 
mediate surroundings in a survey that cultured fun- 
gi from seeds, leaves, leaf litter, roots, root debris, 
and soil. Such a number of fungal associates is 
thought to be typical for a tree species in such an 
environment, but comparisons are uncertain since 
few tree species have been examined so compre- 
hensively. Of these 50 fungal taxa, 9 are thought to 

e undescribed species. Some roots are densely 
filled with endophytic fungi whereas ectophytic 
fungi are found in other cases, an unusual condition 
observed also in other conifers (McGee et al., 
1999). 

Other studies have focused on Wollemia espe- 
cially because it is so rare and vulnerable (Benson, 
1996; Offord, 1996). Wollemia was found to be sus- 
ceptible to some pathogens, including Phytophtho- 
ra cinnamomi, which has been introduced and is 
spreading in southeast and southwest Australia, 
confirming the need for strict protocols for site vis- 
its. Its population genetics was investigated with 
studies of allozymes and DNA (Peakall, 1998), and 
these showed no discernible genetic variation 
among individuals, although more than 800 loci 
were evaluated with AFLP fingerprinting. There 
may be some clonal spread, since plants coppice 
and some have multiple trunks, but the genetic 
findings are interpreted as largely the result of ex- 
tremely low population size over a long time, an 
extended genetic bottleneck. Preliminary data on 
Agathis and Araucaria, while showing some varia- 
tion, indicate that genetic variability is low in the 
amily as a whole (Peakall, 

Was the concentration of attention on Wollemia 
in the media and from scientists justified? This 
question resonated especially when answering 
questions about the scientific significance of the 

nd, while in my view the slender silhouettes of 
other members of the Araucariaceae towered over 
trees of lesser stature on the skyline of Sydney’s 
Royal Botanic Gardens. That question will be con- 
sidered after reviewing a contrasting example. 


Моште 87, Митбег 1 
2000 


Briggs 
Wollemi Pine or Southern Rushes? 


THE SOUTHERN RUSHES 


The Restionaceae and allied families in Austra- 
lia, the southern rushes, are as much in need of 
charisma as Wollemia is blessed with it. The flowers 
are inconspicuous, with small scarious bracts, 
glumes, and tepals (Fig. 3) and with the leaves re- 
duced to sheathing scales (Meney & Pate, 1999). 
They occur exclusively in low-fertility soils and in 
arid or seasonally waterlogged sites, habitats avoid- 
ed for agriculture and therefore of low human pop- 
ulation and often poor access. In one of the allied 
families, Centrolepidaceae, the plants are tiny, 
some species no more than 1 ст tall 

Through such features Restionaceae seem to 
have brought on themselves extreme botanical ne- 
glect over the first half of this century. Before then, 
the three great early figures of Australian botany 
had made a good start on their discovery and clas- 
sification. Robert Brown, naturalist on the first cir- 
cumnavigation of Australia in 1801-1803 de- 
scribed species of 
Restionaceae that are currently recognized (Brown, 
1810). A further 10 species were named by Fer- 
dinand Mueller (1873), whereas George Bentham 
(1878) recognized 71 species. By the early 1960s, 
when L. A. S. Johnson and I started our investi- 
gations, the number of known species had crept to 
86. Around that time also David Cutler of the Royal 
Botanic Gardens, Kew, made extensive anatomical 
investigations (Cutler, 1969). It became clear that 
the generic classification was entirely inadequate 
(Cutler, 1969, 1972; Johnson & Briggs, 1981; 
Briggs & Johnson, 1998a) and that many specimens 
matched no named species 

Clearly this neglected plant group would present 
a fertile field for new discoveries, but it far ex- 
ceeded expectations. The discoveries have been 


seven genera an 


both new species and new understanding of rela- 
tionships, necessitating a radical reclassification. 
In recent decades, great swathes of country had 
become more accessible, especially in the sand- 
plains of Western Australia. Examining the uniden- 
tified collections in herbaria revealed many new 
species; fieldwork brought additional ones. Even 
the largest of all Australian restiads, with flowering 
stems over 2 m tall, is among the recently discov- 
ered species yet to be formally named. Investigation 
of supposedly variable species often showed these 
to be assemblages of several allied species, each 
with a distinctive distribution and ecological range. 
The study brought to light 51 new species, mostly 
from the south of Western Australia. Just when we 
thought that few additional finds could be expected 
in Australian Restionaceae, colleagues in Western 


Australia found a further 10 species (Meney et al., 
1996). There was notably little hybridization among 
the species; the distinctions were sometimes incon- 
spicuous, but they were consistent. 

Restionaceae show an exceptionally high pro- 
portion of newly recognized species, but it is esti- 
mated that about 15% of Australia’s flowering 
plants are still to be discovered (A. Orchard, pers. 
comm.), in addition to many now distinguished but 
awaiting publication. 

The new view of Australian Restionaceae did not 
stop at species. When this study began, 29 Austra- 
lian species were named within the genus Restio, 
but it became clear that Restio was a member of a 

p of genera limited to Africa and Madagascar 
(Cutler, 1972; Johnson & Briggs, 1981; Linder, 
1985, 1991; Briggs & Johnson, 1999). Therefore, 
new genera were required, or old synonyms taken 
into use, to accommodate all the Australian species 


hitherto included there (Briggs & Johnson, 1998a, 


Moreover, Restionaceae showed a pattern similar 
to that in Proteaceae, Fabaceae, Ericaceae, and Po- 
aceae in their post-Gondwanic floristic richness. 
The history of climates, migrations, and survivals 
has left different traces on the African and Austra- 
lian continents, so that Africa has large numbers of 
species in relatively few genera, but Australia has 
a diversity of groups appropriate for recognition as 
genera. This pattern has shown even when the same 
botanist studied groups in both continents, rather 
than being an artifact of different generic concepts 
Peter Weston, pers. comm.; Nigel Barker, pers. 
comm.), although it is not apparent when Protea- 
ceae of the Cape Region are compared with only 
the southwest of Western Australia (Cowling & La- 
mont, 1998). Our case led to the description of the 
rather alarming number of 16 new genera of non- 
African Restionaceae (Briggs & Johnson, 1998a). 

Morphological cladistics (Linder et al., 2000) 
and DNA data both indicate that an early division 
within Restionaceae is between the African clade 
and the Australasian clade (though the DNA data 
give only weak support). This would be consistent 
with an ancient Gondwanic connection. By con- 
trast, the single species in South America, Apodas- 
mia chilensis (Gay) B. G. Briggs & L. A. S. Johnson, 
is extremely similar to the New Zealand A. similis 
(Edgar) B. G. Briggs & L. A. S. Johnson, indicating 
long-distance dispersal. Moreover, Apodasmia (re- 
cently segregated from Leptocarpus; Briggs & John- 
son, 1998a) includes foredune coastal species and 
is the only notably salt-tolerant genus of the family; 
it is singularly well suited to establish successfully 
after dispersal. 


— 


Annals of the 
Missouri Botanical Garden 


Volume 87, Number 1 
2000 


Briggs 77 
Wollemi Pine or Southern Rushes? 


Sequencing of plastid DNA was done in parallel 
with morphological studies and gave a further un- 
expected result, evidence that two new plant fam- 
2000). 


Hopkinsia and Lyginia are small genera, of two and 


ilies should be recognized (Briggs et al., 


three species, respectively (one species of each ge- 
nus undescribed). Their inclusion in Restionaceae 
had never been questioned, even when they were 
the subject of detailed anatomical investigations 
(Gilg, 1890; Cutler, 1969 


sequence data (from rbcL, and from the trnL intron 


ut two sets of 


with the trnL-trnF spacer) are consistent in group- 
ing them (each with 100% jackknife support) with 
Anarthria (Fig. 4) rather than with Restionaceae. 
That grouping is shown in a jackknife consensus 
tree from analysis of the total sequence from these 
DNA regions and is further supported by the pres- 
ence of two distinctive indels (one insertion and one 
deletion) in the trnL intron (Fig. 4). 

though further investigations are needed and 
An- 


arthria, Hopkinsia, Lyginia) is not the sister group 


~ 


proceeding, it appears that the Anarthria clade 


to Restionaceae. That position appears to be held 
by Centrolepidaceae. Such close affinities, or even 
inclusion of Centrolepidaceae within Restionaceae, 
have been suggested on morphological and embry- 
ological grounds (Hamann, 1962, 1975; Kellogg & 
1995; 2000) ie now have 
some support (although not robust) from analyses 
f DNA data (Fig. 4; Briggs et al., 2000). It has 


been suggested that Centrolepidaceae are neoten- 


Linder, Linder et al., 


ous, with mature plants showing some similarities 
to seedlings of related families, although their in- 
florescences are very different. 

Despite the evidence that Hopkinsia, Lyginia, 
and Anarthria form a clade, they share no syna- 
pomorphies of morphology, anatomy, flavonoids, 
pollen, or seeds, except for features that are either 
plesiomorphic within the Poales or widespread in 
the order. Similarly, studying these genera in light 
of t data showed that those features that 
they (бан in common with Restionaceae аге рје- 
siomorphies, although each has distinctive auta- 
pomorphies. Hopkinsia has a reduced carpel num- 
ber and succulent indehiscent fruits. Lyginia shows 
a distinctive arrangement of thick- and thin-walled 
cells interspersed in the chlorenchyma, sloping sto- 
mates, fused stamen filaments, and highly distinc- 


tive seeds ornamented by minute pits and spines. 
Anarthria lacks a sclerenchyma cylinder in the 
culms and has unreduced, ensiform leaves and also 
large chromosomes (Briggs, 1966). Such chromo- 
somes are, in general, associated with large ge- 
nomes, an apomorphic feature (Bennetzen & Kel- 
logg, 1997; Bennett & Leitch, 2000). Without some 
morphological basis there appears to be no case for 
enlarging Anarthriaceae or describing a single new 
family for Hopkinsia and Lyginia. The most logical 
course is the recognition of *Hopkinsiaceae" and 
"Lyginiaceae," and these new families are being 
described (Briggs & Johnson, in press). 

e families mentioned above, Restionaceae, 
Anarthriaceae, — *Hopkinsi- 
aceae," "Lyginiaceae," together with the Ecdeio- 
coleaceae, Joinvilleaceae, and Flagellariaceae, ap- 


Centrolepidaceae, 


the closest relatives of the Poaceae 
(Dahlgren et al., 1985; Chase et al., 1993; Duvall 
1993; Briggs et al., 2000). These are all fam- 
ilies with primarily Southern Hemisphere distri- 


pear to be 
et al., 


bution. Especially notable is their high concentra- 
tion in the southwest of the Australian continent: 
six of the eight families occur in that region and 
four are limited to it. Together with nine other fam- 
ilies they constitute the Poales as recognized by the 
Angiosperm Phylogeny Group (APG, 1998). Three 
of the other families are primarily in the Southern 
Hemisphere (Hydatellaceae, placed here but with 
little evidence, Prioniaceae, and Thurniaceae), 
while six are distributed in both hemispheres (Cy- 
Eriocaulaceae, Juncaceae, Spargani- 
aceae, Typhaceae, and Xyridaceae). The concentra- 


peraceae, 


tion of allied families in the south has led to the 
suggestion that the Poaceae itself had a southern 
origin (Doyle et al., 1992), although these distri- 
butions could also be relictual, as with the Arau- 


Carlacede. 
WHAT IS SIGNIFICANT? 


The examples considered above are two ends of 
a spectrum. In Wollemia nobilis we have a single 
new species and genus in a recognized plant family, 
but an exceptionally charismatic and interesting 
find. In Restionaceae and its allies we have some 


60 new species, many new genera, and two new 


€ 

Figure 3. Australian Restionaceae and Anarthriaceae 
spikelets, each ca. 1.5 em long. 
mm long. —C (bottom left). Meeboldina scariosa, male, s 


Anarthria scabra, female, with prominent stigmas; linear E ca. 6 mm wide; 


—B (top right). je pet ibo ке subsp. metostachyum, female, 


. —A (top left). Lepidobolus preissianus, male le ft) and female 


pikelets ca. 5 
elets ca. 4 mm long: photo B. Fuhrer. —D y si right). 


` photo B. Fuhrer 


Annals of the 
Missouri Botanical Garden 


53 Desmocladus castaneus 
82 Harperia lateriflora 
99 Kulinia eludens 
75 Coleocarya gracilis 
Lepidobolus chaetocephalus 


Dielsia stenostachya 


97 [— — Baloskion gracile 
L—— Baloskion tetraphyllum 


Melanostachya ustulata 
р" у 
Lu Tyrbastes glaucescens 
Guringalia dimorpha 


100 
88 Acion hookeri 


96 Saropsis fastigata 
98 | 
Chordifex stenandrus 
64 100 Chordifex amblycoleus 
Chordifex jacksonii 


Tremulina tremula 


89 Loxocarya gigas 


Meeboldina coangustata 


Australian Restionaceae 


Meeboldina cana 
Leptocarpus tenax 


Dapsilanthus ramosus 


82 Chaetanthus aristatus 
Alexgeorgea ganopoda 
Winifredia sola 
53 93 $ [ Taraxis grossa 
Empodisma minus 
Eurychord planat 


80 Calorophus elongatus 
88 == | 
d onis 
100 Sporadanthus tasmanicus 


Elegia cuspidata African Restionaceae 


80 Centrolepis strigosa Centrolepidaceae 


P 


pl Hopkinsia adscendens 
Lyginia barbata Anarthria 
| аде 


а 
Anarthria polyphylla 


Zea mays 
"LIS Triticum aestivum 
Огуга sativ 
99 | $ a Poaceae 


100 


Ecdeiocolea monostachya Clade 


Georgeantha hexandra 


Carex Cyperaceae 


Figure 4. Jackknife consensus tree for Restionaceae and allied families from parsimony analysis of chloroplast 
ata. Numbers indicate jackknife support for individual nodes. -bars indicate unique (non-homoplasious) 


D 
indels, in the шта, intron ог trnL-trnF spacer, that аге synapomorphies for clades. 


Volume 87, Number 1 
2000 


Briggs 79 
Wollemi Pine or Southern Rushes? 


plant families. Neither of these examples is typical 
of the situation in the Australian flowering plants. 

Wollemia created unprecedented public, media, 
and scientific interest. It raised public enthusiasm, 
awareness, and knowledge of environmental and 
biodiversity conservation issues. Its importance and 
rarity make it a wonderful example in education 
programs and political contexts. It has emphasized 
to the community the need for habitat conservation 
in species survival and given a focus for programs 
to explain the nature of biological research. To in- 
vestigate its significance and conservation there has 
been research in systematics and evolutionary re- 
lationships, palaeontology, ecology, genetics, plant 
pathology, mycology, and plant propagation; this 
has helped to publicize the role of all these disci- 
plines (Hill, 1996). School groups, 
members, and the general community are enthusi- 
astic about seeing the plants, so it has raised the 


government 


profile of our botanic gardens and of their scientific 
and educational programs; it has also been a major 
profile-raiser for the New South Wales National 
Parks and Wildlife Service. Wollemia has contrib- 
uted to understanding of structures in fossil Arau- 
cariaceae and conifer-mycorrhizal associations; its 
survival has added to the picture of long-term re- 
gional floristic change, perhaps even a step in the 
regional replacement of coniferous vegetation by 
flowering plants. After an extended period of small 
population size, it shows no discernible inter-plant 
genetic diversity. 

ur other example, Restionaceae and allies, was 
probably the most neglected of all substantial Aus- 
tralian flowering plant groups and so was the rich- 
est site remaining for new discoveries. Distinguish- 
ing the many new rare species permits a focus to 
be developed on their conservation needs. Better 
knowledge of relationships within the Restionaceae 
clarifies an instance of the distinction between the 
intercontinental links that date from at least Gond- 
wanic times and those that may represent relatively 
recent long-distance dispersal. In addition to great- 
er understanding of Southern Hemisphere floras, 
there is improved understanding of relationships 


demon- 
stration (Fig. 4) of the closeness of ACR mend 
to Poaceae, in agreement with Doyle et al. (199 

Both examples are significant, but in each case 
the significance of the discoveries is only fully re- 
alized in the context of knowledge of organisms and 
their evolution that has been established by re- 
search in many fields. The history and antiquity of 
the Araucariaceae make Wollemia so notable. Sim- 
ilarly, relationship to the Poaceae adds to the rel- 


evance of findings in the Restionaceae. The histor- 
ical biogeography of world floras, especially those 
of the Southern Hemisphere, provides a context for 
discoveries in both these groups; findings in these 
groups, in turn, clarify aspects of the development 
of these floras. 


Literature Cited 


APG (Angiosperm Phylogeny Group). 1998. An ordinal 
classification for the families of flowering plants. Ann. 
Missouri je jue 85: 531-553. 

Bennett, M. D. & I. J. Leitch. 2000. Variation in nuclear 
DNA bane (C-value) in monocots and its significance. 
Pp. 135-143 in K. L. Wilson & D. Morrison (editors), 
Systematics and Evolution of Monocots. CSIRO, Mel- 
bourne. 

Bennetzen, J. L. & E. A. Kellogg. 1997. Do plants have 
a one-way ticket to genomic obesity? Pl. Cell 9: 1509- 
1 


Benson, J. 1996. Threatened by discovery: Research and 
management of the Wollemi Pine, Wollemia nobilis 
Jones, Hill & Allen. Pp. 105-109 in S. Stephens & S. 
Maxwell (editors), Back from the Brink: Refining the 
aes Species Recovery Process. Surrey Beatty, 

E G. 1878. Flora Australiensis, Vol. 7. Reeve, 

э. Chromosome numbers of some Aus- 

tralian monocotyledons: Contr. New South Wales Natl. 

erb. 4 E Pu 
S. John 1998a. New genera and s 
cies of reine Ru. (Poales). Telopea 7 
345-373. 


b. New combinations arising from 
a new classification of non-African Restionaceae. Tel- 
opea 8: 21-31. 

uide to a new classification 
of Restionaceae and allied families. Рр. 25-56 in К. А 
Meney & J. S. Pate (editors), Australian Rushes, Biol- 
ogy, Identification and Conservation of Restionaceae 
and Allied Families. Univ. Western Australia Press, 
Nedlands 

& 


. Hopkinsaceae and Lyginiaceae, two 

new families of Poales in Western Australia, with revi- 
sions of Hopkinsia and Lyginia. Telopea 8 hs press). 

‚ А. D. Marchant, S. Gilmore & С. L. Porter. 2000. 

e Tapes phylogeny of the Restionaceae and allies. 

57 in K. L. Wilson & D. Morrison (editors), 

decis E Evolution of Monocots. CSIRO, Mel- 


urne 
Brooks, A. 1997. The је 4 in the Gardens. Pp. т in 
B. Diekman, E. Higginson, F. Sutton & H. Webb (edi- 


gy at the bine nec Nature Conservation 

Council of New S. Wales, Syd 

Brown, R. 1810 б. Prodtoihus Florae Novae Hollandiae et 
Insulae Van еен. J. Johns don. 

Burrows, G. E. & S. Bullock. 1999. Leaf anatomy of Wol- 
lemi Pine (Wollemia nobilis, Araucariaceae). Austral. J. 
Bot. 47: 61-68. 

Chambers, T. C., A. N. Drinnan & S. McLoughlin. 1998 


ogical features of Wollemi Pine (Wollemia 


Les, E D. Mishler, M. R. Duvall, R. A. Price, 


80 


Annals of the 
Missouri Botanical Garden 


. Hills, Y-L. Qiu, K. A. AE H. Rettig, E 


V. A. Albert. 1993. Phylo- 
An analysis of nucleotide se- 
uences ga the plastid gene rbcL. Ann. Missouri Bot. 
Gard. 80: 528-580 
Cowling, R M & B. B. Lamont. 1998. On the nature of 
Gondwanan species flocks: Diversity of Proteaceae in 
Че шкы ep western Australia and South Afri- 
ca. Austr е. . 46: 335-355. 
Cutler, D. 969. pe ales. Pp. 1-357 in С. R. Metcalf 
(editor), Pi of the Monoc :otyledons IV. Clarendon 
Press, Oxfor 
972. Vicarious species of Restionaceae in Af- 
rica, Australia and South America. Pp. 73-83 in D. H. 
Valentine (editor), Taxonomy, Phytogeography and Evo- 
Academ is "gs London. 
. T. Clifford & P. F. Yeo. 1985. The 


Families of ie Vieh t s Springer- Verlag, Ber- 
lin 


Doyle ‚ Ј. J., J. L. Davis, В. J. Soreng, D. Garvin & M. J. 
Anderson. 1992. Chloroplast DNA inversions and the 
жае of the cq family (Gramineae). Proc. Natl. Acad. 

, U.S.A. 89: 7722-7726. 

Duffy. M. TS Dave Noble and the Wollemi Pine. Pp. 
177—186 in D. Foster et al. (editors), Crossing the Blue 
Mountains: Journeys through two centuries, from natu- 
ralist Charles Darwin to novelist David Foster. Duffy & 
Snellgrove, pu Point, N.S.W. 

Duvall, M. . T. Clegg, M. W. Chase, W. D. Clark, с 
J. Kress, H. G. Hills, L. 
Gaut, E. A. Zimmer & G. H. 
genetic hypotheses for the EEE constructe 
from rbcL sequence data. Ann. Missouri Bot. Gard. 80: 
607—619 

Fensom, G. &C. Offord. 1998. Propagation of the Wollemi 
Pine (Wollemia nobilis). Proc. Int. Pl. Propag. Soc. 47: 


66 Я 
Gilg, Е. 1890. Beitrüge zur vergleichenden Anatomie der 
xerophilen Familie der Restionaceae. Bot. Jahrb. Syst. 
3: 541-606 


Gilmore, 5. & K. D. Hill. 1997. Relationships of the Wol- 
lemi Pine (Wollemia nobilis) and a molecular phylogeny 
of the и ле ел Telopea 7: 275-291. 

Hamann, U. Beitrag zur Embryologie der Centro- 
lepidaceae al Bemerkungen iiber den Bau der Bliiten 
und Blütenstünde und die systematische Stellung der 
rn Ber. isi Bot. Ges. 75: pu dn 

———— 1975. e Untersuchungen zur Embryologie 
und inii is Centrolepidaceae. Bot. Jahrb. Syst. 
96: 154-191. 

Hill, K. D. 1996. The Wollemi Pine: Discovering a living 
fossil. Nat. нчен 32: 20-25 

1997. Architecture of ihe. Wollemi Pine (Wolle- 
mia e Araucariaceae), a unique combination of 
model and reiteration. Austral. J. Bot. 45: 817-826. 

Johnson, L. A. S. & B. G. Briggs. 1981. Three old south- 


ern families—Myrtaceae, Proteaceae and Restionaceae. 
Pp. 429—469 in А. Keast Ка Ecological Biogeog- 
raphy of ок Junk, Utrecht. 
Jones, W. G., . Hill т M. Allen. 1995. Wollemia 
nobilis, a new ling Australian genus and species in 
—176. 
Н. Р. Linder. 1995. Phylogeny of Poales. 
Py ›. 511- 542 in P. J. Rudall, P. J. Cribb, D. F. Cutler 


Humphries (editors), PO гаыа id 


>{ 
= 

ш 

= 

5 

® 

d 

> 

Е 

c 

e 

t 

EX 
о 
= 

Ф 

> 

= 
— 
=! 
w 


African species of 
Restionaceae. Bothalia 15: 387-5 

——. 1991. A review of the ee African Restion- 
асеае. Contr. пи dei p^ 209-264. 

——., В. С. Bri S. Johnson. 2000. Restion- 
aceae—a онсе ей ial У Рр. 639-661 in К 
L. Wilson Morrison (editors), eens and 
Evolution of Per ots. CSIRO, Melbour 

Hill, A. Partridge, E. Truswell & C. Fos- 

ed Pine”—Old pollen records for а 

а: discovered genus of gymnosperm. Geol. Today 


: 48- 
© Р. А 5. Bullock & В. А. Summerell. 1999. Struc- 
ture of mycorrhizae of the Wollemi Pine е по- 
ie and related Araucariaceae. Austral. J. Bot. 47: 


MES K. 1995. Wollemi Pine. Nature Australia 25(2): 
22. 


Mene . Pate. 1999. Australian Rushes, Bi- 
clogs Identific ailion and Conservation of Restionaceae 
and Allied Families. Univ. Western Australia Press, 
Nedlands. 

| г К. W. Dixon. 1996. New species of 

Restionaceae from Western Australia. Telopea 6: 649— 


Miller, C. N. 1977. Mesozoic conifers. Bot. Rev. 43: 217- 


Mueller. К. 1873. о шш > Vol. 

asc. 62. Government | r, Melbou 

me S. 1997. Wollemi Pine 5 ecies Recovery 7 Syd- 
ney Zone, New South Wales. National Parks and Wild- 
life Service, Hurstville. 

Offord, C. A. 1996. Conserving the Wollemi Pine: An in- 
tegrated арр. пи 5(2): 12-1 4. 

.L.P >, К. Meagher & С. Errington. 1999. 
Sexu » г iak ind a plant И of the Wol- 
lemi Pine (Wollemia nobilis), a rare and threatened Aus- 
tralian conifer. Ann. Bot. 84: 1-9. 

Peakall, R. 1998. Exceptionally low genetic diversity in 
an ancient relic, the Wollemi Pine: une cations for con- 
servation theory “У practice. id 1 Genetics Soc. 
Austral. 45th Con зоне Зудп 

Setoguc "hi, T. a, J.-C. Pin aud, T. Jaffré a 

M. Veillon. 1998. Porc rin ships within A 
aucariaceae based on rbcL gene sequences. Amer. у 
Вог. 85: 1507-1516. 

Stefanovic, S., M. Jager, J. Deutsch, J. Broutin & М. Mas- 

selot. 1998. Phylogenetic relationships of conifers т- 
fered from partial 28S rDNA gene sequences. Amer. J. 
Bot. 85: 688-697. 

ine G. A s, J.-Y. Li, E. Ford, J. Sears, R. 

idhu & B. leas 1997. Pestalotiopsis guepinil, 

a x producing endophyte of the Wollemi Pine, Wol- 

lemia nobilis. Austral. J. Bot. 45: 1073-1082. 


FLORISTIC SURPRISES IN 
NORTH AMERICA NORTH OF 
MEXICO! 


Barbara Ertter? 


ABSTRACT 


Contrary to recurring perceptions that the flora of North America north of Mexico has been fully explored and 
ury and shows 


cataloged, the rate of ongoing discov eries has 


remained remarkably: constant for m 


new species and occasional monotypic genera are still coming to lig t, even along highways and n 
rthermore, the same level of ongoing discovery also characterizes other aspects of floristic information, including the 
distribution of rare species and the occurrence of invasive pest plants. The majority of ongoing discoveri t 


ac of floristics as rote data compilation, Wien it i 


The | incompletenes 


that. will сей determine the fate of our national floristic heritage. The cos 


omplex | situations are encoun 
s in fact better understood | in the context n 
model biodiversity. resulting in an intricate suite of nested hypotheses that are constantly being tested and mo 
s of our floristic knowledge takes on critical significance in an era w 


a massive attempt 
dified. 
en decisions are being made 
f this ignorance can cut multiple 


о 
ways, increasing the risk of misplaced mitigation efforts as well as avoidable P: of irreplaceable biodiversity. Although 


the magnitude of the ta 
yield a ane improved floristic knowle 
ey w 


sk is daunting, significant advances are ac 
ge base for informed decision- akin ng. 
t flora. 


ievable in a collaborative framework, which would 


ords: biodiversity, floristics, North America, vascular plant 


Underlying much of our current land-use man- 
agement planning, legislation, funding allocations, 
and hiring decisions is the assumption that the flora 
of North America north of Mexico "Hot brevity's 
sake, hereafter referred to simply as “North Amer- 
ica") has been fully explored, cataloged, and 
mapped, at least to the extent that is needed for 
informed decision-making. Or, to the extent that ex- 
ceptions are allowed, it is assumed that such 
knowledge accumulates in the form of static data 
sets, descriptive rather than truly scientific in na- 
ture, and further that the existing academic infra- 
structure is adequately addressing the gaps in our 
floristic knowledge. This paper sets out to challenge 


diversity of ongoing floristic surprises in North 

merica, whereas the second half addresses the 
factors that influence these discoveries and the re- 
sultant implications 

The majority of statistics and examples that form 
the basis of this paper are derived from the author's 
personal expertise and vascular plant focus. The 
resultant western North American bias should not, 


however, obscure the fact that this region is a rich 

source of ongoing novelties. An effort has never- 

гаң been made to include examples from other 

aphic areas and representing other groups tra- 

ditionally studied by botanists: bryophytes, algae, 
li 


of examples should not 
reflection of actual discoveries among geographic 
areas and plant groups, or of their significance to 
science or land-management issues. 


ASSUMPTION 1: THE FULLY CATALOGED FLORA 
A. HISTORICAL PERCEPTIONS 


The perception that the vascular plant flora of 
North America has been fully explored and cata- 
loged has a surprisingly long history, as analyzed 
from our current state of knowledge. As early as 
1858, Thomas Bridges, an Englishman collecting 
in California, wrote the following to Sir William J. 
Hooker (quoted in Jepson, 1933): 


“I can scarcely describe to you how pleasing and grat- 
ifying it has been to me to learn that in my collections 
you have found some new and rare plants—I was par- 
tially under the impression that from the labours of 
Douglas, Hartweg, Jeffrey, Lobb and other travelers 


! Excerpts from the October 1998 presentation on which this paper is pri have been subsequently highlighted in 


several media publications, notably U.S. News 


and World Report (Tangley, 1998) 
? University and Jepson Herbaria, University p California, Berkeley, California 94720-2465, U.S.A 


and Science News (Milius, 1999). 


ANN. Missouni Bor. GARD. 87: 81-109. 2000. 


Annals of the 
Missouri Botanical Garden 


from Europe with the many United States Exploring Ex- 
peditions that little or nothing remained to be discov- 
ered an only gleanings were left to those of us of the 

present day.’ 


As it happens, the “gleanings” left by Bridges’s 
predecessors comprised the majority of California’s 
flora as currently known. In fact, the number of 
known species increased by one-fourth during the 
subsequent two decades under the auspices of the 
California Geological Survey, primarily due to the 
efforts of William H. Brewer and Henry Nicholas 
Bolander. The two-part botanical report of the Cal- 
ifornia Geological Survey (Brewer et al., 1876; Wat- 
son, 1880), which represented the first comprehen- 
sive flora of California, included full entries for 
approximately 3450 vascular plant taxa. This con- 
trasts both with the initial estimate of 2000 (as not- 
ed in Whitney’s introduction to the first volume) 
and the latest tally of 7036 vascular plant taxa rec- 
ognized as occurring outside of cultivation in Cal- 
ifornia (Hickman, 1993). Not only were there only 
half the number of taxa known in 1880 as in 1993, 
but there is by no means a strict one-to-one cor- 
respondence within the apparent overlap, primarily 
due to misapplied names and non-persisting intro- 
ductions. 

As it happens, Bolander was himself guilty of 
seriously underestimating what still lay waiting to 
be found, when he challenged Alphonso Wood’s 
claim of collecting 1490 species of flowering plants 
on a journey from San Diego up the coast and 
through northern California in 1866. In an address 
to the California Academy of Natural Sciences, 
"Professor Bolander considered it probable that 
there were not over 500 species of flowering plants 
actually existing in that part of California" (Leviton 
& Aldrich, 1997: 87). On the contrary, well over 
4000 taxa of vascular plants are now known to oc- 
cur in the biogeographic subdivisions of California 
that Wood traversed (as calculated from Hickman 
[1993] by R. L. Moe, pers. comm. 1998), though 
how many of these Wood actually encountered is 
admittedly another matter. 

olander's attitude was in full sway several de- 
cades later, when Katharine Brandegee accused 
Edward Lee Greene of conflating California's flora, 
with the statement, “It is safe to say that not more 
than one in ten of [Greene's] species is tenable, and 
probably one in fifteen or twenty would be nearer 
the mark" (Brandegee, 1893: 64). It turns out that 
Brandegee was actually the one who was way off 
the mark, in that a respectable 7096 of Greene's 
taxa, at least those described while he was residing 


in California, have stood the test of time (McVaugh, 
1983) 


A marvelous anecdote relayed by Heller (1908: 
12—13) from one of his correspondents shows just 
how well ensconced was the general belief that the 
North American flora had been fully cataloged by 
the end of the 19th century: 


"[M]y first botanical work was done in California, where 
my teacher was loo 
essence of know 


years old .... 
[in the mountains near San Diego] which probably was 
not searc shed over botanically or qas ally before 
esult I would return with 


usual words whic s took place were 
about as follows on the Voci ina part: *Can't you find 
these in the botany?’ ‘No: Stud of the specimens and 
consulting 


They are not good for anything on that 
account. Throw them away.’ 


As a final example in the botanical lore, one of 
Brandegee's supporters, Marcus E. Jones, is said to 
have commented that “ће felt sorry for all future 
generations of botanists because he [Jones] had 
named all the western American taxa, and there 
would be nothing left for them to do" (S. Welsh, 
pers. comm. 1998). To the contrary, the rate of dis- 
covery of rare plants in Jones's home base of Utah 
remains high, with a significant peak in the 1980s 
(Stone, 1998; Fig. 1) 

In essence, the inclination to believe that the era 
of floristic exploration in North America is over ap- 
parently has an inherent persistency to the point of 
becoming a psychological gris inns worth in- 
vestigating in its own right. In the words of Stan 
Welsh (pers. comm. 1998), * ‘Each major publication 
on western plants has left the impression that a 
of the work has been done, that nothing remains to 
be discovered, that everything worth naming has 
been named.” The perceptions of the 19th century 
have accordingly become the dogma of the 20th 
century, in which the common understanding is that 
the flora of North America has, with the rare ex- 
ception, been fully explored, cataloged, and 
mapped (Reveal, 1991). At its worst, the attitude 
developed that anyone describing new species of 
plants from North America was indulging in species 
conflation for the sake of ego gratification, rather 
than practicing valid science. 

Against this tide, there have admittedly been 
some voices to the contrary. In his introduction to 
the second volume of the botanical report of the 


Volume 87, Number 1 Ertter 83 
2000 Floristic Surprises 
90 
с 
x 
© 
|- 
5 
~ 
8 
Е 
5 
= 
$? S9 <> 39 со q^ се 
S „© $° $° S? S S? S? S 
SSS S os "8 ef Far Re C “OSs СУ 
Decades 
Figure 1. History of rare plant description in Utah (figure and caption prepared by R. D. Stone). Bars represent 


the iim of rare vascular plant taxa (species, subspecies 
each 10- 


. and varieties) in Uta 
year period. ra plants (N = 242) are defined as those taxa “with known or мее range-w 


1 that were formally described within 
wide viability 
is understandable 


ed. This 


ut te dis to be removed from rare 


The data show that most rare p in Utah are recently de 
for t two reasons: (a) ae are often considered rare when they are first described * 
lists a 


they become better known; and (b) after more а. a century of plant exploration and discovery in Utah, the 


& en now being described tend to be “the rarest of the rare.’ 


Geological Survey of California, Watson (1880) in- 
dicated, “There still remains ample opportunity for 
good botanical work at almost any locality among 
the mountains, hills, and valleys of the State, to 
which it is hoped that these volumes may prove 
both an incentive and an aid." And, in a summation 
the Broadening Basis of 
) noted, 


lecture of a symposium on 
Classification, Lincoln Constance (1964 
*Many otherwise informed persons assume that the 
exploratory phase of botany is essentially complete; 
this assumption is, of course, entirely erroneous.’ 


B. STATISTICAL CHALLENGES 
(1) Shevock & Taylor (1987) 


Possibly the first statistical challenge to the com- 
mon perception was that of Shevock and Taylor in 
87, provocatively titled “Plant exploration in 
California: The frontier is still here." In it, the au- 
thors tallied 219 vascular plants described from 


California for the two decades from 1968 to 1986, 
an average of 11 taxa per year. Taylor (pers. comm. 
1998) has continued the analysis, demonstrating 
that the rate of discovery remains constant (Fig. 2). 
He further extrapolates that, if the rate of discovery 
begins to taper off right now and follows the curve 
displayed by more fully cataloged parts of North 
the northeastern United States), a 
minimum of 300 or more undescribed vascular 


America (e.g., 


plant taxa are still waiting in the wings in California 
alone (Fig. 3 


(2) Hartman & Nelson (1998) 


Furthermore, although California clearly leads 
the pack, a recent publication by Hartman and Nel- 
son (1998) demonstrates the pervasiveness of on- 
going floristic discovery throughout North America. 
For the two decades from 1975 through 1994, a 
total of 1197 vascular plant taxa were described as 


84 Annals of the 
Missouri Botanical Garden 
1 OO Ц Т Џ Џ | Џ T T Т 1 т Li Т Т | Џ т LJ т || Т Џ Џ T 
960 
D 
— 80 P "cc 4 
\ ? 
e 
Ф 
О ООСОР РРА 
о 60 
Q 
o) 
5 AQ ИЕ m c n oe a 
um 
Ф 
Q 
я 20 таи алата ви аи И. 
2 
Z 


О 14 
1750 1800 


1850 


1900. 1950 2000 


YEAR 


Yearly description rate of endemic species of plants in California and/or the California Floristic Province 


e 2. 
н by D. W. Taylor, unpublished data 


new to science, ranging from monotypic genera to 
formae and nothotaxa (collectively referred to as 
“novelties”). The 603 full species comprise 3.21% 
of the 18,781 currently estimated to occur in North 
America (1998 estimate provided by Flora of North 
America North of Mexico). The overwhelming ma- 
jority are from the western and southeastern United 
States, but essentially all states and provinces con- 
tributed to the total (including a forma from Rhode 
Island, Lindera benzoin f. rubra R. L. Champlin). 
Most are angiosperms, but 78 pteridophytes and 6 
gymnosperms are represented. 

Other statistics compiled by Hartman and Nel- 
son included: 


Number of holotypes by political unit (excluding 
formae and nothotaxa). Top 10 — California (217), 
Utah (183), Texas (70), Nevada (63), Arizona (57), 
Oregon (42), New Mexico (41), Florida (38), Idaho 
(33), and Wyoming (32). 

Families with the greatest number of novelties 
(excluding formae and nothotaxa). Top 10 = Aster- 
aceae (186), Brassicaceae (91), Fabaceae (84), 
Scrophulariaceae (46), Polygonaceae (46), Poaceae 
(44), Cactaceae (36), Liliaceae (30), Apiaceae (27), 
and Lamiaceae (26). 


Authors of novelties. Top 10 = S. L. Welsh (118), 


R. С. Rollins (62), J. L. Reveal (45), R. С. Ватеђу 
(32), G. L. Nesom (26), N. H. Holmgren (25), W. 
H. Wagner (24), B. L. Turner (23), S. Goodrich (19), 
and B. Ertter (18). 

Taking into consideration that not all published 
novelties are subsequently accepted as worthy of 
taxonomic recognition, Hartman and Nelson ac- 
cordingly calculated the acceptance rate in a va- 
riety of taxonomic works, ranging from 63% to 
98%, with somewhere around 90% apparently be- 
ing the norm. This may in fact be an underesti- 
mation, if a recent study by Windham and Beilstein 
(1998a, b) is any indication, ironically involving the 
two leading authors of novelties. Lest anyone as- 
sume that Welsh’s impressive total (nearly double 
that of Rollins’s) results from a bad case of species 
conflation, Windham and Beilstein give strong ev- 
idence that Welsh erred on the conservative side in 
at least one instance. Not only did the elegant con- 
vergence of micromorphological, molecular, and 
other evidence show that Welsh mistakenly lumped 
some of Rollins’s species of Draba (Brassicaceae), 
but furthermore indicated that Rollins himself had 
confused taxa that were morphologically convergent 
but only distantly related. 

Parallel to Taylor’s analysis, Hartman and Nelson 
showed that the rate of publication of taxonomic 


Volume 87, Number 1 
2000 


Ertter 
Floristic Surprises 


4000 | — n ^ 
É. р — —— с ce 
5 y 
E 2 
= 3000 fo "d 
о 
: ri 
о 
Е 2000 ^ 
z / 
Ф / 
= d 
3 1000 
5 3 
Е 
5 
о 
о "ОЧЕ L 
1700 1800 1900 2000 2100 
YEAR 
Figure 3. Curve depicting the historical trend in dis- 


covery of endemic plants of the California region and an 


through publicatis of The Jepson Manual Ме а 
1993). The data set includes those endemic ta 

(1993) or Skinner a Pav lick 
(1994), plus some 18 taxa found in the Oregon portion 
of the California Floristic Province, 120 taxa found in 


dditional 26 new 
e been desc ribed between 1994 and 1998 (D. 
Taylor, unpublished data). 


novelties has remained relatively constant since 
1955, averaging nearly 60 per year. They predicted, 
however, that the rate of publication of novelties 
will diminish once Flora of North America North of 
Mexico (FNA) is completed. It is accordingly worth 
noting that Taylor’s statistics do not show a com- 
parable drop following the appearance of The Jep- 
son Manual (Hickman, 1993); if anything, the ap- 
pearance of an up-to-date flora has spurred 
essential fieldwork and increased the likelihood of 
recognizing a novelty as such. There is at least one 
example already that the appearance of a generic 
treatment in ЕМА is independent of ongoing dis- 
coveries: Warren H. and Florence Wagner (1994; 
pers. comm. 1998) report that they have already 
accumulated six new species of Botrychium beyond 
the 30 included in their treatment for FNA (Wagner 
& Wagner, 1993). 


Although it is impossible to know exactly how 

much remains unknown, a calculation based on 
Taylors method of extrapolation has at least some 
conceptual validity. If Taylors extrapolation of 
300 + 
ties in California is accurate, and if the 1:6 ratio of 
California-to-North America novelties remains con- 
stant, then at least 1800 more novelties can be ex- 
pected for North America. If half are full-fledged 
species (as in Hartman and Nelson’s analysis), then 
nearly 5% of the North American vascular plant 
flora is still undescribed! 


still-to-be-discovered vascular plant novel- 


(3) Non-vascular plants and fungi 


Comparable statistics have not previously been 
published for non-vascular plants and fungi, but 
Marshall Crosby and Raymond Stotler have com- 
piled the raw numbers on mosses and liverworts, 
respectively, kindly making them available for this 
paper. From to 1998, a total of 63 mosses 
were described from North America, including 42 
full species (M. Crosby, pers. comm. 1998). This 
represents 3.17% of the 1323 species estimated for 
FNA, almost the same percentage as for vascular 
plants (3.21%). Liverworts tell a similar story, with 
19 novelties described in the last 20 years, repre- 
senting 3.44% of the current tally of 553 (R. Stot- 
ler, pers. comm. 

Statistics generated боа the Index Nominum А!- 
garum, maintained by Paul Silva and Richard Moe 
(pers. comm. 1999), indicate that at least 63 marine 
macroalgae have been described from the North 
American coastline since 1980, with an incomplete 
cataloging of pre-1986 publications. This clearly 
indicates that algae are also still being actively dis- 
covered and identified, perhaps at even higher per- 
centages than for vascular plants. Comparable sum- 
maries have not been generated for lichens or fungi, 
which in general lag behind vascular plants in 
terms of systematic research. 

[Note. Although it is now well established that 

plants” are represented by at least three distinct 
lineages (green, red, and brown), and that fungi are 
more closely related to animals than to green plants 
(e.g., the topic of a keynote symposium on “Phy- 
logeny of Life” at the XVI International Botanical 
Congress), there is no evidence that these groups 
are being evicted from herbaria or the research 
realm of botanists. Their inclusion in this paper is 
furthermore justified by the burgeoning move to 
treat bryophytes and lichens, and potentially fungi 
and algae, under the same conservation umbrella 
as currently exists for vascular plants, with the is- 
sues addressed by this paper of relevance to all.] 


86 


Annals of the 
Missouri Botanical Garden 


C. EXAMPLES OF "NEW TO SCIENCE” DISCOVERIES 


Although the statistics cited above are impres- 
sive in their own right, it is not evident to what 
extent strikingly new discoveries are represented, 
as compared to cryptically distinct variants of mar- 
ginal significance. To address this question, a se- 
lection of the most dramatic of the newly discovered 
and/or described taxa are presented here, drawn 
from among the 1197 novelties tallied by Hartman 
and Nelson and numerous others published since 
1994. Choosing among the wealth of riches was one 
of the more challenging parts of preparing this pa- 
per, with new examples constantly coming to the 
fore. The resultant choices are organized among the 
following categories: 


(1) Monotypic genera 


Five species covered by Hartman and Nelson 
were distinctive enough to be described as new 
monotypic genera: Apacheria chiricahuensis C. T. 
Mason (Crossosomataceae), Cochisea robbinsorum 
W. H. Earle (Cactaceae) (subsequently included 
within Coryphantha by Benson [1982], Dedeckera 
eurekensis Reveal & owell (Polygonaceae), 
Shoshonea pulvinata Evert & Constance (Api- 
aceae), and Yermo xanthocephalus Dorn (Astera- 
ceae). Apacheria and Cochisea are from Arizona, 
Shoshonea and Yermo are from Wyoming, and De- 
deckera is from California. Apacheria, found in the 
Chiricahua National Monument in 1973, became 
the second genus assigned to the family Crossoso- 
mataceae (Mason, 1975). whose closest 
probable relatives grow in deciduous forests in 
eastern North America, is known from a single re- 
mote population located along a proposed pipeline 
route in the Wyoming desert (Dorn, 1991). Shosh- 
onea, discovered by retired schoolteacher and rock- 
gardener Erwin Evert in 1979, has more recently 
been found to occur within two miles of downtown 
Cody, Wyoming (Evert & Constance, 1982; R. Hart- 
man, pers. comm. 1998). The distinctive summer- 
blooming shrub Dedeckera was discovered by con- 
servation activist Mary DeDecker (Reveal & 
Howell, 1976; Nilsson, 1994), with one population 
now known within walking distance of a scientific 
research station on the outskirts of Bishop, Califor- 


Yermo, 


nia. 

Subsequent to Hartman and Nelson’s publica- 
tion, two more monotypic genera of vascular plants 
have been described, by coincidence both Brassi- 
caceae from California. The first was Sibaropsis 
hammuttii S. Boyd & T. S. Ross, with three separate 
occurrences encountered in the course of doing an 
environmental impact survey in relatively well-bot- 


anized and well-traveled portions of southern Cal- 
ifornia (Boyd & Ross, 1997). The second was also 
discovered by a consultant in an area proposed for 
development, within earshot of Interstate Highway 
5 in the San Joaquin Valley. Originally suspected 
of being yet one more introduced European annual, 
it was determined instead to represent a unique 

w species and was accordingly described as 


Twisselmannia californica Al-Shehbaz (Al-Shehbaz, 
1999) 


Looking beyond vascular plants, newly discov- 
ered species distinctive enough to be described as 
new genera are particularly common among algae. 
Of the 63 macroalgae cited above, eight were dis- 
tinctive enough to be described as new genera: 
Binghamiopsis caespitosa I. K. Lee, J. A. West & 
Hommers from California; Boreothamnion villosum 
M. J. Wynne and Orculifilum denticulatum S. C. 
Lindstr. from Alaska; Calliclavula trifurcata C. W. 
Schneid. and Nwynea grandispora Searles from 
North Carolina; Chlorojackia pachyclados R. Niel- 
sen & J. A. Correa from Nova Scotia; Rhododra- 
parnaldia oregonica Sheath, Whittick & K. M. Cole 
from Oregon; and Verosphacela ebrachia E. C. Hen- 
ry from Florida. 

Among bryophytes, Ozobryum ogalalense G. L. 
S. Merrill (Pottiaceae) was also recently discovered 
and described as a monotypic genus (Merrill, 
1992). Although it was subsequently transferred to 
Molendoa (Zander, 1993), its distinctiveness as a 
species remains noteworthy. The new moss is fur- 
thermore remarkable in making its appearance in 
the Great Plains, a region otherwise relatively de- 
pauperate in both mosses and new discoveries in 
general. 


(2) Charismatic megaflora 


The award for most publicity for a recent dis- 
covery goes to the Shasta snow-wreath, Neviusia 
cliftonii Shevock, Ertter & D. W. Taylor (Rosaceae), 
whose serendipitous discovery in 1992 in northern 
California made both The New York Times (2 Feb- 
ruary 1993) and the Frankfurter Allgemeine (30 De- 
cember 1992) based on two aspects of particular 
significance (Shevock et al., 1992). First, the Shasta 
snow-wreath was the second species in a genus that 
had previously been known as a single rare species 
in the southern Appalachians, over a thousand 
miles away. This disjunct distribution, indicative of 
a Tertiary relict, was, however, less puzzling than 
the second aspect: namely, that this never-before- 
collected shrub was locally co-dominant (with poi- 
son-oak) along a well-traveled highway, 25 miles 
northeast of Redding, California. Once a focused 


Моште 87, Митбег 1 
00 


Ertter 
Floristic Surprises 


search was undertaken, several additional popula- 
tions were readily located, one adjacent to a de- 
veloped campground. As a further anomaly, the 
currently known range lies largely within one of the 
few areas (Redding quadrangle) whose vegetation 
types were completely mapped and published 
(Weislander et а|., 1939); nevertheless, Neviusia 
was not among the abundant vouchers. 

As an eastern example of “charismatic megaflo- 
ra," a 5—7-m-tall rosaceous shrub or small tree, dis- 
covered in Arkansas in 1970 and initially identified 
as either a Crataegus or an Aronia, was subse- 
quently described as Mespilus canescens Phipps 
(Phipps, 1990). As such, it was a surprising addi- 
tion to a previously monotypic genus known only 
from Europe, the medlar Mespilus germanica L. 
Subsequent isozyme studies supported the inclu- 
sion of the new species within Mespilus (Phipps et 
al, 1991). Described as being *'of exceptional 
beauty," only 25 individuals are known from a sin- 
gle 22-acre grove. 

Another contender for most charismatic recent 
discovery from eastern North America is the Ken- 
tucky lady's slipper, Cypripedium kentuckiense C. F. 
Reed (Orchidaceae), among the tallest and showiest 
species in a genus of showy orchids. Although cur- 
rently known from several southeastern states, it 
was not described until 1981, long after the author 
had first encountered it cultivated in a garden 
(Reed, 1981). The most recently discovered popu- 
lation, in coastal Virginia in 1995, is only 150 km 
as the crow flies from the center of Washington, 


D.C. (Weldy et al., 1996). 


(3) Botanical “hot spots” 


A jackpot of undescribed species is occasionally 
encountered in areas of complex geology and rug- 
ged terrain, which set the stage for an often bewil- 
dering expression of island biogeography in a con- 
tinental setting. In such areas, an unusual substrate 
or isolated mountaintop has the potential of har- 
boring a unique suite of endemic plants, and the 
first botanist to reach the site can reap a bonanza 
of floristic surprises. 

As it happens, my own career as a professional 
taxonomist began when I had the good fortune of 
participating in the discovery of one such botanical 
treasure trove while still an undergraduate, thanks 
to a newly built gravel road in the early 1970s that 
made Leslie Gulch in southeastern Oregon a two- 
hour drive by passenger car from Boise, Idaho. The 
unique ash-flow tuffs of Leslie Gulch have thus far 
yielded a total of five plant taxa new to science: 
Senecio ertterae T. M. Barkley, Mentzelia packardiae 


Glad, lvesia rhypara Ertter & Reveal, Artemisia 
packardiae rimes & Ertter, and Phacelia 
lutea var. mackenziorum J. W. Grimes & P. L. Pack- 
ard (Grimes, 1984). Some of these species are so 
distinctive that their relationships remain unclear, 
whereas variants of several other species might 
prove to be taxonomically distinct upon further 
study. In addition, Leslie Gulch turns out to be the 
*mother lode" for several other species that had 
been known previously from only a handful of pop- 
ulations: Trifolium owyheense Gilkey, iniu, ^ 
sterilis Barneby, and Eriogonum novonudum 
Peck. 

Alas, Leslie Gulch is now overshadowed by the 
latest “hot spot," the Ketona Glades in Bibb Coun- 
ty, Alabama. In 1992, a group of environmental 
consultants undertaking a rare plant survey by ca- 
noe on the Little Cataba River came upon a cal- 
careous glade community harboring at least seven 
undescribed taxa: Spigelia Ay at ре Chapman 
ex A. DC. var. alabamensis ould, Onosmodium 
sp. nov., Erigeron strigosus var. nov., Dalea sp. nov., 
Castilleja sp. nov., Liatris sp. nov., and Silphium sp. 
nov. In addition, several state records for Alabama 
were present, most with some level of formal rarity 
status, along with a new county record for the fed- 
erally Бори Xyris tennesseensis Kral (Allison, 
1994; Gould, 1996). 

The rugged E Sierra Nevada in California 
also continues to be a rich source of novelties, with 
three new taxa discovered in a single day on a re- 
mote marble ridge in 1996: Heterotheca monar- 
chensis D. York, Semple & Shevock, Gilia yorkii 
Shevock & A. С. Day, and a still-undescribed Er- 
iogonum. The ridge also harbors the only known 
California populations of three mosses (J. R. Shev- 
ock, pers. comm. 1998) 


(4) In our backyards 


Although the majority of new discoveries are en- 
countered in relatively remote sites, a surprising 
number appear around significant population cen- 
ters, even those with major herbaria and a long 
tradition of botanical exploration. Among the more 
dramatic of these “in our backyards” examples are 
the following: 
€ Morefield’s leather-flower, Clematis morefieldii 
Kral (Ranunculaceae), was discovered in 1982 b 
a budding botanist who practiced by collecting 
specimens around his neighborhood inside the city 
limits of Huntsville, Alabama (Kral, 1987). Now 
federally listed as Endangered, the species is still 
only known in and near residential areas in the 
Huntsville area. Morefield himself, inspired by this 


Annals of the 
Missouri Botanical Garden 


early experience, went on to become botanist for 
the Nevada Natural Heritage Program. 
@ Not far from Las Vegas, Nevada, in the course 
of a floristic survey of Red Rock Canyon National 
Conservation Area, lonactis caelestis P. J. Leary & 
G. L. Nesom (Asteraceae) was discovered in 1990. 
The highly localized population, within sight of the 
Las Vegas casinos, occurs on an edaphic island of 
sandstone surrounded by limestone. The species is 
distinctive enough that it might justifiably be treat- 
ed as a monotypic genus (Nesom & Leary, 1992). 
Approximately 25 miles west of downtown Los 
Angeles, California, a few miles north of the trendy 
community of Malibu in the Santa Monica Moun- 
tains, Baccharis malibuensis К. M. Beauch. & 
Henr. (Asteraceae) was discovered in 1988 (Beau- 
champ & Henrickson, 1996). Beauchamp had ear- 
lier described another Baccharis, B. vanessae R. M. 
Beauch., from the midst of housing developments 
in central San Diego County (Beauchamp, 1980). 
€ Only about six miles from one of the main her- 
baria in Utah, on cliffs behind some summer cab- 
ins, Viola franksmithii N. H. Holmgren (Violaceae) 
was discovered in 1989 by a “modern-day natural- 
ist” who was monitoring other rare plants known to 
occur in Logan Canyon (Holmgren, 1992). Dis- 
agreement exists as to taxonomic placement of this 
distinctive species (H. J. Ballard, pers. comm. 


€ А short distance outside of Yosemite National 
Park, among the most-visited parks in the United 
States, the showy-flowered Erythronium taylori 
Shevock & G. A. Allen (Liliaceae) was discovered 
in 1996, bringing to three the number of extremely 
local endemic Erythronium in the lower Sierra Ne- 
vada (Shevock & Allen, 1998). In addition to being 
the only New World species of the genus with 
scented flowers, Е. taylori is intriguing in being a 
candidate fer the Erythronium once reported to oc- 
cur in Yosemite Valley (Brandegee, 1891), where 
no representatives of the genus are currently known 
(D. W. Taylor, pers. comm. 1998). 

• In spite of growing less than 10 miles from 
downtown San Francisco, California, in an exten- 
sively botanized county with a recently updated flo- 
ra (Howell, 1970), Calochortus tiburonensis A. J. 
Hill (Liliaceae) was not discovered until 1972. The 
species is so distinctive that “its existence chal- 
lenges the currently accepted infrageneric classifi- 
cation” (Hill, 1973: 104). The protologue further- 
more noted, “That a previously uncollected new 
species ... 
well known area suggests the need for a very care- 
ful look at any areas that are threatened by devel- 
opment or other disturbance, especially near ex- 


was discovered in such a botanically 


panding population centers. Had this species not 
been noticed soon, it might very well have become 
extinct without ever having been recorded.” 

@ Nestled among the Lick Observatory complex 
on Mount Hamilton, within easy reach of the Uni- 
versity of California at Berkeley, the California 
Academy of Sciences, and Stanford University, 
Lomatium observatorium Constance & Ertter (Api- 
aceae) was not described until 1996, too late to be 
included in Hartman and Nelson’s synopsis. Its lo- 
cation on Mount Hamilton was brought to the at- 
tention of Lincoln Constance, the expert on Loma- 
tium, by a wildflower photographer, Nigel Hancock 
(Constance & Ertter, 1996). 


(5) Species-rich genera 


Lomatium, with 10 novelties listed in Hartman 
and Nelson, is also an example of a large genus 
that has undergone extensive speciation, resulting 
in an abundance of highly localized endemic spe- 
cies that are still being discovered at a steady rate. 
Among other significant examples are the following: 
€ Astragalus (Fabaceae) contains the largest 
number of novelties (43) listed in Hartman and 
Nelson, due in large part to the efforts of Rupert 
Barneby. Astragalus is also worth highlighting for 
the potential medicinal value of the new discover- 
ies, given the known value of at least one Old World 
species, A. membranaceus Bunge. This species not 
only has a long history of use in Chinese traditional 
medicine, but it has also entered the American al- 
ternative medicine pharmacopoeia as an immune 
system enhancer, with some clinical evidence of 
activity in cancer patients with impaired immune 
responses (e.g., Chang et al., 1983; Kosuge et al., 
1985). Intriguingly, this eerily approaches a case of 
fact following fiction, in that the hypothetical dis- 
covery of a cure for cancer in the form of Astrag- 
alus, only in this case a North American species, 
plays a key role in Duane Isely’s fictionalized prog- 
nostication of what would happen if all taxonomists 
and their works suddenly disappeared (Isely, 1972). 
€  Penstemon (Scrophulariaceae): Where Astraga- 
lus has potential medicinal value, Penstemon has 
proven horticultural significance. The 29 novelties 
listed in Hartman and Nelson accordingly represent 
valued additions to the existing penstemon palette 
for rock-gardening enthusiasts, at least to the extent 
that they can be brought into cultivation without 
negatively impacting natural populations. In the In- 
termountain Region alone, seven species have been 
discovered and described subsequent to the 1984 
treatment of Penstemon in the Intermountain Flora, 


Volume 87, Number 1 
2000 


Ertter 89 
Floristic Surprises 


averaging one every two years (Holmgren, 1984, 
19 


€ Lesquerella (Brassicaceae): Of the 83 species of 
Lesquerella in the late Reed Rollins’s monumental 
synopsis of the Cruciferae of North America (Rol- 
lins, 1993), nearly half were authored or coauthored 
by Rollins himself over his long career, several in 
the book itself. As massive a compendium as this 
was, three additional species were discovered al- 
most before the ink was dry: L. lesicii Rollins and 
L. pulchella Rollins from Montana к 1995), 
and L. tuplashensis Rollins, К. A. Beck & Caplow 
from the Hanford Nuclear “eset in Washing- 
ton (Rollins et al., 1995). A short two years later, 
a fourth species, L. vicina J. L. Anderson, Reveal 
& Rollins, was published, with the epithet chosen 
in reference to the fact that the type locality was 
behind the home of a neighbor of one of the co- 
authors in Montrose, Colorado, growing in the 
sheep pasture (Anderson et al., 1997; J. Reveal, 
pers. comm. 1998 

€ Arabis (Brassicaceae): The appearance of Rol- 
lins's (1993) compendium also triggered the de- 
scription of a suite of four new Arabis from Canada, 
Alaska, and Greenland, as well as reports of nu- 
merous range extensions from the same region, all 
in the same publication (Mulligan, 1995). This ex- 
ample is significant in demonstrating that the north- 
ern latitudes are also full of floristic surprises, in 
spite of their generally fewer numbers of species. 
Farther south, Arabis hirschbergiae S. Boyd has re- 
cently been described from southern California, a 
stone’s throw from a major highway (Boyd, 1998). 
All of these species are additions to the 17 novel- 
ties listed by Hartman and Nelson. 

© Eriogonum (Polygonaceae): With 38 entries in 
Hartman and Nelson, Eriogonum has also proven 
to be an ongoing source of novelties, with one new 
species discovered on the Hanford Nuclear Res- 
ervation across the river from the new Lesquerella, 
as part of the same botanical survey (Reveal et al., 
1995). An even more recent and dramatic example 
is provided by a pair of Friends of the Jepson Her- 
barium weekend workshops on Eriogonum in Cal- 
ifornia in the summer of 1997, taught by Eriogon- 
um specialist James L. Reveal. Of the 35 
participants, mostly agency botanists and consul- 
tants, 3 ended up providing Reveal with additional 
undescribed taxa, leading to the quip that we 
should schedule a workshop on how to describe 
new species! In addition, Reveal confirmed that the 
variant of E. nudum Benth. that is the host plant 
for a federally listed butterfly, the Lange’s Metal 
Mark (Apodemia mormo langei J. A. Comstock) is 
itself an undescribed taxon, bringing up to three 


the number of plants endemic to Antioch Dunes (a 
badly degraded inland dune complex on the edge 
of the Sacramento River delta in central California, 
less than an hour’s drive from Berkeley; the other 
two are Oenothera deltoides Torrey & Fremont 
subsp. howellii (Munz) У. M. Klein and Erysimum 
capitatum (Douglas) Greene var. angustatum 
(Greene) Rossbach, both federally endangered.) 

€ Carex (Cyperaceae): Carex is noteworthy in that 
the majority of the 21 novelties listed in Hartman 
and Nelson, an average of 2 per year, are found in 
eastern North America. Carex lutea LeBlond, for 
example, was discovered in 1991 in North Carolina, 
where it is a rare endemic of wet savanna underlain 
by limestone. It is furthermore phytogeographically 
interesting in being a southern outlier (by 750 km) 
of a circumboreal species complex, possibly a relict 
from the Pleistocene. Associates of C. lutea include 
numerous other rare species, including Venus fly- 
trap and an undescribed Allium (LeBlond et al., 
1994). 


Another recently described sedge, C. junipero- 
rum Catling, Reznicek & Crins, is known from 
widely disjunct populations in Ontario, Ohio, and 
Kentucky. Although locally а groundlayer domi- 
nant, it was presumably overlooked because the in- 
florescences are nestled at the base of the plant and 
appear unexpectedly early in the season. In the 
protologue, the authors noted, "The recent discov- 
ery of this distinctive new species in a supposedly 
botanically well-known area suggests that even the 
flora of northeastern North America is not as well- 
known as is commonly supposed" (Catling et al., 


1993) 


(6) Scientifically significant discoveries 


On top of the importance of cataloging the com- 
ponents of biodiversity for their own sake, many of 
the recently described species have carried signif- 
icance beyond their intrinsic value. Some, such as 
Neviusia cliftonii and Carex lutea, pose interesting 
biogeographic puzzles. Others, including Dedeckera 
eurekensis and Calochortus tiburonensis, provide the 
key to unraveling phylogenetic questions (Reveal 
& Howell, 1976; Reveal, 1989a; Hill, 1973). De- 
deckera is also significant as an ancient lineage 
postulated to have accumulated such a high segre- 
gational genetic load of heterozygosity that seed set 
is severely depressed (Wiens et al., 1989). Some 
additional examples: 
€ At the time of Keck's revision of ћеча (Rosa- 
ceae) in 1938, 1. shockleyi S. Watson was thought 
to be restricted to the Sierra Nevada of California 
and Nevada. Fieldwork by numerous botanists over 


Annals of the 
Missouri Botanical Garden 


the last two decades, however, has shown not only 
that Z. shockleyi is scattered on mountaintops across 
the Great Basin, with a varietally distinct outlier in 
Utah (var. ostleri Ertter), but that a previously un- 
known complex apparently represents the low-ele- 
vation analog on unusual edaphic sites (Ertter, 
1989). The low-elevation complex consists of sev- 
eral closely related entities with widely disjunct 
distributions: /. rhypara Ertter & Reveal var. rhy- 
para, I. rhypara var. shellyi Ertter, and 1. paniculata 
T. W. Nelson & J. P. Nelson. This example of island 
biogeography in a continental setting is interpreted 
in the context of Pleistocene-driven isolation and 
radiation, with one lineage retreating to isolated 
montane "islands" and the other finding a compa- 
rable niche in unusual edaphic sites at lower ele- 
vations. Molecular investigations with Christopher 
Baysdorfer (California State University at Hayward 
are currently under way to further elucidate the re- 
sultant evolutionary pattern. 

€ Verrucaria tavaresiae R. L. Moe is noteworthy 
not only in being one of the few known marine li- 
chens, but also the only lichen known with a brown 
algal phycobiont. Described in 1997, it is another 
example of "in our backyards," occurring in the 
intertidal zone around San Francisco, California 
(Moe, 1997). 

€ Calycadenia hooveri G. D. Carr (Asteraceae), 
described in 1975, possesses a chromosome аг- 
rangement almost identical to that of C. villosa DC., 
which is unlike that of other species in the genus. 
Reconstructions of chromosomal evolution based on 
molecular phylogenies of Calycadenia show that 
the ancestor of both aneuploid species lineages 
(which comprise the bulk of the genus) had a chro- 
mosome arrangement similar or identical to that of 
С. hooveri or С. villosa. Without С. hooveri, the re- 
construction of chromosome evolution would have 
been equivocal (Carr, 1975; Baldwin, 1993). 

@ Probably the most surprising discovery involv- 
ing North American ferns has been the realization 
that several species of Hymenophyllaceae and Vit- 
tariaceae in the eastern United States exist primar- 
ily as gemmiferous gametophytes, either growing 
north of the range of the sporophytes or, in a couple 
of cases, with sporophytes produced rarely if ever 
(Farrar, 1993a, b). Although vegetatively reproduc- 
ing gametophytes have been known since 1888, 
their relative abundance (10% of all fern species 
worldwide) and significance has only become ap- 
preciated relatively recently (e.g., Farrar, 1974). 
Once gametophytes became the target of attention, 


— 


three new species were discovered in the early 


990s: Hymenophyllum tayloriae Farrar & Raine, 


Trichomanes intricatum Farrar, and Vittaria appa- 
lachiana Farrar & Mickel. 

€ The liverwort genus Pellia йе раке per- 
haps second only to Marchantia in the amount of 
attention previously given to liverwort genera, nev- 
ertheless provides a case where a critical look at 
the “common” species in the field yields unex- 
pected results. Prior to 1981, only four species were 
recognized worldwide; in relatively quick succes- 
sion, however, two new ones were published from 
eastern North America (P. megaspora R. M. Schust. 
and P. appalachiana R. M. Schust. [Schuster, 1981, 
1991]), and one more is currently being described 
from Mississippi (R. Stotler, pers. comm. 

€ The bryophyte Takakia has been a puzzle since 
its discovery in 1951, at which time it was consid- 
ered to be a liverwort. However, only vegetative and 
archegonial material was known, and attempts to 
induce fertile structures in cultivation met with fail- 
ure. Sporophytic plants of T. ceratophylla (Mitt.) 
Grolle were finally encountered in 1990, in the 
course of fieldwork in the Aleutian Islands, firmly 
establishing Takakia's identity as a moss (Smith & 
Davison, 1993). In essence, floristic discovery re- 
sulted in the transfer of a genus from one division 
(Hepatophyta) to another (Bryophyta)! 


D. OTHER KINDS OF FLORISTIC SURPRISES 


Although newly described novelties capture the 
imagination, they represent only the tip of the ice- 
berg of floristic surprises, only the starting point for 
the comprehensive information that is truly needed 
for making difficult decisions in a scientifically in- 
formed manner. Even more incomplete than our 
knowledge of what species exist is our knowledge 
of where they occur, what their habitat require- 
ments are, and similar questions that can only be 
answered by extensive fieldwork coupled with crit- 
ical taxonomic analysis. 

An excellent example of the incomplete and non- 
static nature of floristic information is provided by 
the recently revised flora of Missouri, in which the 
number of plants known to occur in a relatively 
well-studied state (with one of the oldest and largest 
herbaria in the country) has increased by nearly 
12% since 1963, two-fifths of them native (Yats- 
kievych, 1999). Current research on the flora of 
Mount Diablo, an isolated mountain and popular 
state park situated 25 miles east of San Francisco, 
California, shows an even more dramatic increase. 
In spite of the high quality of the original floristic 
effort (Bowerman, 1944), a recent update (Bower- 
man & Ertter, in press) has increased the known 
taxa by 25%, approximately half of which are na- 


Volume 87, Number 1 
2000 


Ertter 
Floristic Surprises 


tive. Furthermore, several species in the 1944 
treatment have been deleted or replaced, as a result 
of misidentified vouchers or changed circumscrip- 
tions. On the other hand, locally occurring variants 
of Eriogonum, Lomatium, Calystegia, and Gilia 
have the potential of being undescribed novelties 
(Ertter & Schultheis, 1998). 

Some of the main categories of “floristic surpris- 


s" 


other than novelties are the following: 


(1) "Presumed extinct" 


Nearly as dramatic as the discovery of new spe- 
cies is the rediscovery of species that had been 
thought to be extinct. А recent example in Califor- 
nia is that of the Ventura Marsh milk-vetch, As- 
tragalus pycnostachyus А. Gray var. lanosissimus 
(Rydb.) Munz (Fabaceae), which was recently foun 
in Ventura County, California, after being presumed 
extinct for 40 years. Local newspaper coverage re- 
ferred to *A botanical resurrection" (Ventura Coun- 
ty Star, 21 Aug. 1997) and “The Elvis Presley of 
flowering plants" (Santa Barbara News-Press, 15 
Aug. 1997). The wildlife biologist who found the 
plant, Kate Symonds, was quoted in one account as 
noting, “It is more common to realize something is 
gone that used to be around, rather than finding 
something thought to be gone that is still in exis- 
tence. It feels like a second chance for the species" 
(Sacramento Bee, 15 Aug. 1997). Ironically, the site 
was a former oil field waste dump, dispelling any 
notion that significant discoveries occur only in 
pristine habitats. 

Coincidentally, another recently rediscovered 
Californian Astragalus, A. agnicidus Barneby, was 
also associated with disturbance. In this case, a 
plant that had been deliberately eliminated be- 
of its perceived toxicity to livestock (agni- 
cidus = “lamb-killing”) reappeared when logging 
activity apparently triggered the germination of 
seeds that had lain dormant for decades (Hiss & 
Pickart, 1992). This example also serves to illus- 
trate the difficulty of determining presence versus 
absence of a species at a site, let alone globally, 
even when no mature individuals are evident. 

The systematic search for selected subsets of the 
416 plants and animals that are considered poten- 
tially extinct in the United States was given a major 
boost recently by the Canon Exploration Grants 
Program directed by The Nature Conservancy (Stol- 
zenburg, 1998; Anonymous, 1998). Although a de- 
pressing majority have not been relocated to date, 
there have been enough satisfying success stories 
to justify the program, in more ways than one. As 
evidence, consider the following story transmitted 


by the director of the program, Bruce Stein (pers. 
comm. 1998): 


I was jotting my note to you, a second 1998 find 
from the Canon vex d was Beppe ad into my box. This 
just in from South Tex Corpus Christi: Paro- 
сы [Caryophyllaceae], 
which was last collected in 1958. As Bill Carr, the guy 
who refound it says (after finding it 9 paces from where 
he parked his car on his first stop), ‘For me the expe- 
rience was just another reminder of how few active 
anists there are in Texas and how far behind the rest of 


species that, give otanists in this part 
of the continent, might otherwise have remained enig- 
matic for who knows how long." ' 


Not quite as exciting as the rediscovery of glob- 
ally “extinct” species, but of potentially equal im- 
plications for land management, is the rediscovery 
of globally rare species that had been considered 
regionally extinct (i.e., extirpated). Excitement on 
he Mendocino National Forest in California has 
centered around the 1996 discovery of several pop- 
ulations of the federally threatened Howellia aqua- 
пса А. Gray (Lobeliaceae), previously known from 
California only on the basis of a single fragment 
collected in 1928 (Isle, 1997). Interpopulational 
genetic studies are currently under way to compare 
the California plants with those in Washington, Ida- 
ho, and Montana. In addition, another plant that 
had been thought extinct in California, Ophioglos- 
sum pusillum Raf. (Ophioglossaceae), was recently 
located adjacent to one Howellia population (D. 


м 


Isle, pers. comm. 1998 

Even the reappearance of a not-so-rare species 
in a part of its range where it had seemingly dis- 
appeared can be newsworthy, as evidenced by the 
attention given to a population of Mimulus tricolor 
Lindl. found on the outskirts of Corvallis, Oregon 
(Holden, 1999). Although this species remained 
relatively common in the Central Valley of Califor- 
nia, it had been assumed to be locally extinct in 
Oregon. As with Astragalus agnicidus, the reap- 
pearance of Mimulus tricolor after nearly 10 years 
demonstrates how long a species can persist in the 
seed bank, and accordingly how difficult it is to 
verify absence from a site. 


(2) Distributional discoveries 


More prosaic but gaining significance through 
sheer weight of numbers is the constant stream of 
distributional discoveries: major extensions in the 
known ranges of native species. Only the most dra- 


matic are published (e.g., new state records); the 


92 


Annals of the 
Missouri Botanical Garden 


bulk accumulate in the form of herbarium speci- 
mens. A recently verified, curiously overlooked ex- 
ample in the Jepson Herbarium (JEPS) is a speci- 
men of Luzula piperi (Coville) M. E. Jones 
(Juncaceae) from northwestern California (Ferlatte 
349), over 600 km south of the nearest previously 
reported occurrence in northwest Washington 
(Hitchcock & Cronquist, 1973). A good example 
from eastern North America is Schizandra glabra 
(E. P. Bicknell) Rehder (Schizandraceae), the only 
American representative of an otherwise Asiatic ge- 
nus. A population found in 1991, clambering over 
a sandstone cliff in southeastern Kentucky, is 250 
km from the nearest of the previously known lo- 
calities scattered across the coastal plain of the 
southeastern United States (D. D. Taylor, 1994). 

More problematic is the recent discovery of Lim- 
nanthes macounii Trel. (Limnanthaceae) in a sea- 
sonally fallow field in west-central California (Bux- 
ton & Ornduff, 1998). Previously known only as a 
rare endemic of southeastern Vancouver Island in 
Canada, L. macounii was at one point presumed 
extinct (Hitchcock, 1961). What is currently under 
debate is whether this represents a surprising dis- 
persal event, a previously overlooked natural range 
disjunction, or evidence that additional populations 
might exist in intervening sites (A. Ceska, pers. 
comm. 1998). Ornduff (pers. comm. 1998) supports 
the dispersal hypothesis, citing the reverse example 
of Lasthenia minor (D rnduff (Asteraceae) be- 
ing found in northwestern Washington, over 1000 
km north of the nearest naturally occurring popu- 
lation in central California (Vasey et al., 1994). The 
field in which the California population was found, 
which was probably significantly larger than the 
British Columbia population, was subsequently 
plowed prior to planting cabbage (Buxton & Orn- 
duff, 1998). 

Distributional discoveries are not restricted to 
single species within North America, but can occur 
as unexpected suites, as evidenced by the follow- 
ing: 

• А special category is that of continental-level 
range extensions: species previously known only 
from Eurasia that are determined to occur in Nort 

America as well, not as introductions but as natu- 
rally occurring populations. William Weber (pers. 
comm. 1998) addressed the large number of Asi- 
atic-Rocky Mountain disjunctions, many recently 
located, with the comment: “J. 
tainly right when he was shocked to see some of 


OOKer Was сег- 


his Asiatic things on his five days in the Rockies 
[in 1877]; sadly, Asa Gray evidently was on a va- 
cation/picnic and didn't recognize that there might 


be a high latitude component to his Tertiary dis- 
coveries in eastern North America." 
Continental-level range extensions are not re- 
stricted to the arctic and alpine regions, however, 
but can also be found farther south. For example, 
specimens from Texas and Arizona previously con- 
fused with Ophioglossum engelmannii Prantl 
(Ophioglossaceae) turned out to be conspecific with 
the widespread Old World species O. polyphyllum 
А. Br. (Zech et al., 1998). More recently, Eleocharis 
mamillata H. Lindb. (Cyperaceae) has been deter- 
mined to be native and widespread in boreal North 
America (S. G. Smith & T. Gregor, in prep. 1998). 
While continental-level range extensions are 
noteworthy in vascular plants, they are more rou- 
tine in bryophytes and lichens (B. Murray, pers. 
comm. 1998). Even here, however, some examples 
stand out from the crowd, such as Aspicilia moen- 
ium (Vainio) Thor. Described in 1986 from Scan- 
dinavia, where it often occurs on the mortar of old 
churches, this lichen was unexpectedly encoun- 
tered on an old retaining wall connected to the 
building that houses the COLO herbarium in Boul- 
der, Colorado (Weber, 1996). The account of the 
discovery of this population amusingly addressed 
the quandary of how to obtain a decent specimen 
rom an intact structure, solved with the coopera- 
tion of Facilities Management staff. 
© Conifers are perhaps the best-mapped group of 
plants in North America (e.g., Little, 1971), being 
both conspicuous and economically significant. 
Nevertheless, recent fieldwork by David Charlet 
has determined that 4396 (90) of the 207 conifer- 
bearing mountain ranges in Nevada harbor at least 
one more conifer than previously reported, and 
12% (24) have had two to four species added to 
the known complement (Fig. 4). Approaching the 
same data from a different angle, of the 22 species 
of conifer known to occur in Nevada, 14 occur on 
at least one more mountain range than had previ- 
ously been reported, resulting in 15 new county 
records in Nevada's 13 western-size counties (Char- 
let, 1996, pers. comm. 1998). 
€ On the Pacific Coast, ongoing surveys of пеаг- 
shore banks that rise to within 30 m of the surface 
are revealing a hitherto unsuspected and remark- 
ably uniform assemblage of around 40 species of 
marine macroalgae (seaweeds), extending from Pu- 
get Sound to northern Baja California. Included in 
the assemblage are noteworthy range extensions 
such as Pleurophycus gardneri Setch. : 
Saunders (Laminariales), before 1970 осоо 
only as far south as Oregon but now known to ђе 
dominant at depths of 40 m off the central Califor- 
nia coast (Kjeldsen, 1972; P. Silva, pers. comm. 


Volume 87, Number 1 
2000 


Ertter 
Floristic Surprises 


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Figure 4. Proportional representation of mountain ranges in Nevada in which additional species of conifers beyond 
е еме и recorded were located during the Charlet survey, 1988—1998 (fig. prepared by D. Charlet). 


1998). Although the kelp is readily identifiable, the 
existence of these southern populations was not ap- 
parent because the plants grow at depths that are 
undisturbed by even violent storms and are accord- 
ingly seldom cast ashore. 

€ Probably the biggest distributional surprise in 
fungi is the discovery that the occurrence of mush- 
rooms above ground and the fungal species diver- 
sity below ground, as determined by molecular 
analysis of hyphal fragments in the soil, can b 
completely independent (Gardes & Bruns, 1996). 
As a result, determining the distribution and rarity 
of various fungal species presents a challenge well 
beyond that posed by vascular plants and bryo- 
phytes. 


i 


(3) Declines and invasions 


Whereas all of the preceding examples involve 
changes to our knowledge of the distributions of 
species, there are also actual changes in the dis- 
tributions themselves. On the one hand are signif- 
icantly diminished ranges, in which historical oc- 
currences documented by herbarium vouchers no 
longer reflect current distributions. An example is 
afforded by Horkelia cuneata Lindl. subsp. puber- 
ula (Greene) D. D. Keck (Rosaceae), in which a 
significant portion of the historically documented 
range has disappeared under Greater Los Angeles 
(Ertter, 1995). This kind of distributional attrition 
has obvious conservation implications, but is ex- 


tremely difficult to become cognizant of, such that 
it is theoretically possible for a species to go extinct 
before its endangered status has even been noted. 

On the flip side, and often contributing to the 
decline of native species, is the spread of species 
into areas where they did not historically occur. The 
explosion of aggressive non-natives is of increasing 
concern due to the various negative impacts such 
invasions can have on both natural and economic 
systems. The sheer numbers of newly reported non- 
natives can be mind-boggling, though it is difficult 
to determine which are new occurrences and which 
have simply been overlooked, given that natural- 
ized species are historically undercollected. Vin- 
cent and Cusick (1998) documented 70 additions 
to the Ohio flora, and also emphasized the fact that 
the non-native component of floras is dynamic, with 
species appearing, flourishing, and occasionally 
disappearing. Even in the region around the United 
States’ national capital, which has been rather sys- 
tematically collected since the 1690s, recent sur- 
veys have resulted in the discovery of seven new 
plant records for Maryland (including two native 
Carex), five of which came from the grounds of the 
Agriculture Research Center in Beltsville (J. Re- 
veal, pers. comm. 1 1 

In California, over 70 non-native species are cur- 
rently known to have become naturalized beyond 
those included in The Jepson Manual (Hickman, 
1993; F. Hrusa, pers. comm. 1998) 


. Аз dramatic 


94 


Annals of the 
Missouri Botanical Garden 


evidence of how difficult such new occurrences are 
to keep abreast of, at least 19 occur within an 
hour’s drive of the building in which The Jepson 
Manual was edited. Five are even fully naturalized 
in the Berkeley campus natural areas (Geranium 
rotundifolium L., Geranium lucidum L., Geranium 
purpureum Vill., Hypericum androsaemum L., an 
Hedera helix L. subsp. canariensis (Willd.) Cout.), 
and at least three others have already achieved sig- 
nificant pest status in local parklands (Dittrichia 
graveolens (L.) W. Greuter, Limnobium laevigatum 
Willd., and Maytenus boaria Molina). The dynamic 
nature of California’s non-native flora, as well as 
the difficulty of obtaining reliable information on 
current occurrences, has been addressed by Re- 
jmanek and Randall (1994) 

E. WHY NOT FOUND BEFORE? 

The preceding examples should serve to empha- 
size that the era of significant floristic discoveries 
in North America north of Mexico is far from over, 
despite perceptions extending back to the mid-19th 
century. Even the initial cataloging of novelties is 
incomplete, to the extent that conspicuous shrubs 
along highways are still being discovered and de- 
scribed as distinctive new species. The comprehen- 
sive mapping of known species, including newly 
invasive pest plants, is equally erratic, at a time 
when such information is sorely needed to make 
sound science-based land-management decisions. 

The inevitable question arises as to why so much 
of our floristic heritage has remained unexplored, 
uncataloged, and unmapped. The principal answer 
is relatively straightforward: it's a big job! As a re- 
sult, and as many of the previous examples testify, 
a primary factor contributing to ongoing floristic 
discoveries is the number of people who are ac- 
tively scouring the field. Fortunately, this is by no 
means limited to professional scientists in academ- 
ic institutions, but instead depends heavily on the 
collective efforts of agency biologists, environmen- 
tal consultants, and native plant enthusiasts (Ertter, 
1995; Yatskievych, 1999). Representatives of this 
diverse group are highlighted in a later section of 
this paper. 

However, the number of people actively search- 


dresses other assumptions that have influenced the 
cataloging of North American plants, both histori- 
cally and currently. 


ASSUMPTION 2: HYPOTHESIS-FREE DESCRIPTIVE 
SCIENCE 


One key assumption is the common and recur- 
ring one that taxonomy and floristics, as examples 


of “descriptive” science, are not intrinsically sci- 
entific, at least as contrasted to the more overtly 
experimental sciences. This assumption has direct- 
ly influenced hiring, funding, and promotional de- 
cisions, which in turn determines research priori- 
ties. Although the full structure of my argument is 
beyond the scope of the current paper, I will nev- 
ertheless posit that science is most definitely in- 
volved in all aspects of taxonomy and floristics, 
complete with the full panoply of falsifiable hy- 
potheses and scientific methodology, even when 
these are not explicitly expressed. 


A. DESCRIPTIVE HYPOTHESES 


The assumption that recognizing and “describ- 
ing" novelties is a simple descriptive process re- 
flects an outdated understanding of biodiversity as 
consisting of discrete, pre-Darwinian quanta, lack- 
ing significant internal variation and separated from 
one another by inviolate boundaries (Ertter, 19972). 
This was noted by Constance (1971: 22) over a 
quarter-century ago: “Although the doctrine of 
‘Special Creation’ of species has lacked any sci- 
entific status for a hundred years, many people 
seem still to be thinking in terms of a finite number 
of objects created once and for all, and which mere- 
ly have to be recognized, described, and named." 

Much of the confusion has a semantic underpin- 
ning, in that “describing” a species is by no means 
equivalent to “describing” a concrete individual 
item. To place species “description” in the explicit 
framework of set theory and hypothesis generation, 
the standard phrase: 


A description of new species Alpha beta, which е 
from other species of Alpha in characters X, Y, апа Z" 


can be expanded into the complex hypothesis: 


"There exists а previously undiscerned component of 
nano diversity that falls within the pedcs param- 
eters of the current species concept, which is hereby 
one species-set A 
and др и of this hypothesis, all members of species- 
Alpha beta are hypothesized to possess biological 
geben X, Y, and Z, whereas all members of other 
species-sets in genus-set Alpha аге hypothesized to lack 
this combination of biological attributes.’ 


Furthermore, not only are species and their cir- 
cumscriptions best understood as complex hypoth- 
eses, but so also are such seemingly "factual" state- 
ments as “Leaves (2.4)3—5(6.1) cm long," which is 
in actuality shorthand for the predictive statement: 
"Based on a measured subset, leaf-length for ALL 
leaves for ALL members of species-set Alpha beta, 
past, present, and future, is predicted to be at least 


Volume 87, Number 1 
2000 


Ertter 95 
Floristic Surprises 


2.4 cm long but no more than 6.1 cm long, with 
the majority falling in the 3-5 cm range.’ 

In both cases, the first phrase is obviously much 
less cumbersome, but the expanded version more 
clearly expresses the fact that nested hypotheses 
are involved, all of which are subject to subsequent 
testing and modification whenever new data are ob- 
tained, most often in the form of new collections of 
plants that “haven’t read the book.” Even the iden- 
tification of an individual specimen can be worded 
to reflect the complexities of set-assignment, to wit: 
“The specimen in hand possesses the diagnostic 
biological attributes that characterize members of 
the set Alpha beta.’ 

B. PARSING SPECIES 

In other words, rather than being routinely sim- 
ple and straightforward, the task of parsing bio- 
diversity into taxonomic components can be a sig- 
nificant intellectual challenge. As а result, 
although blatantly distinct species are still being 
encountered, the majority of recently described 
novelties are determined to be such only after an 
extended and detailed comparison with other spe- 
cies, often requiring a wholesale re-thinking of ex- 
isting taxonomic frameworks. In these cases, it is 
not at all intuitively obvious what qualifies as a 
“previously undiscerned component of natural di- 
versity” that falls within “the biological parame- 
ters of the current species concept,” based on 
some yet-to-be-determined suite of diagnostic bi- 
ological attributes. 

An excellent example is provided by Potentilla 
morefieldii Ertter (Rosaceae), in which multiple col- 
lections had accumulated and been variously iden- 
tified as (= assigned to species-sets) P. pseudoser- 
‚ P. pensylvanica L., or P. breweri S. 
Watson (Ertter, 1992). The non-obvious nature of 
the taxonomic hypothesis that an undescribed spe- 
cies was involved is evident from the fact that an 
earlier numerical analysis of phenetic variation 
failed to uncover the novelty (Johnston, 1980). The 
species is actually quite distinct, once the appro- 
priate diagnostic attributes are highlight 

n fact, it is more the norm than the exception 
for the first few collections of a species to be shoe- 
horned into existing species-sets, generally with 
modifications to the “biological attributes" hypoth- 
eses. Shevock and Taylor (1987), for example, not- 
ed a range of 1 to 121 years between earliest her- 
barium specimen and publication in their analysis 
of California novelties, with an average of 41 years! 
Ап even greater span is noted by Hartman and Nel- 
son (1998), with the oldest holotype over 200 years 


old, and 6096 of the novelties having type speci- 
mens over 10 years old. However, Hartman's and 
Nelson's statistics underrepresent the actual range 
between initial collection and date of publication, 
in that the earliest collection is not always chosen 
as holotype. Monardella beneolens Shevock, Ertter 
& Jokerst (Lamiaceae), for example, was typified 
on a 1986 collection (Shevock, Bartel & X 
11727), but included among the paratypes was an 
1896 collection (Purpus 1866) that had languished 
in the undetermined-to-species folder for nearly a 
century (Shevock et al., 1989). This example also 
illustrates that the distinction between a novelty 

ased on a new discovery and one resulting from a 
novel analysis of existing specimens is not always 
clear-cut, in that the collection of an undescribed 
Monardella on a 1986 “Inter-Institutional Haybal- 
ing Expedition" is what triggered the herbarium 
search that uncovered the older specimen. 

Nor does the proposing, testing, and rejection of 
alternate hypotheses end once new species are de- 
scribed. For my doctoral work, I essentially tested 
the hypothesis proposed by my advisor that a series 
of annual Juncus (Juncaceae) did not meet the cri- 
teria for recognition as distinct species, as had been 
previously proposed (Hermann, 1948), but rather 

“appear to be mere technical variants, often locally 
constant as in self-pollinated groups in other gen- 
era, but with widely overlapping ranges and similar 
habitat requirements” (Cronquist, 1977: 64). As it 
turned out, my doctoral work not only provided sup- 
port for all of Hermann’s hypothesized species ex- 
cept one, but gave evidence of three additional nov- 
elties (Ertter, 1986). Although I enjoy the notoriety 
that comes with being able to say that I proved Art 
Cronquist wrong and got him to адти it, I will also 
submit that his was a perfectly legitimate hypoth- 
esis based on the information available to him at 
the time. 


C. FLORISTIC MODELS 


The last example introduces the concept of 
monographs and floras as representing complex 
models encompassing multiple species, whose in- 
dividual identities depend on the larger context. 
As a result, the binomial Juncus kelloggii Engelm. 
codes for three very different entities, depending 
on whether it is in the context of Hermann’s, Cron- 
quist’s, or Ertter’s model. In this example, suffi- 
cient evidence has been accumulated to support 
one model over the alternatives, but this is not 
always the case. A contrasting example is pre- 
sented in Table 1, а partial list of corresponding 
units of the taxonomically challenging genus Po- 


96 Annals of the 
Missouri Botanical Garden 
Table 1. Concordance of selected к from alternate taxonomic models of Potentilla (Rosaceae) in the Inter- 


mountain West (= portions of Arizona, Californ 
authors: N. H. Holmgren (1997 


a, Idaho, Nevada, Utah, and Wyoming), as proposed by co 
), B. Ertter шашы (ог Flora of North America, in prep 


mporaneous 


nte 
p.), J. Soják (unpublished 1995 


synopsis of North American tribe Potentilleae), and S. L. Welsh (1993; Utah only, “п/а” indicates entities not occurring 
in Utah). Table includes some unpublished combinations used by Soják. 


Holmgren (1997) 


Ertter (in prep.) 


Soják (unpublished) 


Welsh (1993) 


Potentilla pensylvanica 
Potentilla bipinnatifida 


(= pensylvanica) 
Potentilla pseudosericea 


(= rubricaulis) 
Potentilla ed aulis 
Potentilla concinna 


(7 rubricaulis) 
(= concinna) 


Potentilla bicrenata 
Potentilla diversifolia 
var. diversifolia 
(= concinna) 
(= diversifolia) 
var. perdissecta 


Potentilla gracilis 
var. fastigiata 


var. flabelliformis 
var. elmeri 


var. pulcherrima 


Potentilla pensylvanica 
ar. strigosa 

Potentilla bipinnatifida 
var. bipinnatifida 
var. ovium 

Potentilla pseudosericea 


[to be determined] 
[to be determined] 
Potentilla concinna 
var. concinna 
[to be determined] 
— concinna) 


Potentilla bicrenata 
Potentilla diversifolia 


Potentilla glaucophylla 


[to be determined] 
Potentilla gracilis 
var. fastigiata 


var. permollis 
var. brunnescens 
var. flabelliformis 
var. elmeri 


Potentilla pulcherrima 


Potentilla pensylvanica 
ar. ylvanica 


Potentilla litoralis 
var. litoralis 


var. paucijuga 
P. rubricaulis 
Potentilla concinna 
var. concinna 
2 


Potentilla X concinnaeformis 


var. concinnaeformis 
var. beanii 
P. concinna var. bicrenata 
Potentilla X diversifolia 
var. diversifolia 


раје Е 


Potentilla fastigiata 

var. fastigiat 

var. hallii 

var. jucunda 

var. permollis 
Potentilla nuttallii 
Potentilla flabelliformis 
Potentilla pectinisecta 

var. pectinisecta 


var. comosa 
Potentilla X pulcherrima 
var. pulcherrima 
var. wardii 
Potentilla filipes 
ar. filipes 


Potentilla X lupina 


Potentilla nica 
r. pensylva 


p n/a) 
ru dicen 


var. 
E лит 
Potentilla concinna 


var. modesta 

var. proxima 

var. bicrenata 
Potentilla diversifolia 


var. diversifolia 
P. concinna var. proxima 


Potentilla gracilis 
var. glabrata 


var. brunnescens 


var. elmeri 


var. pulcherrima 


tentilla (Rosaceae) occurring in the Intermountain 
West, as proposed by four different specialists, all 
with access to the same data. The lack of consen- 
sus is not an indication of an inability to agree on 
standards, of taxonomists not being able to “get 
their acts together,” but is rather a reflection of 
four equally valid models for which insufficient 
evidence currently exists to strongly support one 
over the others. 

An even larger-scale example of a floristic model 
is provided by the numerous differences between 


the comprehensive list of California taxa as sum- 
marized in The Jepson Manual (Hickman, 1993) 
and the contemporaneous Inventory of Rare and 
Endangered Plants of California (Skinner & Pavlik, 
1994). As analyzed by Skinner and Ertter (1993), 
the differences result not from one or the other be- 
ing intrinsically “wrong,” but from legitimate phil- 
osophical differences in the rationales behind th 


maximize the likelihood of unequivocal identifica- 
tion, while that of the /nventory was to highlight 


Volume 87, Number 1 
2000 


Ertter 97 
Floristic Surprises 


units of plant diversity that merited conservation 
attention. These different goals resulted in different 
models; in those situations where there was legiti- 
mate room for alternate taxonomic hypotheses, the 
Manual tended to lump where the /nventory tended 
to split, so as to avoid “lamentation over taxa that 
are shown to be distinct only after their disappear- 


ance" (Skinner & Ertter, 1993: 27) 


D. NOVELTIES IN WAITING 


Nevertheless, even within an /nventory-type 
model emphasizing the smallest defensible units as 
worthy of taxonomic recognition, the requirements 
for scientifically legitimate, peer-reviewed publi- 
cation of novelties demand rigorous support for the 
proposed taxonomic hypothesis. For example, the 
Draba study by Windham and Beilstein (1998a, b), 
discussed in an earlier section, clearly demon- 
strates how sophisticated an analysis is often re- 
quired even for the recognition of unequivocally 
distinct species. In addition, although many of the 
highlighted novelties prove that radically different 
species are still being discovered, the truth is that 
the majority of blatantly distinct and/or readily en- 
countered taxa have already been described. As a 
result, ferreting out the remainder will require not 
only continued exploration, but also increasingly 
rigorous scientific analysis 

Because of this, there оне exists ап un- 
known number (50? 200? 500?) of potential nov- 
elties from North America that members of the tax- 
onomic community are collectively aware of, but 
which need to be extensively tested before being 
written up for publication. 1 am personally aware 
of several possibilities, in Juncus, Rosa, Potentilla, 
Horkelia, Eriogonum, Montia, and Lomatium, and 
in fact have as a rule of thumb that any complex 
group that has not been intensively monographed 
recently is likely to harbor undescribed novelties. 
However, all of these possibilities are just that, pos- 
sibilities, and will require a significant investment 
of research effort to determine if they are rigorously 
supportable as taxonomic hypotheses. In other 
words, the limiting factor for many novelties is not 
whether they have been encountered or not, but the 
existence of persons with sufficient expertise, mo- 
tivation, and time to undertake the necessary sci- 
entific analysis. 


ASSUMPTION 3: ACADEMIC PARTICIPATION 


To recapitulate, although the initial discovery of 
novelties does not require professional training, the 
analysis of potential novelties is another matter, in 
which scientific expertise plays a crucial role. Most 


collectors accordingly rely on the network of taxo- 
nomic specialists, who in turn rely on the analytical 
resources represented by herbaria and botanical li- 
braries, as well as established and innovative tech- 
nologies. These resources, along with the custodi- 
чине н апа transmission of the extensive legacy of 
axo ic knowledge, skills, and techniques, have 
sas fallen within the domain of plant tax- 
onomy in an academic setting, including research 
museums and botanical gardens. 


A. THE ROLE OF REGIONAL FACULTY 


Although university-based faculty are only one 


sionals), they are highlighted here on the grounds 
that they are generally assumed to provide the 
backbone of the rigorous taxonomic analysis de- 
scribed in the previous section, especially faculty 
at those universities with large herbaria that occur 
in the regions where most novelties are being dis- 
covered. In this context, it is illuminating to analyze 
the current status of persons in table 4 in Hartman 
and Nelson (1998): “Individuals who authored six 
or more novelties of North American plants during 
the past two decades,” according to the categories 
in Table 2. Of the 56 individuals listed, the two 
largest categories, both in number of individuals 
and number of novelties, are “Emeritus (or nearly 
so)” and “Deceased.” Together, the two categories 
account for 60% of the novelties described from 
1975 through 1994. In contrast, faculty who are 
currently mid-career account for only 6% of the 
novelties. 

To pursue the specific question of novelty de- 
scription by regionally based faculty further, I 
polled plant systematists at universities who 
matched all of the following criteria: 


(1) Located in the contiguous western United 
States (Arizona, California, Colorado, Idaho, 
Montana, Nevada, New Mexico, Oregon, Texas, 
Utah, Washington, Wyoming), a region with a 
high rate of ongoing discoveries. 

(2) Located at a university or college with an her- 
barium of at least 20,000 specimens, repre- 
senting the equivalent of a fully equipped lab- 
oratory for doing taxonomic research on the 
ocal flora 

(3) Self-defined as vascular plant systematist (ver- 
sus ecologist, plant population geneticist, etc.) 
OR serving as director/curator of the depart- 
mental herbarium. 

(4) Department-based (versus adjunct) faculty ap- 
pointment, excluding emeriti, as those persons 


98 Annals of the 
Missouri Botanical Garden 


Current status of individuals listed in table 4 of Hartman and Nelson (1998), “Authors of 6 or More 
Гће first column gives the total number 


Table 2 
Vascular Plant Taxa North America North of Mexico from 1975 through 1994. 
of individuals in each category, and the second column is the sum total of novelties described by these individuals. 
Authors describing fewer than 6 novelties are not included in the tally, nor are novelties described by these individuals. 


No. (percent) 
of novelties 


No. (percent) 


Current status of persons 


Deceased 13 (23%) 174 (21.4%) 
Faculty 

Emeritus 

(or within several years) 12 (21%) 316 (38.8%) 

Early to mid-career 5 (9%) 50 (6.1%) 
Non-faculty academic staff 5 (996) 65 (8.0%) 
Museum staff 9 (16%) 81 (9.95%) 
Government agency biologist 4 (7%) 46 (5.65%) 
Environmental consultant 2 (4%) 29 (3.6%) 
Private individual 2 (4%) 22 (2.7%) 
Unknown 4 (796 ЗІ (3.8%) 
Total 56 814 


whose hiring, promotion, and tenure are deter- 
mined by current departmental expectations. 


The specific question addressed was whether 
each respondent had described (a) zero, (b) one, or 
(c) more than one vascular plant novelty from any- 
where within the contiguous western United States. 
The number of responses to this survey was grati- 
fying, but the collated results (Table 3) are thought- 
provoking. Of the 56 persons included in the sur- 
vey, over half had not described a single novelty 
from the region, and over half of the remainder had 
only described a single novelty (or at least had one 
in press). In several cases, this solitary western 
novelty was described during the course of graduate 
work but not since attaining faculty status. Only 10 
qualifying faculty members in the entire region 
have described more than one novelty from the re- 
gion, and several of these persons are within a 
handspan of years from retiring. 

Furthermore, of the 48 western universities with 
significant herbaria, 5 currently lack faculty-level 
vascular plant systematists, including 2 that house 
the largest herbaria for their respective states (Uni- 
versity of Montana, Missoula; University of Nevada, 
Reno). In Oregon, the two primary herbaria were 
recently свиле. eliminating the position of plant 
systematist at the University of Oregon, Eugene 
One state (Colorado) currently lacks a faculty-level 
plant systematist who has published any novelties 
from the region, while four others (Arizona, Mon- 
tana, Nevada, Washington) can claim only one fac- 
ulty systematist who has described a single novelty 


from the target area. This in a region in which 813 
taxa were described from 1975 through 1994, 
around 41 per year, with no evidence of tapering 


off (Hartman & Nelson, 1998) 


B. ACADEMIC SELECTION PRESSURES 


The purpose of this survey was not to call into 
question the scientific productivity of the respon- 
dents, who are all actively pursuing a commendable 
diversity of significant research in plant systemat- 
ics, including describing novelties from other parts 
of the world. Nor is it intended to slight the signif- 
icant contributions of individuals outside the ad- 
mittedly narrow survey criteria, professional and 
otherwise. The survey does, however, underm 
any assumption that faculty-level plant а 
at the best-equipped western universities comprise 
the major pool of expertise in ongoing efforts to 
analyze and describe the unknown elements in a 
novelty-rich regional flora. 

Moreover, there is evidence that this is not a 
statistical curiosity, but rather an indication that the 
current academic infrastructure actually discour- 
ages such participation. Several respondents indi- 
cated that they knew of undescribed regional nov- 
elties, but could not justify the research time and 
effort required to publish them. In the words of one 
such respondent, “the value of new species de- 
scriptions in terms of professional prestige and sat- 
isfaction of university administrators (who control 
raises and promotions) seems low relative to other 
publications that could be generated in a similar 


Моште 87, Митбег 1 
2000 


Ertter 
Floristic Surprises 


Table 3. 


Participation of current vascular plant systematist faculty in the contiguous western United States in the 


description of regional novelties (Ertter, unpublished data). Column 1 is the number of universities or colleges in each 


state with significant herbaria (defined her 


e as at least 20,000 spec 


cimens). Column 2 is the number of department- 


based (vs. apii vascular plant systematists, and non-systematists actively serving as herbarium director. Columns 


3, 4, an аге t er of persons in column 


Percent representation among these three categori 


ies is given i 


who have described, respectively, (a) zero, (b) one, or (c) more than 
one жаш ке ннн Кот ey within the rone western United States, i 


including novelties in press. 
n parenthesis after summary totals. Column 6 is the total 


number of novelties from each state published 1975 to 1994, “tallied i in table 8 of Hartman and Nelson (1998). 


Novelties described 


Total 
Institutions Faculty 1 >1 novelties 
Arizona 3 3 1 0 60 
California 16 14 2 4 223 
Colorado 3 3 0 0 33 
Idaho 3 4 2 0 33 
Montana 2 1 1 0 12 
2 1 1 0 64 
New Mexico 2 3 2 1 41 
Oregon 2 3 2 0 44 
Texas 8 14 || 1 2 75 
tah 3 3 1 2 183 
Washington 3 5 1 0 13 
Wyoming 1 2 1 1 32 
Totals 48 56 31 15 10 813 
(percentages) (55%) (27%) (18%) 


period of time.” In effect, the publication of re- 
gional novelties is not only of little value, it is ac- 
tually counterproductive to career development in 
the current academic environment. Paradoxically, 
the fact that the amount of time and effort it takes 
to publish a novelty can be equivalent to that need- 
ed for other research activities in itself provides 
evidence that describing novelties is not the trivial 
activity it is routinely perceived to be. 

it illuminating to compare the preceding 
quote with another, from nearly a half century ago: 
“If taxonomy and taxonomists are to regain some of 
their lost prestige—and they have lost a great 
deal—it seems obvious that mastery of a local flora, 
an ability to recognize characteristic members of 
the more common plant families, a familiarity with 
the rules of nomenclature, and the capacity to write 
descriptions are bound to prove woefully inade- 
quate” (Constance, 1951: 229). We have apparently 
come full circle, where the skills that were once 
the sine qua non of a practicing taxonomist have 
apparently gone from being “inadequate” to being 
irrelevant. Or, at best, these former skills are as- 
sumed to come as part of the “systematist pack- 
age,” overlooking the tenet otherwise well known 
to biologists that “you get what you select for.” Ad- 
mittedly, the above analysis is only a single slice 
in time, but it nevertheless strongly suggests that 


the activities that result in the publication of re- 
gional novelties are NOT among those currently be- 
ing selected for within academia, and as a conse- 
quence are de facto being selected against. 


C. WHO IS DOING THE WORK? 


The question then arises: If not faculty-level sys- 
tematists at the best-equipped regional universities, 
who is responsible for generating the 41 novelties 
per year in the contiguous western United States? 
Obviously there are numerous people who are dis- 
covering and describing western novelties other 
than those targeted here, who I have neither sur- 
veyed nor otherwise statistically analyzed. The ma- 
jor categories, however, would include the follow- 
ing: emeriti plant systematists; museum-based 
plant systematists, often with adjunct appointments 
at nearby universities; faculty-level plant systema- 
tists at less well-equipped regional university and 
colleges (i.e., with herbaria having less than 20,000 
specimens); plant systematists outside of the region; 
non-faculty research and curatorial appointments; 
non-systematists (e.g., ecologists, population genet- 
icists); government agency biologists; biologists 
working for the private sector, mostly as environ- 
mental consultants; and amateur enthusiasts. 

Certainly the academic and museum-based cat- 


100 


Annals of the 
Missouri Botanical Garden 


egories play significant roles, which should not be 
underestimated. What I wish to draw attention to 
at this point, however, is the high degree of partic- 
ipation by professionals and amateurs outside of 
academia, many of whom have an exceptional eye 
for novelties and a serious commitment to floristic 
undertakings. As representative examples, some of 
the more outstanding are spotlighted below: 


(1) Government agency biologists 


Beginning with his stint in 1979 as botanist for 
the Sequoia National Forest in California, James R. 
Shevock has now tallied 6 vascular plants and 1 
moss named in his honor, 12 others that he has 
authored, and several undescribed novelties in var- 
ious stages of publication. Many of his earlier nov- 
elties were encountered by his being the first bot- 
anist each year on newly constructed portions of 
the Pacific Crest trail, which ended up bisecting a 
population of Allium shevockii McNeal that is still 
one of the only populations known. While retaining 
a focus on the southern Sierra Nevada, Shevock’s 
botanical interests have subsequently expanded to 
include mosses and lichens, with Orthotrichum 
shevockii Lewinsky-Haapasaari & D. H. Norris (Or- 
thotrichaceae) being the most recent addition to his 
eponymous tally. The protologue credits Shevock 
with “opening the eyes of the junior author to the 
bryophyte riches of the southern Sierra," a signifi- 
cant accomplishment considering Norris's extensive 
expertise with the California bryoflora (Lewinsky- 
Haapasaari & Norris, 1998). Shevock's agency ca- 
reer has likewise expanded; as Regional Botanist, 
he prepared the status report on rare and endemic 
plants for the Sierra Nevada Ecosystem Project 
(Shevock, 1996), and he has recently moved on to 
become Associate Regional Director of the National 
Park Service. Shevock's botanical explorations are 
accordingly now confined to weekends and vaca- 
tions, but have not noticeably slowed as a result. 

n a 1996 foray (a.k.a. “death march") to an iso- 
lated marble ridge, he and protégé Dana York (bot- 
anist for Death Valley National Park) discovered 
three novelties in a single day: Heterotheca mon- 
archensis D. A. York, Shevock & Semple, Gilia yor- 
kii Shevock & A. G. Day, and a still-undescribed 
Eriogonum. 


~ 


(2) Environmental consultants 


The tragic death of James D. Jokerst, who 
drowned while trying to retrieve the family canoe, 
cut short the career of one of the persons who did 
the most to convert environmental consulting into 
a legitimate career for skillful, well-trained bota- 


nists. While working full-time for the environmen- 
tal consulting firm Jones & Stokes Associates, Jok- 
erst nevertheless found time to develop expertise in 
the Lamiaceae, preparing treatments of several gen- 
era in The Jepson Manual (Hickman, 1993), in- 
cluding the notoriously difficult Monardella (Mag- 
ney, 19 e also authored or coauthored three 
novelties: Acanthomintha obovata subsp. cordata 
Jokerst, МопагаеЏа beneolens Shevock, Ertter 4 
Jokerst, and Pogogyne floribunda Jokerst. While 
doing a botanical survey in 1985, Jokerst discov- 
ered an unusual gold-flowered Trifolium, which was 
posthumously named in his honor (Vincent & Mor- 
). According to Vincent (pers. comm. 


Heller had collected at several times, but never 
early enough in the season!” 

Environmental consultants in general are playing 
an increasingly significant role in discovering nov- 
elties, as the persons most likely to have access to 
poorly botanized areas. As prime examples, the dis- 
coveries of Yermo xanthocephalus, Twisselmannia 
californica, and Neviusia cliftonii were connected 
to environmental survey efforts. Unfortunately, a 
great many biological consultants lack the training 
or orientation needed to recognize potential novel- 
ties, and may in fact be discouraged from taking 
note of anything but a mechanically generated list 
of rare species determined to be potentially present 
at a given site. This practice is based on the dan- 
gerously flawed assumption that previously existing 
knowledge is an accurate indication of likely oc- 
currence, an assumption at odds with the theme of 
“floristic surprises.” As summarized by S. Boyd 
pers. comm. 1998), discoverer of Sibaropsis and 
other novelties in the course of doing environmental 


~ 


surveys: 


"There is the strong possibility that other ipii 
d some 


der how many other undescribed taxa have been over- 
looked and к кайыга lost to ae шш 


(3) Amateur enthusiasts 


Among the more unexpected of the amateur en- 
thusiasts is Lowell Ahart (Geary, 1978), a sheep 
rancher who started out cataloging the plants of his 
ranch and has since moved on to county floras, col- 
laborating with retired zoology professor Vern Os- 

(e.g., Oswald & Ahart, 1994). Two plants from 


his ranch have been anes after him (Juncus leio- 


Volume 87, Number 1 
2000 


Ertter 101 
Floristic Surprises 


spermus var. ahartii Ertter, Paronychia ahartii Ert- 
ter), and an Eriogonum is also being named in his 
honor (J. Reveal, pers. comm. 1997). When Ahart 
brought the undescribed Paronychia to my atten- 
tion, begging that someone provide a name for it so 
he could complete the checklist of his ranch, it had 
actually been known for some years but had been 
assumed to represent yet one more introduced Eur- 
asian annual. However, by then a worldwide mono- 
graph of the Paronychiinae was available (Chau- 
dhri, 1968), making it evident that an anomalous 
undescribed species was involved, whose affinities 
are still unclear (Ertter, 1985). 


(4) Other para-academics 


A final example of expertise outside of academia 
is provided by Amold (Jerry) Tiehm, who has an 
advanced degree in botany and previous profes- 
sional experience (e.g., curatorial staff at The New 
York Botanical Garden). For the last several years, 
however, Tiehm has earned his living as bell cap- 
tain and limousine driver at the Peppermill Casino 
in Reno, Nevada, doing his botanizing on his days 
off. At last count he has nevertheless made the type 
collections of 19 species (Holmgren, 1998), ap- 
proximately one per year, several of which are 
named after him. Probably the most significant is 
Stroganowia tiehmii Rollins, the single Nort 
American representative of a genus otherwise con- 
fined to central Asia (Rollins, 1982). It also qual- 
ifies as another “in our backyards” discovery, not 
encountered until 1980 even though occurring only 
a few miles off a well-traveled highway 20 airmiles 
east-southeast of Reno 


D. IS THE POOL SUFFICIENT? 


These highlighted individuals are only a sam- 
pling from a large pool of talented and dedicated 
individuals operating outside of an academic set- 
ting, including some who not only discover but an- 
alyze and describe their own novelties. When these 

ara-academics are combined with museum-based 
systematists, faculty at smaller institutions, and 


universities should appropriately be encouraged to 
address avenues of research that cannot be handled 
by others 

While confirming my enthusiastic support for a 
diversity of individual research interests within ac- 
ademia, and likewise for the active participation of 
individuals outside of the academic mainstream, I 
will nevertheless argue (as have others, such as 


Kruckeberg, 1997) that a core of professional plant 
systematists will continue to play an indispensable 
role in the task of discovering, analyzing, and de- 
scribing the remaining unknown element in the 
North American flora, as well as critically evalu- 
ating new information accumulated about previous- 
ly described species. In other words, rather than 
being made redundant by the para-academic net- 
work, an active core of professional systematists is 
integral to the proper functioning of the network. 

urthermore, a significant percentage of this pro- 
fessional core needs to be housed at the large re- 
gional herbaria, especially in the West and South- 
east where the majority of floristic discovery is 
occurring. 

The obvious argument for academic participation 
is, of course, to provide the formal systematic train- 
ing for all other participants in the network, 
cluding agency biologists and environmental con- 
sultants. Perhaps even more critical, however, is the 
reality that regionally based professional systema- 
tists represent the essential source of quality con- 
trol and accessible scientific expertise to turn to 

hen non-systematists encounter "plants that 
haven't read the book." Furthermore, para-academ- 
ics who analyze and describe their own novelties 
generally do so only after a period of *apprentice- 
ship" with a regionally based, practicing taxono- 
mist. In this regard, it is unsettling to realize how 
many of the regional professionals who provided 
early encouragement and training to the current 
crop of active para-academics are now retired or 
deceased. А prime example of the latter is the late 
John Thomas Howell of a California Academy of 
Sciences, who provided s 
to most of the individiula highlighted above. 

Howell also represents the category of museum- 
based research staff that is becoming increasingly 
important in maintaining the role of the profession- 
al core. As significant as this contribution is, how- 
ever, museums and botanical gardens are too few 
in number to provide complete regional coverage, 
and are also less likely to be involved in formal 
training. At the same time, the suggestion that de- 
scriptive systematics should be relegated to smaller 
universities. and colleges runs counter to the fact 


ificant encouragement 


regional universities is therefore essential to the on- 
going task of discovering and analyzing the regional 
flora. If this task is in fact incompatible with aca- 
demic realities currently facing faculty systematists 
(and not just a matter of erroneous perceptions), 
then it is imperative that alternate ways to ensure 
such participation be investigated, perhaps involv- 


102 


Annals of the 
Missouri Botanical Garden 


ing direct funding or collaborative programs with 
state or federal land-management agencies. 


E. THE TAXONOMIC LEGACY 


Approaching the situation from a different angle, 
there is the question of the taxonomic legacy, a term 
used here to refer to the material resources (1.е., 
regional herbaria and associated libraries); the fun- 
damental knowledge, skills, and techniques for flo- 
ristic analysis; and the setting of scientific stan- 

ards, generally in a peer-reviewed context. 
Custodianship of this legacy, developed over sev- 
eral centuries, has traditionally fallen within the 
domain of plant taxonomy in an academic setting: 
universities, research museums, and botanical gar- 
dens. The question that needs to be asked is not 
only to what extent this legacy is being maintained, 
but to what extent the components are current 
available to whomever is actually doing the bulk of 
analyzing and describing regional novelties, what- 
ever their professional credentials and self-identi- 
ties might be. 

A prime example is the situation referred to 
above, in which large universities that house major 
herbaria struggle to justify their upkeep while si- 
multaneously relegating descriptive systematics to 
less well-equipped or well-situated institutions. The 
transfer of floristic survey work to the domain of 
ecologists, another apparent trend, is also a poten- 
tial problem if it is not accompanied by the transfer 
of associated skills and techniques (e.g., critica 
taxonomic analysis and an understanding of the 
role played by vouchers). Paradoxically, it is pos- 
sible that, in the name of increasing the scientific 
respectability of systematics, one outcome might 
actually be a net decrease in the scientific standards 


= 
~ 


underlying the analysis and description of new spe- 
cies, the very foundation of our knowledge of bio- 
diversity. 


ASSUMPTION 4: THE PROPERTY RIGHTS CONFLICT 


A. SOCIOLOGICAL RAMIFICATIONS OF FLORISTIC 
SURPRISES 


Furthermore, what are the broader ramifications 
of allowing taxonomy and floristics to take place 
largely as a collective avocation, a labor of love 
even for those in professional positions, rather than 
an academically supported undertaking? For better 
or worse, the days when new taxonomic and floristic 
discoveries were of concern only to professional 
botanists and amateur enthusiasts are far behind 
us. Although there are numerous exceptions (e.g., 
Juncus tiehmii Ertter, which is widespread in west- 


ern North America), the majority of newly de- 
scribed species qualify for some level of sensitive 
species status, with immediate implications for 
land-management activities on public and private 
lands. Problems can arise if negatively affected 
landowners develop the impression that so-called 
"amateurs" are behind newly described rare spe- 
cies without the backing of socially sanctioned ex- 
pertise. 

Even for those species that aren't novelties, a 
lack of sufficient information on their taxonomy and 
distribution interferes with effective conservation 
efforts. The magnitude of this problem can be seen 
in the list generated by Skinner et al. (1995) of 182 
rare California plants for which further taxonomic 
resolution is needed, and another 44 that require 
additional distributional information before their 
conservation status can be properly assessed. The 
implications of this knowledge gap become appar- 
ent in the face of decisions that are being made 
now which will irrevocably determine the fate of 
much of our natural heritage, representing a vast 
resource containing both pragmatic and aesthetic 
values. 


B. IN FRONT OF THE BULLDOZER 


In this context, it is unsettling to realize how 
many plants (and other organisms) that would qual- 
ify for some level of management activity, possibly 
representing 546 of the North American flora (based 
on Taylor's extrapolations as previously discussed) 
and including some of the rarest of the rare, are 
currently receiving NO attention because they have 
not yet been discovered, analyzed, and described. 
In other words, we risk losing a significant per- 
centage of our floristic heritage out of sheer igno- 
rance of its existence, not just in the tropics but in 
our own backyards. 

In support of this statement, an increasing num- 
ber of novelties in North America, as in other parts 
of the world, are being discovered “in front of the 
bulldozer." The type locality of Neviusia cliftonii, 
for example, is threatened by a limestone quarry, 
and the monotypic genus Yermo was discovered as 
part of a survey along a proposed pipeline route. 
The narrow endemic Ivesia aperta (J. T. Howell) 
Munz var. canina Ertter (Rosaceae) was still in 
manuscript when plans to build a dam that would 
have flooded almost the entire population came to 
light (Ertter, 1988). Ceanothus ophiochilus S. Boyd, 

oss & L. Атпзећ (Rhamnaceae) was found 
during the environmental impact study of a pro- 
posed development in southern California (Boyd et 
al., 1991). Even more recently, the type population 


Volume 87, Number 1 
2000 


Ertter 103 
Floristic Surprises 


of an undescribed Pseudostellaria (R. Hartman & 
R. Rabeler, in. prep.) was found adjacent to the 
staging area for an active logging site, potentially 
surveyed for sensitive species prior to approval for 
timber harvest, but not for undescribed taxa. A new 
tarweed currently being described from Livermore, 
California, also falls into this category, occurring as 
it does in an area of some of the heaviest devel- 
opment pressure in the San Francisco Bay Area 
(Baldwin, 1999). 

Unfortunately, there is also evidence of potential 
novelties being eliminated before they could even 
be described. This may be the case with an unde- 
scribed Eriogonum mentioned in the protologue of 
E. capistratum Reveal var. welshii Reveal (Reveal, 
1989b), whose only known population in south- 
western Idaho has possibly been eliminated by the 
construction of communication towers. 


C. LANDOWNER RESISTANCE TO SURVEYS 


In-front-of-the-bulldozer discoveries, exciting as 
they might be to the botanical community, can be 
a decidedly rude surprise for the landowner, rep- 
resenting an unexpected and potentially very ex- 
pensive complication in what might otherwise have 
been a relatively straight-forward and profitable un- 
dertaking. One newspaper covering the rediscovery 
of the Ventura Marsh milkvetch (Astragalus pyc- 
nostachyus var. lanosissimus) noted how the plant 
was “causing trouble" and had “thrown a kink” in 
the developer's plans to build $300,000 homes sur- 
rounding a man-made lake (Sacramento Bee, 15 
Aug. 1997). Fanned by negative publicity and 
property rights advocates, the fear that property val- 
ues and development options will be severely cur- 
tailed by the discovery of such unwelcome surpris- 
es has unfortunately led to a significant polarization 
between private landowners and advocates of bio- 
diversity protection. This in turn has often resulted 
in a refusal to allow floristic surveys on private 
lands, which can contain significant portions of rel- 
atively unexplored areas that could harbor novelties 
and populations of other significant plants. 

The scale of the fear and distrust has even led 
to the paradoxical situation in which local land- 
owners insist that “there are thousands" 
called rare plant on their properties, while simul- 


of а 50- 


taneously refusing to allow the scientific surveys 
needed to justify less stringent management op- 
tions. Frank testimony to exactly this situation, and 
to the massive amount of distrust, fear, and outrage 
that can build up in the absence of trustworthy 
sources of reliable information to the contrary, is 


provided by Janssen and Williamson (1996). In 


summarizing her efforts to gain access to reported 
populations of Frankenia johnstonii Correll (Fran- 
keniaceae) on the private ranchlands of Zapata 
County, Texas, state botanist Gena Janssen (in Jans- 
sen & Williamson, 1996: 3) shared these insights: 


I began to meet and get to know more and more 
el I began to notice that most of them did 
basic € the same thing when they met me: They yelled 
atm nd then one day it рам ћи me as to why they 


just needed to vent, so I let them. They ha 
there for them. There was no one there to say, "Мо, that's 
not true,’ or ‘Yes, that was a very difficult situation for 
“i or “Well, only part of that is true,’ etc., until 
no 


Encouragingly, this stage was the prelude to a 
particularly noteworthy success story. Janssen’s pa- 
tience, honesty, and willingness to listen compas- 
sionately paid off, first in obtaining the access 
needed to acquire critical distributional and other 
biological data on a plant that occurs almost exclu- 
sively on private land, and subsequently in working 
with the landowners to develop a voluntary conser- 
vation plan. As a result, Ё johnstonii is currently 
being removed from the endangered species list, 
with many of the ranchers now taking legitimate 
pride in "their" rare plant (Janssen, pers. 

8; see also http://www.tpwd.state.tx.us/news/ 
news/980518a.htm). 

n this context, there is a distinct irony in the 
fact that the fundamental floristic work in Kern 
County, California, currently a property-rights 
stronghold, was undertaken by a local rancher, Er- 
nest C. Twisselmann. His contributions to Califor- 
nia botany have recently been acknowledged in the 
form of a newly discovered genus named in his hon- 
ia (Al-Shehbaz, 1999). Twissel- 
mann’s interest in S triggered by an outbreak 
of nitrate poisoning in his cattle (McClintock, 
1973), eventually led to the в 9 of two flo- 
ras (Twisselmann, 19 and the discovery 
of several new species le g., Nemacladus twissel- 
mannii J. T. Howell, Eriogonum temblorense J. T. 
Howell & Twisselm.). The acknowledgments to his 
1967 flora provide insight into the respect for pri- 
vate property that lay behind the success of his 
undertaking 


mm. 


or, Twisselmannia 


“In a time when malicious trespass and vandalism are 
form of outd 


gave me access to their property, and whose friend- 


104 


Annals of the 
Missouri Botanical Garden 


ships have been one of the quite unexpected dividends 


of the fieldwork.” 


D. FLORISTIC SURPRISES OR “NO SURPRISES”? 


The Frankenia johnstonii example is only one 
among many in which an increased floristic infor- 
mation base, sometimes paid for by the private 
stakeholders, worked in their favor, either by pro- 
viding sufficient scientific evidence for reduced 
protection status (e.g., downlisting) or by increasing 
the mitigation options. These examples need to be 
brought together for impact, but currently exist only 
in scattered documents and word-of-mouth reports. 
Granted, there is a difference between a ranch fam- 
ily that wishes to continue a way of life requiring 
large open spaces, and a developer who needs to 
subdivide and build in order to realize an invest- 
ment. Even in the latter case, however, the negative 
consequences of floristic ignorance can cut both 
ways, increasing the risk of misplaced mitigation 
efforts as well as the unintentional extinction of 
species. 

As a society, we have acknowledged that the per- 
petuation of our biodiversity heritage is a highly 
desirable goal, for pragmatic, aesthetic, and ethical 
reasons. Within this context, the key question be- 
comes how to accomplish this goal as fairly and 
effectively as possible. Unfortunately, instead of 

making the necessary hard decisions on a solid ba- 
sis of complete scientific knowledge of all elements 
involved, we are forced to face the tragic fact that 
the “best available scientific evidence” 
woefully inadequate reflection of the actual data 
needed for the kind of far-reaching decisions that 
are currently mandated. 

A common quandary, for example, is determining 
whether a species is truly as rare as existing evi- 
dence indicates. In these circumstances, it is some- 
times argued that, if the scientific evidence is in- 
complete, then no land-management constraints 
can be justified. This argument, however, runs 
counter to the fact that all legal decisions, includ- 
ing those addressing environmental issues, are 
based solely on best evidence available at the time 
of the decision, with neither hearsay nor supposi- 
tion having a legitimate role. One can speculate 
that a species is more widespread than the cur- 
rently available scientific evidence indicates, but a 
decision based on m speculation without hard ev- 
idence to back it up is no more justified than is 
ruling on a ыш. guilt strictly on speculation 
that the person might have done the crime. Given 
this, it is readily apparent that operating from a 
maximally comprehensive and accurate information 
base is vastly preferable to acting in ignorance, and 


is often a 


willful ignorance becomes inexcusable, if not out- 
right foolish. When all is said and done, the best 
guarantee of “No Surprises” (the nickname for a 

ey landowner incentive in regional conservation 
plans) is complete information up front. 


ASSUMPTION 5: THE OVERWHELMING CHALLENGE 


Within this framework, the significantly prefer- 
able option to isolated, development-driven surveys 
would be a proactive, comprehensive effort to ad- 
dress the existing gaps in our floristic information 
base. It works to no one’s benefit for an undescribed 
plant or a significant population of a sensitive spe- 
cies to be discovered after significant funds have 
already been expended on a proposed project [e.g., 
a newly discovered Draba that is “complicating 
Olympics preparations” for the 2002 Winter Games 
in Utah, having been found at the site of the men’s 
downhill race course (Deseret News, 22 Aug. 1998; 
Windham & Bellstein, 1998b)]. There is further- 
more a distinct sense of unfairness in having the 
short straw fall to the landowner(s) of the last refuge 
of a once-common species, which only became en- 
dangered when neighboring landowners had devel- 
oped their parcels first. 

Avoiding such situations is in fact a primary goal 
behind the current focus on developing regional 
conservation plans on which to base land-manage- 
ment decisions. Although excellent in principle, in 
reality such efforts have often been deficient in ad- 
dressing species-specific information, in large part 
because of the assumption that obtaining the rele- 
vant species-specific floristic information is too for- 
midable a challenge to pursue. This in turn leads 
to the argument that alternate information (e.g., sat- 
ellite imagery, umbrella species) serves as an ad- 

equate substitute to field-based, species-specific 
floristic data. A dramatic counter to this argument 
is provided by the Shasta snow-wreath (Neviusia 
cliftonii), in which a relatively conspicuous shrub 
was completely overlooked by one of the most com- 
plete vegetation mapping projects ever undertaken 
(Weislander et al., 1939). As a bottom line, large- 
scale land-management plans that address only 
dominant and formally listed species have the po- 
tential of allowing the incremental disappearance 
of all other species in the region, including any 
undescribed novelties, without even leaving a rec- 
ord of their previous existence. 

The challenge of comprehensively addressing the 
species-specific gaps in the floristic information 
base is indeed formidable, but the assumption that 
it is an overwhelmingly unrealistic goal is based in 
large part on the assumptions previously addressed. 


Volume 87, Number 1 
2000 


Ertter 105 
Floristic Surprises 


In particular, it can hardly be said that the as- 
sumption has ever been put to the test, given the 
low level of support that floristic efforts have his- 
torically received. For such an undertaking to be- 
come a reality, however, the following would ac- 
cordingly need to be addressed: 


• acknowledge incompleteness of existing floristic 
knowledge base; 

assign value to floristic information commensu- 
rate with the effort required to generate it and its 
value to society at large; 

• ensure that essential academic resources аге 


available at regional level; 
• foster the network of professional and рага-рго- 
fessional expertise; 
promote the training and participation of 
para-professionals within a framework of accept- 
able scientific standards; 
depolarize relations with private landowners, with 
academic participation providing an essential 
agenda-neutral framework; 
disperse floristic information in a framework that 
addresses the particular needs of all participants. 


Several possible prototypes incorporating one or 
more of these elements have already been devel- 
oped. The Rocky Mountain Flora Project, for ex- 
ample, demonstrates the scale that can be accom- 
plished by focused floristic surveys within an 
academic setting (Hartman, 1993). In contrast, the 
Oregon Flora Project depends less on graduate stu- 
dent projects and more on existing and newly gen- 
erated information from an extensive network of ac- 
ademic, agency, and native plant society sources, 
critically analyzed by herbarium-based professional 
systematists (Sundberg, 1997). The 1980 peak of 
novelty description in Utah, as shown in Figure 1, 
resulted in large part from the collaborative activ- 
ities of regional academics, agency biologists, and 
environmental consultants, and similarly collabo- 
rative *haybaling expeditions" have taken place in 
Idaho (Big Horn Crags) and the southern Sierra Ne- 
vada. In the San Francisco Bay area, a regional 
checklist was specifically designed to facilitate and 
encourage the participation of para-academics in 
floristic inventory efforts (Ertter, 1997b). In that all 
of these efforts, and the discovery of “floristic sur- 
prises" in general, have proceeded with minimal 
institutional support in an increasingly avocational 
network, one can only speculate as to what could 
potentially be accomplished within the framework 
of a well-coordinated, seriously supported floristic 
undertaking, taking full advantage of both profes- 
sional and para-academic networks. 


THE Bic PICTURE 


In conclusion, I propose that what taxonomists 
have been up to is nothing less than one of the 
most massive scientific endeavors ever undertaken: 
namely, a centuries-long, internationally collabo- 
rative effort to model global biodiversity. If this 
does not qualify as “Big Science,” I don’t know 
what does! The significance of this undertaking 
takes multiple forms, starting with the fundamental 
desire to know what other forms of life share this 
planet with us, the only island of life we know for 
certain exists in the universe. The resultant model 
also forms the foundation underlying other branch- 
es of biological knowledge, and it follows that the 
more complete and accurate the model is, the stron- 
ger the foundation is (cf. the “taxonomic impedi- 
ment” of R. W. Taylor, 1983). Most important, as 
we now find ourselves in an era when crucial de- 
cisions are being made that will determine the face 
of life on the planet, it is imperative that these de- 
cisions be made with the most comprehensive in- 
formation possible. 

Furthermore, the challenge of obtaining the spe- 
cies-specific floristic information needed to make 
science-based land-management decisions in North 
America north of Mexico, although formidable, is 
not beyond our grasp. However, the viability of the 
essential professional taxonomic infrastructure 
needs to be ensured, and the undertaking ap- 
proached as a seriously supported collaborative ef- 
fort combining academic and para-academic re- 
sources at the regional level. If not, then we risk 
losing 5% of the floristic diversity in the North 
American “backyard” by ignorance alone, as well 
as unfairly allocating the conservation costs for bio- 
diversity in general. 

A quote by Thomas Bridges opened this paper, 
expressing his amazement that there were still flo- 
ristic surprises in North America in 1858. I will 
end with a more accurate perception by Dieter 
Wilken, expressing his delight in the Colorado flora 
in 1984, over a century after Bridges’ visit to Cal- 
ifornia (transmitted by R. Patterson, pers. comm 

998): “I am continually amazed at the things that 
are yet to be discovered.” 


ACKNOWLEDGMENTS 


This paper would have been significantly depau- 
perate, if not outright impossible, without the open- 
handed sharing of favorite examples, a. 
personal anecdotes, survey responses, and other 
critical information by a multitude of persons, to 
whom I am accordingly indebted and deeply grate- 
ful. In particular I must express my deep appreci- 


106 


Annals of the 
Missouri Botanical Garden 


ation for the generosity of those individuals who so 
willingly lent me a fantastic selection of their beau- 
tiful slides of photogenic novelties, and/or provided 
me with unpublished figures that provided a critical 
statistical underpinning for this paper: James Af- 
folter, Steve Boyd, Thomas Bruns, Roy Buck, Adolf 
eska, David Charlet, Beth Corbin, Raymond 
Cranfill, Marshall Crosby, Charmaine Delmatier, 
Ronald L. Hartman, Noel Holmgren, Fred Hrusa, 
David Isle, Gena Janssen, Patrick Leary, David 
Magney, Richard L. Moe, James D. Morefield, Jan 
Nachlinger, Elizabeth Neese, James L. Reveal, An- 
ton A. Reznicek, James R. Shevock, Paul Silva, 
Frank (Buddy) Smith, R. Douglas Stone, Raymond 
Stotler, Dean W. Taylor, John Taylor, Michael Vin- 
cent, and Carol W. Witham 
contributed in various ways are Lowell Ahart, 
James R. Allison, Kelly W. Allred, Ihsan Al-Sheh- 
baz, Bonnie Amos, Tina Ayers, Bruce Baldwin, 
Mary Barkworth, Kathryn A. Beck, Peter Bowler, 
Gregory Brown, Richard Brummitt, Steven Bruns- 
feld, Thomas Carlson, William Carr, Terri Charlet, 
Curtis Clark, Lincoln Constance, Allyson Davis, El- 
len Dean, Dennis Desjardin, Heidi Dobson, David 
L. Dyer, Wayne Ferren, Stuart Garrett, Arthur 
son, Ann Halford, Richard Halse, Gary SOM 
alter Holmes, Larry Hufford, C. Eugene Jones, 
David Keil, Sylvia (Tass) Kelso, Les Landrum, Matt 
Lavin, Aaron Liston, Timothy Lowrey, Don Mans- 
field, Michael Mancuso, Niall McCarten, Lucinda 
McDade, Dale W. McNeal, Brent Mishler, Joseph J. 
Molter, David Morgan, Walter (Tony) Morosco, Bar- 
bara Murray, David Murray, Kathleen Nelson, Bry- 
an Ness, Wesley Niles, Richard Olmstead, Brad Ol- 
son, Robert Ornduff, Vernon H. Oswald, Jose L. 
Panero, Robert Patterson, James B. Phipps, Jackie 
Poole, Daniel Potter, Jerry Powell, Teresa Prendusi, 
Richard Rabeler, Thomas Ranker, Peter H. Raven, 
Monique D. Reed, Andrew Sanders, Michael San- 
derson, Kristina Schierenbeck, Lisa Schultheis, 
Leila Shultz, Beryl B. Simpson, Michael G. Simp- 
son, Alan R. Smith, S. Galen Smith, Douglas Soltis, 
Brian Speer, Bruce Stein, Kingsley Stern, Scott 
Sundberg, David Tibor, Arnold (Jerry) Tiehm, Catol 
A. Todzia, Billie Lee Turner, Florence Wagner, War- 
. Wagner, Phil S. Ward, William A. Weber, 
Steve Weller, Stanley L. Welsh, Jun Wen, Charlie 
Werth, Dieter Wilken, Hugh Wilson, Michael Win- 
dham, Lindsay Woodruff, Richard Worthington, Mi- 
chael J. Wynne, Vern Yadon, James Zarucchi, and 
anonymous reviewers. 


. Other individuals who 


Literature Cited 


Allison, J. R. 1994. A botanical “Lost World" 
Alabama. Pp. 323—327 in J. S. Fralish, R. C. 


in central 
Anderson, 


J. E. Ebinger & R. Szafoni (editors), Proceedings of the 
North American C Bii ге on Barrens and Savannas. 

Illinois State University, ral. 

Al-Shehbaz, 1. 1999. ied (Brassicaceae), a re- 
markable new genus from California. Novon 9: 132- 
135. 

Anderson, J. L., J. L. Reveal & R. C. iis 1997. Les- 
querella vicina (Brassicaceae), a species from the 
Ms 'ompahgre River Valley in western d, olorado. Novon 

: 9—1: 

mun 1998. 5 аге redis- 

covered. BioScienee 48: 

у 93. Molecular а netics of Caly- 

sadenia (С mpi) based on ITS jences of nu- 

clear ribosomal DNA: Chromosomal ме morphological 

evolution reexamine 5 Amer. J. Bot. 80: 222-238. 

. 1999, кши ара (Compositae—Ma- 
diinae), a new tarweed f astern Alameda County, 
California. Модгоћо E 55-5 

een 'hamp, R. M. 198 ) Бакса гапеѕѕае, а пем ѕре- 

es from San Diego Ca California. Phytologia 46: 

216-2 22. 

& J. Henrickson. 1996. 
(Asteraceae): А new species from the San 
TOUR ез Aliso 14: 197-203. 

Benson, L. . The Cacti of the United States and Can- 
ada. iram Lini a 'ss, Stanford, California. 

Bowerman, M. L. 1944. The Flowering Plants and Ferns 
of Mount Diablo, ^ud Their Distribution and As- 
sociation into Plant Communities. Gillick Press, Berke- 
ley, Саш гта. 

. Ertter. The Flowering Plants and Ferns of 
Mount Diablo, гиков (revised edition). California 
Native Plant Society, Sacramento (in press 

Boyd, S. 1998. duis hirshbergiae (Brassic scene), а паг- 
row endemic from the Cuyamaca Mountains, San Diego 
County, ( Баша; Aliso 17: 203-205 

T. S. Ross. 1997. Sibaropsis (Brassicaceae), a 
new monotypic genus from southern California. Madro- 


fio 44: 29-47. 


spec ies “missing in action” 
4. 


Baccharis eager ie 5 


ta Monic 


, а L Amseth. 1991. Ce ibd ңа ophio- 
PM (Rhamnaceae): A distinctive, narrowly endemic 
species from Riverside County, California. Perieg 
0: 28— 
Brandegee, K. 1891. 


The flora of Yo Semite. Zoe 2: 155- 
167 
1893. The botanical writings of Edward Г. 
Greene. Zoe 4: 63-103. 
Brewer, W. H., S. pe & A. Gr 


5 . 1876. Botany ad 

California], Vol. re h, Bigelow, "Univ. Press, Ca 
ridge, Massac des 

Buxton, E. & R. Omduff. 1998. Noteworthy collections— 
California. Limnanthes macounii Trel. (LIMNANTHA- 

CEAE). Madroño 45: 184. 

Carr, G. D. 1975. Calycadenia hooveri pe raceae), a new 
tarweed from California. Brittonia 27: 136-141. 

Catling, P. M., A. A. Reznicek & W. J. Crins. 1993. Carex 
msi de (Cyperaceae), a new species from north- 
eastern North America, with a key to Carex sect. Phyl- 
lostac jus Syst. Bot. 18: 496—501. 

и & Е-М. Хи. 1983. Effect of As- 

tragalus membranaceus on enhancement of mouse nat- 


activity. C hung- Кио I Hsueh K’O Hsueh 
234. 


SE: 
т 


Chang, С.-У., Y.-D. 


. 1996. "Айаз of Nevada Conifers: А Phyto- 
орао іс Кек rence. Univ. 
де 


Chaudhri, M. N. 1 


Nevada Press, Reno. 


8. А revision of the Paronychiinae. 


Volume 87, Number 1 
2000 


Ertter 107 
Floristic Surprises 


Meded. Bot. Mus. Herb. Rijks Univ. Utrecht 285: 1– 


Constance, L. 1951. The versatile taxonomist. Brittonia 7: 
225-231 


64. Systematic botany—An unending synthe- 
sis. bro 13: 257-273. 
———. 1971. The uses of diversity. Pl. Sci. Bull. 17: 22— 


еее & Ertter. 1996, Post-Manual Pipe pd : 
Californian soy (Apiaceae). Madrofio 4: 515-52 


Cronquist, 7. Juncaceae. de 47-67 in A. Fie 
quist, A. A n, J. L. Reveal 
& P. K. Holmgren (editam doses Flora: Vas- 


cular Plants of the M er es West, U.S.A., Vol. 6. 
Columbia Univ. Press, New Yor 

Dorn, R. D. 1991. Yermo xanthocephalus (Asteraceae: Se- 

ae) A new genus and species from Wyoming. 
Madroiio 38: 198-201. 

Ertter, В. 1985. Paronychia ahartii dug кар пар за а 
new species from California. Ma —90. 

. 1986. The Ju He informs К Mem. New 
York en е 39: 

—— . Ives rta var. canina (Rosaceae), new 
from cinia Brittonia 40: 398-399 

. Revisionary studies in hesia (Rosaceae: 

Ree Syst. Bot. 14: 231-244. 

— ———. 1992. A re- исо of Potentilla drummondii 

and P breweri (Rosaceae), with the new species Poten- 
tilla Ear Brittonia 44: 429—435. 
e changing face of California botany. 

TES 22. 

а. Taxonomic grays vs. black and white ex- 
em ee Implications for conservation management of 
diversity. Pp. 11-13 in T. N. Kaye, A. Liston, R. 

Love, D. L. Luoma, R. J. Meinke & M. V. Wilson (ed 
bors), Conservation and Management at Native Plants 
and Fungi. Native Plant Society of Oregon, Corvallis. 

——. 1 


Madrone 42: 
997 


tra Costa Counties, California. Special Publication No. 

3, California Native Plant Society, East Bay Chapter, 
Berkeley. 

& L. Schultheis. 1998. [Abstract] The Mount Dia- 

blo flora update: A case study in dynamic floristics and 

floristic hypotheses. Amer. J. Bot. 85(6) supplement: 
126. 


Evert, E. F. & L. Constance. 1982. Shoshonea = 
а new genus D species of Umbelliferae from Wyo 
ming. Syst. Bot. 7: 471-475 

Farrar, D. R. 1974. “Genes f ee жае tophytes—Vit- 
tariaceae. Amer. J. Bot. 61: 1 155 

—— ———,. |993a. Vittariaceae. Pp. - 187- 189 in Flora of 

North America North of Mexico, Vol. 2. Oxford Univ. 
Press, Oxford. 

. 1993b. Hymenophyllaceae. Pp. 190—197 in Flora 
of North America North of Mexico, Vol. 2. Oxford Univ. 
Press, Oxford. 

Gardes, M. & T. D. Bruns. 1996. gae. structure of 
por fungi a Pinus muricata bn 

below- pastus views. Canad. J. 
1572-1583. 
ш e I. 1978. Lowell er rancher botanist of Yuba 
au] Fremontia 6(2): 15-18. 

Gould, K. 1996. A new, di inel variety of Spigelia aii 
bus ^: oganiaceae) from Bibb County, Alabam 
Sida 17: 417—421. 


Grimes, J. 1984. Notes on the flora з ган Gulch, 
Malheur Coun. оа, Madroño 3 
Hartman, R. L. 3 [1992]. The dn pot Her- 


floristic inventory, and the Flora of 
the Rocky Mountains iae J. A Acad. Sci. 28(2): 
22-43. Gee. also http://www.rmh.uw 
& ?. Nelson. 1998. Taxonomic Novelties from 
North merida North of Mexico: A 20-Year Vascular 
Plant vibes | Baseline. Monogr. Syst. Bot. Missouri 
Bot. Gard. – 
Heller, A. А. 1908. A botanical experience. Muhlenbergia 
4: 12-1 


Hermann, F. J. 1948. The Juncus Poner group in North 
America. Le . Bot. 5: 109-1 

Hickman, J. C. (editor). 1993. The rm Manual: Higher 
Plants of California. Univ. California Press, Berkeley 

Hill, A. J. 1973. A distinctive new Calochortus (Liliaceae) 
rom acs County, coe Madroño 22: 

Hiss, A. & A. Pickart. update on — rediscov- 


E 


axifragaceae to Ericaceae. Univ. Washington 
Press, sate 

А. Санды ist. 1973. Flora of the Pacific North- 
m Illustrated Manual. Univ. Washington Press, 


Holden, c. bes ‘Extinct’ Oregon flower reappears. Sci- 


Mese dae Pp. 3 in n 
Ee T L. R 
veal & P. K. Holmgren, emunt Flora: и 
Plants of the qe ar oe ‚ U.S.A., Vol. 4. New 
York Botanical Garden, Bro 

о new i ог Viola (Violaceae) from 

the Intermountain West, U.S.A Tis ота 44: 300-305. 

1 osaceae. Pp. n A. Cronquist, N. 

mgren & P. K. Lea n, Ге tie Flora: 
ahem ти of the Intermountain West, U.S.A., Vol. 
3(A). New York Botanical ar Bronx. 

. 1998. Two new species of Penstemon Seed 
ariaceae: sect. Sacca dea Eos Nevada, U.S.A. B 
tonia 50: 159-164. 

Howell, J. T. 1970. Marin Flora: Manual of the Flowering 
Plants and Ferns of Marin County, California, 2nd ed., 
with wir Univ. California Press, Berkeley. 

Isely, D. 2. The disappearance. Taxon 21: 3-12. 

Isle, D. x dps Rediscovery of water howellia for Cali- 
fornia. Fremontia Tu 9—32. 

Janssen, G. K. & Williamson. 1996. Encouraging 
conservation of endangered | set on private lands: A 

n’s Frankenia (Frankenia johnsto- 


Johnston, В. 980. Studies of пи Variability 
Leading to a ran Classification of Potentilla sect. e 
пјирае (Rosaceae). Ph.D. Dissertation, Uni y of 
Colorado, Boulder. [Publ. no. 8021589, University Mi 

crofilms, Ann Arbor, Michigan.] 

Keck | р. 1938. Revision of Horkelia and Ivesia. Lloy- 

: 75-142. 


108 


Annals of the 
Missouri Botanical Garden 


Kjeldsen, C. K. 1972. Pleurophycus gardneri Setchell & 
Saunders, a new alga for northern California. Мадгоћо 
21: 416. 

о М. Yokota, К. Sugiyama, Т. Yamamoto, M.-Y. 

‚ S.-C. Yan & Z. ушу аме 1985. Studies оп anti- 
балоо activities and а inciples zhi 
herbs. I. Antitumor Activities of Chinese Herbs 105: 
791-795. 

Kral, js ias A new "Viorna" се from northern 

a. Ann. Missouri Bot. Gard. 5—669. 

Kruckeberg, A. R. 1997. Essay: гле би plant taxonomy 
in the 21st reci Syst. Bot. 22: 181-182. 

LeBlond, R. J., . Weakley, A. A. Reznicek & W. J. 
Crins. 1994. с. lutea (Cyperaceae), а rare new 
coastal plain endemic from North Carolina. Sida 16: 
153-161. 

Leviton, A. E. & M. L. Aldrich (Editors). 1997. Theodore 
Henry Hittell’s “The California Academy of Sciences. 
A Narrative History: 1853-1906.” Men. Calif. Acad. 
Sci. 22: 1-623. 

Lewinsky-Haapasaari, J. & D. H. Norris. 1998. Orthotri- 
chum shevockii RC uin a new moss species 
from the southern Sierra, California. Bryologist 101: 


435—438. 

Little, E. L., Jr. 1971. Atlas of United States Trees, Vol. 
1. Conifers and Important Hardwoods. U.S. Dept. of Ag- 
riculture, Forest Service. Misc. Publ. 1146, Washington, 
D.C 


Magney, D. L. 1996. Remembering Jim Jokerst, 1956— 
995. digi. 24(2): 30-31. 

Mason, C. T. 1975. Apacheria chiricahuensis: А new genus 
and species from Arizona. Madrofio 23: 105-168. 

McClintock, E. 1973. Ernest oe Twisselmann 
(1917-1972). Fremontia 1(1): 3 

McVaugh, К. 1983. Edward Lee a An appraisal of 
his Eu to botany. Pp. 54—84 in F. N. Egerton 
(editor), Landmarks of Botanical History: Edward Lee 
Greene, Part 1. Hunt Institute for Botanical Documen- 
tation. Stanford Univ. Press, California. 

Merrill, G. L. S. 1992. Ozobryum ogalalense (Pottiaceae), 
a new moss genus and species from the American Great 
Plains. Novon 2: 255-258. 

Milius, S. 1999. Unknown plants under our noses: How 
much режу кы remains to be disc avered? Sci. 
News 155: 8-1( 

Moe, R. 1997. ни tavaresiae sp. a mari 
lichen with a Bd algal phycobiont. Bull Calif. Li- 
chen Soc. 4: 

Mulligan, G. A. d Synopsis of the genus Arabis (Bras- 
sicaceae) in Canada, Alaska, and Greenland. Rhodora 
97: 109-163. 

Nesom, С. L. & P. J. Leary. 1992. A new species of Jon- 
actis (Asteraceae: Astereae) from southern Nevada an 


Flower by Any Other Name: 
Sketches of Pioneer Naturalists Who Named Our West- 
ern Plants. Yosemite Association, Yosemite National 
Park, California. 

Oswald, V. H., in collaboration with L. Ahart. 1994. Man- 
ual of the Vascular Plants of Butte County, California. 
California Native Plant Society, Sacramento. 

Phipps, J. B. 1990. Mespilus canescens, a new Rosaceae 
endemic p Arkansas. Syst. Bot. 15: 26-32. 

F. Weeden & E. E. Dickson. 1991. Isozyme 

pude eM the naturalness of три К Peconi 

subfam. Maloideae). Syst. Bot. 16: 546-55 


Reed, C. F. 1981. yir aaa kentuckiense Reed, a new 
species of orchid in Kentucky. Phytologia 48: 426—428. 
.M.& . 1994. Invasive m 

plants in California: 1993 summary and compar 
with other areas in North America. Madrofio 41: 161- 


Reveal, J. L. 1989a. Notes on selected genera related to 
И" (Polygonaceae: Eriogonoideae). Phytologia 
66: 236-245. 


1989b. New combinations and novelties in Er- 
Е (Polygonaceae: Eriogonoideae). Phytologia 66: 
251-265. 


1991. Botanical explorations in the American 
West—1889— 1989: An essay on the last centüty of a 
floristic frontier. Ann. Missouri Bot. Gan Ч 
———— & J. Т Howell. 1976. Dedeckera (Polygonaceae), 
a new genus from California. Brittonia 28: 245-251. 
Caplow & K. Beck. 1995. Eriogonum codium 
( Puls gonacsae: Eriogonoideae), a new species from 
southcentral Washington. Rhodora 97: 350—356. 
Rollins, R. C. 198 new species of the Asiatic genus 
Stroganowia (Cruciferae) from North America and its 
biogeographic планина Syst. Вог. 7: 214–220 
——. 1993. ;ruciferae of Continental North Amer- 
ica: Sepala of the Mustard Family в the Arctic 
to Panama. Stanford Univ. Press, Califo 
Two зеца а (Cruciferae) " south cen- 
tral and western burn s 5: T 75. 
ес . Caplow. 1995. Ап unde- 
seribed apet ies of Lori Cruciferae) e the 
жите Кћодога 97: 201– 
Schuster, R. M. 1981. Evolution and speciation in Pellia, 
with special i eee to the Pellia megaspora- dii: 
ifolia complex а) I. Taxonomy and distri- 
bution. 1, Вгуој. 11: 1. 
—— а. of new 
сетити Hattori Bot. Lab 70: 
Shevock, J. R. . Status of rare and endemic plants. 
Chapter 24 in ese Nevada Ke SS Project Final 
Report to Congress, Vol. essments and Sci 


xa of Hepaticae. I. Jun- 


143- 150. 


. Taylor. 1987. Plant exploration i in zoa 
fornia, the frontier i is still here. Pp. 91–9 T. S. Elia 
(editor), Conservation and M i: Rare and En- 
dangered Plants. California Native Plant Society, Sac- 
ramento. 


Ertter & J. D. Jokerst. 1989. Monardella be- 
men (amics ae), a new species from the crest of the 
southern Sierra Nevada, California. Madroño 36: 271- 
279. 


— ——, B. Еп жез р. W. Taylor. 1992. Neviusia oe 
(Rosaceae: Kerrieae), an intriguing new relict s 
from California. poen 2: 285-289. [See also a series 
of popular related articles in Fremontia 22(3): 3-13, 
1993 


Skinner, M. W. & B. Ertter. 1993. vei pis plants in 
The di Manual? Fremontia 22(3): 2 

. Pavlik (editors). 1994. a | aoe of 

Rare e Endangered Vascular Plants of California, 5th 

ed. California Native Plant Society Special Publication 

No. 1 7. 


, D. P. Tibor, R. L. Bittman, B. Ertter, T. S. Ros 
S. Boyd, J. R. Shevock & D. W. Taylor. 1995. Research 


Моште 87, Митбег 1 
2000 


Ertter 109 


Floristic Surprises 


needs for conserving California’s rare plants. Madrofio 
42: 211-241. 

Smith, D. K. & P. G. Davison. 1993. Antheridia and spo- 
rophytes in Takakia ceratophylla (Mitt.) СтоПе: Evi- 
dence for reclassification among the mosses. J. Hattori 
Bot. Lab. 73: 263-271. 

Stolzenburg, W. 1998. The seekers P the rare. Nature 
ишкс сй Йй 1998: 12— 

Stone, R. D. emic and rare iua. of Utah: An 
overview * ves distribution and status. Unpublished 


Natural Heritage Pro- 

gram, Division of Wildlife Resources, Salt Lake City. 

а ћете 5. D. 1997. The Oregon Flora Project: Steps 
toward a new flora of Oregon. Pp. 133-138 in T. N 
Kaye, A. Liston, R. M. Love, D. L. Luoma, R. J. Meinke 
& M. V. Wilson (editors), Conservation and Management 
of Native Plants and Fungi. Native Plant Society of 
Oregon, Corvallis. (See also http://www.orst.edu/dept/ 
boli леа ироа eot ORB Iola. html.) 

Tangley, L. 1 flowering of finds for American bot- 
anists: New species challenge the common wisdom. 
U.S. News & World Report 125(19): 64. 

Taylor, D. D. 1994. Schisandra E (Schisandraceae) 
new to Kentucky. Sida 16: 2 

Taylor, R. W. 1983. Descriptive ER Past, present, 
and future. Pp. 93-134 in E. Highley & R. W. Taylor 
(editors), Australian Systematic EU Ниве A Bicen- 
tenary Perspective. odia Melbourne. 

Twisselmann, E. C. 1956. A flora of the Temblor Range 
and the neighboring part sth the San Joaquin Valley. 
a Biol. 14 

. 1967. A flora. Р С. p ounty, California. Was- 
mann J. p 25: 1-395. [Reprinted by the California 

Native Plant Society, 1995.] 

ff & E. N. Kozloff. 1994. 


Note- 


Ornduff (Asteraceae). Madrofio 41: 332— 

Vincent, M. A. & A. W. Cusick. 1998. "a records ha 
alien species in the Ohio vascular flora. Ohio J. 5 
98(2): 10-17 

— & R. Morgan. 1998. Trifolium jokerstii (Legumi- 


nosae, in aac a new species from Butte Coun- 
ty, California. Novon 8: 91–93. 
1994. Another widely disjunct, rare 


106 in Flora of North America North of Mexico, vol. 2. 
Oxford Univ. Press, Oxf 

Watson, S. О. Botany ji California], Vol. 2. Little, 
rown, Boston 

Weber, W. A. 1996. Е moenium іп the Western 
Hemisphere. Evansia 13: 159-160. 

Weislander, А. Е. (in s Forest Service Staff. 1939. 
Vegetation Types of California: Redding Quadrangle. 


.S. Forest Service. 
Weldy, T. W., H. pei zeniec, L. E. Wallace & * А. 
Саве. 1996. The current status of Cypripe sod kentuck 
iense (Orie including a morphological уа? is 


of a newly discovered population in eastern Virginia. 
Sida 17: 423-435. 
. 1993. Rosaceae. Pp. 587-616 in S. L. Welsh, 
N. p. үүн 5. Goodrich & L. С. Higgins, А Utah 
Flora, : onte L. Bea n Life Science Museum, 
Brigham Young University, Provo, kya h. 
ib D., D. L. Nickrent, rn, C. L. Calvin & 
N. J. Vivrette. 1989. VONT питам udo and loss of 
repiddus tive capacity in the rare JC REN shrub 


Dedeckera eurekensis. Nature 338 
Wilkinson 990. Systat: The ы pe Satisi Ev- 
anston, Illi 


M. Beilstein. 1998a. [Abstract] Some 
rare, endemic ‘drabas of Utah prove to be rarer and more 
narrowly endemic than originally thought. Amer. J. Bot. 
85(6) supplement: 167—168. 
1998b. A taxonomic E ud of Draba maguirei and 
allied pu (Brassic aceae). Report on file with Wasatch- 
Cache onal Forest, Salt Lake City, Utah. 
Valens dur: 1999. Steyermark' s Flora of ный. Vol. 
Revised ed. Missouri Department of Conservation, 
Doc City, in pe odi AUN Missouri Boni | 
Garden Press, St. Loui 
E 1993. Mer. E Pottiaceae. Bull. Buffalo Soc. 


Zander, 
Nat . 32: i- 
Zech, i C. P. R. e dit W. H. Wagner. 1998. A n 


adder’s-tongue (Ophioglossum: Ophioglossaceae) for 
North America. Sida 1: 307-313. 


A NEW AGE OF DISCOVERY! 


Michael J. Donoghue? and William S. 


Alverson?’ 


ABSTRACT 


Whereas past ages of discovery have 
marked by exploration at smaller spatial scales an 


been associated with the exploration of new continents, the current episode is 
d in more extreme 


habitats, some of which are surprisingly close at 


and. This new age of discovery is richer and more illuminating by virtue of the attention being paid to phylogenetic 


relationships and geographic ranges. Although it is difficult to quantify progress in understanding the tree 


of life or 


biogeographic patterns, a series of examples illustrates that the magnitude and significance of recent discoveries are 
unquestionably great. Extraordinary discoveries gab an obvious, yet underutilized, mechanism to capture the imag- 


ination of the scientific community and the public 
Key words: 


age of discovery, biodiversity, а exploration, new species, phylogeny. 


e tend to regard “the age of discovery” as 
something long since past. Whether the phrase re- 
fers to the colonization of North America by trans- 
Beringian migrants, arrival of the Moa Hunters and 
Maoris in New Zealand, or (most likely) the forays 
of European explorers into the New World from the 
15th century onward, depends on one’s sense of 
history and focus. However, there may be general 
attributes of all previous epochs with which we can 
evaluate whether any particular period really qual- 
ifies as an age of discovery. We will argue that the 
present era most certainly deserves this title. 

Briefly consider the most recent of these ages of 
discovery, which began in the late 1400s. European 
exploration of the Americas, Australia, Africa, and 
the South Pacific brought to light an extraordinary 
number of new species. Even more importantly, 
these were species quite unlike any known before: 
the rhinocerous (Fig. 1), the kangaroo, and the 

odo, to name only a few. David Quammen (1996) 
helps us imagine what it must have been like to 
hear about or see such animals for the first time: 


. the first report of a hummingbird, . . . of a toucan, 
firs 


almost too majestic to be real. Imagine how these prod- 


= must have Вы a сотрјасеп у pious six- 
enth-century Europe 


How can the present day possibly compare either 
in terms of the number of novelties being discov- 


эе 


ered or their "surprisingness"? 


NEW SPECIES 


In most major lineages the number of new spe- 
cies being described is not falling off. Instead, the 
numbers have actually been rising over the last de- 
cade to levels comparable to those of the mid-18th 
to late 19th century (before which comparisons are 
difficult, since formal rules for naming species had 
not been developed). Surprisingly, perhaps, this in- 
cludes the groups that we tend to think of as being 
the best known. For example, the description of 
ew mammal species is on the rise (Wilson & 
Reeder, 1993), and this is not, as one might initially 
suppose, primarily due to the splitting of known 
species as a consequence of using molecular mark- 
ers. Instead, it appears to be directly correlated 
with serious exploration of new areas (Morell, 
1996). For instance, fieldwork over the last decade 
in the Annamite mountains on the border of central 
Vietnam and Laos has brought to light several large 


5 


! We owe more than the usual amount of thanks to the many colleagues who brought examples to our attention, and 


who provided additional information and illustrations. We are especially grateful to Steve Solheim, who helped inspire 
this work by introducing us to the Museum of Jurassic Technology, = who opened his own bibliographic Wunder- 
ammern for our use. In addition to the other speakers in the symposium, who generously shared materials in advance 
of and following their talks, we are indebted to David Ackerly, Elena ripe Buylla, Joe Ammirati, ee Boom, John 
Cadle, Roy Caldwell, Lisa DeCesare, Dennis Desjardin, Andrew Douglas, Jim Doyle, Thomas Eisner, Frank Glaw, Ken 
Halanych, David Hibbett, David Hillis, Kathy Horton, Andy Knoll, Joyce Longcore, Santiago Madriñán, Jody Martin, 
Russ Mittermeier, Mark Moffett, Hal Mooney, Debby Moskovits, Darlyne Murawski, Dan Nickrent, Ronald Nussbaum, 
Akiko Okusu, Jim Patton, Dan Perlman, Peter Raven, Scott Redhead, Rick Ree, Mick Richardson, Robert Ridgely, 
Gustavo Romero, George Schatz, David Wake, Doug Wechsler, Mark Wilkinson, Bill Willers, David Wilson, Catherine 
ja wpe Wright, and Liz Zimmer. Kandis Elliot generously helped with the preparation of the figures 
University Herbaria, 22 Divinity Avenue, Cambridge, Massachusetts 02138, U.S.A. 
cira address: Environmental and Conservation Programs, The Field Museum, 1400 5. Toledo Drive, Chicago, 
Illinois 60605-2496, U.S.A. 


ANN. Missouni Bor. GARD. 87: 110-126. 2000. 


Donoghue & Alverson 111 


New Age of Discovery 


Volume 87, Number 1 
2000 


а: 
Т ДЕЖ, 
иг 


УРЫЙ 
ISIS i 


RH Tom 


DA 


£u 


Men vint(e te coope by Hendrick Hondius Plaet(nijder ins Gravenhage. 


The fantastic rhinoceros, as conceived by Albrecht Diirer at | time these animals were first discovered 
Agnes Mongan Center, Fogg Art Muse 877 
ove уп ге marked, * 


Figure 


by Europeans [ Reprinted, with и from ап original print at the 
ies later, upon see ing a shino 'erous, John Е 


Harvard University Art Museums. | Diei) 
more дине а huge enormous s 


extravagant as the Skin of the bea 


like Armor.. 
(Evelyn, 1684). 
mammals—a species of bovid, Pseudoryx nghetin- 


hensis (Dung et al., 1993), and at least three species 
of muntjac deer (Schaller & Vrba, 1996; Giao et 
al., 1998; MacKinnon, 2000 this issue). Moreover, 
the rate of description of new mammal species has 
risen despite decreased attention to taxonomy and 
a documented increase in the time between discov- 
ery and description (Patterson, 1994). 

Amphibians provide another well-documented 
example of the rising rate of species description 
(Glaw & Kohler, 1998; see Fig. 2). Again, this re- 
flects, to a considerable extent, the discovery of 
novelties in the wild, rather than the application of 
molecular methods or changes in species concepts. 
Hanken (1999) documented this point, citing David 

(1996) discovery of a new lungless sala- 
mander from the San Gabriel mountains, less than 
30 miles from Los Angeles, as well as species de- 


other B 

st. wbich hung ын on h 

and these lappe ts ч stiff skin, began to be studdied with impenetrable Se ien like a Target of coate of maile, loric ated 
was certainly а very wonderful creature, of immense strength in the neck and nose especially’ 


. but in my opinion nothing was so 
Ee like so much Coach leather . 


scribed in 1998 from Brazil, Mexico, Nepal, and 
New Zealand. 

The same trends in discovery (if not formal nam- 
ing) doubtlessly apply in less well studied groups, 
such as plants, fishes, insects, mites, nematodes, 
and especially Bacteria (eubacteria) and Archaea 
(archaebacteria, e.g., Pace, 1997; Fuhrman & 
Campbell, 1998; Madigan, 2000 this issue), al- 
though reliable estimates are harder to come by 
e.g., see Gaston, 1991, on insects; Prance et al., 
2000 this issue, on vascular plants). If there were 
more taxonomists, and if the lag time between the 
discovery and naming of new taxa were shorter, 
then the rate of description would surely be far 
higher. Many new species have already been col- 
lected, or are already known to be new, but remain 
undescribed simply for lack of expertise and time. 
Plant monographers, for example, can show any cu- 


~ 


112 Annals of the 
Missouri Botanical Garden 


700 == 


600 T 


500 + 


400 + 


200 + 


o 
" 


о 
E 


1758-59 
1760-69 
1780-89 
1790-99 
1800-09 
1810-19 
1820-29 
1830-39 
1840-49 
1850-59 
1860-69 
1870-79 
1880-89 
1890-99 
1900-09 
1910-19 
1920-29 
1930-39 
1940-49 
1950-59 
1960-69 
1970-79 
1980-89 
1990-99 


Amphibian species descriptions since 1758 (synonyms not included), clustered by decade, from Glaw 
» po (1998). The arrow indicates the end of 1995; descriptions for the period 1996-1999 were extrapolated. 


rious visitor a a specimen cabinet containing think about the diversity and evolution of life. Un- 
undescribed specie fortunately, this is much harder to quantify than 
By the criterion “of the number of new species counting the number of published descriptions of 
being described, we believe that the present com- new species. 
pares favorably with any time since the mid-1700s. 
But how surprising are the new species that аге yew PHYLA. ЕТС. 
being discovered in the late 20th century? Are we 
finding anything significantly different, or are we One possible indicator of the distinctiveness of 
just filling gaps in an already well-known range of newly discovered life forms, or perhaps of their in- 
variation? John Horgan, author of The End of Sci- — formativeness with respect to our understanding of 
ence (published by Helix Books, in 1996), presents evolution, is the rate of description of new higher 
the latter view: taxa. As Brusca (2000 this issue) shows, the de- 
are down to the ет now. Every now and then seiiption of new листе. phyla Goes not Appear 10 
something interesting will turn up. We will find a new ave diminished in recent years. Species are still 
nein Madagasc ar, or weird bacteria living being discovered that are Пинк by the геје- 
in deep-sea vents. But at this point we are unlikely to vant taxonomic communities to be as distinct as the 
discover something reall surprising..." (J. Horgan, familiar phyla аге from one another. The major dif- 
Ө). ference is that these new organisms are, оп the 
This will sound absurd to those actively engaged іп whole, smaller, which means we are having to look 
the discovery of new life forms, but such a “dotting more closely. Sometimes, as in the case of Nana- 
the i's and crossing the ts” assessment of the sit-  loricus mysticus of the recently described metazoan 
uation is widespread both within the scientific com- phylum Loracifera (Kristensen, 1983), this has en- 
munity and the public at large. tailed careful scrutiny of an obscure (though cer- 
To counter Horgan’s naive but evidently persua- tainly widespread!) habitat—between sand grains 
sive claim, we need to consider criteria whereby а оп the ocean floor. In other cases new life forms 
new species would qualify as “really surprising.” have been found more or less right under our noses. 
Perhaps the primary criterion should be whether а Symbion pandora, a metazoan animal referred to 
new species fundamentally expands the way we the new Cycliophora, was described in 1995 from 


i 


Volume 87, Number 1 
2000 


Donoghue & Alverson 113 


New Age of Discovery 


a specimen collected on the mandible of a Nor- 
wegian lobster (Funch & Kristensen, 1995). 

New “classes” of animals, sometimes with enor- 
mous significance for our understanding of major 
lineages, are also still being described at a regular 

ace. Examples include the Remipedia within 
Crustacea, discovered in a marine cave in the Ba- 
hamas (Yager, 1981), and the Concentricycloidea 
within echinoderms, found on waterlogged wood in 
deep waters off the coast of New Zealand (Baker et 
al., 1986). 

In angiosperms there are excellent examples, 
mainly in the form of new families erected to ac- 
commodate extraordinary new species. For exam- 
ple, recall the excitement generated by Ticodendron 
incognitum Gómez-Laur. and L. D. Gómez (Ticod- 
endraceae) from Costa Rica (Gómez-Laurito & Gó- 
mez P., 1989, 1991), and the subsequent realization 
that these trees occupy wet montane forests from 
southern Mexico to central Panama (Hammel & 
Burger, 1991). But the most spectacular case is 
probably Lacandonia schismatica E. Martínez and 
Ramos. This species of Triuridaceae-like monocot 
was described in 1989 in the new family Lacan- 
doniaceae from material collected in the Lacandon 
region of Chiapas, Mexico (Martínez & Ramos, 
1989). Astonishingly, its stamens are on the very 
inside of the flower (at the apex of the receptacle), 
surrounded by the carpels (Márquez-Guzmán et al., 
1989; see Fig. 3a). This discovery, only a few years 
ago, came as a complete surprise—nothing about 
the other ca. 250,000 species of angiosperms pre- 
dicted this brand new flower architecture. It also 
provides a unique opportunity to test the *ABC 
model" for genetic determination of flower organ 
identity (Coen & Meyerowitz, 1991; Vergara et al., 

9) 


NOT PHYLA, ETC. 


If the description of higher taxa were our only 
means of assessing the surprisingness of recent dis- 
coveries, we would miss many of the most exciting 
finds. The several examples below highlight radi- 
cally new life forms (as different, we believe, as 
those discussed above), which were described only 
as new genera or as species within existing genera. 
These cases provide a dramatic reminder of the ar- 
bitrary nature and inadequacy of the Linnaean 
ranks—even in gauging the degree of disparity, 
which ranks are most often thought to represent. 

In 1995, with very little fanfare, Dennis Desjar- 
din and his colleagues described *an unusual psy- 
chrophilic aquatic agaric from Argentina" within 
the existing fungal genus Gloiocephala (Tricholom- 


ataceae). This remarkable new species, С. aquati- 
ca, develops its fruiting bodies and spores under- 
water, often beneath a layer of ice covering lakes 
in a zone between the Patagonian steppes and the 
Andes. Аз it appears to be the only known agaric 
to sporulate underwater, the term “unusual” is 
clearly an understatement—this is a brand-new 
way of being a mushroom. А similar discovery was 
also made in gastromycetes. In 1976, Escobar et 
al. described the genus Limnoperdon to accommo- 
date the only known "aquatic" (actually, floating) 
puffball, which they found in marshes adjacent to 
the University of Washington in Seattle. 

Amphibians also provide excellent examples of 
recently discovered lifestyles. In 1973, a new genus 
and species of leptodactylid frog, Rheobatrachus 
silus, was described on the basis of animals found 
in streams near Brisbane, Australia (Liem, 1973). 
Shortly thereafter, Corben et al. (1974) documented 
that Rheobatrachus exhibited a unique form of pa- 
rental care. Brooding of the embryos and young 
takes place inside the stomach until the tadpoles 
or juvenile frogs are eventually (some eight weeks 
after ingestion) “propulsively ejected” through the 
mouth. This brooding mechanism is otherwise com- 
pletely unknown in vertebrates. Furthermore, it re- 
quires special physiological mechanisms to turn off 
the secretion of gastric juices, which otherwise 
would kill the young. In 1983, Tyler et al. reported 
that prostaglandin secreted by the larvae inhibit 
acid production in the female's stomach *in a man- 
ner not seen elsewhere in the Animal Kingdom." 
Despite the potential medical significance of this 
remarkable phenomenon, further study will not be 
possible. Rheobatrachus silus appears to be extinct 
in the wild (Laurance et al., 1996). 

In 1996, Nussbaum and Wilkinson described a 
new species and genus of caecilian (Amphibia: 
Gymnophiona), Atretochoana eiselti. As they docu- 
mented (also see Wilkinson & Nussbaum, 1997), 
this is the largest lungless terapod, by far! It is 74— 
80 cm in length, while the next largest, a pletho- 
dontid salamander, is about % this size, at around 

cm. The mouth is not connected to the nasal 
cavity in this animal, and its many other weird cra- 
nial features allow an exceptionally wide gape (see 
ig. 4). These modifications are presumably asso- 
ciated with a novel feeding mechanism, but the nat- 
ural history ef these organisms is completely un- 
known. In fact, this extraordinary beast was 
"discovered" in a museum in Vienna and was 
known from only a single specimen collected from 
an unknown South American locality sometime in 
the last century. Now, another specimen has been 
located, this time in a jar in a Brazilian collection 


Annals of the 
Missouri Botanical Garden 


Counterclockwise, from upper left. 


Figure 3. 


with three кейш ек stamens surrounded p partially obscure 


Mexico, in July 1 )89. "The bell diameter is ca. 1 m 


Meyerowitz, and F. Vergara. —b. Chry 


—a. Flower of Lacandonia schismatica (Lac i eae, Triuridales), 


X 
Martin and iru Hall а = 


photograph, are чаша а н 6 m long. Photo used with permission of J. 
Squamanita contortipes parasitizing a host mushroom, probably Galerina cerina. Photo by 5. А. Redhea 
| | 


(Wilkinson et al., 1998). Ironically, it, too, lacks a 
specific collection locality. 

ther newly discovered species highlight how 
much we still have to learn. From Bacteria, consid- 
er Prochlorococcus, discovered only about a decade 
ago. Of great interest owing to the presence of both 
chlorophyll a and b (as in green plants), it is known 
now to be a major component of the phytoplankton 
in tropical and subtropical seas, and among the 


world's most important primary producers (see 
Fuhrman & Campbell, 1998). Moreover, the ability 
of Prochlorococcus to thrive in a wide range of light 
conditions seems to be explained by the existence 
of closely related but quite distinct species, living 
i 1998). Also of 


potentially huge ecological significance are new 


at different depths (Moore et al., 


disease-causing bateria. For example, a newly dis- 
covered species of proteobacterium (related to 


Volume 87, Number 1 
2000 


Donoghue & Alverson 115 


New Age of Discovery 


Figure 4. 


relaxed constraints on aspects of skull architecture involvec 


Lateral view of the head of the lungless caec et Atretochoana eiselti. Its odd skull shape may reflec 


ith dicen and selection for increased gape size for 


prey capture. Photo by R. Nussbaum, iin Wilkinson and аи (196 


Sphingomonas) seems to be causing massive coral 
die-offs in Florida’s reef communities (Richardson 
1998). 

One of our favorite metazoan examples is the 
scyphozoan jellyfish Chrysaora achlyos (see Fig. 


et al., 


3b). This species was described by Joel Martin and 
colleagues in 1997, based on four specimens col- 
lected off the coast of southern California in the fall 
of 1989, and some photographs from northern Baja 
California (Martin et al., 
cies in this genus, C. achlyos is remarkable by vir- 


1997). One of several spe- 


tue of being enormous. In fact, with a bell measur- 
ing over 1 m in diameter, this is the largest 
invertebrate described in this century, rivaled in 
size only by the pon MU and an arctic jellyfish 
described in the 1800s (J. Martin, pers. comm.). 
Although thousands э ashore in 1989, it has 
not been seen since. 

From vertebrates comes the story of the sala- 

mander Eurycea sosorum. This species was 
scribed in 1993 from specimens collected in "a 
Barton Springs Aquifer, within the city of Austin, 
1993). Known only from 


a single popular swimming spot, and therefore in 


Texas (Chippendale et al., 


grave danger of extinction, this species continues 
to attract considerable attention in Austin. Subse- 
quent exploration of nearby springs, some located 
within Austin housing developments, has turned up 
dozens of new species (D. Hillis, pers. comm.). De- 
scription of these species, from a relatively small 
area well endowed with herpetologists, will increase 
the number of described salamanders of the world 
by about seven percent! 


— 


ust as numerous new amphibians are coming to 
light, so, too, are novel pathogens that prey upon 
them (Hanken, 1999). 
chytrid 
described very recently (Longcore et al., 1 


new genus and species of 


„—. 


ungus, Batrachochytrium | dendrobatidis, 
999). ap- 
pears to be a major proximate cause of mass mor- 
tality events in frogs around the world (Berger et 
al., 1998; Morell, 1999). Being the only chytrid 
known to prey on vertebrates, this new species also 
greatly expands our understanding of a major and 
ubiquitous i" group (Longcore et al., 1999; 
Pessier et al., 

Seed plants ne provide fine examples. Wollemia 
nobilis W. С. Jones, К. D. Hill & J. M. Aller 
sister group of Agathis (Gilmore & Hill, 1997) ог 


, the 


116 


Annals of the 
Missouri Botanical Garden 


possibly all other Araucariaceae (Setoguchi et al., 

8), and closely resembling a fossil known from 
the Cretaceous through the Oligocene, was discov- 
ered in 1994 growing in the Wollemi National Park, 
ca. 200 km northwest of Sydney, Australia (Jones 
et al., 1995; Briggs, 2000 this issue). How could a 
large tree, with such distinctive architecture, grow- 
ing near the largest city in Australia, have escaped 
detection until 1994? 

At about the same time, Andrew Douglas and 
colleagues were realizing that collections of an odd 
proteaceous tree made in the 1960s and 1980s from 
northeastern Queensland represented a new spe- 
cies, Eidothea zoexylocarya A. W. Douglas & B. 
Hyland. These 40-m-tall plants, seemingly endemic 
to rainforests in the Bloomfield River watershed, 
are the sole representatives of an apparently basal 
lineage of Proteaceae, the Eidotheoideae (Douglas, 
1995; Douglas & Hyland, 1995). Its fruits are near- 
ly identical to 60-million-year-old fossils of Xylo- 
caryon lockii F. Muell. (Holden, 1995). 

Neviusia cliftonii Shevock, Ertter & D. Taylor 
(Rosaceae, Kerrieae) provides a similar example 
(Ertter, 2000 this issue). This species was de- 
scribed in 1992 from material collected south of 
Mount Shasta in northern California (Shevock et al., 
1992). Although it is a rather conspicuous and lo- 
cally abundant shrub and grows at several localities 
in an area where the vegetation had previously 
been mapped, it seems never to have been collect- 
ed before 1992. As the authors speculated, the 
abundance of poison-oak in the vicinity may have 
been enough to discourage thorough exploration. 
Judging by its resemblance to an Eocene fossil from 
British Columbia, N. cliftonii is probably a relict in 
this area (Shevock et al., 1992). Its closest living 
relative, Neviusia alabamensis Gray, is known 
from only a few localities in Alabama and Arkan- 
sas. 


ATTENTION TO PHYLOGENY 


The ongoing age of discovery differs importantly 
from earlier episodes by virtue of the attention be- 
ing paid to phylogeny. The phylogenetic notion of 
relationship was nonexistent before the mid-1800s, 
and not until recently have new methods and sourc- 
es of evidence allowed rigorous and (we trust) in- 
creasingly accurate phylogenetic inferences. А]- 
though biodiversity is still commonly equated with 
the enumeration of species (and genetic diversity 
within species), the idea has now expanded. Bio- 
diversity is not just about species, at the tips of the 
tree, but about the whole tree of life. Thus, finding 
any branch of the tree, external or internal, consti- 


tutes a genuine act of discovery. Viewed in this 
light, it is counterproductive to draw too strong a 
boundary between discovering species and discov- 
ering clades. These activities are pieces of the same 
underlying project, namely, understanding the en- 
tire tree of life. 

Addition of this phylogenetic dimension makes 
the ongoing age of discovery far richer, and cer- 
tainly far more powerful in connection with prac- 
tical problems such as, for example, the choice and 
design of nature reserves and the search for useful 
natural products. In general, knowledge of phylo- 
genetic relationships brings with it the power of 
prediction. Phylogenetic research should therefore 
be viewed as a welcome addition to, rather than a 
competitor with, the study of species diversity. 

Consequently, it is of great interest to consider, 
as we have for species, both the rate of discovery 
of new clades and the surprisingness of these new 
discoveries. Unfortunately, we know of no straight- 
forward way to quantify these things. There is little 
baseline information regarding the rate of discov- 
ery, and databases such as TreeBASE (http://phy- 
logeny.harvard.edu/treebase), which could eventu- 
ally be a ready source of such information, have 
grown too unevenly to provide an accurate measure 
of clade discovery. Nevertheless, the rate appears 
to be growing exponentially, and shows no signs of 
tapering off (Sanderson et al., 1993). At loast ihis 
is the case if the number of clades being d 
grows in proportion to the rapidly on num- 
ber of phylogenetic analyses being published, 


which we have no reason to doubt. 

In theory, one might approximate the rate of dis- 
covery by tallying new names applied to clades, but 
at present this information is not being databased 
consistently enough. However, it is important to ap- 
preciate that even if this were now possible, the 
number of clades actually being named is far less 
than the number of clades being discovered, or 
even the number of clades identified with great 
confidence. Reluctance to formally name clades 
stems in part from the traditional Linnaean nomen- 
clatural system's emphasis on the assignment of 
taxonomic rank, and the potential consequences of 
such assignments on the application of names at 
other levels (Baum et al., 1998; Hibbett & Dono- 
ghue, 1998). Related to this is the realization that 
there are not enough familiar ranks in the Linnaean 
system to do justice to the many nested clades be- 
ing discovered even in average-sized phylogenetic 
analyses. In practice, the current nomenclatural 
codes (e.g., Greuter et al., 1994) encourage formally 
naming clades only after relationships throughout 
the group in question have been rather well estab- 


Volume 87, Number 1 


Donoghue & Alverson 
New Age of Discovery 


lished, as opposed to encouraging the naming of 
well-supported new clades as they are discovered, 
usually one or a few at a time (Hibbett & Dono- 
ghue, 1998). 

The surprisingness of recent discoveries is also 
hard to judge because prior views on relatedness 
have often not been expressed unambiguously 
enough. Nevertheless, it is clear that many entirely 
unexpected discoveries have been made. We now 
appreciate, for example, that “prokaryotes” are not 
a single branch, but two (or more) major branches, 
with lineages of Archaea probably more closely re- 
lated to eukaryotes than they are to Bacteria (e.g., 
Pace, 1997). Likewise, we can say with growing 
confidence that animals and fungi (Baldauf & 
Palmer, 1993; Wainright et al., 1993), and possibly 
also slime molds (Baldauf & Doolittle, 1997), are 
more closely related to one another than they are 
to plants or any other eukaryotes. 

While these are obviously major advances in our 
understanding of life, such discoveries may not 
seem terribly surprising since relationships of the 
major lines of life have always seemed obscure. 
What about within metazoan animals and green 
plants, where phylogenetic theories have been bet- 
ter developed—have ni really surprising results 
emerged from recent stu 

e answer is an шы каш yes! Relation- 
ships among major metazoan lineages have come 
into much sharper focus, and a number of tradi- 
tional views have been overturned. For example, 
whereas it has long been assumed that annelids are 
more closely related to arthropods than they are to 
molluscs (as evidenced by segmentation and the 
existence of seemingly intermediate onycophorans), 
much recent evidence points instead to the exis- 
tence of a euthrocozoan or lophotrochozoan clade, 
which includes annelids and molluscs, to the ex- 
clusion of arthropods (Eernisse et al., 1992; Hal- 
anych et al., 1995) 

More astonishing, perhaps, is the possibility of 
an ecdysozoan clade, which includes all of the 
molting animals, and therefore unites arthropods 
with such groups as onycophorans, Ен Са, and 
even nematodes (Aguinaldo et al., iribet & 
Ribera, 1998; Knoll & Carroll, 1999). And within 
arthropods (Fortey & Thomas, 1997; Brusca, 2000), 
there is mounting evidence for the union of crus- 
tacea and insects and even the paraphyly of crus- 
tacea with respect to insects (and possibly the other 
major arthropod groups as well). Among other 
things, this implies convergent evolution of adap- 
tations to life on land (tracheal systems, Malpighian 
tubules, etc.) in hexapods and myriopods (tradition- 
ally the Atelocerata). 


Even within mammals, where one supposes that 
many relationships have long since been estab- 
lished with certainty, there have been some major 
surprises in the last few years (de Jong, 1998). Our 
favorite case concerns the position of elephant 
shrews and golden moles. Once considered insec- 
tivores, these are now seen to be united with aard- 
varks, hyraxes, sea cows, and elephants, which im- 
plies extraordinary morphological radiation within 
a basically African clade (Springer et al., 1997; 
Stanhope et al., 1998). 

Within green plants, molecular phylogenetic 
studies have tended to confirm earlier suggestions 
based on morphology (Donoghue, 1994), b 
are also some very surprising results, of which we 
mention only a few (see Doyle, 1998, for others). 
The whisk-ferns, Psilotaceae, now appear to be re- 
lated to Ophioglossales (e.g., Hasebe et al., 1995; 
Manhart, 1995; Wolf, 1997), as opposed to being 
remnant rhyniophytes or related to some group of 
leptosporangiate ferns, as suggested previously. 
Within ferns, the aquatic and heterosporous Salvi- 
niaceae and Marsileaceae, long viewed as unrelat- 
ed, now appear to be directly linked (e.g., Pryer et 
al., 1995; Wolf et al., 1999), a connection also sup- 
ported by fossil evidence (Rothwell & Stockey, 
1994). Within angiosperms, the tricolpate or eudi- 
cot clade, recognized first in morphological analy- 
ses (Donoghue & Doyle, 1989; Doyle & Hotton, 
1991), has been rather consistently upheld in mo- 
lecular studies (e.g., Chase et al., 1993; Rice et al., 
1997; Soltis et al., 1998; and even the highly un- 
resolved parsimony jackknife analysis of Källersjö 
et al., 1998). This represents a real departure from 
the traditional view that caryophylids, hamamelids, 
and other major lineages were derived indepen- 
dently from magnoliids (e.g., Cronquist, 1988). Oth- 
er surprises include Drosera and Nepenthes being 
related to caryophillids (Albert et ini 1992), a 
clade including almost all plants with mustard oils 
(Rodman et al., 1996), and a connection of woody 
Hamamelidaceae and Cercidophyllum with herba- 
ceous Saxifragaceae and relatives (Soltis & Soltis, 


Some of the most interesting early results, such 
as the basal position of Ceratophyllum within an- 
giosperms in rbcL analyses (e.g., Chase et al., 
1993), have not been consistently recovered as ad- 
ditional data have been brought to bear. But other 
seemingly weird results have been upheld. Of 
hese, we are most excited about the possibility that 
Proteaceae, Platanaceae, and Nelumbo are posi- 
tioned near the base of eudicots, and may form 
clade (Chase et al., 1993; Rice et al., 1997; Soltis 
1997; Hoot et al., 1999). Based on outward 


co 


et al., 


118 


Annals of th 
Missouri Saree Garden 


appearances, one would never guess that these 
plants are closely related, and a direct relationship 
of the three had never been suggested. On closer 
consideration, this arrangement is not illogical from 
a morphological standpoint, though it has proven 
quite difficult to identify any morphological syna- 
pomorphies (Hoot et al., 1999; A. Douglas, pers. 
comm.). 

Although many phylogenetic problems have 
been convincingly resolved in recent years, the re- 
lationships of many groups remain unclear. In most 
cases, this reflects nothing more than limited atten- 
tion, and we can expect straightforward solutions. 
Yet, in some other 
resolved despite considerable attention. For exam- 


r cases, relationships remain un- 


ple, it is fair to say that relationships among major 
mammal lineages are still up in the air (de Jong, 
). Likewise, relationships among major seed 
plant groups have still not been satisfactorily re- 
vini Е 1998; Hansen et al., 1999; 
999). Within flowering plants, perhaps the 
most ыо. confusion surrounds Rafflesiales. 
While the position of most of the other parasitic 
groups has fallen into place, the relationships of 


inter 


Rafflesiales remain highly uncertain, despite great 
art of Nickrent and colleagues 
‚ 1998). In fact, at this stage it is 
still incisa whether Rafflesiales are eudicots, or 
are instead more e related to one or another 
monosulcate grou Hydnoraceae now appear to 
be related to Aristolochiales/Piperales). A remark- 
ably accelerated rate of evolution of the nuclear 
ribosomal genes of these plants may be creating 
artifacts in phylogenetic analyses (e.g., placement 
of Rafflesiales at the very base of angiosperms; 
Nickrent & Starr, 1994; Nickrent & Duff, 1996). 

What can we expect in the near future? First, 
and most obviously, the discovery of clades is still 
in its infancy, and many more breakthroughs are on 
the horizon. We expect that even the most recalci- 
trant. problems, including those just highlighted, 
will eventually yield to more data and better meth- 
ods of analysis. 

Second, we trust that more attention will be paid 
to the naming of newly discovered clades, so as to 
facilitate communication about exciting results as 
rapidly, unambiguously, and with as little nomen- 
clatural disruption as possible. For this purpose we 
believe that a new phylogenetically oriented code 
of nomenclature is needed (see above). Fortunately, 
such a phylogenetic code is being developed as a 
result of a workshop held at the Harvard Herbaria 
in August 1998. This will soon be available via the 
internet for evaluation and, subsequently, for formal 
use. In this system there are no categorical ranks, 


and names are applied to clades by means of phy- 
logenetic definitions (e.g., a “node-based” defini- 
"the least inclusive clade that con- 
tains sp s X and species : de Queiroz & 
Cane. 1992 1994). This, we believe, represents 
a major and very positive shift away from arbitrary 
decisions about rank and the need to appeal to au- 
thority (e.g., Cronquist, 1988; APG, 1998) concern- 
ing the circumscription of taxa. One would imme- 
whether or not 


tion, such as 


diately know, for example, 
“Bombacoideae” is included in “Malvaceae,” if 
both names were given phylogenetic definitions and 
if a particular phylogenetic hypothesis were ac- 
cepted (Baum et al., 1999); 
without phylogenetic definitions, botanical author- 


lverson et al., 


ities could and probably would disagree about this, 
even if they accepted the same underlying phylog- 
eny for their nomenclatural decisions. 

inally, as emphasis shifts toward phylogenetic 
knowledge, the need to database such information 
will become increasingly obvious, both to track pro- 
gress and to render phylogenetic discoveries acces- 
sible to a very wide variety of potential users. Sev- 
eral complementary efforts have been initiated to 
keep track of and synthesize this information, in- 


cluding Ter PASE (pee above) and the Hee of Life 


ny. html). However: ink to the éxponsntial “ 
crease in phylogenetic research, and limited sup- 
port for these projects, they have not been able to 
keep pace and are therefore still inadequate for 
many purposes. We hope that this situation will im- 
prove as journals move toward electronic submis- 
sion of phylogenetic data sets as a co-requisite of 
publication (see, e.g., Mycologia), perhaps also 
linked with the registration of clade names. In any 
case, this is a problem that the systematics com- 
munity will need to address in order to capitalize 
on the major investments being made in phyloge- 
netic research. 


ATTENTION TO GEOGRAPHY 


The present age of discovery also differs in the 
attention being given to geographic ranges. Collec- 
tion localities were barely a consideration until 
quite recently. Recall, for example, Darwin’s diffi- 
culties in reconstructing (after the fact) the islands 
from which his Galápagos finches were actually col- 
lected (Sulloway, 1982). In contrast, we now place 
great value on detailed, accurate specimen labels 
and on databasing geographic information. Many of 
the most important questions about the organization 
of biodiversity depend heavily on such information, 
such as the distribution and relative “hotness” of 


Моште 87, Митбег 1 Donoghue & Alverson 119 
2000 New Age of Discovery 


Number of species 


123 45 6 7 8 9 1011121314 15 16 17 18 19 20 
Number of specimens 


Figure Number of specimens for species of woody tropical plants. The dita represented in this graph were 
obtained by Madrifián-Restre po (1996) from recent ан а of neotropical plants: 317 species were scored in 15 
genera from 8 families: Rollinia (Annonaceae), Licania (Chrysobalanceae), рока, Се епа (Епсасеае), Агоиеа, 
Aniba, Mezilaurus, Nectandra, ши КУ in, tae (Lauraceae), Eschweilera, Gustavia (Lecythidaceae), 
Guarea (Meliaceae), Pouteria (Sapotaceae), and Arytera (Sapindaceae). 


= 


diversity hotspots (Reid, 1998), or the existence of neither of the two collections of this remarkable 
refugia in Amazonia (Nelson et al., 1990). More- species provides locality data more specific than 
over, many critical practical applications of taxo- “South America." 


nomic knowledge, such as estimating the impact of Data collected by Madrifién-Restrepo (1996) 
human disturbance on species loss (e.g., Pimm et from monographie treatments of 15 tropical plant 
al., are impossible without reasonable genera (Fig. 5) illustrate a situation that is undoubt- 
knowledge of geographic distributions. edly « common. Of the 317 species he considered, 


ыы 


ere, then, do we stand in our knowledge of 8.5%) were known from just one specimen, 
geographic ranges? This question is difficult to ап- 52 (16.4%) from two specimens, and 26 (8.2%) 
swer, since baseline data have not been compiled from three specimens. That is, more than half were 
in an appropriate fashion. However, there are sev- represented only by one or two specimens, and al- 
eral reasons to believe we are still very far from most two-thirds by three or fewer. Few of the spe- 
having an accurate picture. One clear indication is cies he scored were known from more than 10 spec- 
the fact that many species are known from one ог imens, and none from more than 20. In such cases 
a few museum collections, and these are often old there are simply insufficient data to draw reason- 
and lack sufficiently detailed locality data. The able inferences about geographic range. 

lungless caecilian, Atretochoana eiselti, which we It is possible, of course, that species known from 
highlighted above, provides a striking example: one or two specimens or localities are truly very 


120 


Annals of the 
Missouri Botanical Garden 


restricted in occurrence. However, in most cases 
subsequent fieldwork indicates otherwise. Detailed 
data on this point have not been assembled, but we 
know that in most groups of organisms every field 
trip yields range extensions, sometimes major ones. 
A good example is provided by recent fieldwork in 
a small corner of Qinghai, China’s fourth largest 
province, with an area of around 720, k 
(about 6% larger than the state of Texas). Before 
an expedition in 1995, only four bryophyte species 
were reported in the literature for all of Qinghai. 
Three weeks of moss collecting by Benito Tan in 
Yushu Prefecture alone yielded 57 genera and 109 
species (Tan & Yu, 1997). Two of these species 
were new to science, three were previously un- 
known in China, and ten collections represented 
major range extensions within Chin 

Even in groups of organisms and i in places that 
have been better collected, key elements have of- 
ten been overlooked. In 1998, our colleague in the 
Harvard Herbaria, Gustavo Romero, collected Er- 
isma japura Spruce ex Warm. (Vochysiaceae) from 
the Isthmus of Pimichin (between the Orinoco and 
Amazon basins) in Venezuela. The first collection 
of this species from Venezuela had been made 
only the year before, from the same area; previ- 
ously, it was known from a few collections along 
the Rio Negro in Colombia and from Brazil. What 
makes this remarkable is that Alexander von 
Humbolt and Aimé Bonpland, Richard Spruce, 
Alfred Wallace, Llewelyn Williams, Julian Stey- 
ermark, Bassett Maguire, and others, all worked 
here, but none of them collected Erisma japura, 
despite the fact that it appears to be the dominant 
tree in the non-flooded forests of the area. Indi- 
viduals range to 35 m in height and over 1 m in 
diameter, and have conspicuous winged fruits that 
are used as a source of starch by local people (G. 
Romero, pers. comm.). 

Two other discoveries, also of large neotropical 
trees, emphasize that our knowledge of the distri- 
bution of many clades is still rudimentary. Rupti- 
liocarpon caracolito Hammel & N. Zamora was de- 
scribed in 1993, primarily on the basis of 
collections from Costa Rica’s Osa Peninsula (Ham- 
mel & Zamora, 1993). Its closest relative appears 
to be Lepidobotrys (Lepidobotryaceae) of the Ga- 
bon-Cameroon region of Africa. In a similar case, 
Pseudomonotes tropenbosii A. С. Londofio, E. Al- 
varez & Forero was described in 1995 from the 
Colombian Amazon (Londofio et al., 1995). This is 
only the second New World species of the otherwise 
Old World Dipterocarpaceae, which dominate the 
tropical forests of Asia. The other neotropical spe- 
cies, Pakaraimaea dipterocarpacea Maguire & Ash- 


ton, was discovered less than 20 years earlier (Ma- 
guire & Ashton, 1977). 

Examples like these are surely common, but are 
they really very surprising? After all, one could ar- 
gue that these are not organisms about which we 
thought we had good knowledge to begin with. Have 
we learned anything really new about the distri- 
butions of organisms that we thought we knew well, 
organisms of great enough interest that conscious 
efforts had been made to find them and establish 
their ranges? 

Again, the answer is a definitive yes! The most 
striking recent case concerns the coelacanth, La- 
timeria chalumnae. Thought to have been extinct 
since the Upper Cretaceous, and of great evolution- 
ary interest from the standpoint of the origin of tet- 
rapods, a specimen was caught off the coast of 
South Africa in 1938. After an intensive search, a 
second specimen was found in 1952 near the Com- 
oro Islands, northwest of Madagascar (Thomson, 
1989). Since then, more than 100 additional spec- 
imens have been taken of the now endangered fish 
in the vicinity of the Comoro Archipelago, but no- 
where else. It therefore came as a great shock 
when, on the 30th of July, 1998, a coelacanth was 
caught in a gill-net off the coast of north Sulawesi, 
Indonesia, almost 10,000 km away from the Com- 
oros (Erdmann et al., 1998; see note p. 126). Other 
populations may exist between the two sites, but in 
any case this discovery reminds us how little we 
know about the distributions of even large and well- 
publicized marine organisms. 

Similar lessons are provided by the rediscovery 
of species thought to have become extinct in recent 
time. Botanists will be aware, for example, of the 
rediscovery of Takhtajania perrieri (Capuron) Bar- 
anova & J-F. Leroy in northeastern Madagascar, 
about 150 km east of the spot where the original 
and only collection was made 85 years earlier 
(Schatz et al., 1998). Being of great interest from 
the standpoint of angiosperm evolution (as the only 
modern African-Madagascar representative of the 
vesselless Winteraceae), botanists from the Missou- 
ri Botanical Garden and their Malagasy colleagues 
had tried in vain since 1974 to relocate Takhtaja- 
nia. A Malagasy collector stumbled upon a popu- 
lation of over 250 adults 
work in 1994, and study material has been made 
available to botanists around the world. Molecular 
phylogenetic analyses now suggest that Takhtajania 


while doing inventory 


is the sister group of the rest of the Winteraceae 
(E. Zimmer, pers. comm.). 

Even better known rediscoveries involve verte- 
brates, especially mammals. Again from Madagas- 
car comes the story of the rediscovery of the greater 


Volume 87, Number 1 
2000 


Donoghue & Alverson 121 


New Age of Discovery 


bamboo lemur, Hapalemur simus (Wright, 1988), 
which was thought to have become extinct in 1900. 
A living animal was purchased in a market in 1964, 
but soon escaped from captivity; not until 1972 was 
a small population found in the wild. In 1986, two 
expeditions, one led by Patricia Wright from the 
United States and the other by Bernhard Meier of 
Germany, were successful in locating it again. This 
set the stage for detailed studies of behavior and 
ecology, which led in turn to the realization that 
there were actually two separate species involved. 
The new species, which they called the golden 
bamboo lemur, H. aureus (Meier et al., 1987), ap- 
parently specializes on new shoots of the bamboo 
plant, exceptionally rich in the poison cyanide. 

o fungal examples further emphasize the po- 
tentially great importance of such discoveries. The 
mushroom Squamanita contortipes, first described 
in 1957 (in the genus Cystoderma), was rediscov- 
ered in 1992 during the course of systematically 
sampling the macrofungi of old-growth spruce-hem- 
lock forests on the Olympic Peninsula of Washing- 
ton (Redhead et al., 1994). As shown in Figure 3c, 
S. contortipes fruit bodies were found growing from 
the cap of another agaric, a species of Galerina. 
This observation provided the crucial clue to the 
interpretation of the lifestyle and morphology of an 
entire group of fungi. Squaminita species were ini- 
tially described as having “protocarpic tubers,” 
sometimes mysteriously resembling the tissues of 
distantly related mushrooms. Owing to the Olympic 
Peninsula specimens, these structures are now un- 
derstood to be deformed hosts, and Squaminita is 
now recognized to be only the second agaric genus 
in which all species are obligate parasites of other 
basidiomycetes. 

In 1994, a group of Cornell students taking a 
field mycology course in upstate New York en- 
countered unusual fruiting bodies growing out of 
the backs of dead beetle larvae. These turned out 
to be the ascomycete Cordyceps subsessilis. Prop- 
agation of this material in the laboratory revealed 
it to be the formerly unknown sexual stage (teleo- 
morph) of the mold Tolypocladium inflatum, which 
produces the immunosuppresive compound cyclo- 
sporin, a billion-dollar drug used in preventing the 
rejection of transplanted organs (Hodge et al., 

6). This knowledge makes possible, for the 
first time, the search for other pharmaceutically 
important chemicals (possibly of use in treatment 
of autoimmune diseases) in related Cordyceps spe- 
cies. 

What developments do we anticipate with re- 
spect to geographic ranges? Most obviously, it will 
be critical to continue databasing geographic infor- 


mation associated with museum specimens and 
making this accessible to potential users. This will 
give a much better picture of the distributions of 
species and clades, but will still be insufficient for 
many purposes, both for lack of collections, and 
because, as several of our examples illustrate, a set 
of collection localities is not the same as a range, 
and may even be a poor approximation of the entire 
distribution. A critical step will, therefore, be the 
development and application of models for esti- 
mating real geographic ranges from sets of collec- 
tion sites. Fortunately, great strides have been made 
in this area with the development of Geographic 
Information Systems (GIS) and models such as 
those underlying BIOCLIM/BIOMAP, for example 
(see Austin, 1998; Scott & Jennings, 1998). 
Another exciting prospect entails improved con- 
nections between geographic and phylogenetic in- 
formation. This would not only revolutionize the 
study of historical biogeography, but would also 
provide a genuinely new perspective on the distri- 
bution of diversity as this relates to conservation 
and management issues (e.g., Vane Wright et al., 
1991). Already there are concrete examples, in- 
volving a range of organisms and spatial scales. For 
instance, phylogenetic analyses point to Papua New 
Guinea as a hotspot of genetic diversity in shiitake 
mushrooms, and therefore a conservation priority 
(Hibbett & Donoghue, 1996). Analyses in Astera- 
ceae and curculionid beetles highlight the signifi- 
cance of southern temperate regions in relation to 
1996). Similarly, 


in the north temperate zone the Pacific Northwest 


global biodiversity (Morrone et al., 


harbors a variety of species of tremendous phylo- 
genetic significance (D. Wake, pers. comm.), such 
as Ascaphus truei, the sister group of all modern 
frogs (Ford & Cannatella, 1993). Ultimately, we en- 
vision a concatenation of databased geographic in- 
formation (especially predicted ranges along the 
lines of BIOCLIM/BIOMAP) with databased phy- 
logenetic information (along the lines of Tree- 
BASE). Among many other things, this would en- 
able the wide-scale application of Dhylogenetis 
measures is diversity (e.g., Williams et al., 1 


Faith, 199 


CONCLUDING THOUGHTS 


To systematists, it is probably already clear that 
we live in an age of discovery and that this one is 
richer and more profound by virtue of the attention 
being paid to both phylogeny and geography. Yet, 
this is not at all obvious to much of the scientific 
community or to the general public, who tend to 
regard the discovery of diversity as a tedious filling- 


122 


Annals of the 
Missouri Botanical Garden 


in of minor gaps in our already comprehensive 
knowledge. Whether or not we have failed, our- 
selves, to fully appreciate the number and signifi- 
cance of recent discoveries is debatable. What is 
not debatable is that we have failed to effectively 
deliver news of these stunning discoveries to those 
outside of our community. As we have suggested, 
there are a variety of reasons for this, including our 
limited ability to compile information on the num- 
ber of newly discovered species and clades, or to 
convey the surprisingness of new discoveries. We 
also are limited by a system of nomenclature that 
tends to discourage rather than facilitate a full elab- 
oration of the tree of life. These issues need to be 
confronted directly, and soon. 

n the meantime, there are several avenues 
along which we can advance public awareness and 
appreciation of biodiversity. Perhaps most obvi- 
ously, we can highlight exactly how it is that un- 
derstanding and protecting biodiversity is in our 
own best interest, even economically. It is hard to 
imagine a more compelling statement of this con- 
nection than the “Teaming with Life” document 
prepared recently by the Biodiversity and Ecosys- 
tems Panel of the President’s Committee of Advi- 
sors on Science and Technology (PCAST; Raven 
et al., 1998). Big and bold new biodiversity dis- 
covery projects are another mechanism to capture 
the public’s imagination. For example, we see 
enormous educational potential in the All Taxa 
Biodiversity Inventory (ATBI) being undertaken 
by the Great Smoky Mountains National Park 
(www.discoverlife.org). Several of the projects to 
be sponsored as part of the DIVERSITAS Inter- 
national Biodiversity Observation Year (IBOY) in 

www.icsu.org/diversitas) have similar poten- 
aon for example, retracing key voyages of past 
ges of discovery (by Darwin, Banks, von Hum- 
boldt. and Wallace), or assembling an enormous 
tree of life and somehow physically displaying its 
grandeur to the public, or launching major inter- 
national expeditions to assess diversity in little 
known regions or in extreme environments 

New discoveries of the sorts we have highlighted 
have enormous potential to inspire commitment to 
the project of discovering the diversity of life. How- 
ever, to fully capitalize on this potential, we will 
need to pay more attention to specific mechanisms 
for awakening wonder. Is it possible to induce the 
sense of awe, amusement, and even befuddlement 
that remarkable new organisms inspired during the 
last great age of discovery? What is it that inspires 
wonder in six-year-old children visiting a natural 
history museum for the first time? Along these lines 
we are encouraged by several recent developments. 


In a fine analysis of the aesthetics of biological di- 
versity, Kiester (1997) drew attention to the exis- 
tence of a wide range of biodiversity experiences, 
contrasting those centered on the immediate, tan- 
gible beauty (or bizzareness) of individual organ- 
isms with those invoking a sense of the vastness or 
power of nature (what Immanuel Kant called “sub- 
lime” experiences). If we pay closer attention to 
such distinctions, the range of experiences trig- 
gered by biodiversity discoveries could surely be 
expanded, and their impact could be far greater on 
the public's perception of nature and the discovery 
process itself. 

In view of this, we should renew our attention to 
how new discoveries are brought forward in natural 
history museums and elsewhere. We are delighted 
by recent experimentation with museum exhibits, 
especially those inspired by the “wonder cabinets" 
of the past (Hutchinson, 1965; Weschler, 1995). 
These collections of rarities—natural objects often 
commingled with works of art and assembled in 
seemingly haphazard fashion—help put the observ- 
er slightly, but very constructively, off balance. Per- 
haps the premier modern experiment of this type is 
David Wilson's Museum of Jurassic Technology in 

s Angeles, California (Weschler, 1995). By play- 
fully straddling the dynamic boundary between re- 
ality and fiction, Wilson seems to have hit upon an 
unusually successful way of stimulating doubt and 
wonder. Today's natural history museums, which 
sometimes convey an unwarranted certainty or fi- 
nality about our knowledge of the world, would ben- 
efit from careful scrutiny of the Wilson model. 
There is great power, we believe, in the mystery 
and strangeness of life, and in deeply appreciating 
how little we still know about it. The torrent of bi- 
ological novelties now coming to light may "abus 
the very best way to tap that power. 


Literature Cited 


Aguinaldo, A. ‚ J. M. Turbeville, 
Cien, LR Carey, R. A R Raff & 


L. S. а is 
& J. А. Lake. 


Evidence for a clade of nematodes, arthropods, gr ite 
moulting m Nature 387: 489. 
йен. V. A., S. E. Williams & M. W. Chase: 1992. Car- 


nivorous bu. Phylogeny and structural evolution. 
Science 257: iar 1495. 
d W. S., B. A. Whitlock, R. Nyffeler, C. Bayer & 
Baum. о. Phylogeny of the core peg 
ш from ndhF sequence data. Amer. J. 
1459-1471 

APG Жар брен Phylogeny Group). 1998. Ап ordinal 
classification for the families of flowering plants. Ann. 
Missouri Bot. Mud 85: 531-543. 

Austin, M. P. 1998. An ecological perspective on biodi- 
versity investigation Examples from eer е euca- 
lypt forests. Ann. Missouri Bot. Gard. 85: 

Baker, A. N., F. W. E. Rowe & H. E. S. Clark. Ж А 


~ 


Моште 87, Митбег 1 
2000 


Donoghue & Alverson 
New Age of Discovery 


123 


new class of Echinodermata from New Zealand. Nature 
321: 862-864. 
Baldauf, 5. L. & W. Е. Doolittle. 1997. Origin and evo- 
lution of Hi т vius (Mycetozoa). Proc. Natl. Acad. 
: 12007-12012. 


1 a e pes Animals and fungi are each 
other's dod relatives: Congruent evidence from mul- 
ros m Proc. Natl. Acad. Sci. U.S.A. 90: 11558- 


E P А., W. 5. Alverson & R. Nyffeler. 1998. A du- 
rian by any other name: Taxonomy and nomenclature of 
the core Malvales. Harvard Pap. Bot. 3: 315—330. 

Berger, L., R. Speare, P. Daszak, D. E. Green, A. A. Cun- 
ningham. C. L. Goggin, R. Slocombe, M. A. Ragan, A. 
D. Hyatt, K. R. McDonald, H. B. Hines Lips, G. 
Maranteli & H. Parkes. 1998. Chytridiomycosis causes 


. Natl. Acad. Sei. U.S.A. 95: m –9036. 


Briggs, B. С. 000. What is significan e Wollemi 
pine or э southern rushes? Ann. ты bot Gard. 
87: 

Brusca, R. ri 20 


OO. Unraveling the history of arthropod 
biodiversification. Ann. Missouri _ Gard. 87: 13—25. 


Chase, M. hi. D. E. Soltis, R 


Olmstead, D. Mores, 


D. . B. D. Mishler, ^ R. Duvall, R. A. 

H. С. Hills, Y-L. Qiu, A ‚ J. H. Rettig, | 

Conti, J. D. Palmer, J. R. Manhart, K. J. Sytsma, H. J. 

Michaels, J. Kress, K. G. Karol, D. Clark, M. 

a В. 5. Gaut, R. K. Jansen, K-J. Kim, С. Е Wim- 
SJ T G. R. uek > Pa Strauss, Q-Y 


E. Golenberg, G. H. ed ТЕ: 5. W. prea 

‚А. ‚ 1993. Dc 
genetics of seed plants: An analysis of quielestide se- 
quences from the plastid gene rbcL. Ann. Missouri Bot. 
Gard. 80: 528—580. 

Chippindale, P. T., A. H. e & D. M. Hillis. 1993. A 
new species of Adi ено is salamander (Eurycea: 
Plethodontidae) from Austin, Texas. Herpetologica 49: 

259. 


Coen, Е. 5. & E. М. Meyerowitz. 1991. The war of the 
whorls: Genetic interactions controlling flower devel- 
2 E 353: 31-37. 

Corben, С. J. Ingram & M. Ј. Tyler. 1974. Gastric 
duni due Вијена form of parental care in an Australian 
frog. Science 186: 946—947. 

Cronquist, A. 1988. The Evolution and Classification of 
Flowering Plants. New York Botanical Garden, Bronx. 
New York. 

Desjardin, D. E., L. Martinez-Peck & M. Rajchenberg. 
1995. An ия psychrophilic aquatic agaric from Ar- 
gentina. Mycologia 87: 547—550 

M. . Progress сой ects in recon- 

structing plant огр Апп. Missouri ue Gard. 81: 

5-418. 


& J. A. Doyle. 1989. Phylogenetic analysis of an- 
giosperms and the relationships of Hamamelidae. Pp. 
17-45 in Р. Crane & S. Blackmore (editors), Evolution, 
Systematics and Fossil History of the Hamamelidae, 

Vol. 1. Clarendon Press, Ox 

Douglas, A. W. 1995. Proteaceae, Мене. Flora of Aus- 
tralia 16 aes eae, Proteaceae 1): 6-10. 

& B yland. 1995. Proteaceae, Eidotheo- 
ideae. In: Fora м Australia 16: 127-129. CSIRO Aus- 
tralia, Melbou 


BS 
а. 


Поуе, Ј. А. 1998. ор of vascular plants. Annual 
Rev. Ке :ol. Syst. 29: 567-5 
Hotton. 1991. i? versification of early an- 
69— 


giosperm pollen: in a cladistic context. Pp. 195 in 
S. Blackmore & < . Barnes (editors), Pollen and 
rd. 


Spores. Clarendon Dux Oxfo 
.. P. M. Giao, N. N. Chin, D. Tuoc, P. Arctander 

; . А new species of living bovid 
hom Vietnam. purs 363: 443—445. 

Eernisse, D. J., J. S. Albert & F. E. ne i An- 
nelida and Arial are not sister e 
netic ПАО of spiralian metazoan ea Silas Syst. 
Biol. sig 330. 

Erdmann, К. L Caldwell & M. K. Moosa. 1998. 


Indonesian ы of the sea” discovered. Nature 395: 


fie B. 2000. Floristic surprises in North America north 
of Mexico. Ann. Missou ri Bot. Gard. =з 81-109. 

Escobar, McCabe & C. W. Harpel. 1976. 
соет а floating _Eastromyeete isolated from 

rshes. Мус eee 68: 874—880 
Evelyn, J. 1684. Diary of Toh: Плеј Vol. IV. Pp. 389- 
390, 22 October; quote from K. Whittaker, 1996, га 

мрке of а . Рр. 75-90 т N. Jardine, J. А. 
cord & К. oe (editors), Cultures of Natural tw 

tory. | bre Univ. Cambridge 

Faith, 1992. Conservation iius and phyloge- 
Big: prie Biol. Conservation 61: 1-10 

Ford, L. 5. & D. С. Cannatella. 1993. The major clades 
of 2 Herp. Monogr. 7: 94—117. 

Fortey, R. A. & R. H. Thomas ок 1997. Arthropod 
ни Chapman and Н 

кина . J. D. & L. Campbell. 
> 303: 410-411. 

.M. 


Kristensen. 199 


Press, 


London. 


1998. Microbial micro- 


Func h. P. & ycliophora is a new 
phylum vit affinites to Е ie ta pu Ectoprocta. Na- 
ture 378: –714. 

Gaston, K. " 1991. The pad s a с De spe- 
cles richness. Conservation Biol. 5 

us vanayake, 

. Amato, P Arctander & J. R. MacKinnon. 1998. 
sc itin 1 of Muntiacus truongsonensis, а new s spec ies ~“ 
muntjac (Artiodactyla: Muntiacidae) from central Viet- 
nam, and аат for conservation. Animal Соп- 
servation 1: 

Gilmore, 5. 4 к. D. - Hill 1997. Relationships of the Wol- 
lemi pine d ipu A a molecular phylogeny 
of the bu gc Telop 275-291. 

Ribera. TR "The position of arthropods 
in the i kingdom: A search for a reliable outgroup 
for oS eM phylogeny. Molec. Phylogenetics 
X Evol. –488. 

ac F. & T pina 1998. Amphibian species diversity 
exceeds that of Side Herpetological Rev. 29: 11— 
12. 

Gómez-Laurito, J. & L. D. Gómez P. 1989. Ticodendron: 

A new tree from Central America. Ann. Missouri Bot. 

Gard. 76: 1148-1151. 

& . 1991. Ticodendraceae: A new family 

of flowering plants. Ann. Missouri Bot. Gard. 78: 87— 

88. 


кее W., F. R. Barrie, H. M. Burdet, W. G. Chaloner, 


International. Code of Botanical Nomenclature (Tokyo 
Code). Regnum Veg. 1 
Halanych, K. M., J. D. Hae lies A. M. A. Aguinaldo, S. 


124 


Annals of the 
Missouri Botanical Garden 


, D. M. Hillis & J. A. Lake. 1995. Evidence 
from 188 ribosomal DNA that ih ы аге 


ba nearly: The Aap о Ticodendraceae. Ann. Mis- 
uri Bot. Gard. 78: 89-9 


. А. Zamora. iva о (Lepido- 

botryace ae): A new arbor genus and tropical 

American link. to Africa, ih a reconsideration of the 
Vin 


e there so many new amphibian 
spec ү о а are declining? Trends Ecol. 
Evol. 
Hansen, А. A Haaman T. Samigullin, A. Antonov & 
. Gnetum and the angiosperms: Molec- 
ular evidence that their shared morphological charac- 
ters are convergent, rather than homologous. Molec. 
Biol. Evol. 16: 1006-1009. 
Hasebe, M., P. G. Wolf, K. M. Pryer, K. Ueda, M. Ito, R. 


xin Fern phylogeny based on rbcL nucleotide se- 
ces. Amer. Fern J. 85: 134—181. 
нец, р. 5. & М. Ј. Donahue. 1996. Implications of 
phylogenetic merus for conservation of genetic diver- 
sity in shiitake mushrooms. Conservation Biol. 10: 
1321-1327. 


. 1998. Integrating phylogenetic analy- 

sis ie classific ation in оте, neh ologia 90: 347-356. 

Hodge, K. T., K . А. Humber. 1996. To- 
lypocladium inflatum i is the i of Cordyceps sub- 
sessilis. Mycologia 88: 715-719 

Holden, C. 1995. Ancient trees down under. Science 267: 
334. 


Hoot, S. B., S. Magallon & P. B. Crane. 1999. Phylogeny 
of basal аш ГЕ based on th пее molecular data sets: 
atpB, rbcL, and 185 nuclear ribosomal DNA sequences. 
Ann. Missouri Bot. Gard. 86: 1-32. 

Horgan, J. 1998. Quoted in "Resolved: science is at an 
end. Or is it?" by D. Overbye (editor), The New York 
Times, 10 Nov. 1998 

Hutchinson, G. E. 1965. The dias E ddr ud =. Ше 

ее Play. Yale Univ. w Hav 

Jones, W. . D. Hill & J. м. o 1995. Wollemia 
nobilis, a new living Australian epe and species in 
the Araucariaceae. Telopea 6: 17 


Jong, W. e 1998. Molecules remodel the mammalian 
tree. Trends Ecol. Evol. 13: bn 
Eun б, M Farris, M м ‚В. Bremer, М. Е. 


Humphries, С. Petersen, 0. Seberg & K. 
а 1998. Simultaneous parsimony jackknife anal- 
ysis of 2538 rbcL sequences reveals support for major 
clades of green plants, land plants, Heg plants, and 
flowering plants. Pl. Syst. Evol. 213: —287. 

Kiester, А. В. 1997. Aesthetics of Te de diversity. Hu- 
man Ecol. Rev. 3: 151-157. 

Knoll, A. H. & S. B. Carroll. 1999. Early animal evolu- 
tion: Emerging views from comparative biology and ge- 
ology. Science 284: 2129-2137. 

Kristensen, R. M. 1983. Loracifera, a 
Asc Regan characters 
Z _Evolutionsforsch. 21: se 80. 

Laurance McDonald à Speare. 1996. 
Ера dua her the catastrop ir darling of Aus- 

tralia an rain forest frogs. Conservation Biol. 10: 406– 


ew phylum e 
from vis ыш Шы 7. 


bien: D. 5. 1973. А new genus of frog of the family Lep- 


todactylidae Кот SE Queensland, У enis Mem. 
ueensland Mus. 16: 459-470, pl. 3 
C., E. М Е. Еогего p C. M. 


Morton. 
and species of Dipterocarpaceae 
from the Neotropics: | Bee ‘tion, taxonomy, ecology, 

and distribution. Bri а 4T: ov 

Longcore, J dens er & D. K. Nichols. 1999. 
Batrachochytrium dendrobatidis Aus Pl sp. nov., a chy- 
trid n to amphibians. Mycologia 91: 219-227. 

rcx . 2000. New iw e in the 21st century? 

"Missouri Bot. Gard. 8 

2000 м Бас Ape and micro- 

Ann. Missouri Bot. Gard. 87: 3-12. 

Madrifüán-Restrepo, S. 1996. Systematic studies in the 
Lauraceae. Monograph of Rhodostemonodaphne. Ph.D 
Dissertation, Department of Organismic and Evolution- 
ary Biology, Harvard University. 

Maguire, B. & P. S. Ashton. 1977. Pakaraimoideae, Dip- 
terocarpaceae of the Western Hemisphere. II. System- 
atic, geographic and phyletic considerations. Taxon 26: 

41-385. 

Manhart, J. В. 1995. Chloroplast 16$ rDNA sequences 
and phylogenetic ara of fern allies and ferns. 
Amer. Fern om 85: 182-19 

‚М. Enslo eman, А. Martínez-Mena, E 

7 & С. бон 1989. Anatomía reproductiva de 

Lacandonia е (Lacandoniaceae). Апп. Mis- 
souri x» : 124-127. 

Martin, I Gershwin, J. W. Burnett, D. G. Car 
D. a 1997. Cheyanne ti a НЕ 
пем species of Scyphozoan from the Eastern Pacific. 
Biol. Bull. 193: 8-13. 

Martínez, E H. Ramos. 1989. Lacandoniaceae 
(Triuridales): Una nueva familia de México. Ann. Mis- 
souri Bot. Gard. 76: 128—135. 


Meier, B., R. Albignac, A. Peyr iéras, Y. Rumpler & P. 
Wright. 1987. A new species of Hapalemur eae 
from South East Madagascar. Folia Primatologica 48: 

1-2 
Moore, L. G. Rocap & S. W. Chisholm. 1998. Physi- 


ology A mx ular ста of coexisting Prochlo- 
е ecotypes. Nature 393: 

orell, 1996. New mammals disc overed by biology's 
new explorers. Science 273: 146 

Are pathogens felling frogs? Science 284: 


728-731. 

Morrone, J. J., L. Katinas & J. V. Crisci. 1996. On tem- 
perate areas, basal clades and biodiversity conservation. 
Oryx 30: 187-194. 

Nelson, В. W., С. A. С. Ferreira, M. Е da Silva & M 

x i . Endemism centres, refugia and bur 

ical collection densis in Brazilian Amazonia. Natur 
345: 714-716. 

Nickrent, D. L. & R. J. Duff. 1996. Molecular studies of 
parasitic plants haing ribosomal RNA. P 2 in M. 
Moerno et al. (editors), Advances in Parasitic Plant Re- 
search. Junta de Andaluci ма, Dirección General de In- 


каше 1бп ч Согдођа, Зрат. 


. E. Colwell, A. D. Wolfe, N. D. Young, 
a C. W. deP amphilis. de Molecular 
es of parasitic 
‚ D. n Soltis & J. J. 
; Molecular | шшен of Plants Il. 
ГА | Kluwer Academic, Bosto 

& . Starr. 1994. High rates of m sub- 
stitution in nuclear small-subunit (185) rDNA from hol- 
oparasitic flowering plants. J. Molec. Evol. 39: 62-70. 


Volume 87, Number 1 
2000 


Donoghue & Alverson 125 


New Age of Discovery 


Nussbaum, R. A. & M. Wilkinson. 1996. A new genus of 
lungless tetrapod: A radically divergent caecilian (Am- 
phibia: ee Proc. Roy. Soc. London, B, Biol. 
Sci. 261: 

Face, N. R. em ^ анге ular view of microbial diversity 

and the ж е 276: 734—140. 

i B. D. 4. Accumulating knowledge on the 
dimensions of Rn Systematic perspectives on 
Neotropical mammals. Biodiversity Lett. 2: 79—86. 

Pessier, A. P., D. K. Nichols, Longcore & M. 
Fuller. 1999. Cutaneous chytridiomycosis in poison e 
frogs (Dendrobates spp.) a nd White's tree frogs (Litoria 
caerulea). J. Veterinary Diagnostic Investigation 11: 


Pimm, S. L., С. J. Russell, J. L. Gittleman & T. M. Brooks. 
1995. The future of biodiversity. Science 269: 347-350. 

Prance, G. T., H. Beentje, J. Dransfield & R. Johns. 2000. 
The tropical flora а undercollected. Ann. Missou- 
ri Bot. Gard. 87: 67-71. 

Pryer, K. M., A. R. anb & J. E. Skog. 1995. Phyloge- 
netic relationships of extant ferns based on evidence 
from morphology and rbcL sequences. Amer. Fern J. 85: 

28: 


Quammen, D. 1996. The Song of the Dodo. Island Bio- 
eography in an Age of Extinction. Touchstone, Simon 

= Schuster, New York. 

Queiroz, K. de & J. Gauthier. 1992. oo taxon- 
omy. Annual Rev. yen Syst. 23: 449-4, 

Toward a i ae system 

of biological тен Trends Ecol. Evol. 9: 27 

31. 


Raven, P., F. J. Ayala, С. C. Bowker et al. 1998. бина 
with life: Inv vesting in science to understand and us 
America’s living capital. The President’s Committee of 
Advisors on Science and аи (PCAST). Office 
of the President, Washington, D.C. 

Redhead, S. A., J. F. Er G. R. Walker, L. L. Nor- 
vell & M. B. Puccio. 1994. Squamanita eontortipes, the 
Rosetta Stone of : ын agaric genus. Canad. 
J. Bot. 72: 1812-1824. 

Reid, W. V. 1998. о hotspots. Trends Ecol. 
Evol. 13: 275-280 

Rice, K. А., M. J. Donoghue & R. С. Olmstead. 1997. 
Analyzing е ов sets: rbcL 500 revisited. Syst. Biol. 
46: 554—56 

Richardson, m W. M. Goldberg. K. G. pe» R. B. 
Aronson, G. W. даш, К. B. Ritchie, J. C. Halas, J. S. 
Feingold & 5. L. Miller. 1998. BÉ. енн coral- 
killer identified. Nature 392: 557 

Rodman, J. | . С. Karol, R. A. Pric ce i K. J. Svtsma. 

6. Males las, morphology, and Dahlgrens expanded 
iie C ОТ m Bot. 21: 289-307. 

Rothwell, H. A. Stockey. 1994. The role of Hy- 
dri зет cee gen. el sp. nov. in reconstructing the 
cladistics of iM ad Amer. J. Bot. 81: 479— 
492. 

nm M. J., G. Baldwin, С. Ваш Нап, C S 

mpbell, D. ferguson J. M. Porter, C. Von Dohlen, 
LE Wojciechowski & M. J. Donoghue. 1993. The 

и of phylogenetic а апа the need for а 

poe — Syst. . 42: 562-5 

Schaller, G. Vrba. 1996. Description of the 
giant mun са (мешати vuquangensis) in Laos. 
J. ada 77: 675-683. 

Schatz, G I & A. Ramisamihantanurina. 
1998. Takhtajania perrieri rediscovered. Nature 391: 


133-134 


Scott, J. M. & M. D. Jennings. 1998. Large-area ‚ mapping 
of biodiversity. Ann. Missouri Bot. Gard. 85: : А 
Setoguchi, H., T. A. Osawa, J-C. Pintaud, Т. Qs & J- 


sequences. Amer. J 


516 
‚В. Ertter & D. W. Taylor. 1992. Neviusia 
cliftonii ива Kerrieae), an intriguing new relict 


species from vy fornia. Novon . 
Soltis, D. E. & P. S. Soltis. 1997. Phylogenetic relation- 
ships in odium eae sensu lat mparison of to- 


to: 

pologies "ud on we _ and hel sequences. 

Amer. J. B 4: 504. 

Soltis, D. E., Е 4 Soltis, gd L. Nickrent, L. A. Johnson, 
W 


re, R. K. Kuzoff, K 


somal DNA sequences. Ann. Missouri Bot. Gard. 
1-49. 

Soltis, D. E., P. S. Soltis, M. E. Mort, M. W. e, V. 
Savolainen, S. B. Hoot & C. M. Morton. 1998. pr 
complex phylogenies using parsimony: Ап empirical ap- 
proach using three eoe rs data sets for angio- 
sperms. Suit Bio 1.4 

Springer, M. S., С.С Xx D Madsen, м W. de Jong. 
V. С. Wa ddell, H. M. Amrine & M. J. Stanhope. 1997. 

Badan mic African је а the ee tree. 
Nature 388: 61–64. 

Stanhope, M. Ј., О. Madsen, V. С. Waddell, С. C. peras 

W. W. de Jong & M. S. Springer. 1998. Hi ghly co - 
ы molecular support for a diverse пи ейде 
of n African mammals. Molec. Phylogenetics 
Evol. 9: —508. 

Sulloway, E » 1982. Darwin and his uc The evolu- 
tion of a legend. Т IM Biol. 15(1): 1 

Tan, B. 2 à J. Yu. 7. Mosses of Qinghai: Tibetan Pla- 
teau, C ee Manon Bot. Lab. 82: 305-320. 

Thomson, ra 89. The second Coelacanth. Amer. Sci. 
77: pr 

Tyler, M. J., D. C. Shearman, R. Fra ni пренет 
Seamark & R. Kelly. 1983. Inhibition of g a 
secretion in the gastric brooding frog, ааган 
silus. Science = Ages 0. 

Vane-Wright, R. . Humphries & P. H. Williams. 
1991. What to His o Tar a and the agony of 
choice. Biol. Conservation 55: 235-254. 

Vergara, F., C. Ferrándiz, E. Meyerowitz & E. R. Alvarez- 
Buylla. 1999. Molecular basis and evolution of the in- 
side-out flower of Lacandonia schismatica. Abstract 
15.13.2, р. 188 of Pare of the XVI International 
Botanical Congress, St. Louis, Missouri. 

O., G. Hinkle. У a Sogin & S. К. Stickel. 


tionary link with fungi. Science 260: 340— 

Wake, D. B. 1996. A new species of rado (Am- 
phibia: Plethodontidae) from the San Gabriel Moun- 
tains, southern California. Contr. Sci. © Hist. Mus. 
Los Ange n Bad 463: 1-12. 

es hler, L. . Mr. Wilson's _ о of Wonder. Vin- 

ge cls, her House, New York. 

Wilkinson М. А. & В. "Nussbaum. 1997. је ny ol 
morphology and ev к ungless caecilian Atre- 
tochoana eiselti (Taylor) (Amphibia: Gymnophiona: Ty- 
phlor oe Biol. J. Linn. Soc. ба: 39-109. 

en, z & C. A. Schwartz. 

1998. The largest lungles chat Report on a second 


d 


126 


Annals of the 
Missouri Botanical Garden 


specimen of the lungless caecilian Atretochoana eiselti 
(Amphibia: Gymnophiona: И poc E from Brazil. 
J. Nat. Hist. d ) 32: 617-6 

Williams, P. H., C. J. Humphries "i R. I. Vane-Wright. 
1991. aa biodiversity: Taxonomic relatedness 
for conservation priorities. Austral. Syst. Bot. 4: 665 
79. 

Wilson, D. E. & D. M. Reeder (editors). 1993. Mammal 
Species of the World: A Taxonomic and Geographic 
Reference, 2nd ed. Smithsonian Institution Press, 


ker, T. Münster, J. T. Kim, H. Saedler 

& G. Theissen. 1999, MADS-box genes reveal that gne- 

tophytes are more closely related to чеш than to 

е pes Proc. Natl. Acad. U.S.A. 96 
1342-134 

€ Ф on Lemurs lost and fona. Nat. Hist. 97: 


и =. a 1997. Evaluation of atpB nucleotide seq 


for phylogenetic studies of ferns and other pterido- 
phytes. Amer. J. Bot. 84: 1429-1440. 
‚ К. M. Pryer. ‚ A. В. Smith & M. sebe. 1999 
Phylogenetic studies of extant e be Pp. 
556 in P. S. Soltis, D. E. Soltis, & J. J. Doyle (editors), 
Molecular РН of Plants Il. DNA Sequencing. 
Kluwer Academic, Boston. 
Yager, J. 1981. Remipedia, a new class of Crustacea de 
the Bahamas. J. Crustacean Biol. 1: 


marine cave in 
33 


Note added in proof. The coelac ~ specimens = 
Indonesia have been named as a new species, Latimeria 
menadoensis, a conclusion edic well ene by 
molecular data (see M. a S P. 
a Caldwell & D. M. Hillis. 
Acad. Bu. U. S.A. 96: I os and L. 
НА 1. Rachmatika, А. Tjakrawidjaja, В. Had- 

ау & adie. 1999, Compt. Rend. Acad. Sci. Paris, 
sér. 3, 322: 261–267. 


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Our Unknown Planet: Recent Discoveries and the Future, the 45th Annual Systematics 
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Extremophilic Bacteria and Microbial Diversity Michael T. Madigan 3 

Unraveling the History of Arthropod Biodiversification Richard C. Brusca 13 
Many Fishes, So Little Time: An Overview of Recent Ichthyological Discovery in 
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New Mammals in the 21st Century? John MacKinnon 63 
The рее Flora Remains Undercollected —__ Ghillean T. Prance, Henk Beentje, 


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What is Significant —The Wollemi Pine or the Southern Rushes? __ Barbara С. Briggs 72 - 
Floristic Surprises in North America North of Mexico ______- _ Barbara Ertter 81 
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Volume 87 Annals 
Number 2 of the NZ 
2000 Missouri 


Botanical 


Garden 


REPRODUCTIVE BIOLOGY Leonardo Galetto,? Gabriel Bernardello,” 


Irene C. Isele,? José Vesprini,* 
СЕН CRISTA-GALLI Gabriela Speroni,? and Alfredo Berduc? 


ABSTRACT 


Flowering phenology, floral morphology, nectar features (chemical composition, secretion pattern, standing crop. 
removal effects), breeding system, and flower visitors were analyzed in seven populations of Erythrina crista-galli from 


chemical composition and concentration (ca. 2296) constant across all flowering stages. Most of the total nectar was 
secreted by buds. When the flowers first opened, most (> 50%) of the a nectar was available to pollinators. As 
flowers faded, a resorption period began. T The overall sugar Ми си was not affected by nectar removal. Hand crosses 
showed that this species is self-compatible. Crossed fruits showed significant differences from hand-selfed ones (autog- 
amy and geitonogamy). Xenogamous fruits and seeds showed the highest values for many traits (fruit mass, total seeds 

er fruit mass, mean seed mass, and seed germination percentage). Approximately 6% of the flowers set seeds in natural 
populations. Hymenopterans (carpenter bees and honeybees) and hummingbirds (four species) assiduously visited the 
trees in all the areas sampled and can be assumed pollinators. Almost 93% of recorded flowers were visited by bees, 
with the rest visited by hummingbirds. Phylogenetically, this species was included in the basal clade for the genus and 
characterized as passerine/hummingbird pollinated. However, we found that not only birds but bees functioned as major 
pollinators. This observation may indicate that this basal clade may represent an intermediate step from entomophily 
(typical of tribe Phaseoleae) to ornithophily (typical of Erythrina). 

y words: breeding system, Erythrina, Fabaceae, flower visitors, nectar features, nectary. 


Ecological and evolutionary success of the le- bees, birds, and bats (Kalin Arroyo, 1981; Schrire, 
umes has been strongly related to highly success- 1989). Although much is already known about the 
ful biotic pollination mechanisms, utilizing mostly pollination biology of Fabaceae, as indicated by 


! This work was supported by funds from CONICET, CONICOR, ANPCYT, and SECYT (UNC). Two anonymous 
reviewers improved previous versions of this manuscript with their comments and ideas. We are especially grateful to 
Victoria Hollowell for her hard work as scientific editor on early versions of this paper. Carolina Torres and Cecilia 
Eynard are thanked for helpful comments on the manuscript. Nidia Flury and Diana Abal Solís kindly prepared Figures 
4 and 5 (Flury), 2 and 7 (Abal Solís). Claudio Sosa identified the insects. 

? Instituto Multidisciplinario de Biología Vegetal, Casilla de Correo 495, 5000 Córdoba, Argentina 

3 Escuela de Biología, Fac. Cs. Exactas, Físicas у Naturales, Universidad Nacional de Cordoba, 5000 Córdoba, 
Argentina. 

* Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Santa Fe 2051, 2000 Rosario, = Fe, Argentina. 

5 Facultad de Agronomía, Universidad del Uruguay, Av. Garzón 780, 12900 Montevideo, Urugu 


ANN. Missouni Bor. Савр. 87: 127-145. 2000. 


128 


Annals of the 
Missouri Botanical Garden 


Kalin Arroyo (1981) and Schrire (1989), their data 
are too narrowly focused to enable a detailed syn- 
thesis. Thus, we emphasized a thorough analysis of 
several aspects of the pollination biology in several 
populations of one legume species. 

Erythrina crista-galli L. was chosen because this 
genus is well known from a number of studies (see 
references following). Erythrina comprises about 

12 species, mostly arborescent and pantropical, 
but also extending into warm-temperate areas, such 
as Argentina (Krukoff & Barneby, 1974). It is as- 
signed to the large, economically significant papi- 
lionoid tribe Phaseoleae. Recent cpDNA restriction 
site analyses of this tribe suggest that Erythrina is 
highly derived (Doyle & Doyle, 1993; Bruneau et 
al., 1995). The diversity of floral morphology may 
be associated with differences in pollination sys- 
tems (Bruneau, 1997). Among the primarily ento- 
mophilous Phaseoleae, Erythrina is atypical in be- 
ing ornithophilous (Neill, 1987; Bruneau, 1997), 
providing good examples of adaptation to different 
types of birds (Proctor et al., 1996). It shares with 
other bird-pollinated groups the red or orange, 
odorless flowers with copious nectar, and diurnal 
anthesis (Proctor et al., 1996; Bruneau, 1997). Most 
American species are pollinated by hummingbirds 
(Neill, 1987; Bruneau, 1997), although some are 
passerine pollinated. Both guilds of birds tend to 
be species specific (cf. Bruneau, 1997). 

Phylogenetic hypotheses considering morpholog- 
ical and cpDNA restriction site characters suggest 
that shifts from passerine bird to hummingbird pol- 
lination have occurred several times in the genus 
(Bruneau, 1997). Erythrina crista-galli is included 
in a paraphyletic assemblage of South American 
species that are basal in the genus (Bruneau & 
Doyle, 1993; Bruneau, 1996). Thus, a detailed 
study of the reproductive biology of this species 
may provide clues to the evolution of the pollination 
biology within the genus. 

Erythina crista-galli is native to southern Brazil, 
Bolivia, Paraguay, Uruguay, and Argentina as far as 
the Rfo de La Plata, having the southernmost dis- 
tribution for the genus in the Americas. The species 
consists of trees to 10 m tall, with non-tubular, 
large, red flowers of ornamental, economic, and me- 
dicinal interest (Burkart, 1987). It is cultivated 
throughout the world in frost-free climates (Krukoff 
& Barneby, 1974) and is the national flower of Ar- 
gentina and Uru 

Scattered reports are found on its reproductive 
biology, mainly reporting floral visitors, breeding 
system, or nectar chemical composition (Knuth, 
1906; Schrottky, 1908; Werth, 1915; Pickens, 
1931; Ali, 1932; Fryxell, 1957; Faegri & van der 


Pijl, 1971; Raven, 1974; Toledo, 1974; Parodi, 
1978; Toledo & Hernández, 1979; Kalin Arroyo, 
1981; I. Baker & H. G. Baker, 1979; H. G. Baker 
& I. Baker, 1983, 1990; Neill, 1987, 1988), but no 
study has been comprehensive. Herein, we address 
the following questions for several populations of 
E. crista-galli, broadly spanning its distribution: 


(1) What is its reproductive phenology? 

(2) What are the flower duration, different floral 
phases, and the functional structure of the 
flower in relation to pollination? 

(3) What is the nectary structure as well as the 
chemical composition of the nectar? 

(4) What is the secretory pattern, the sugar com- 
position of nectar for a single flower’s duration, 
with respect to the nectar standing crop? 

(5) What is the floral response to nectar removal? 

(6) What is the breeding system of this species? 

(7) Are there fruit differences produced by the 
different pollination treatments? 

(8) Who constitutes floral visitors, what is the fre- 
quency of the visits, and what is their polli- 
nation role? 


MATERIAL AND METHODS 


Fieldwork was conducted on a total of 158 adult 
individuals from four natural populations and three 
cultivated ones (Table 1, Fig. 1). Cultivars were 
considered because there is high fruit and seed 
predation by beetles (Curculionidae, Araptus spar- 
sepunctatus; Viana, 1965) in natural populations, 
obscuring data. Vouchers for all seven populations 


are deposited in CORD, UNR, and MVFA (Table 
1) 


Phenology data were recorded in 1994—1995 for 
five of the seven populations (see Results). Data 
included periods of bud production, major and ad- 
ditional flowering events, fruit production, and seed 
In population ARG-ER1, a detailed 
study of 10 trees was performed at the same time. 

Flowers from population АКС-ЕК] were fixed in 
FAA, dehydrated through an ethyl alcohol/xylol se- 
ries, and embedded in paraffin to study their ana- 
tomical structure (Conn et al., 1960). Sections in 
cross and longitudinal views were cut at 10-рт 
intervals and stained with safranin and astral blue 
(Johansen, 1940). To localize stomata or starch 
grains, nectary tissue was cleared with NaOH (10% 
aqueous solution), washed with acetic acid : water 
(1 : 3), spread on a slide, and stained with an aque- 
ous I,-IK solution (Johansen, 1940). Drawings were 
made using a camera lucida attachment to a Zeiss 
microscope. Photomicrographs were taken on a 


dispersal. 


Volume 87, Number 2 
2000 


Galetto et al. 
Erythrina crista-galli 


129 


Table 1. 
МУКА (Speroni). 


Study sites for E. crista-galli populations. Vouchers are housed at CORD (Galetto), UNR (Vesprini), and 


Population Localities, vouchers 
abbreviation (number of trees studied) Latitude and longitude 
ARG-ERI Argentina. Entre Ríos Province, Dept 32°55'40"S 
Victoria, Isla Charigüe, Galetto 274, Vesprini s.n. (41) 60°36'67"W 
ARG-ER2 Argentina. Entre Rfos Province, Dept. 32°05'36"5 
Paraná, National Park Pre-Delta, Galetto s.n. (18) 60?40'48"W 
ARG-Chaco Argentina. Chaco Province, Dept. 27°30'03"S 
Capital, Resistencia, Galetto 382 (7) 58°56'48"W 
URU Uruguay. Dept. Colonia, Balneario 34725'31"8 
Astilleros, Speroni 5.п. (80) 57°33'59"W 
CULT-1 Cultivar. Argentina. Cérdoba Province, 31?19'12"S 
Dept. Capital, Argüello, Galetto 381 (3) 64°18'36"W 
CULT-2 Cultivar. Argentina. Córdoba Province, 31°22'12"5 
Dept. Capital, Alta Córdoba, Galetto 79 (2) 64?09'02"W 
CULT-3 Cultivar. Argentina. Córdoba Province, 31°27'00"S 
Dept. Capital, Observatorio, Galetto 380 (7) 64°12'01"W 


Zeiss Axiophot microscope, using Kodak T-max 
film, 100 ASA 

Pollen and ovule numbers from 15 randomly 
chosen flowers from population ARG-ER1 were 
counted (1 to 3 flowers from seven trees). All ten 
anthers belonging to a single preanthesal flower 
were softened in 1 N H 
temperature, transferred to a known volume of lac- 
tic acid: glycerin (3 : 1) in a test tube, and mac- 
erated with a glass rod. The mixture was homoge- 


or 12 hours at room 


nized using a vortex mixer, and a sample of known 
volume of this solution was placed in a hemocytom- 
eter. The grains within six randomly chosen squares 
were counted, and the total number of grains per 
flower was then calculated. The ovary was directly 
dissected under a Zeiss stereo microscope and the 
ovules counted. Pollen stainability, as an indicator 
of pollen viability, was calculated with aniline blue 
(1% mass/vol) in 100 grains per flower, correspond- 
ing to sampling for pollen/ovule. 

Experimental fieldwork was done from October 
to April for three reproductive seasons, 1993 to 
1996. In all cases, flowers in bud stage were tagged 
for identification. They were bagged using paper 
bags to prevent pollinator visits and nectar robbers. 
Flower longevity was determined in three popula- 
tions (ARG-ER1, URU, and CULT-1) for randomly 
chosen bagged flowers by following their develop- 
ment until they began to fade. 

Nectar was extracted with capillary glass tubes 
without removing the flowers from the plant, avoid- 
ing damage to the nectaries. Two variables were 
immediately taken: volume (in pl) using graduated 
micropipettes and sugar concentration (%, mass/to- 


tal mass) with a pocket refractometer (Atago, Ja- 
pan). The amount of sugar produced was expressed 
in milligrams and was calculated after Kearns and 
Inouye (1993). In Table 3, dates, population iden- 
tification, samples, individuals, and those flowers 
analyzed were recorded. Separation of sugars was 
by gas chromatography following methodology in- 
dicated in Bernardello et al. (1994). Sugar ratio (r) 
was calculated as sucrose/fructose + glucose (H. 
G. Baker & I. Baker, 1983) and the hexose ratio 
hr) as glucose/fructose. Nectar samples for sugar 
analysis were collected from the four anthesal stag- 
es for flowers, from different populations, and from 
different periods within the flowering season. Tests 
for amino acids, lipids, phenols, alkaloids, and re- 
ducing acids were also performed on nectar drops 
placed on Whatman #1 chromatography paper (H. 
G. Baker & I. Baker, 1975). A histidine scale was 
used to quantify amino acids (H. G. Baker & I. 
Baker, 1975). 

Nectar secretion pattern was determined during 
one reproductive season (1994—1995) in population 
ARG-ER1 (4 trees) and population CULT-1 (3 
trees), using eight bagged-flower sets of 5 to 8 flow- 
ers in each population. Data were taken once for 
each set, allowing the nectar to accumulate until 
the measurement. The studied period covered the 
four days of the flower lifetime. Measurements were 
performed twice a day (two untouched new sets 
were used each day, at 8:00 and at 19:00 hr re- 
spectively). 

The effect of nectar removal on total nectar pro- 
duction per flower was estimated on sets of 7 to 10 
flowers within a population and on the same trees 


— 


130 


Annals of the 
Missouri Botanical Garden 


Tee 


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У 
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Y E31 187 f Y Ё 

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BUENO $ AIRES 


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igur 
Victoria; A entina, Entre Ríos, Paraná; 
ULT-1 to in 


as for nectar secretion. Nectar was removed and 
measured from the same flower twice at 8:00 and 
19:00 hr throughout the first two days after flower 
opening. Set 1 was used as a control (flowers not 
yet involved within the resorption period). Thus, in 
order to compare the results accurately, sets were 
subjected to 1 to 4 measurements according to the 
scheme suggested by Torres and Galetto (1998). 
Data on the total mg of sugar produced by sets 1 


ano 


to 4 were compared by a one-way analysis of var- 
iance (ANOVA), at the 0.05 significance level. 

e standing crop of nectar was evaluated by 
measuring accumulated nectar (volume, concentra- 
tion, and mg of sugar in the way described above) 
from individual flowers that had been fully exposed 
to pollinators. Data were randomly collected from 
open flowers (anthesal stages 3 and 4) from Feb- 


1. ae P is studied populations of E. crista-galli. oe ARG-ERI = 
RG-Chaco = Arge 
іт а Argentina, Córdoba. For more details, see Table 


Argentina, Entre Ríos, 
U 


ns Chaco; URU = Uruguay, Colonia; 


ruary to March 1996, sampling seven trees (N — 
10 flowers/tree) from population ARG-ER1. 
Different hand-pollinating treatments were ap- 
plied to sets of l- or 2-day-old flowers (N = 10- 
19). Data were taken in зүр ARG-ER1 (8 
trees, Ее 996), роршаһоп ОКО 
995), 2 population CULT-1 (3 
= the number of flowers used is 
included in Tables 5-7. Flowers were treated as 
follows: (1) autonomous self-pollination (buds 
bagged without treatment); (2) autogamous crosses 
(hand-pollinated flowers with pollen from their own 
dehisced anthers); (3) geitonogamous crosses 
(emasculated hand-pollinated flowers with pollen 
from flowers of the same ramet); (4) xenogamous 
crosses (emasculated hand-pollinated flowers with 
pollen from flowers belonging to a different genet 


Volume 87, Number 2 
2000 


Galetto et al. 
Erythrina crista-galli 


131 


Table 2. Flowering, fruiting, and seed dispersal periods of Е. crista-galli. Data taken in November 1994 to April 
1995. 
F ri = 1 

жей (cites! period) Fruiting Seed dispersal 

Population Major Additional (dates/period) (dates/period) 
ARG-ERI 11/02-12/01 12/30–03/11 12/01–03/11 01/21-03/11 
URU 11/05-11/30 01/10-03/15 12/10-04/12 01/29-04/12 
CULT-1 to 3 11/10-12/08 01/09-03/06 12/20-03/27 01/20-03/27 


located at least 200 m away); (5) tests for apomixis 
(stigmas and anthers were clipped at flower open- 
ing); and (6) controls (flower sets were tagged and 
left open to flower visitors). 

Fruit set was recorded 6 to 7 weeks after treat- 
ment with the following features measured: fruit 
length and mass, total seed mass per fruit, seed 
number, mean seed mass, and percentage of seed 
germination. Seed samples were placed in Petri 
dishes lined with filter paper and watered regularly 
at room temperature for two months. To obtain the 
seed germination percentage, the germinated seeds 
were counted and divided by the number of seeds 
used. Pre-emergent reproductive success or PERS 
(Wiens et al., 1987) was calculated as (# of fruits 
obtained/# of treated flowers) X (mean # of seeds 
per fruit/mean # of ovules per flower). 

Flower visitors were identified during 1 to 3 flow- 
ering periods (1993-1996) in five populations 
(ARG-ER1, ARG-ER2, ARG-Chaco, URU, CULT- 
1) with an average of 49.8 hours of observation per 
population. Visitors were collected and/or photo- 
graphed for identification. Relative abundance was 
quantified in population ARG-ER1 on one day. The 
number of individual visitors and flowers visited 
were recorded. The inflorescences were observed 
for a total of 7 hr., in 30-min. periods beginning at 
06:00. All observations were made from a fixed spot 
from which 42 flowering branches of three trees 
could all be monitored. 

Fruit quality traits were compared by t-tests or 
one-way analyses of variance (ANOVA) combined 
with Bonferroni test at P < 0.05. Analogous non- 
parametric methods (Mann-Whitney, Kruskal- Wal- 
lis, and Wilcoxon rank tests) were used when the 
assumptions required for parametric methods were 
not met (Sokal & Rohlf, 1995). The statistical pro- 
gram package SPSS (1992) was used for these anal- 


yses. 
RESULTS 


PHENOLOGY 


This species displayed a major flowering burst 
in November for all the studied populations (Table 


2). Additionally, most of the trees presented one or 
two flowering episodes of lower amplitude in Jan- 
uary through March. This second flowering was 
very variable among individuals and even among 
branches of the same tree. Detailed analysis on 
population ARG-ER1 showed that, although there 
is a general synchrony in flowering, fruiting, and 
seed dispersal periods within each population, 
there were some differences among the trees at the 
beginning and the peak of flowering (Fig. 2). The 
observed phenology corresponds to the sub-annual 
frequency class of Newstrom et al. (1994), with at 
least two cycles per year seen in spring and sum- 
mer. Within the latitudinal boundaries of the study, 
this pattern appeared stable, although there were 
minor differences among the individuals within a 
population (Fig. 2). 


INFLORESCENCE AND FLOWER FEATURES 


Flowers in Erythrina crista-galli are resupinate, 
red, odorless, diurnally open, lasting for 3 or 4 
days. They were borne in racemes (pictured in Fig. 
3A) with a mean of 51.67 + 13.06 flowers per ra- 
ceme (N = 46, 17 trees, populations ARG-ER1, 
ARG-Chaco, CULT-1, CULT-2). Raceme axes are 
generally erect, but may be pendulous or interme- 
diate in position, principally on the lower branches. 
Anthesis proceeds acropetally and, as a result, 
there are many open flowers simultaneously in each 
raceme. 

The calyces are bowl-shaped and fleshy. The 
standard is broad, elliptic, and rigid; the wings are 
asymmetric, very small, and concealed by the ca- 
lyx, while the keel is falcate and acute at apex. 
Anthers and stigma may remain exposed at floral 
summit (Figs. 4, 5A). The androecium is pseudo- 
monadelphous with 10 stamens, arranged as 9 lon- 
ger + 1 shorter. The anthers аге dorsifixed and 
introrsely dehiscent. The ovary is stipitate and pu- 
bescent with a glabrous style and capitate, moist 
stigma (Fig. 5A). Flowers average 15.25 + 1.61 
ovules and 878,485 + 144,770 pollen grains (a 
pollen/ovule ratio of 57,605). Pollen viability is 
high (96.45% + 2.70). 


132 Annals of the 
Missouri Botanical Garden 
TREE # NOVEMBER | DECEMBER JANUARY FEBRUARY MARCH 
2 12 19 26 1 10 17 24 30 7 16 21 284 11 25 3 11 
1 1 1 | | | l 1 1 1 | | 1 | 1 1 1 
1 
2 
3 
4 
9 
6 
7 
8 
9 
pem e mo оно 


wwwwws Seed dispersal 


ure 2. Flowering, fruiting, and seed dispersal periods in E. crista-galli, taken for 10 trees of population ARG- 
ERI (November 1994 to March 1995). Dots indicate qualitative flowering peak. 


Floral ontogeny has been subdivided into four 
stages, as illustrated in Figure 4: (1) 4–5-ст prean- 
thesal bud ready to open, with the standard com- 
pletely folded and occluding floral parts (time: 0 
hr); (2) corolla standard beginning to unfold from 
its base, initiating the anthesis (first day, time: 3 to 
12 hr); (3) fully open flower, 5—6 cm long with stan- 
dard entirely spread (the degree of spreading of the 
standard is variable, reaching its maximum when it 
is perpendicular to the keel; 2nd and 3rd days, 
time: 24 to 36 hr); and (4) 3- to 4-day-old flowers 
beginning to fade (3rd and 4th days, time: 36 to 64 
hr) Flowers of most populations typically went 
through all these stages. However, many flowers 
from population АКС-ЕК], and many of population 


URU, remained in stage 2, i.e., the standard did 
not open spontaneously; in this case, the flower life- 
time was also 3 to 4 days. 


NECTARY 


A structural floral nectary is located in the re- 
ceptacle between stamens and ovary (Fig. 5), and 
is supplied by vascular bundles having both phlo- 
em and xylem (Fig. 6). It is a small ring, slightly 
divided at the apex into 10 lobules. Histologically, 
the external part of the ring is composed of 10 to 
14 layers of secretory tissue with small cells highly 
stained. In contrast, the internal ring portion is 
composed of 22 to 26 parenchymatous layers with 


Galetto et al. 


Volume 87, Number 2 133 
2000 Erythrina crista-galli 

Figure 3. Inflorescence and fruit photographs of E. crista-galli from pupa URU. —A. Partial view of an 
inflorescence. —B. Dehiscent fruit showing seeds. Both bars are equivalent tc 


larger, less stained cells, and the vascular bundles 
(Fig. 6). Stomata are found mainly in the upper part 
of ile lobules. The basal part of the fused staminal 
filaments delimits a nectariferous chamber where 
the exudate accumulates, protected from evapora- 
tion. 


NECTAR CHEMICAL COMPOSITION 


Flowers produced nectar with a mean sugar con- 
centration of 21.896 (Table 3). Nectar concentration 
was consistent among the different populations 


sampled (Table 3). Nectar sugars were glucose, 
fructose, and sucrose. Mean percentage of sucrose 
was lower than 296 or absent, whereas the per- 
centage of hexose sugars was higher than 9896, and 
always with more glucose than fructose (Table 3). 
All samples had amino acids in variable concen- 
trations (from traces to 1.9 mg/ml, i.e., 9 in the 
histidine scale). Samples from Uruguay (population 
URU) had the lowest amount, while the ones from 
population ARG-ER1 had the highest (Table 3). 
Phenols were always detected, whereas alkaloids, 
reducing acids, and lipids were not. In population 


134 


Annals of the 
Missouri Botanical Garden 


1cm 


е 4. Flower stages of E. crista-galli as indicated in the text. —1. Preanthesal bud. —2. Bud initiating anthesis 
(1- dud flower). —3. Open flower (2- to 3-day-old flower). —4. Old flower beginning to fade (3- to 4- -day-old flower). 


CULT-1, we analyzed the sugar composition 
throughout the flower lifetime (Table 4). The sugar 
proportions data obtained for the different flower 
ages also showed a remarkable constancy through- 
out flower lifetime (Table 


NECTAR PRODUCTION AND RESORPTION 


This was studied in two populations from differ- 
ent habitats: a natural one growing in a very humid 
environment (ARG-ER1), and a cultivated one from 


e 5. Open flower of Е. crista-galli. —A. чиш showing Der location (arrow head). —B. Flower 
andard k = = 


Fi 
transection at the nectary level. Abbreviations: с = calyx, s 
tary, о = ovary. 1-ст bar for А; 3-mm bar for B. 


rd, w = wing, k = keel, a = androecium, n = 


Volume 87, Number 2 Galetto et al. 135 
Erythrina crista-galli 


igure Optical е photomicrographs showing nectary structure іп Ё. crista-galli. —А. Flower abe 
longitudinal section. —B. Detail of the central nectary tissue with the к bundle indicated by arrow. —C. Flov 


partial transection at the level. of nectary base. Abbreviations: n = nectary, o = ovary 


136 Annals of the 
Missouri Botanical Garden 
Table 3. Nectar chemical саг и of Е. crista-galli. N = number of individuals (number of flowers) from which 


nectar was sampled. C 


ectar concentration (mass/total mass). HS = histidine scale to quantify amino acids 


G. Baker & I. Baker, 1975), ИЕ + С) = sugar ratio, G/F = hexose ratio, t = traces. 


Н. 


— 


Ratios 
Sugars (mean + sd; %, mass/mass) S/ 

Population Date N Conc. % HS Sucrose Fructose Glucose (F + G) G/F 
ARG-ERI Dec-94 1 (9) 220-11 8 1.0 = 0.2 31.5 + 2.8 67.5 + 3.0 0.009 2.14 
ARG-ERI Mar-95 2 (10) 21.4 + 0.7 6 0.0 41.9 + 1.1 58.1 = 1.1 0.0 1.41 
ARG-ERI Mar-96 3(13 195-08 8 0.2 + 0.1 36.0 + 1.1 63.8 + 1.0 0.001 1.77 
ARG-ER2 Nov-95 1(10 243+16 8 1.2 + 0.3 361+ 1.0 62.7 + 1.4 0012 1.74 
ARG-ER2 Dec-95 1(15) 21.5 + 12 9 0.0 36.0 = 2.6 64.0 + 2.6 0.0 1.78 
ARG-ER2 Jan-96 1 (7) 202-03 7 0.0 35.2 + 1.3 64.8 + 1.3 0.0 1.84 
АСЕ-Сћасо Јап-96 4(22) 225+04 6 0.9 = 0.1 30.7 = 0.7 684+0.7 0.009 2.23 
URU Dec-96 1 (10) 212 = 11 t 0.0 30.6 = 27 694 + 27 0.0 2.27 
URU Jan-95 1(15) 234+18 9 0.0 28.2 + 2.9 71.8 + 2.9 0.0 2.54 
URU Jan-95 1 (19) 199+1.1 1 0.0 374+ 1.1 626+1.1 0.0 1.67 
CULT-1 Dec-95 3 (12) 24.0 + 0.6 7 0.0 38.1 + L7 61.9 +17 0.0 1.62 
CULT-2 Jan-96 2 (16) 23.2 + 0.5 7-8 O1+01 369-22 63.0 + 21 0.001 1.70 
CULT-3 Feb-96 4 (17) 198 +09 7 0.8 + 0.5 35.1 = 0.3 63.9 + 0.6 0.008 1.82 
Mean of means 218+16 64 0.3+0.5 349+ 3.7 648+ 3.7 0.003 1.88 


a drier habitat (CULT-1). Most of the total nectar ЕК1, most of the nectar was secreted in bud stage 


was secreted by bud 


s. As soon as 


the flowers 


opened, nectar was available to pollinators (Fig. 7). 
Some differences were seen between the flowers of 
the two analyzed populations. In population ARG- 
ER1, the flower nectar volume was higher (Fig. 7A), 
the nectar concentration lower (Fig. 7B), and ac- 
cordingly, the amount of sugar secreted by a flower 
was comparable (Fig. 7C). With respect to nectar 
secretion pattern, concentration remained un- 
changed with a little decrease at the end of the 
flower lifetime in both populations. In contrast, 
some differences between the populations were 
found for flower nectar volume and sugar mass pat- 
terns. In population CULT-1, there was a short nec- 
tar cessation period after flower opening which last- 
ed for 12 hours, followed by a new nectar secretion 
period on the first night. Only then did the resorp- 
tion phase begin (Fig. 7A, C). In population ARG- 


CULT-1 


NECTAR REMOVAL 


mg/h 


(Fig. 7A, C). Then flowers showed two alternate 

short periods of resorption and secretion until the 

third day, whereupon a final resorption period be- 

gan (Fig. 7A, C). The nectar resorption rate was 

similar in both populations: —0.3 
3 


ulation ARG-ERI, 


r. in pop- 


1 mg/hr. in population 


Figure 8 shows the amount of sugar produced by 


sets subjected to different numbers of removals in 
the two studied populations. Very little or no nectar 
was removed at consecutive intervals (Fig. 8). One- 
way ANOVA results indicate that removal had no 
effect on nectar production, since there were not 
significant differences between the treated flower 
sets (population ARG-ERI: Fa 4, = 


2.22, 


Nectar sugar composition throughout flower lifetime for bagged flowers not visited by pollinators of E. 
(F + G) 


able 4. 
crista-galli. Data taken on sets of flowers (n = 


Hexose ratio — G/F. 


8 flowers each) from population CULT-1. Sugar ratio — S/ 


Sugars (mean = sd; 96, mass/mass) Ratios 

Flower stage Sucrose Fructose Glucose S/(F+G) G/F 
Bud 1.2 + 0.3 37.3 + 1.0 61.6 + 1.3 0.010 1.65 
lst day 0.0 34.5 + 1.5 65.9 + 0.5 0.0 1.9] 
2nd day 1.9 + 08 40.6 + 1.3 57.5 = 0.9 0.020 1.41 
3rd day 1.8 + 0.6 37.1 + 1.2 61.1 + 1.9 0.019 1.64 
4th day 1.7 = 12 35.4 + 0.9 62.9 + 2.3 0.017 1.77 
Mean of means 1.3 + 0.7 36.9 + 2.1 61.8 + 2.8 0.013 1.67 


Volume 87, Number 2 
2000 


Galetto et al. 
Erythrina crista-galli 


137 


200 


= = ule е В 
Е 
5 10) 
о 
> 
0 : 
: 15 27 39 51 63 75 87 
В « 
ја ] 
с 
.Q 
= 
iz 
Е 20) ---¢ ---@ ~~ и "ИД 
о + t---2 
6 
о 10- 
0 5 | 
3 15 27 39 51 63 75 87 


Sugar mass (mg) С) 


Flower lifetime ( h ) 


ure 7. Nectar production of Е. crista-galli through- 
out "he lifetime. Data correspond to nectar volume per 

flower (pl), sugar concentration in nectar 
and sugar mass (mg) per flower. oo ARG-ERI = 


(96. mass/mass). 


broken line, black circles; population CULT-1 = dips 
line, white circles. Plotted values represent ће mean 

l s.d. and correspond to a sample size of 5 to 8 bag sued 
flowers each. 


0.08; population CULT-1: КЕ, = 0.45, P = 0.13). 
Concomitantly, there were no significant differences 
in mean total amount of nectar secreted per flower 
at the population level (t = 1.36, P = 0.18). 


NECTAR STANDING CROP 


Flower average values of nectar standing crop 
were obtained on seven trees from population ARG- 
. Опе-мау ANOVA comparisons showed no 
significant differences among the sampled trees (mg 
of nectar sugars: Ке ал = 0.79, Р = 0.58; nectar 
volume: Ё, а = 0.24, Р = 0.96). Nectar data from 


open flowers significantly differed from that ob- 
tained from bagged 1- or 2-day-old flowers: volume 
(36.4 + 39.8 and 158.3 + 36.0, respectively; U = 
126.5, P < 0.00001) and milligrams of sugar (7.4 
= 8.6 and 31.1 = 7.8, respectively; U = 184, P 
< 0.00001). However, they did not differ with re- 
spect to concentration (17.7 = 5.8 and 18.3 + 1.4, 
respectively; U — 1543, P — 0.06). 


BREEDING SYSTEM 


Hand-pollination tests confirmed Erythrina cris- 
NS to be self-compatible (Table 5). Artificial 
autogamous crosses were more successful than gei- 
tonogamous or xenogamous ones (Table 5). How- 
ever, spontaneous autogamy very rarely occurred: 
data from two populations showed that less than 196 
of the flowers set fruit in this way (Table 5). Natural 
fruit set was always lower than fruit set from hand 
crosses, except for population АВС-ЕВ1 (Table 5). 
FRUIT AND SEED CHARACTERISTICS 

These legume pods are brown, arcuate, and co- 
riaceous to subligneous, dehiscing by the two mar- 
gins (Fig. 3B), and presenting 1 to 9 seeds. The 
seeds are brown mottled with black, dry, and hard. 
In spite of low natural fruit set (< 6%, Table 5), 
individuals often yielded thousands of seeds be- 
cause of the large number of both flowers produced 
per plant, and seeds produced per fruit (Table 6). 

Fruit set obtained by geitonogamous hand-polli- 
nation was the lowest and significantly different 
from autogamous hard-pollination, but not from 
fruit set obtained by hand cross-pollination (Table 
6). Reproductive success for geitonogamy was also 
the lowest but the differences from the other pol- 
lination treatments were not significant. Fruits ob- 
tained by hand-pollination treatments from popu- 
ation CULT-1 were compared in their quantity 
(Table 6). All variables showed significant differ- 
ences, except for seed number per fruit. Highest 


values for fruit length and mass, seed number and 
seed mass per fruit, mean seed mass, and seed ger- 
mination percentage were observed in xenogamous 

ruits and seeds (Table 6). In contrast, the lowest 
mean seed mass and the lowest seed germination 
percentage were seen for autogamous seeds (Table 


When unbagged-pollinated fruits were com- 
pared, natural populations were variable. Signifi- 
cant differences were found for fruit length and 
mass, seed number and seed mass per fruit, mean 
seed mass, and seed germination percentage (Table 
7). Fruits from cultivated trees (populations CULT- 
1 to 3) showed the highest values for most variables 


138 Annals of the 
Missouri Botanical Garden 


Nectar production per flower (mg) 
~ 
сл 


20- 
15- 
10 Population 
5 Ё сит-1 
| ee oe ris [. ]ARG-ER1 
1(4) 2(3) 3(2) 4(1=control) 
Flower sets (# of nectar removals performed) 
e 8. Histogram of nectar production per flower in sets (n = 5 to 8 flowers each) subjected to periodic nectar 


mui in two populations of E. crista-galli. Values represent the mean + 1 s.d. Divisions within bars e odes 
to the amount of nectar removed (mg) after each measurement. Set 4, with one removal, was the control. Abbr 
ations: АКС-ЕК] = Argentina, Entre Ríos, Victoria, natural population; CULT-1 — Argentina, Córdoba, ы 
cultivar. 


of fruits and seeds (i.e., fruit mass, seed number тепоріегапѕ and hummingbirds consistently visit- 
and seed mass per fruit, mean seed mass, and seed ed the populations of these trees in most of the 


germination percentage; Table 7). areas sampled (Table 8, Fig. 9B-H). The most com- 
Germination percentages of seeds from open-pol- mon insects were carpenter bees (Xylocopa nigro- 
linated fruits were similar to those obtained for ex- cincta, Fig X. ordinaria, Fig. 9D, and other 


perimental autogamous fruits, but were consider- Xylocopa) as well as honeybees (Apis mellifera, Fig. 
ably lower than those of geitonogamous and 9G, H). Four species of hummingbirds were noted 
xenogamous fruits (Tables 6, 7). (Table 8), with Hylocharis chrysura and Chlorostil- 
bon aureoventris (Fig. 9B) most frequently observed. 
One species of perching birds (the epaulet oriole, 

Animals visiting the flowers for nectar were doc- Emberizidae, Icterus cayanensis, Fig. 9A) was oc- 
umented only during daylight hours. Visitor activity casionally seen in population ARG-ER2. Flies, 
was noted as low or absent in rainy, windy condi- butterflies, wasps, ants, and beetles were occasion- 
tions, or during the heat of midday (> 30°C). Hy- ally seen as incidental flower visitors (Table 8). 


VISITORS 


Table 5. Fruit sets by hand and naturally pollinating treatments in E. crista-galli. Numbers in parentheses give the 
number of flowers treated. 


Population 
Total 
Treatment ARG-ERI URU CULT-1 fruit set 
Autogamy (by hand) 0.024 (166) 0.108 (37) 0.308 (120) 0.139 (323) 
Geitonogamy (by hand) 0 (104) 0.086 (35) 0.100 (80) 0.050 (219) 
Xenogamy (by hand) 0.026 (115) 0.081 (37) 0.197 (76) 0.092 (228) 
Autonomous self-pollination 0.006 (452) no data 0.014 (145) 0.008 (597) 
Natural pollination 0.126 (174) 0.036 (2683) 0.132 (1245) 0.054. (4102) 


Volume 87, Number 2 
2000 


Galetto et al. 139 
Erythrina crista-galli 


Table 6. Reproductive success and fruit and seed parameters from different iw Se AA treatments in Е. crista- 


galli from population CULT-1. Except for fruit set and reproductive success, data are means + s.d 


Lowercase letters 


as superscripts indicate a posteriori test results. Reproductive success = (# of "йй ae “ treated flowers) Х 
(mean # of seeds per fruit/mean # of ovules per flower), ns = not significant, ** = P < 0.01 0.001. 


*= P< 


Variable Autogamy Geitonogamy Xenogamy Statistical tests 
ruit set (flowers treated) 0.308: (120) 0.100" (80) 0.197» (76) x? = 10.76** 
Fruit length (cm) 178 + 2.9* 14.4 + 3.5 22.8 + 1.9 Fre, 36, = 13.39*** 
Fruit mass 25309 2.2 + 1.0 4.3 + 0.6^ Fro, за 10.29**9 
Total seeds per fruit mass (2) 1.2 + 0.6" 1.1 + 0.68 2.0 + 0.7 Fou = 4.79** 
Оуше пштђег 15.1 + 2.0 14.0 + 0.8 13.3 + 1.3 Fre зв = 1.73 ns 
Seed number per fruit 4.5 + 1.8 3.3 = 1.6 5.9 = 2.1 Е». зы = 3.12 ns 
Mean seed mass (mg) 26.3 + 6.2 334 = 11.3 34.2 = 10.1 Fo зы = 6.08** 
Seed germination (%) 49.50 + 17.2* 88.2 + 12.0 OS Set г Fo, а = 12.58*** 
Reproductive success 0.092 0.023 0.087 = 4.38 ns 


Monkeys (Cebidae, ee caraya) were observed 
in population AGR-Chaco breaking and eating 
flowers. Although фм ue these actions may 
produce some accidental pollinations. 

Visitor frequency was recorded only for popula- 
tion ARG-ER1. Almost 93% of the flowers regis- 
tered were visited by hymenopterans (42.9 and 
49.9% for honeybees and carpenter bees, respec- 
tively) with the remainder by hummingbirds 
(7.2%). Considering flower number of a tree visited 
by a single individual, carpenter bees and hum- 
mingbirds averaged a mean of 5 flowers per tree 
(4.7 + 2.9 and 5.0 + 2.5, respectively). By con- 
trast, honeybee visits were more numerous, aver- 
aging over 21 flowers per tree (21.3 + 7.5 

Hummingbirds and bees can be considered ef- 
fective pollinators, because they contacted both an- 
thers and stigma while taking nectar (Fig. ЭВ-С). 
However, we did not compare their effectiveness. 
Regardless of the flower visitor, the average time 
elapsed in each visit was 10 to 30 seconds per 
flower (range: 1—50 seconds). They usually visited 
many open flowers of several inflorescences in each 
tree before going to the next one. The remaining 


visitors were nectar thieves, because they did not 
contact both fertile whorls due to their small size. 
In population ARG-ERI, holes made by unknown 
primary nectar robbers were detected; these holes 
were used by small wasps to take nectar. 

Our observations indicate that hummingbirds 
were widespread visitors while orioles were rare 
and local, but bees were even more frequent than 
hummingbirds. Hummingbirds usually foraged in 
the upper branches of the trees, taking nectar by 
inserting their bill between the standard and keel 
while touching anthers and stigma with their heads 
(Fig. 9B). Chlorostilbon aureoventris showed terri- 
torial behavior in population ARG-ER2 

Carpenter bees displayed a particular behavior 
recorded for all populations of Е. crista-galli. They 
usually forced open preanthesal flowers (stage 2, 
Fig. 4) with their proboscide. This was of signifi- 
cance in the flowers of populations ARG-ERI and 
URU that did not display natural flower opening. 
When these insects visited the flowers, they first 
opened them moving downward between the stan- 
dard and the keel. Then, while looking for the nec- 


Reproductive characters from open-pollinated experiments in E. crista-galli populations. Nd = no data. 


able 
Data represented by mean = s.d. Lowercase letters as superscripts indicate a posteriori test results. *** — P « 0.001. 


Population Population Population 
ARG-Chaco CULT-1 to 3 
Variable (n = 33) (n = 40) (n = 35) Statistical tests 
Fruit length (cm) 20.6 + 42: 16.7 + 12 18.0 + 3.0% Ер on = 716*** 
Fruit mass (g) 1.6 = 0.8 Ма 29 + 10 t = 5.6*** 
Total seeds per fruit mass (g) 0.7 + 0.4 Nd 1.6 + 0.7 | = 7.49% 
Ovule number 16.9 + 1.7 15.2 + 1.4 t = —4.3*** 
Seed number per fruit 3.4 + 1.6 2.4 + 2.6" 4.5 + L8' Еро = 1223*9* 
Seed mass (mg) 21.7 + 5.5" 26.6 + 8.3" 38.5 + 8. Коло = 92.63*** 
Seed germination (46) 37.0 = 22.5 52.1 + 15.0 t= 2.70% 


140 


Annals of the 
Missouri Botanical Garden 


isitors of Ё 


Figure Floral v 
B. не [Chlorostilbon а aureoventris (D'Orbigny & 
ith 


(Xylocopa 

CULT-1. 
populations URU phe CUL 
Entre Ríos, Paraná; URU 


nigro- DA icta 


tively. The 1-ст ы in 
= Uruguay, Colonia; CUI 


I-1, respect 


tar, they touched the anthers and the stigma with 
their back, transferring the pollen. 

Apis mellifera (Fig. 9G, H) was one of the most 
frequent visitors in all the populations, mainly dur- 
ing peak blooming. They occasionally forced prean- 
thesal flowers. Honeybees are the only pollinators 
that collected pollen, in addition to nectar, mainly 
early in the morning from 7:00 to 9:00 hr. Landing 
on the apex of the androecium, they collected pol- 
len, moving around the anthers and touching the 
stigma with their pollen-laden heads and forelegs 
(Fig. 9G). Then, they moved downward in the flower 


rom population URU. —D. Cz 
mide be зе (Bombus morio Swed) from population CULT-1 


ta-galli. —A. Oriole |/стетих cayanensis (Viellot)] from pp ARG-ER2. — 
.ULT-1 


Lafresnaye)| from population ( Carpenter bee 
arpenter bee didis pa ordinaria Said) i rom population 
—G, H. Honeybee ee mellifera L. from 
1 Cis сс. Abbreviations: ARG-ER2 = Argentina, 
= Argentina, Córdoba, Argüello, cultivar. 


valid for € 


to take nectar (Fig. 9H). The average time they 
spent in each flower was 4 to 10 seconds. 


DISCUSSION 


The flowers of Erythrina crista-galli present a 
nectary type characteristic of Papilionoideae (e.g., 
Fahn, 1979; Davis et al., 1988, and references 
therein). Nectar secretion possibly occurs via the 
modified stomata of the nectary through which nec- 
ar 


— 


ows. The presence of stomata is a common 
feature of floral nectaries of legumes (Davis et al., 


Volume 87, Number 2 
2000 


Galett al. 
Es is pow. 


141 


Table 8. Flower visitors observed at E. crista-galli during the day. 


Population 


ARG-ERI 


ARG- 
ARG-ER2 Chasd URU Mor 


Sample 1 


3 4 5 6 7 


Visitors Hours (days) 18 (6) 


7(0 4 (10 


4 (2) 105(3) 60(10) 18 (6) 


Trochilidae 
Chlorostilbon aureoventris 
(D'Orbign 
Hylocharis eis (Shaw) 
Heliomaster furcifer (Shaw) 


Leucochloris albicollis (Viellot) 


y & Lafresnaye) 


= ж 


Hymenoptera 
Anthophoridae 
Xylocopa nigro-cincta Smith X 
Xylocopa ordinaria Smith 
Xylocopa sp. 
Apidae 
Bombus morio Swed 
Apis mellifera L. Х 
Vespidae 
Polybia scutellaris White 
Polybia sericea Olivier 
Brachygastra lecheguana Latreille X 
Vespoidea (n° species 
Formicidae (n° species) 3 
Lepidoptera 
Pieridae (n° species) 3 
Diptera 
Muscidae (n° species) 1 
Syrphidae (n° species) 3 
Coleoptera 


Diabotrica speciosa Germ. X 


= Xx 


~ ж 
~ ~ 
= ж 


рб < 


» ~ 


1988, and references therein). Studies on the пес- 
tary stomata of Vicia faba L. demonstrated that their 
major functions seem to be those of assisting nectar 
escape from the gland and perhaps enhancing re- 
absorption of uncollected nectar (Davis & Gunning, 
1993). Erythrina crista-galli secreted dilute and 
hexose-dominant nectar, with a remarkable con- 
stancy in its sugar proportions within the sampled 
area. Nevertheless, when comparing our data with 
previous studies, some differences appear in the 
nectar sugar composition. I. Baker and H. G. Baker 
(1979) noted traces of melezitose and maltose, 
whereas Gottsberger et al. (1984) found a much 
higher amount of sucrose (ca. 4596). This would 
indicate that there is some intraspecific variation in 
nectar sugar proportions, particularly when data 
from the northern Brazilian population are consid- 


ered (22°45'S, 48°25'W; cf. Gottsberger et al., 


m 
O 


With respect to nectar secretion pattern, there 
were some differences between the analyzed pop- 
ulations. Habitats are assumed to play a role in the 
differences found in nectar secretion between the 
populations, since trees from Entre Ríos (popula- 
tion ARG-ER1) live near the Paraná river, a more 
humid habitat than that of trees from Córdoba (pop- 
ulation CULT-1). The fact of presenting most nectar 
at flower opening is unusual and unknown for other 
Erythrina species. As far as we know, it had been 
recorded only for some Indian bird-pollinated Lor- 
anthaceae (Davidar, 1983) and for the South Amer- 
ican bee-pollinated Mandevilla pentlandiana (A. 
DC.) Woodson (Apocynaceae, Torres & Сајено, 
1998). On the other hand, the observed decrease 


142 


Annals of the 
Missouri Botanical Garden 


in nectar volume and solute in 3-day-old bagged 
flowers can be inferred as resorption, based upon 
the constancy of nectar concentration throughout 
anthesis. There are some species of other families 
with flowers that last 4 days, with a similar nectar 
secretion-resorption pattern, and which are visited 
by a great variety of flower visitors (Combretaceae: 
Bernardello et al., 1994; Loranthaceae: Rivera et 
al., 1996; Apocynaceae: Torres & Galetto, 1998). 

Most previous data on E. crista-galli report or 
suggest bird pollination, either by hummingbirds 
(Knuth, 1906; Schrottky, 1908; Werth, 1915; Pick- 
ens, 1931; Ali, 1932; Gottsberger et al., 1984) or 
by hummingbirds and passerine birds (Toledo, 
1974; I. Baker & H. G. Baker, 1982; H. G. Baker 
& I. Baker, 1983, 1990). In particular, orioles have 
been frequently recorded as visitors and pollinators 
of Erythrina species from the Americas (Feinsinger 
et al., 1979; Morton, 1979; Toledo & Hernández, 
1979). In contrast, Hymenopterans have been rare- 
ly mentioned as possible visitors for the genus (To- 
ledo & Hernández, 1979) or for E. crista-galli (Ali, 
1932; Faegri & van der Pijl, 1971; Duncan in Ra- 
ven, 1974; Parodi, 1978). Monkeys have been re- 
ported elsewhere as visitors in a few New World 
Erythrina taxa. Jaeger (1961) saw howling monkeys 
on Erythrina species from Paraguay, and Janson et 
al. (1981) indicated that flowers of two species from 
Peruvian forests were an important food source for 
monkeys. 

Studies on flowers and their animal visitors have 
led to the assumption that there are coevolutionary 
relationships between nectar traits and pollinator 
type. Several authors have discussed nectar volume 
and concentration of flowers attracting different 
guilds of pollinators. In general, they found that 
hummingbird and honeyeater flowers present large 
amounts of dilute nectar, especially relative to nec- 
tars of bee flowers (e.g., Baker, 1975; Pyke & Was- 
er, 1981, and references therein; Cruden et al., 
1983; Opler, 1983). Nevertheless, and under lab- 
oratory conditions, hummingbirds, honeyeaters, and 
sunbirds given a choice of sugar solutions have 
been found to prefer the highest sugar concentra- 
tions offered at an equal volume presentation 
(Hainsworth & Wolf, 1976; Stiles, 1976; Tamm & 
Gass, 1986; Mitchell & Paton, 1990). On the other 
hand, studies on the sugar composition of nectar 
showed that hummingbird flowers produce a su- 
crose-dominated nectar while those associated with 
passerine birds produce nectar dominated by glu- 
cose and fructose (H. G. Baker & I. Baker, 1983, 
1990). Sugar preferences of hummingbirds have 
been examined in previous studies, which have 
found that they preferred sucrose solutions instead 


of equivalent monosaccharide solutions (Stiles, 
1976; Martínez del Río, 1990a; Stromberg & John- 
sen, 1990). Physiological studies on New World 
nectarivorous passerine birds have shown a corre- 
lation between the sucrose aversion with a relative 
enzymatic lack of sucrase activity (Martínez del Río 
et al., 1988, 1989; Martínez del Río, 1990b). How- 
ever, recent studies on Old World passerine birds 
have shown that they possess high efficiency in su- 
crose absorption, and Old World counterparts do 
not reject sucrose in favor of hexose sugars (Downs 
& Perrin, 1996; Lotz & Nicolson, 1996; Downs, 
19972, b). 

In addition to nectar traits, red flowers can be 
largely considered as bird-adapted because insects 
have little or no sensibility to red (e.g., Proctor et 
al., 1996; Vogel, 1996). However, recent literature 
suggests that red flowers are not invisible to bees 
as previously thought (Chittka & Waser, 1997). In 
particular, for Papilionoideae a conspicuous UV re- 
flectance/absorbance patterning was demonstrated 
for mellitophilous species but not for ornithophilous 
ones (Kay, 1987). Because flowers of E. crista-galli 
lack UV patterns and showed little or no UV re- 
flectance (Kay, 1987), red flowers of this species 
should be seen not only by hummingbirds but also 
by carpenter bees, the most frequent flower visitor. 

The two woody subfamilies Caesalpinioideae and 
Mimosoideae and primitive woody tribes of the 
Papilionoideae have mostly maintained self-incom- 
patibility, whereas self-incompatibility has been 
lost on numerous occasions in herbaceous legumes 
(Kalin Arroyo, 1981). It is possible that its loss 
occurred as a consequence of the peculiarities of 
tripping in papilionoid flowers, because this mech- 
anism permits the maintenance of high levels of 
outcrossing in many self-compatible legumes (Kalin 
Arroyo, 1981). Nevertheless, the level of outcross- 
ing will be primarily correlated with the size of 
flower display, enhancing the possibilities for gei- 
tonogamous selfing. In large plants such as trees, 
much selfing will result from inter-flower transfer- 
ence of pollen. 

Previous data on some species indicate that both 
self-compatibility and self-incompatibility are pre- 
sent in Erythrina (cf. Kalin Arroyo, 1981; Neill, 
1988). According to Fryxell (1957), E. crista-galli 
is self-incompatible, but later reports (Raven, 1974; 
Neill, 1988), confirmed herein, indicated that it is 
self-compatible. According to Cruden and Lyon 
(1989), facultatively xenogamous species are self- 
compatible, adapted for cross-pollination, present- 
ing a single flower type, with delayed autogamy, and 
most show high fruit and seed set. Facultative xe- 
nogamy is considered a mixed mating system in 


Volume 87, Number 2 
2000 


Galetto et al 


\ 143 
Erythrina crista-galli 


which the level of pollinator activity is the primary 
determinant of the balance between self- and cross- 
pollination (Cruden & Lyon, 1989). Facultative xe- 
nogamy has been correlated with a moderately high 
pollen/ovule ratio (i.e., ca. 700; Cruden, 1977). Al- 
though E. crista-galli presented a pollen/ovule ratio 
(> 57,000) that coincides with Cruden’s (1977 
class for obligate xenogamy, we considered this 
species as facultatively xenogamous because it is 
self-compatible, the flowers are adapted for cross- 
pollination, and both pollinator behavior and inflo- 
rescence size affect selfing (i.e., mainly geitonoga- 
my). Empirical evidence suggests that numerous 
plant species have evolved a mixed reproductive 
strategy (Richards, 1997). An evolutionarily stable 
strategy can develop for mixed mating, because self- 
ing as well as outcrossing can confer fitness benefits 
onto offspring (Holsinger, 1991). 


~ 


Pollinators preferentially visit large inflorescenc- 
es because they provide a larger signal to attract 
them and/or offer greater pollen and nectar rewards 
(e.g., Stephenson, 1979; Augspurger, 1980). When 
a plant displays numerous flowers simultaneously, 
geitonogamy may occur if the pollinator moves be- 
tween flowers. Although plant attractiveness to pol- 
linators often increases with the number of flowers 
open at one time, display size also bears mating 
costs to the plant. Aspects of floral design and dis- 
play that mitigate the mating costs of autogamy but 
that promote the benefits of enhanced pollinator at- 
traction may be widespread, given that most ani- 
mal-pollinated plants display several to many flow- 
ers each day (Harder & Barrett, 1996). Some 
deductions on the E. crista-galli reproductive strat- 
egy can be made when fruit set as well as fruit and 
seed parameters from hand- and natural-pollina- 
tions are compared, particularly when these results 
are related to inflorescence size, to the number of 
simultaneously open flowers per tree, and to the 
foraging behavior of the flower visitors. The possi- 
bility of regularly occurring natural selfing would 
explain the low natural fruit set and low seed ger- 
minability observed іп Ё. crista-galli. That quan- 
titative traits from fruits and seeds obtained by nat- 
ural pollination are more similar to those from 
selfed flowers than from crossed ones can be relat- 
ed to significant pollen flow within a tree mediated 
by the flower visitors. Results indicate that E. cris- 
ta-galli shows reproductive plasticity, which allows 
this species to reproduce successfully despite var- 
iation in the quality and quantity of the pollinator 
guild. 

Cladistic analyses of Erythrina based on mor- 
phological and cpDNA characters suggest that Е. 
crista-galli has a basal position and lies sister to 


the remaining species within the genus (Bruneau, 
1996, 1997). Within the primarily entomophilous 
tribe Phaseoleae, there is some agreement that Er- 
ythrina is bird-pollinated (Raven, 1974, 1977; Bru- 
neau, 1997). Most Erythrina species in the Amer- 
icas are pollinated by hummingbirds, although 
perching birds are also common visitors and pol- 
linators of several taxa (Neill, 87; Bruneau, 
1997). In an article on the coevolution of birds with 
Erythrina (Bruneau, 1997), E. crista-galli was in- 
cluded within a basal clade with E. fusca Lour. and 
E. falcata Benth. with this clade characterized as 
passerine/hummingbird-pollinated. However, for Е. 
crista-galli, we observed not only birds, but also 
bees as significant visitors and pollinators. 


CONCLUSIONS 


Erythrina crista-galli, like the other species in 
the genus (Neill, 1988), is self-compatible, showing 
low fecundity and high rates of flower and fruit 
abortion. Its showy red flowers produce hexose- 
dominant nectar and are assiduously visited pri- 
marily by hymenopterans (carpenter bees and hon- 
eybees) and also by hummingbirds (four species), 
all of which can be assumed as pollinators. In nat- 
ural populations, several traits related to pollinator 
behavior, e.g., high number of simultaneously open 
flowers per tree, high nectar production per flower, 
would favor pollen flow within trees. If this is true, 
a reproductive consequence such as inbreeding de- 
pression could occur. An outcome of this situation 
might be the low quality of selfed seeds compared 
to seeds derived from outcrossed flowers. 

Knowledge of directional changes in pollination 
and breeding systems can provide valuable clues 
for reconstructing phylogenies (Kalin Arroyo, 
1981). As the genus Erythrina seems to be wholly 
self-compatible (Neill, 1988), information on shifts 
in pollinators may be valuable. This highly derived 
genus was previously considered to be ornithophil- 
ous within the primarily entomophilous Phaseoleae 
(Doyle & Doyle, 1993; Bruneau et al., 1995). Cla- 
distic analyses in Erythrina included E. crista-galli 
in the basal clade (Bruneau, 1997). However, our 
data indicate that flowers were primarily visited by 
bees. This may indicate that the pollinators for this 
basal clade have departed from entomophily, typi- 
cal of Phaseoleae, to ornithophily, more typical of 
Erythrina. Further studies on the reproductive bi- 
ology and frequency of visitors are needed on the 
other members of this basal Erythrina clade for 
confirmation of this. Our results suggest that the 
pollination biology of the Erythrina species is com- 
plex, that the species have a wide array of visitors, 


144 


Annals of the 
Missouri Botanical Garden 


and that more detailed field studies are needed to 
clarify their evolution. 


Literature Cited 


Ali, S. А. 1932. Flower-birds and bird- = in India. 
J. Bombay Nat. Hist. Soc. 35: 573—60: 

Mass- eae of a tropical 
shrub (Hybanthus prunifolius); Influence on pollinator 
attraction and movement. Évolution 34: 475—488. 

Baker, H. G. 1975. Sugar concentrations in nectars from 
hummingbird flowers. Biotropica 7: 37 

— а 1. Baker. 1975. Studies of nec барара инн 
and pollinator-plant Pulp ues Pp. 126-152 in L. E. 
Gilbert & P. H. Rave DN abes of Animals and 
Plants. Columbia Uni p д ‚ New York. 

& —— — ‚А brief PA review of the 
eps of Ru nec tak Pp. 127-152 in B. Bentley 

S. Elias, The pene of ie "laries. C nbn Univ. 
ee New Yor 


. The predictive value of nectar 

chemistry to the Ки of pollinator types. Israel 
Bot. 39: 157-166. 

Baker, I. & H. G. Baker. 1979. Chemical constituents of 
the nectars of two Erythrina species and their hybrid. 
Ann. Missouri Bot. Gard. 66: 446—450. 

1982. Some chemical constituents of 
floral nectars of Erythrina i in Dow to pollinators and 
systematics. Allertonia 3: 25— 

Bernardello, L., L. Galetto & I. E одначе, 1994. Re- 

productive biology: variability of nectar features, and 
pollination of Combretum fruticosum een in 
Argentina. pa J. Linn. Soc. 114: 293-3 

Bruneau, A. 1996. Phylogenetic and biogeographical pat- 
terns in a (Leguminosae: oleae) as in- 
ferred from morphological and a DNA char- 
acters. Syst. Bot. 21: 587—605. 

1997. Evolution and homology x bird pollina- 

tion ijndmines à in Erythrina. Amer. J. 54-71. 

. J. Doyle. 1993. Cladistic ipd of chlo- 
roplast DNA A restriction site characters in Erythrina (Le- 

guminosae: Fhaseolae), Syst. Bot. 18: 229-247. 

. L. Doyle. 1995. Phylogenetic re- 

lationships of Pinselen Evidence from Жа 

DNA restriction e characters. Pp. 309—330 ; à 

‚ А deg in Legume Sy ~ 

. Royal Botanic Gardens 

бый. А. 1987. Flora stad de Entre Rfos (Argen- 
tina). Tomo VL Parte III: cales a Rosales (incluso 
Leguminosas). Colección нем del INTA, Buenos 

ires. 

Chittka, L. & N. M. Waser. 1997. Why red flowers are 
not invisible to pe là РІ. Sci. 45: 169-183. 
Conn, H. w & V. M. Emmel. 1960. Stain- 
ing Procedures. Willan & Wilkins, Baltimore. 
Cruden, R. W. 1 Pollen-ovule ratios: A conservative 
indicator of тае systems in flowering plants. Еуо- 

lution 31: 32-46. 


— 


D. L. Lyon. 1989. Facultative ПА 
amination of а mixed mating system. Pp. 171—207 in i 
H. Bock & Y. B. Linhart, The van een Ecology of 
Plants. а Press, Boulder, Colorado. 

— S. M. Hermann & $. Peterson. atterns 
of nectar production and pilin: us convulution. 
Pp. 80-125 in B. Bentley & T. S. E X Biology of 
Nectaries. Columbia Univ. Press, New 

Davidar, Р. 1983. Similarity between fe a fruits in 


some flowerpecker pollinated mistletoes. Biotropica 15: 
32-37. 


Davis, A. R. & B. E. S. Gunning. 1993. The modified 
stomata of the floral nectary of Vicia faba L. 3. Physi- 
ological aspects, including co вы with foliar sto- 
mata. Bot. Acta 106: 241.253 

— ———, R. L. Peterson & В. W. Shuel. 1988. Vasculature 
and marae ‘ture of the floral and Fue (rein of 
Vicia faba (Leguminosae). Canad. J. 66: 14 

48. 


Downs, C. T. 1997a. Sugar digestion efficiencies of Gur- 
ney's sugarbirds, malachite sunbirds, and black sun- 
birds. Physiol. Zool. 70: 93-99, 

— 1997b. Sugar preferences and apparent sugar 
assimilation in the red lory. Austral. J. Zool. 45: 613- 
619. 


M. R. Perrin. 1996. Sugar preferences of some 
southern African nectarivorous birds. Ibis 138: 455- 
459. 


Doyle, J. J. & J. L. Doyle. 1993. Chloroplast DNA phy- 
logeny or the papilionoid legume tribe Phaseoleae. Syst. 
Bot. 18 —327. 

Faegri, K. & L. van der Pijl. 1971. The Principles of 
Pollination Ecology, 2nd ed. Pergamon Press, Oxford. 
Fahn, A. 79. Secretory Tissues in Plants. Academic 

Press, London. 
a P., Y. B. Linhart, A. Swarm & J. A. Wolfe. 
979. Aspec ts of the pollination biology of three Ery- 
дна species on Trinidad and Tobago. Ann. Missouri 


1957. Mode of reproduction in higher 
Bot. Rev. 25: 135-233 


ant 
Gottsberger, G., J. Schrauwen & H. Е. Linskens. 1984. 
А ; 


Hainsworth, F. R. & L. L. Wol Nectar character- 
istics and food selection by hummingbirds. Oikos 25: 
101-113 

Harder, L. D. & S. C. H. Barrett. 1996. Pollen dispersal 
and mating patterns in animal- -pollinated plants. Pp. 
140-190 in D. G. Lloy . H. Barrett, Floral Bi- 
ology: Studies on Floral Evolution’ in Animal-Pollinated 
Plants. Chapman & Hall, New Yor 

Holsinger, K. E. 1991. Mass-action models of plant mat- 
d gia The evolutionary m of mixed mating 

s. Amer. Naturalist 138: 606 

i P 1961. The Wonderful Life e Fio. Harrap, 
London. 

Janson, C. H., J. Terborgh & L. H. Emmonds. 1981. Non- 
flying mammals as detis agents in the Amazonian 
forest. aps 13 (Supp 

Johansen, D. 1940. Plant o Й McGraw Hill, 
New York. 

Kalin Arroyo, M. T. 1981. Breeding Рада and polli- 
nation biology in vica mi Pp. 723-771 in R. M. 
Polhill & P. H. Raven, Advances in Legume System- 
atics 2. Royal Badio: Ganda ens, Kew. 

Kay, Q. O. N. 1987. Ultraviolet heus and ultravio- 
let-absorbing pigments in flowers of the Leguminosae. 

3 in C. H. Stirton, Advances in Legume 
Systematics 3. Royal Botanic Gardens, Kew. 

Kearns, C. A. & D. W. Inouye. 1993. ike cud for 
Pollination Biologist s. Univ. Press of Colorado, Niw 

Knuth, P. 1906 ` 


Flowers by Insects,” 


Vol. 1. Clarendon: Press, Oxford. 


Volume 87, Number 2 
2000 


Galetto 


al. 145 
nma р. 


Krukoff, B. A. & В. C. Ватеђу. 1974. А gus of 
the genus Erythrina. Lloydia 37: 332-4 

Lotz, C. N. & S. W. Nicolson. 1996. sun preferences 
of a nectarivorous passerine bird, the lesser double-col- 
а, нв (Nectarinia chalybea). Funct. Ecol. 10: 


У oes y? Río, 1990a. Sugar preferences in hum- 
mingbirds: The nw menm of subtle d cal differences 
on food choice. Condor 92: 1022-103 

—. | Dietary, ie навија 
сасне of intestinal т ап т 
birds. as шы 63: 9 : 

. В. ‚ D. E Da aneke & P. T. Andreadis. 

1988. Physiological ‘comebies of preference and aver- 

sion for sugars in three species of birds. Physiol. Zool. 

61: 222-229. 


ad ecolo gical 
altase activity in 


arasov & D. J. Levey. 1989. Physiolog- 
ical hus and ecological consequences of sugar pref- 
erences in es dre Auk 106: 64—71. 

Mitchell, R. J. & D. C. Paton. —— Effects of nectar 
volume and concentration on sugar intake rates of Aus- 
tralian honeyeaters (Meliphagidae). Oec ologia 83: 238— 
2 


Morton E. S. 1979. Effective pollination of Erythrina fus- 
a by the мек oriole (Icterus spurius): Coevolved be- 
Баха! manipulation? Ann. Missouri Bot. Gard. 66: 


482 . 

Neill, D. A. 1987. Trapliners in the trees: Hummingbird 
pollination of Erythrina sect. Erythrina (Leguminosae: 
Papilionoideae). Ann. Missouri Bot. Gard. 74: 27-41 

—_. "xperimental studies on species relation- 
ships 1 in Erythrina (Leguminosae: Papilionoideae). Ann. 

d. 75: 886—969. 


Newstrom, L. E., С. W. Frankie & Н. С. Baker. 1994. А 
new classification for plant phenology based on flower- 
ing patterns in lowland tropical rain forest trees at La 
Selva, Costa Rica. Biotropica 26: 14 £s 

Opler, P. 1983. Nectar production in a tropic ; 
Pp. 30-79 in B. Bentley & T. S. Elias "ће Biology ol 

ia Univ. Pena. New Yor 

Parodi, 1978. Enciclopedia Argentina de Agricul- 
tura 1 шенин» 3“ ed., Tomo I. Editorial ACME, 

Buenos Air 

Pickens, А |. “1931. 
dor 3 

Proctor, ers Yeo & A. J. Lack. 1996. The Natural 
apes of PE Timber Press, Portland, Oregon. 

Pyke, G. H. & N. M. Waser. 1981. The production of 
dilute nectars by н апа т кым flowers. 

27( 


Some flowers visited by birds. Con- 


974. Ery do о Achievements 
opportunities. Lloydia 37: 321-3: 


. 1977. Erythrina (Fabaceae: erring Intro- 
duction to D II. Lloydia 40: 401-4 

Richards, A. J. 1997. Plant Breeding Systems. о ИА 
& Hall, Cambridge. 

Rivera, G. L., L. Galetto & L. Bernardello. 1996. Nectar 
secretion pattern, removal effects, and breeding system 
of Ligaria cuneifolia (оосо села. Canad. J. Вог. 74: 

2001. 


3-242 i in C. 
nces in Бренан Bi- 
ology. Monogr. Syst. Bot. Missouri ih Gard. 29. 
Schrottky, C. 1908 

Wiss. Insekten-Biol. 4: 22-26, 3-78. 

Sokal, R. R. & F. J. Rohlf. 1995. pom W. H. Free- 

San Francisco. 

‚. 1992. SPSS for Windows: 29 system user's 

uide, Release 5.0. SPSS Inc., Chica 

Stephenson, A. G 79. An ct pne d examination of 
the floral display of gag speciosa (Bignoniaceae). 
Evolution 33: 1200-12 

Stiles, F. G. 1976. Taste nere color pre. 
and flower wwe " humm s. Condor 78: 10-26 

Stromberg, M P. В. musca ummingbird 
sweetness ien Taste or viscosity? Саодат 92: 

+612. 

Tamm, S. & C. L. Gass. 1986. Energy intake rates and 

r concentration preferences by hummingbirds. 
бега ТО: 20—23. 

Toledo, У. М. 1974. Observations on the relationship be- 
tween oe and Erythrina species. Lloydia 37: 
482-487 

& H. M. Hernández. 1979. Erythrina oliviae: А 

new case of oriole pollination in Mexico. Ann. Missouri 

—511. 


›. 1998. Patterns and implications 

of foes! nectar secretion, chemical composition, remov- 

: effects, and standing crop in Mandevilla pentlandi- 
na ы a Bot. J. Linn. Soc. 127: 207-223 


Via iana, M. J. dioi ecológicos de Scolytidae — 
me ic ntomol. Argent. 27: 119-130 
Vogel. S. 1996. C Eu Konrad Sprengel's rua of the 


flower: The cradle of floral ecology. Pp. 44—62 in D. 

Lloyd & S. C. H. Barrett, Floral Biology: Studies on 

Floral Evolution in Animal-Pollinated Plants. Chapman 
sa New Yor 


Werth, 1915. | em Überblick vo p Gesamtfrage 
доли Вог. Jahrb. 53: 378. 

Wiens, D., ( I Calvin, С. A. t А rn, D. 
Frank . R. Seavey. 1987. Reproductive success, 
usn embryo abortion, and genetic load in flow- 
ering plants. Oecologia 71: 501-509. 


THE LONG-PROBOSCID FLY 
POLLINATION SYSTEM IN 
SOUTHERN AFRICA! 


Peter Goldblatt? and John C. Manning? 


ABSTRACT 


Orchidaceae, across southern Africa. Lo 


in the genera Prosoeca, Moegistorhynchus, and Stenobasipteron (Nemestrinidae) and Philoliche (Tabanidae). Flies in the 


A close association between the form and color 
of flowers and pollination by a particular pollinator 
is well known. Convergence in floral morphology 
among species that rely on the same pollinator class 
led to the recognition of floral syndromes (e.g., Vo- 
gel, 1954; Faegri & van der Pijl, 1979). Moreover, 
species with morphologically similar flowers that 
share the same pollinator species constitute a par- 
ticular pollination guild, an extension of the term 
describing a group of species that exploit the same 
class of resources in a similar way (Root, 1967). 
Likewise, insect species using a particular group of 
plants as a food resource in a similar way may also 
be regarded as a guild. A guild is thus a functional 
unit independent of taxonomic considerations, as 
are floral syndromes. Although a number of polli- 
nation syndromes have been identified in the south- 
ern African flora (Vogel, 1954), relatively few guilds 
have been described. The most striking of those 
that have been documented is the association be- 


tween the satyrid butterfly, Aeropetes (Meneris) tul- 
baghia, and late-summer-flowering species with 
large, bright red blossoms (Johnson & Bond, 1994). 
Others include the association of several plant spe- 
cies with magenta to violet-colored flowers and the 
fly Prosoeca peringueyi (Manning & Goldblatt, 
1996), the suite of plant species with cream to pale 
pink flowers blooming in autumn that depend on P. 
longipennis for their pollination (Manning & Gold- 
blatt, 1995), and the Moegistorhynchus longirostris 
guild of the west coast of South Africa. 
Pollination by long-proboscid flies is a relatively 
unusual phenomenon, first recorded in southern Af- 
rica by Marloth (1908) and later described in some- 
what more detail by Vogel (1954). In their review 
of insect pollination in the Cape Flora of South Af- 
rica, Whitehead et al. (1987) were the first to really 
recognize long-proboscid fly pollination as a unique 
pollination system, although very little was then 
nown about either the flies or what plant species 


his research was supported by National Geographic 


Society Grants 4816-92, 5408-95, and 5994-97. We grate- 


T 
fully acknowledge the work of B.-E. van Wyk, Rand Afrikaans University, Johannesburg, who provided the analyses of 
sugar nectars. We also thank Peter Bernhardt and Dee Paterson-Jones for helpful comments during the preparation of 


the paper, and Mervyn Lotter and Cameron and Rhoda McMaster for their help and hospitality in the fielc 
* B. A. Krukoff Curator of African Botany, Missouri Botanical Garden, РО. Box 299, St. 1 


0229, U.S.A 


0015, Missouri 63166- 


* Compton Herbarium, National Botanical Institute, Private Bag X7, Claremont 7735, South Africa. 


ANN. Missouni Вот. Савр. 87: 146-170. 2000. 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


147 


they pollinated. Thus, long-proboscid fly pollination 
has only been regarded as a class of pollination 
system comparable with that of the major pollinator 
groups or syndromes, bird, bat, bee, butterfly, and 
muscid/carrion fly, since the late 1980s. Muscid/ 
carrion fly pollination, also called myophily, is clas- 
sically associated with actinomorphic, bowl- or sal- 
ver-shaped flowers of pale or dull colors, readily 
accessible nectar, and well exposed sex organs 
(Faegri & van der Pijl, 1979). Pollination by Dip- 
tera is, in fact, diverse and cannot be usefully re- 
garded as a single pollination syndrome. Sapro- 
myophily is already distinguished from general 
myophily because the flies in the syndrome have 
lapping mouth parts, are attracted by unpleasant 
odors of decay or fermentation, and are associated 
with flowers with dull colors, often with mottled pig- 
mentation, and hairy or frilled petals or tepals. 
Long-proboscid fly pollination (or rhinomyophily, 
viz. Rebelo et al., 1985) may readily be distin- 
guished from classical myophily and its specialized 
derivative, sapromyophily, and differs in all criteria 
enumerated by Faegri and van der Pijl for this my- 
ophily. Long-proboscid fly flowers are typically zy- 
gomorphic, normally have an elongate floral tube, 
and have bright coloration. The sex organs are pre- 
sented in a wide range of orientations, sometimes 
concealed within or above the mouth of the tube, 
or are elongate and held distant from the source of 
the nectar reward in a unilateral, arcuate (adaxial) 
or declinate (abaxial) disposition. 

Although pollination by long-proboscid flies has 
been described from various parts of the world, the 
system as we define it here is restricted to the Hi- 
malayan Region (Fletcher & Son, 1931; Dierl, 
1968) and southern Africa, where the system has 
received a fair amount of attention since 1990 (e.g., 
Goldblatt et al., 1995; Manning & Goldblatt, 1996, 
1997; Johnson & Steiner, 1995, 1997). So-called 
long-proboscid fly pollination described in the lit- 
erature, for example, by Grant and Grant (1965) in 
California, refers to bombyliid flies with probosces 
less than 15 mm long and differs from the polli- 
nation system we regard as long-proboscid fly pol- 
lination. Here we review long-proboscid fly polli- 
nation, report additional examples of pollination by 
long-proboscid flies, and assess the importance of 
the system in the southern African flora relative to 
other pollination systems. 


REVIEW OF LONG-PROBOSCID FLY POLLINATION 
LONG-PROBOSCID FLIES 


Definition characteris- 
tics. We define long-proboscid flies here as those 


and morphological 


insects that have mouth parts at least 15 mm long 
and a body length of more than 15 mm. Fifteen 
species in two families, Nemestrinidae and Taban- 
idae, are known to have mouth parts this long, 14 
of them restricted to the southern African region, 
Lesotho, Namibia, South Africa, and Swaziland 
(Fig. 1, Table 1), and one to the Himalayas (Dierl, 
1968). Adult morphs of these flies depend largely 
or exclusively on floral nectar for their nutrition and 
are avid foragers of nectar-rich flowers (female Ta- 
banidae also require a blood meal). Their visits to 
the flowers of some plants result in the passive ac- 
cumulation of pollen or pollinaria as they brush 
against anthers, and in turn, the passive transfer- 
ence of pollen or pollinaria to stigmas during visits 
to other flowers of the same species. Most other 
Nemestrinidae and Tabanidae have substantially 
shorter mouth parts and, although they also feed on 
nectar and pollinate plants, they are not known to 
be the only pollinator(s) of any plants. Instead they 
share pollen resources with other insect taxa in- 
cluding long-tongued bees, Lepidoptera, hopliine 
beetles, and bee flies (Bombyliidae) (Goldblatt et 
al., 1995, 1998b, in prep.). The tabanid, Philoliche 
aethiopica, and the acrocerid flies, Psilodera spp. 
(Goldblatt et al., 1997; Potgieter et al., 1999), have 
mouthparts of intermediate length, mostly 12-15 
mm long, and they are provisionally excluded from 
consideration here: their shorter mouthparts pre- 
vent them from foraging effectively on flowers of 
plant species that have exclusively long-proboscid 
fly pollinators. 

Long-proboscid flies are large-bodied insects, 
typically measuring 15—24 mm from the tip of the 
abdomen to the base of the proboscis. Mouth parts 
are as long as, or often substantially longer than, 
the insect's body, the most extreme example bein 
Moegistorhynchus longirostris—individuals along 
the Cape west coast have been recorded with pro- 
bosces up to 100 mm long (Fig. 2). Foraging be- 
havior is similar in all species, irrespective of fam- 
ily or genus, and although flies have been described 
as hovering while foraging (Struck, 1997), this is 
not the usual pattern. Our observations show that 
flies firmly grasp tepal or petal lobes or other floral 
organs as they forage for nectar and while doing so 
they continue to vibrate their wings rapidly (Gold- 
blatt et al., 1995; Goldblatt & Manning, 1999) (Fig. 
3A-D). 

Foraging patterns vary, but our observations 
show that long-proboscid flies are seldom flower 
constant. While flies sometimes forage for a time 
on a particular floral form and may visit a particular 
species more frequently than any other, more often 
their foraging appears to be random, and foraging 


148 


Annals of the 
Missouri Botanical Garden 


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Figure 1. 
—1. 


The Prosoeca peringueyi guild. —2 


Southern Africa, showing ri geographical extent of each of the three о fly pollination guilds. 
e Moegistorhynchus—Philoliche guild (shaded). . The 
glbaueri guild. The ranges of guilds 1 and 2 correspond almost exactly to the bens un winter-rainfall zone 


Note the limited geographic overlap that does not coincide temporarily. 


bouts may include flowers of several different spe- 
cies, some of which even have different shapes, siz- 
es, and colors. This emphasizes that long-proboscid 
flies can and do forage on a wide range of flowers. 
Long-tubed flowers, however, are the only ones that 
offer a secure reward, and because of their size, a 
significantly larger reward, and one that is not 
available to most other nectarivorous insects. 


Zoogeography. The southern African long-pro- 
boscid flies have variable geographic ranges (Table 
1). All but one of the species are, however, restrict- 
ed to one or the other of the two major climatic and 
biotic zones of southern Africa, the winter-rainfall 
zone in the southwest, and the summer-rainfall 
zone, which covers the rest of the subcontinent (Fig 
1). Prosoeca ganglbaueri has the widest range and 
extends from the Northern Province of South Africa 
through eastern southern Africa to the Kleinswart- 
berg in the south (at the interior edge of the winter- 
rainfall zone), a distance of over 1500 km. In con- 
trast, and despite pollination research in the area, 
P. rubicunda is known from one specimen from the 
southwestern Cape, while P. nitidula is restricted 
to the Cape Peninsula at the extreme southwestern 


edge of the subcontinent and the center of the win- 
ter-rainfall zone. Evidently also rare, Moegistorhyn- 
chus sp. is known from two high-mountain sites in 
the southwestern Cape. 


PLANT SPECIES 


Plants that depend on long-proboscid flies for 
their pollination comprise a varied group taxonom- 
ically and morphologically (Table 2). They range 
from seasonal perennials, mostly geophytes with 
corms, bulbs, or tubers (Amaryllidaceae, Iridaceae, 
Orchidaceae, some Geraniaceae), to s ri- 
caceae, some Geraniaceae, Lamiaceae, Scrophular- 
iaceae). No annuals or trees have so far been found 
with this pollination system. 


Floral characteristics. Flowers of most species 
have in common a long floral tube (Figs. 4, 5) (we 
use the term here to include a corolla or perianth 
tube, tepal spurs of orchids, as well as the recep- 
tacular tube of Pelargonium), usually exceeding 20 
mm, and usually produce ample nectar. Notable ex- 
ceptions are Aristea spiralis (Iridaceae) and species 
of Brunsvigia and Nerine (Amaryllidaceae), which 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


149 


Table 1. 


Data are taken from 


*Note 


synonyms of M. longirostris. 


rainfall zone. that we ard M. b 


m our observations, the literature, and museum collections. So little is known 
cunda that its inclusion in the Moegistorhynchus—Philoliche guild is tentative. Taxonomic affiliation: Nemestrir 
Moegistorhynchus, Prosoeca, | Tabanidae—Philoliche. WRZ = 


Proboscis length, peak months of activity, and geographic ranges of long-proboscid flies in southern Africa. 


about Prosoeca rubi- 


1 
паас 


winter-rainfall zone, SRZ = summer- 


raunsii and M. perplexus (each known only from their type ж л" as 


Proboscis length Months on 
Fly species nge mm ( the wing Geographic range 
Prosoeca peringueyi guild 
P. peringueyi (15-)25—40 (15) July-Sep. WRZ: S Namibia to N Western Cape 
P. sp. nov. 32-48 (8) Aug.—Sep. WRZ: Northern Cape (Calvinia District) 
Moegistorhynchus—Philoliche guild 
M. longirostris* (35—)42-80 (14) Sep.-Nov. WRZ: Western and Northern Cape coast and 
interior 
M. sp. nov. ca. 21 (1) Јап. WRZ: Western Саре 
Philoliche gulosa 18-33 (9) Sep.-Nov. WRZ: Western Cape, mainly interior 
> rostrata 21-27 (7) Oct.—Nov. WRZ: Western Cape, coast and interior 
Prosoeca nitidula 18-28 (5) Oct.—Jan. WRZ Western Cape (only Cape Peninsula) 
P. rubicunda ca. 21 (1) Jan. WRZ: Western Cape (only Caledon District) 
Prosoeca ganglbaueri guild 
P. ganglbaueri (17-)25-42 (12) Jan.-Apr. SRZ & WRZ: Northern Province to Е Western 
Cape incl. Lesotho 
P. longipennis 38-40 (3) Mar.- Apr. WRZ: Southern Cape 
P. robusta 20–46 (7) Feb.—Apr. SRZ: Mpumal 
Stenobasipteron wiedmannii 18-30 (5) Feb.-Apr. SRZ: Mpumalanga and KwaZulu-Natal 


have floral tubes less than 10 mm long. Species 
with short floral tubes have elongate stamens so that 
the body of a foraging fly will brush against anthers 
even when its proboscis much exceeds the length 
of the tube 

Flower colors fall into two major groups (Gold- 
blatt et al., 1995; Manning & Goldblatt, 1996, 
1997). In northern Western Cape Province and Na- 
maqualand flowers are typically intensely dark red 
to purple or violet with pale nectar guides (Fig. 4). 
In the rest of southern Africa, however, these colors 
are rarely associated with long-proboscid fly flow- 
ers. Instead, flowers are usually shades of cream to 
pink, with pink to red nectar guides (Fig. 5). A few 
species may have pale blue or mauve flowers, but 
species of Nivenia (Iridaceae) are exceptional in 
having deep blue perianths. Again with few excep- 
tions, the nectar guides consist of longitudinal 
streaks. Flower form is usually zygomorphic and 
bilabiate with unilateral, arcuate stamens and style 
(Figs. 4G—L, 5F, С) or with declinate stamens and 
style (Figs. 40-Е, 5В-Е). A few species of Erica, 
Hesperantha, Ixia, Nivenia, Romulea, and one of 
Lapeirousia have actinomorphic flowers (Figs. 4A— 


Anthers and pollen are often unconventionally 
pigmented and frequently match the color of the 


perianth, or are simply a dull blue-gray to mauve 
(Goldblatt et al., 1995). Some Gladiolus and Triton- 
ia species with cream flowers, however, have dark 
purple pollen (Goldblatt & Manning, 1999). It has 
been suggested that unusually colored pollen is an 
example of crypsis, making the pollen less con- 
spicuous to pollen-collecting insects (Manning & 
Goldblatt, 1996). In Pelargonium species, anthers 
and pollen are bright red to orange and may con- 
tribute to the floral signal to flies (Goldblatt et al., 
1995), especially if the petals are weakly marked. 

Scent production is rare (Table 2). In the Irida- 
ceae only Babiana sambucina has strongly scented 
flowers, while B. framesii often has lightly fragrant 
flowers. In the Amaryllidaceae, species of Bruns- 
vigia typically have a light sweet fragrance. 

А particularly notable aspect of long-proboscid 
fly pollinated plants is the range of stamen orien- 
tation and length, which are directly related to the 
site of deposition of pollen on the body of a fly. 
Actinomorphic flowers have symmetrically dis- 
posed stamens either held close to the mouth of the 
floral tube, or in Erica, within the tube. In Aristea 
spiralis the anthers are held 16-20 mm from the 
vestigial floral tube. Nivenia species either have the 
anthers held at least 10 mm from the mouth of the 
tube, or in the distylous N. argentea and N. steno- 


150 


Annals of the 
Missouri Botanical Garden 


1ст 


Figure 2. Moegistorhynchus longirostris, the fly with 
the longest proboscis of all southern African a 
cid flies. The tongue measures between 35 and 100 mm 
long. Note the scattering of pollen over the dorsal part of 
the fly’s body. 


siphon stamens of the pin (long-styled and short- 
stamened) morph are held just above the apex of 
the floral tube. In zygomorphic flowers, the stamens 
are unilateral and either arcuate (arching above the 
mouth of the tube), as in most Iridaceae, or decli- 
nate (arching below the mouth of the tube), as in 
Pelargonium (Fig. 40-Е), Geissorhiza (Fig. 5B), and 
the amaryllids Brunsvigia and Nerine. In the latter 
two genera, the filaments are extended forward for 
35—40 mm, so that the anthers are held at least this 
distance from the nectar source. Stamen length is 
notably variable in Pelargonium, species of which 
have anthers held close to, or up to, 15 mm from 
the mouth of the floral tube. Pollen deposition on 
foraging flies is dorsal in species with arcuate sta- 
mens and ventral in species with d 
When anthers are held close to the mouth of the 


eclinat 


floral tube, pollen deposition is on the frons and/or 
the base of the proboscis. 


COMPARATIVE POLLINATION ECOLOGY OF THE 
THREE GUILDS 


SUBDIVISION OF THE LONG-PROBOSCID FLY 
POLLINATION SYNDROME 


We propose recognizing three separate pollina- 
tion guilds or systems within the long-proboscid fly 
pollination strategy. The flies that belong to each 
system show little or no overlap with those of other 
systems in plants visited or season on the wing, 
although there is some overlap in geographic range 
(Fig. 1). Within each system there are well-defined 
guilds of plant species that have one or occasionally 
two fly species as their sole pollinator. A few plant 
species may be pollinated by different fly species 
in different parts of their ranges. Thus, Lapeirousia 
silenoides is pollinated only by Prosoeca peringueyi, 
but L. jacquinii may be pollinated by P. peringueyi 
or, on the Bokkeveld Plateau, by Prosoeca sp. nov. 
Likewise, L. fabricii has been recorded as pollinat- 
ed by Moegistorhynchus longirostris at some sites, 
by Philoliche gulosa at others, and by both flies at 
one site (Manning & Goldblatt, 1997). The occa- 
sional presence of two long-proboscid fly species at 
a few sites has also been documented by Goldblatt 
and Manning (1999) both in Western Cape and 
Mpumalanga Provinces, but this appears to be a 
relatively infrequent situation. 

The Prosoeca peringueyi guild includes two fly 
species, P. peringueyi and Prosoeca sp. nov. (Table 
1). With complementary ranges in southern Namib- 
ia and Namaqualand to the Olifants River Valley 
and the Western Karoo in South Africa (Fig. 1), 
these two flies are active from July to late Septem- 
ber. The range is thus restricted to the western half 
of the southern African winter-rainfall zone and ac- 
tivity to the cooler months of the year. The two flies 
pollinate separate suites of species with similar flo- 
ral presentation, but at least the widespread Babi- 
ana framesii, two varieties of B. sambucina, and 
Lapeirousia jacquinii share both flies as their sole 
pollinators (Goldblatt et al., 1995; Manning & 
Goldblatt, 1996). 

Plants in the guild (Table 2) stand out in having 
flowers intensely pigmented in shades of magenta, 
deep purple, or violet, the lower, or all the petal or 
tepal lobes in the case of actinomorphic flowers, 
with cream to yellow markings and areas of darker 
pigmentation (Fig. 4). Tepal lobes are often rela- 
tively small, 12-15 mm long (e.g., Lapeirousia јас- 
quinti, L. silenoides, Romulea hantamensis), but 
some species of Babiana belonging to the guild 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


have relatively large flowers with tepal lobes up to 
28 mm long. Anthers and pollen are often shades 
of mauve to violet or cream. 

The floral tube in most species of the Prosoeca 
peringueyi guild is at least 20 mm long, and usually 
in the 30-45 mm range. All members of the guild 
flower in the late winter to late spring, late July to 
mid September, and are geographically restricted to 
the southern African west coast and near interior 
(Namaqualand and the northwestern Cape, South 
Africa, and southwestern Namibia). Some plant 
species in the guild have somewhat wider ranges 
than their pollinators and today reproduce sexually 
only by autogamously produced seed in the parts 
of their ranges where no pollinating flies occur (e.g., 
Lapeirousia jacquinii, Sparaxis metelerkampiae) 
(Manning & Goldblatt, 1996; Goldblatt et al., in 
prep.). 

2. The Moegistorhynchus—Philoliche guild, 
named for the most conspicuous fly genera in the 
system, includes six or seven fly species. The two 
tabanids, P. rostrata and P. gulosa, have the widest 
ranges, collectively extending from southern Na- 
mibia to the southeastern Cape (Fig. 1). The three 
Moegistorhynchus species and the one or two Pro- 
soeca species of the system have narrower ranges 
in the southwestern Cape (Table 1). Flies of this 
system are mostly active from late spring to early 
summer, mid September to November, but are still 
on the wing until January locally at higher eleva- 
tions. Species pollinated by this group of flies most- 
ly have a similar floral presentation, and the par- 
ticular pollinator depends on geography. The same 
species may be pollinated by up to three different 
flies over its entire range. Although the geograph- 
ical ranges of the Pr. peringueyi and Moegistorhyn- 
chus—Philoliche systems overlap in western south- 
em Africa, they overlap very little in period of 
activity of fly species and, as far as is known, not 
at all in plants pollinated. 

Plants in the Moegistorhynchus—Philoliche guild 
mostly have flowers in shades of white to cream, 
usually with pink undertones, or pale to deep pink 
(Fig. 5). Nectar guides are deep pink or red, oc- 
casionally with a white streak in the center of each 
dark mark. On the southern African west coast, 
flowers often have relatively large tepal lobes, 30— 
50 mm long (e.g., Gladiolus angustus, G. undulatus, 
Pelargonium longicaule), but Babiana tubulosa, 
Lapeirousia anceps, and P. appendiculatum have 
fairly small, inconspicuous tepal or petal lobes, up 
to 15 mm long. The longest floral tubes (60-110 
mm long) are found in west coast populations of G. 
angustus, L. anceps, and P. appendiculatum, and 
these species are associated exclusively with a sin- 


gle pollinator, Moegistorhynchus longirostris. There, 
this fly has a proboscis at least 65 mm long, where- 
as in central Namaqualand, to the north, it has a 
proboscis only 37-45 mm long. Populations of G. 
angustus and L. anceps elsewhere have shorter 
tubes and are pollinated by Philoliche species, 
which have correspondingly shorter probosces. 

Several plant species from the Roggeveld and 
Hantam areas of the western Karoo including Ba- 
biana spathacea, Disa karooica, and Romulea syr- 
ingodeoflora (white-flowered race), as well as sev- 
eral species of Pelargonium, all of which have 
cream to white flowers with red markings and elon- 
gate floral tubes, present a problem in the context 
of long-proboscid fly pollination. Although they 
bloom from October to early December, when Phil- 
oliche gulosa and P. rostrata are on the wing, and 
have flowers that appear to be adapted for polli- 
nation by long-proboscid flies, no long-proboscid 
species have been captured or recorded in this 
area. Can the legitimate pollinator(s), whether one 
of these Philoliche species or another fly species, 
be extinct locally, as suggested by S. D. Johnson 
(pers. comm.)* 

Too little is known about the Western Cape spe- 
cies, Prosoeca rubicunda, to confidently place it in 
one of the long-proboscid fly guilds, but its appar- 
ent period of activity, January, suggests that it may 
be a local extension of the Moegistorhynchus—Phil- 
oliche guild, which is centered in Western Cape 
Province. Several plant species from the Caledon- 
Bredasdorp area of the winter-rainfall zone that 
flower in the summer, from November to January, 
including Nivenia concinna, N. stokoei, and Trito- 
niopsis flexuosa (Iridaceae) and Pelargonium cau- 
califolium (Geraniaceae), have no known pollinator 
although they have flowers that conform closely to 
the long-proboscid fly type. 

3. The Prosoeca ganglbaueri guild, named for the 
most widespread and common fly species in the 
system, operates in the southern summer and au- 
tumn, from January to April, and largely in eastern 
southern Africa in areas of summer precipitation 
(Fig. 1), but also in the eastern half of the winter- 
rainfall zone that receives appreciable summer pre- 
cipitation. As treated here, the P. ganglbaueri guild 
includes four fly species (Table 1). While P. gan- 
glbaueri extends from Northern Province southward 
to the Kleinswartberg Mountains in Western Cape 
Province, P. longipennis is restricted to the southern 
Cape coastal belt. Because of the opposed season- 
ality there is virtually no overlap in plant species 
with the P. peringueyi and Moegistorhynchus—Phil- 
oliche systems. The ranges of the flies Prosoeca lon- 
gipennis and P. ganglbaueri, in the south of its 


Annals of the 
Missouri Botanical Garden 


Figure 3.  Long-proboscid flies foraging on their flowers. —A. Prosoeca peringueyi hovering above a cluster of 
Lapeirousia silenoides flowers. —B. Moegistorhynchus аи about to forage on a flower of Lapeirousia anceps. 


Volume 87, Number 2 
2000 


Goldblatt & Manning 153 


Long-Proboscid Fly Pollination 


range, overlap the eastern half of the range of flies 
belonging to the Moegistorhynchus—Philoliche 
guild, but the months of activity of the flies in the 
two systems differ. 

Guilds of plant species pollinated by Prosoeca 
ganglbaueri, P. longipennis, and P..robusta overlap 
to some extent. For example, populations of Trito- 
niopsis revoluta occur within the ranges of P. gan- 
glbaueri and P. longipennis; Disa amoena pollinaria 
have been found on both Р ganglbaueri and Pr. 
robusta, and both flies have been captured while 
foraging on Gladiolus calcaratus and Watsonia 
wilmsii. Likewise, P. ganglbaueri and P. longipen- 
nis have been observed and captured pollinating 
Pelargonium dipetalum in different parts of its 
range. No overlap has so far been encountered in 
the species pollinated by Stenobasipteron wiedman- 
nii, and further investigation may show this fly rep- 
resents a guild separate from the Р. ganglbaueri 
guild of flies. 

Flowers pollinated by flies in the Prosoeca gan- 
glbaueri guild are usually pink with dark pink to 
red markings, but Zaluzianskya microsiphon has 
cream petals, pink on the reverse, and Gladiolus 
calcaratus has white flowers. Nivenia stenosiphon is 
unusual in having actinomorphic, uniformly deep 
blue flowers, at least to the human eye. Thus, in 
general flowers pollinated by flies of the P. gan- 
glbaueri guild resemble those pollinated by flies of 
the Moegistorhynchus—Philoliche guild. Species 
pollinated by P. longipennis, a fly restricted to the 
southern Cape coastal belt, have flowers with rel- 
atively small petal or tepal lobes, mostly ca. 10 mm 
long, and a corolla or perianth that ranges from 
cream to pale pink or salmon, usually with darker 
pink or red on the lower (Gladiolus, Tritoniopsis), 
or upper (Pelargonium), or all the lobes (Cyrtanthus 
leptosiphon) (Manning & Goldblatt, 1995). Flowers 
pollinated by Stenobasipteron wiedmannii аге 
shades of pale blue, mauve, or pink (Goldblatt & 
Manning, 1999; Potgieter et al., 1999) and usually 
have small tepal ог petal lobes, but species роШ- 
nated by P. ganglbaueri and P. robusta often have 
large lobes. 


FLORAL REWARD—NECTAR 


Nectar volume. Nectar production is usually 
ample, and quantities mostly range from 1.1 to 5 


Ш per flower in a standing crop (unbagged flowers) 


(Table 3). Large-flowered species like Gladiolus an- 
gustus and С. undulatus may produce up to 10 pl 
of nectar per bloom. Aristea spiralis is unusual in 
having flowers that offer less than 0.5 р of nectar. 
Disa draconis, D. harveiana, and D. oreophila, Pel- 
argonium sericifolium, and Hesperantha scopulosa 
produce no nectar (Table 3) and evidently depend 
for their pollination on deceit, their flowers closely 
resembling those of other members of their respec- 
tive pollination guilds (Goldblatt et al., 1995; John- 
son & Steiner, 1995, 1997; Manning & Goldblatt, 
997) 


— 


Nectar sugar chemistry. In species of the In- 
daceae, Lamiaceae, and Orchidaceae, nectar is su- 
crose-rich to sucrose-dominant (ratio of sucrose to 
hexose sugars greater than 1—see Table 3), but 
nectar in most species of Pelargonium examined is 
hexose-rich or hexose-dominant. Nectar sugar con- 
centrations are mostly in the 20-30% sucrose 
equivalent range, but may be as low as 19% (Ro- 
mulea hantamensis) or as high as 38% (P. incras- 
satum) (Table 3). Two species of Hesperantha, H. 
grandiflora and H. galpinii, also produce nectar of 
unusually low sugar concentration, less than 18% 
sucrose equivalents. 


POLLEN PLACEMENT 


Pollen placement on the body of a fly appears to 
be an important consideration in long-proboscid fly 
pollination systems. At all sites that we have in- 
vestigated, the number of plant species utilizing a 
particular long-proboscid fly for their pollination 
appears to be closely correlated with the number 
of pollen deposition sites (Manning & Goldblatt, 
1996, 1997). Typically the frons and base of the 
proboscis, the dorsal part of the head and thorax, 
and the ventral part of the thorax and abdomen are 
used by different plant species for pollen deposition 
(Fig. 6). In the case of some Orchidaceae, pollinaria 
are usually deposited near the base of the proboscis 
(Johnson & Steiner, 1995, 1997). The need for spe- 
cific sites for pollen or pollinarium deposition is 
presumably related to the behavior of long-probos- 
cid flies, which are not flower constant. Instead, 
they visit flowers of different species in an appar- 
ently random pattern on foraging bouts (Goldblatt 
et al., 1995; Goldblatt & Manning, 1999). The de- 


position of more than one pollen species at the 


ae 


— С. Prosoeca sp. nov. about to insert its proboscis into the long perianth tube of Lapeirousia oreogena. —D. Philoliche 
rostrata about to forage on Tritonia flabellifolia; note the heavy deposit of purple Tritonia pollen on the dorsal part of 


the thorax 


154 Annals of the 
Missouri Botanical Garden 


Table 2. Floral characters of plants pollinated by long-tongued flies arranged by guild. Abbreviations: b = blue 
with light throat; cr = pale cream with red markings; p = pale pink or deep pink with red or purple nectar guides; v 
= vivid (red, purple, or violet shades with contrasting pale nectar guides); wh = white. Scent indicated by — = absent; 
+ = present; ++ = present and strong. For tube length we give only the functional length, not total length—in B. 
curviscapa, В. dregei, and В. framesii, all acaulescent species, the lower part of the tube is closed and only serves to 
raise the flower above the foliage. Additional species inferred on the basis of floral жөн to belong to шаш 
guilds, but with no insect visits recorded, are listed in parentheses. References, column eight, are as follow = 
Goldblatt et al. (1995) and Manning & Goldblatt (1996); 2 = Manning & Goldblatt (1997) 3 — Goldblatt & sabe 
(1999); 4 = Goldblatt et al. (1999); 5 = Johnson & Steiner (1995); 6 = Johnson & Steiner (1997); 7 = Struck (1997); 
8 — Manning & Goldblatt (1995); 9 — Vogel (1954); 10 — Goldblatt & Bernhardt (1990); 11 — Potgieter et al. (1999); 
12 = Goldblatt & Manning (new data; for methodology see Goldblatt et al. (19982), Goldblatt & Manning (1999)). 


~ 


Flower 


Mouth Floral 


Sym- 
Species metry Color Scent Pollinator partt mm tube mm Reference 


Prosoeca peringueyi pollination system 


Geraniaceae 
Pelargonium 
echinatum Curtis z wh = Pr. peringueyi n/a n/a 7 
incrassatum (Andr.) Sims 2 у = Pr. peringueyi 28-33 30-40 1 
magenteum van der Walt 7 v -= Pr. peringueyi 30-35 33-39 1 
sericifolium van der Walt 7 у – Pr. peringueyi 35—40 35—60 1 
Iridaceae 
Babiana 
curviscapa G. J. Lewis Z у = Pr. peringueyi 25-28 26—36 1, 12 
dregei Baker z у = Pr. peringueyi 25-28 30-35 1 
ecklonii Klatt Z у = Pr. peringueyi 27-32 40-50 12 
flabellifolia С. J. Lewis 7 у — r. 40-45 0-6 1 
Јтатези L. Bolus 2 v + Pr. sp. nov. and 40-45 45-50 1 
Pr. peringueyi 30-35 45-50 1 
суна С. Ј. Lewis 7 у = Pr. peringueyi 30–35 35-45 1 
pubescens С. J. L 2 у ~ Pr. peringueyi 32-35 44—50 1 
ius 
var. longibracteata С. J. Lewis z у ++ Рг. sp. nov. 40—45 40-50 12 
var. unguiculata G. J. Lewis 7 у + Pr. peringueyi 40-45 40-50 12 
Hesperantha 
latifolia (Klatt) M. P. de Vos Z у = Pr. peringueyi 15-25 20-25 1 
Lapeirousia 
dolomitica subsp. 
dolomitica Dinter 7 у + Pr. peringueyi 33-35 35—40 1 
јасашти N. Е. Br. Z у — Pr. sp. nov. and 33-40 33-35 1 
Pr. peringueyi 30-35 33-35 1 
oreogena Schltr. a у = Pr. sp. nov. 40-45 53-70 1 
pyramidalis subsp. regalis 7 у — Pr. peringueyi 32-34 40—47 1 
Goldblatt & J. С. Manning 
silenoides (Jacq.) Ker Gawl. д у = Pr. peringueyi 35—40 43-55 1 
violacea Goldblatt 7 у - Pr. peringueyi 32-35 34—40 1 
Котшеа 
hantamensis (Diels) Goldblatt a у = Pr. sp. nov. 40-45 50-70 1 
Sparaxis 
metelerkampiae (L. Bolus) 7 v - Pr. peringueyi 30-35 40-45 1 


Goldblatt & J. C. Manning 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


155 


Table 2. Continued. 


Species 


Flower 


Sym- 
metry Color 


Scent 


Pollinator 


Mouth 
part mm 


Floral 


tube mm Reference 


[Inferred members of the guild—Geraniaceae: Pelargonium cortusifolium UHér., P. crassicaule UHér. Iridaceae: Lap- 


eirousia dolomitica subsp. lewisiana (B. No 


gantha Diels, H. reds Goldblatt, Romulea kamiesensis M. P. de Vos, nih roxburghii (Baker) Goldblatt, 


Tritonia marlothii 


fruticosa (Benth.) e 


Campanulaceae 


Lobelia 


coronopifolia L. (pink form) 


Geraniaceae 
Pelargonium 
articulatum (Cav.) Willd. 
barklyi Scott Elliot 
elongatum (Cav.) Salisb. 
(also as P. zonale) 


laevigatum (L.f.) Willd. 
longicaule Jacq. 


myrrhifolium (L.) UHer. 
oxyphyllum D 

patulum Jacq. 

peltatum (L.) LHer. 


praemorsum (Andr.) Dietr. 
Iridaceae 
Aristea 
spiralis (L.f.) Ker Gawl. 
Babiana 


tubulosa (Burm. f.) Ker Gawl. 


Geissorhiza 
bonaspei Goldblatt 


exscapa (Thunb.) Goldblatt 
confusa Goldblatt 


Gladiolus 
angustus L. 
carneus D. Delaroche 


floribundus Jacq. 


monticola G. J. Lewis ex 


Goldblatt & J. C. Manning 


rhodanthus J. C. Manning 
& Goldblatt 

undulatus Jacq. 

vigilans Barnard 

virgatus Goldblatt 
& J.C. Manning 


Moegistorhynchus —Philoliche pollination system 


N N N WN 


. P. de Vos, Xenoscapa uliginosa Goldblatt & J. C. Mann 


Ph. gulosa 


Ph. rostrata 


and Ph. gulosa 
M. longirostris 


Ph. gulosa 


M. longirostris 


Ph. rostrata 

and Pr. nitidula 
M. longirostris 
Ph. rostrata 

and Ph. gulosa 


M. longirostris 


Ph. rostrata 


and/or Pr. nitidula 


Ph. rostrata 

and Ph. gulosa 
Ph. rostrata, 

Pr. nitidula 

. Sp. nov. 


Ph. rostrata 
Ph. rostrata 
Ph. rostrata 


18-20 


ca. 20 


— — 
мю МЮ ~ М о щл лю ы м 


©з Q2 Q2 Q2 о ш м 


w ~ 


rd.) Goldblatt, Geissorhiza kamiesmontana Goldblatt, Hesperantha oli- 


g. Scrophulariaceae: Jamesbrittenia 


22 


» 


„ 


ке - 
~ 


w 


156 Annals of the 
Mun Botanical Garden 


Table 2. Continued. 


Flower 
Sym- Mouth Floral 
Species metry Color Scent Pollinator рай mm tube mm Reference 
Ixia 
bellendenii R. C. Foster a p Е Ph. gulosa 18-22 30-32 4 
paniculata D. Delaroche a ег = М. longirostris 67—70 65-75 2, 6 
and Ph. rostrata n/a 12 
paucifolia G. J. Lewis a ег = Ph. gulosa 18-22 26-28 4 
and Ph. rostrata ca. 16 16-18 4 
Lapeirousia 
anceps (L.f.) Ker Gawl. 2 р = M. longirostris 52-63 65-76 1,2 
and Ph. gulosa 29-34 2 
fabricii (D. Delaroche) Ker 
z erp > M. longirostris 42-70 38-605 1,2 
and Ph. gulosa 29-34 n/a 2 
Tritonia 
crispa (L. f.) Ker Gawl. z p = М. longirostris 42-46 40-48 2 
and Ph. gulosa 29-34 40-48 2 
flabellifolia (D. Delaroche) z p x: Ph. rostrata 27-31 45-55 12 
G. J. Lewis and Ph. gulosa 17-18 45-55 12 
pallida Ker Gawl. 2 р - Ph. rostrata 30–34 45-55 12 
Watsonia 
borbonica (Pourr.) Goldblatt zZz p + Ph. rostrata, 19-22 14-18 12 
Pr. nitidula, 18-19.5 1418 12 
pd and 
megachilid 
bees, Ph, lateralis 12 
paucifolia Goldblatt 2 p = M. sp. nov. 18-20 22-30 12 
Orchidaceae 
isa 
draconis (L. f.) Sw. 2 cr = M. longirostris, ca. 57 ca. 48 6 
Ph. rostrata са. 35 57-72 6 
harveiana Lindl. z pb = Ph. rostrata 23-27 32-38 6 


[Inferred e of the guild—Ericaceae: Erica aristata Andr., E. embothriifolia Salisb., E. jasminiflora Salisb., Е. 
јипота Bolus, E. praecox Klotzsch, E. shannoniana Andr. Geraniaceae: jon uic alchemilloides (L.) UHer., P. 
про ufum (L.f.) Willd., P. articulatum (Cav.) Willd., P. denticulatum Jacq., P. moniliforme E. Mey. ex Harv., 
P. oblongatum Harv., P. punctatum (Andr.) Willd., P. radiatum (Andrews) Pers., P. они (L.f.) Willd., P. 
suburbanum Cliff., P. tetragonum (L.f.) L'Hér. Iridaceae: Babiana brachystachys С. J. Lewis, B. spathacea (L.f.) Ker 
Gawl., Geissorhiza callista Goldblatt, С. longifolia (С. J. Lewis) Goldblatt, С. schinzii m Goldblatt, G. stenosi- 
phon Goldblatt, G. tenella Goldblatt, Gladiolus eS F. Bolus, G. — из LL . Lewis) Goldblatt & J. C. 
rie Ixia fucata Ker Gawl., I. splendida С. Lewis, Lapeirousia macro: Bak., L. реке Goldblatt & J. 
С. g, L. verecunda Goldblatt, Nivenia еви Goldblatt, Romulea unis Goldblatt & J. C. Manning. 
Thereianthus elongatus (Schltr.) С. J. Lewis, Tritonia bakeri Klatt, Т. cooperi (Baker) Klatt, 7. lancea (Thunb.) N. Е. 
Вг., T. tugwelliae L. Bolus, Watsonia dubia Klatt, W. strictiflora Ker Gawl. Orchidaceae: Disa karooica Johnson & 
Linder, D. longicornu L.f., D. marlothii Bolus, D. porrecta Sw., D. salteri С. J. Lewis.| 


= 


Prosoeca ganglbaueri—Pr. robusta pollination system 


Amaryllidaceae 
Brunsvigia 
grandiflora Lindl. 2 р + Pr. ganglbaurii 31-42 >10 12 
gregaria К. A. Dyer 7 р + Pr. ganglbaurii 31-42 >10 12 


striata (Jacq.) Aiton zZz p + Pr. ganglbaurii 34-36 >10 12 


Volume 87, Number 2 Goldblatt & Manning 157 
2000 Long-Proboscid Fly Pollination 
Table 2. Continued. 
Flower 
m- Mouth Floral 
Species metry Color Scent Pollinator part mm tube mm Reference 
Nerine 
cf. angustifolia (Baker) Watson z р = Pr. robusta ca. 33 >5 12 
bowdenii S. Watson 2 р Е Pr. ganglbaueri 27-30 ca. 5 12 
Geraniaceae 
Pelargonium 
carneum Jacq. z р = Pr. ganglbaueri 34—36 55-60 12 
Pr. Side 38-40 55—60 8 
dipetalum UL Her. z crip - Pr. long 38—40 ca. 60 8 
gracillimum Fourc. z p = Pr. ganglbauri 34—36 50-60 12 
pinnatum (L.) L'Her. z cip — Pr. longipennis 38—40 50—60 8 
Iridaceae 
Gladiolus 
bilineatus G. Lewis z p == Pr. longipennis 38—40 50—60 8 
calcaratus G. J. Lewis ® w Е Pr. robusta 20-23 28-40 3 
engysiphon G. Lewis Zz сг = Pr. longipennis 38-40 52—60 3 
таспеци Oberm. z р = t. wiedmannii 23-29 40-45 3 
microcarpus G. J. Lewis РА р = Pr. ganglbaueri 27-30 35—40 3 
mortonius Herbert 2 р = Pr. ganglbaueri 31—42 36-52 3 
oppositiflorus J. D. Hook. 2 p = Pr. ganglbaueri 28-32 40-50 12 
varius F. Bolus 2 p = Pr. ganglbaueri 33-35 45-55 3 
Pr. robusta 20-30 45-55 12 
Hesperantha 
brevicaulis Vas: G. J. Lewis a p = St. wiedmannit 18—30 25-37 12 
grandiflora G. J. Lewis 2 p = Pr. ganglbaueri 23-25 33-36 12 
scopulosa Hilliard a p – Pr. ganglbaueri 32-35 32-40 12 
cf. woodii R. C. Foster a p = Рг. ganglbaueri 27-31 35-38 12 
Nivenia 
stenosiphon Goldblatt a b = Pr. ganglbaueri 23-25 32-38 10 
Tritoniopsis 
revoluta (Burm.f.) Goldblatt zZz р = Pr. ganglbaueri 23-25 ca. 40 12 
Watsonia 
wilmsii N. E. Br ® p = Pr. ganglbaueri 33-35 40-45 12 
and Pr. robusta 6—45 40—45 12 
Lamiaceae 
Orthosiphon 
tubiformis R. Good 2 р = St. wiedmannii 23-29 28-35 12 
Plectranthus 
ambiguus (Bol.) Codd z b E St. sp. 20-33 22-290 11 
ecklonii Benth. 7 та = St. wiedmannii 16-21 24—26 11, 12 
МШагай Сода z b = St. sp. 21-29 22-290 ll 
Orchidaceae 
Disa 
oreophila Bolus 2 p = Pr. ganglbaueri 19.6(2.3) 20.6(1.1) 5 
amoena Linder z p = Pr. ganglbaueri 33-35 25-30 12 
and Pr. robusta 20-30 25-30 12 
Brownleea 
coerulea Harv. ex Lindl. z m = St. wiedmannii ca. 23 20-24 12 
macroceras Sond. 2 p = Pr. ganglbauert 19.6(2.3) 38.9(6.4) 5 


158 


Annals of the 
Missouri Botanical Garden 


Table 2. Continued. 


Flower 


Sym- 
metry Color 


Mouth Floral 


Species Scent Pollinator рай mm tube mm Reference 
Scrophulariaceae 
Zaluzianskya 
microsiphon (Kuntze) К. Schum. z w = Pr. ganglbaueri 33-35 30-42 12 


[Inferred members of the guild—Acanthaceae: d glabratum Vollesen, [soglossa cooperi С. B. Cl., Rhinacan- 
e (C. B. СІ.) T. 


thus gracilis Klotzsch, icq natalens 
Amaryllidaceae: Brunsvi, 
krigei W. F. Barker. C И UNA Dianthus basutic 
Geraniaceae: Pelargonium acraeum R. A 
& Zeyh.) St 
um Oberm., G. saxatilis Goldblatt & J. C. Man 
(Backh. & Harv.) Goldblatt & J. C. Mann 


а зрр., би ан жаы Snij., С. sp 
us Burtt 
. Dyer, Р. caucalifolium Jacq. subsp. caucalifolium, P. ionidiflorum (Eckl. 
eud., P. transvaalense Knuth. Iridaceae: Geissorhiza fourcadei (L. Bolus) С. Lewis, Gladiolus 
ning, G. scabridus Goldblatt & J. C. Man 
ing, H. curvula Hilliard & Burtt, H. ee Hilliard & Burtt, H. 


. J. Edwards, od a (Lindau) C. B. Cl. 
Nerine filam a W. F. Barker, N 


t Dao: Ericac eae: pon neues L. (pink bul: 


cataractar- 
g. Hesperantha coccinea 


huttonii (Baker) Hilliard & Burtt, H. pulchra Baker, Nivenia concinna N.E. Br., N. stokoei L. Guthrie, Radinosi- 


phon leptostachya (Baker) N. E. Br., Tritoniopsis flexuosa (Thunb . Br. 
iflorus Benth., S. macranthus (Garke) Ashby, S. rotundifolius E. Mey., Thorncroftia longiflora N. 
Br., T. media Codd, T. succulenta (Dyer & Bruce) L. E. Codd 


Syncolostemon dens 


.) G. Lewis, Watsonia occulta N. E. Br. Lamiaceae: 


Orchidaceae: Disa rhodantha Schltr., D. saxicola 


Schltr., Satyrium hallackii Bolus. NOTE—Hesperantha pubinervia Hilliard & Burtt may belong here, but although 
it does have an elongate perianth tube, the tube itself is extremely narrow, accommodating only the style, and it 


does not produce nectar. 


Anecdotal MN of long-proboscid flies visiting flowers but no claims for pollination or evidence of pollen 


transfer to stigma 
Ericaceae. Erica ju јипота Bolus (?Ph. rostrata —E. 


Oliver, pers. comm.). 


Scrophulariaceae. Jamesbrittenia fruticosa (Benth.) "iiu (Pr. peringueyi—Museum record); Zaluzianskya microsi- 
d). 


phon (Pr. peringueyi—Museum recor 


rbanum 


Geraniaceae. Pelargonium alchemilloides (L.) UHer. (Ph. rostrata—Vogel, 1 
ubu (M. longirostris—Johns 


McDonald in Struck, 1997); P. s 
fly—van Jaarsveld in Struck, 1997). 


same site on the insect's body would frequently re- 
sult in insect visits failing to accomplish pollination 
as a result of stigma clogging by foreign pollen. 
In the Prosoeca peringueyi pollination guild pol- 
len deposition on the ventral head or thorax is ef- 
fected by Pelargonium species, on the frons or dor- 
sal thorax by Lapeirousia species (normally one or 
two species of a genus is present at any site), and 
on the dorsal thorax by Babiana species (also nor- 
mally only one species present at any site) (Man- 
ning & Goldblatt, 1996). Pollen of Hesperantha la- 
пра, occasionally part of the guild using P. 
peringueyi as pollinator, is du eus on the lateral 
and upper ventral thorax. In P. incrassatum and P. 
sericifolium, which are таран С at some sites in 
Namaqualand and are both pollinated by P. perin- 
gueyi, pollen contamination is avoided by place- 
ment of their respective pollen on the ventral head 
or ventral thorax of Р peringueyi, the result of 
shorter or longer filaments in these two species. 
This pattern is repeated in the Moegistorhynchus 
longirostris—Philoliche pollination guild (Manning 
& Goldblatt, 1997). Pollen deposition on the distal 


954); P. denticulatum (unidentified fly— 
on & Steiner, 1997); P. tetragonum (unidentified 


ventral thorax is effected by Pelargonium spp. (ei- 
ther P. praemorsum or P. longicaule) or Geissorhiza 
spp. (either G. confusa or G. exscapa) and on the 
proximal ventral thorax or lower head by P. tabu- 
lare. Deposition on the frons is effected by Lapei- 
rousia anceps, the dorsal head or thorax by Tritonia 
crispa at some sites, and by Gladiolus undulatus, 
G. angustus, or L. fabricii at others. Ixia paniculata 
is unusual in having short stamens held within the 
mouth of the tube, and its pollen is deposited on 
the frons around the base of the proboscis. Species 
of Orchidaceae that have stalked pollinaria are 
probably not directly involved in competition for 
pollen deposition, but the number of species of Or- 
chidaceae at any site is usually limited to one. 
Pollen deposition in the Prosoeca ganglbaueri 
guild follows this general pattern. For example, an- 
thers of Brunsvigia, Nerine, and Pelargonium spp. 
brush different parts of the ventral head, thorax, or 
abdomen, depending on stamen length, while pol- 
len of Gladiolus and Watsonia species is deposited 
on the dorsal thorax. Orchid pollinaria are placed 
near the base of the proboscis. Pollen placement 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


159 


by Hesperantha species, which mostly have sym- 
metrically disposed, divergent anthers, is less well 
defined but is always on the lateral or ventral parts 
of a fly’s body. Pollen of the important long-probos- 
cid fly plant, Zaluzianskya microsiphon, is depos- 
ited on the ventral head as the anthers are held at 
the abaxial side of the mouth of the floral tube. 


EVOLUTION OF THE SYSTEM 


Associated with butterfly pollination by Vogel 
(1954) in his keystone work on pollination systems 
in the flora of southern Africa, long-proboscid fly 
pollination appears to have no direct connection 
with that strategy. Butterfly and long-proboscid fly 
pollination have some similarities, notably includ- 
ing plant species with odorless, long-tubed flowers. 
The two systems are, however, independent of one 
another, and no species of either group of insects 
share the flowers of any plant species. Moreover, 
butterfly pollination in southern Africa (excluding 
the highly specialized Aeropetes butterfly system— 
Johnson & Bond, 1994) is frequently a part of gen- 
eralist systems that use other insects, including 
bees, hopliine scarab beetles, and settling moths 
(Goldblatt et al., 1995). In the Iridaceae, phyloge- 
netic studies in Gladiolus, Lapeirousia, and Spar- 
axis show that long-proboscid fly flowers are nearly 
always most closely related to ancestors pollinated 
by long-tongued bees (Apidae) (Goldblatt et al., 
1995, in prep.; Goldblatt & Manning, 1999). Spe- 
cies of Gladiolus pollinated by long-proboscid flies 
fall in taxonomic sections where long-proboscid bee 
pollination is ancestral and in which butterfly pol- 
lination may not occur. In the only section of Glad- 
iolus in which butterfly pollination (by the special- 
ist pollinator, Aeropetes) is significant, section 
Blandus, butterfly flowers most likely evolved from 
long-proboscid-fly-pollinated ancestors (Goldblatt 
& Manning, 1998). Pollination by the Aeropetes but- 
terfly does not occur in Babiana or Lapeirousia, 
genera in which long-proboscid fly pollination is 
especially common, nor in [xia or Sparaxis, or Tri- 
tonia. 

In most families in which long-proboscid fly pol- 
lination occurs, a shift from bee to long-proboscid 
fly pollination appears to be relatively straightfor- 
ward, involving shifts in perianth color and marking 
pattern, elongation of the floral tube, and loss of 
floral odor if present in the ancestor. For example, 
in most Iridaceae, long-proboscid bee flowers are 
zygomorphic and have a funnel-shaped perianth 
tube of moderate length, with the upper flared por- 
tion about as long as the cylindric basal half. Uni- 
lateral arcuate stamens are held above the mouth 


of the perianth tube where they readily brush 
against the body of a large bee as it forces its upper 
body into the upper part of the floral tube while 
extending its proboscis into the lower part of the 
tube. The perianth may be almost any color, but is 
not often red or cream, and the flowers are fre- 
quently scented (Goldblatt et al., 1998а). Long-pro- 
boscid fly flowers of the Moegistorhynchus—Philol- 
iche and the Prosoeca ganglbaueri pollination 
systems merely require a shift in flower coloring 
toward the pale pink to cream end of the color 
spectrum, the acquisition of darker, linear nectar 
guides, and an elongation of the narrow part of the 
floral tube to exclude nectar feeders with mouth 
parts less than 15 mm long. Nectar quality, espe- 
cially sugar constituents, is much the same as is 
found in long-proboscid bee flowers, but nectar vol- 
ume is typically much greater and sometimes more 
dilute. For example, flowers of bee-pollinated spe- 
cies of Gladiolus have nectar volumes between 0.5 
and 4 wl compared with 1.8 to over 10 pl in long- 
proboscid fly pollinated species (Goldblatt et al., 
1998a; Goldblatt & Manning, 1999). 

It is noteworthy that fly diversity is higher in the 
geographically much smaller winter-rainfall zone. 
Three fly species range over all of eastern southern 
Africa, whereas there are 10 fly species in the two 
long-proboscid fly guilds in the winter-rainfall zone. 
Some of these flies are relatively widespread, but 
Moegistorhynchus sp. nov. and Prosoeca sp. nov., P. 
rubicunda, and P. nitidula have very narrow ranges 
and moreover, except for Р. sp. nov., they appear to 
be rare, at least as far as one can judge from the 
few specimens known. 


TERMINOLOGY 


There is unavoidable confusion when comparing 
long-proboscid fly and long-proboscid (or long- 
tongued) bee pollination. Long-proboscid fly polli- 
nation, according to our definition, includes flies 
with probosces in excess of 15 mm and usually 
much more. Few long-proboscid bees have probos- 
ces longer than 12 mm. Moreover, acrocerids, ta- 
banids, and nemestrinids with probosces 10—15 
mm long are frequently referred to as long-probos- 
cid flies to contrast them with short-tongue flies that 
lap fluid. Rebelo et al. (1985) coined the term rhi- 
nomyophily, which is useful but not favored by 
many biologists who prefer more direct terms (e.g., 
bird pollination vs. ornithophily, etc.). Struck 
(1997) favored the term hoverfly pollination, but 
that has the disadvantage of misrepresenting the 
typical behavior of the flies, which grasp floral or- 
gans whenever possible while foraging although 


160 


Annals of the 


Missouri Botanical Garden 


227 


“SRN 


С SS р 
`` 
ES 


SN 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


161 


they do hover while inserting the proboscis into a 
floral tube. We have no solution to offer and suggest 
long-proboscid fly pollination for the syndrome in- 
volving flies with probosces usually exceeding 15 
mm, and short-proboscid fly pollination for the syn- 
drome involving flies with true sucking mouth parts 
usually less than 15 mm long. Flies with lapping 
mouth parts would be excluded from this definition. 
The confusion between long-proboscid fly and long- 


proboscid bee syndromes as regards length of 


mouth parts seems unavoidable. 


NECTAR CONSIDERATIONS 


Although most data available for flowers polli- 
nated by Diptera suggest that flies favor nectar with 
hexose sugars predominant (Baker & Baker, 1983, 
1990), these authors did not distinguish between 
flies with short, lapping mouth parts and those with 
long, sucking mouth parts. Recent studies, howev- 
er, show that nectar of plants with flowers adapted 
for pollination by long-proboscid flies is often su- 
qnd to sucrose-dominant (Goldblatt & Bern- 
hardt, 1990; Goldblatt et al., 1995; Manning & 
Calla, 1996, 1997; Goldblatt & Manning, 

). The broader survey here in general confirms 
ii: observation, but the data indicate that this may 
simply be the result of the taxa involved. Most Ir- 
idaceae belonging to subfamily Ixioideae (which in- 
cludes most species pollinated by long-proboscid 
flies) have sucrose-rich nectar as do all species of 
the family with flowers adapted for pollination by 
bees. Significantly, nectar of Aristea spiralis, the 
only species of Iridaceae with long-proboscid fly 
flowers and not a member of Ixioideae, has hexose- 
rich nectar. It also produces nectar from perigonal 
nectaries (Goldblatt & Manning, 1996), unlike spe- 
cies of Ixioideae, which have septal nectaries 
(Goldblatt, 1990, 1991). Flowers of some other fam- 
ilies with long-proboscid fly flowers (e.g.. Lami- 
aceae, Orchidaceae) also have sucrose-rich nectar. 
However, Geraniaceae does not follow this pattern; 
species of Pelargonium that have been examined 
(Manning & Goldblatt, 1996, 1997) have, with the 
exception of P. cortusifolium, hexose-rich to hexose- 
dominant nectar. Among long-proboscid fly flowers 
then, nectar sugar chemistry may simply be a re- 
flection of systematic affiliation or nectary type, and 
not the result of selection by long-proboscid flies 


for a preferred type of nectar. This reflects the con- 
clusions of van Wyk (1993) and van Wyk et al. 
(1993) that the nectar sugar characteristics of flow- 
ers are highly conserved and that nectar sugar pat- 
terns often reflect taxonomic affinities rather than 
pollination systems. 

ectar concentration is relatively constant, al- 
though it seldom rises above 32% sucrose equiva- 
lents and seldom dips below 2096. The two species 
of Hesperantha in the Prosoeca ganglbaurii guild 
are exceptional in having nectar concentrations be- 
low 2096. Concentrations above 3296 may make the 
nectar too viscous to be easily sucked into the pro- 
boscis. 


A Co-EVOLVED SYSTEM 


As suggested by Feinsinger (1983) for sphinx 
moths, it seems that long-proboscid flies and their 
flowers have probably evolved through reciprocal 
selection. Frequent visits from long-proboscid flies 
select for long-tubed flowers, which in turn select 
for longer-proboscid flies capable of reaching the 
nectar within the tubes. This pattern fits the Red 
Queen effect (van Valen, 1973; Futuyma, 1979), in 
which species may coevolve indefinitely, some be- 
coming extinct in the process, or else arrive at a 
static, nonevolving equilibrium. Evolution within 
this system may not, however, be all that simple. 
Population densities of long-proboscid flies appear 
to be highly erratic (Goldblatt et al., 1995; Gold- 
blatt & Manning, 1999), a situation comparable to 
that for sphinx moths, as noted by Gregory (1963— 
1964). But whereas shorter-proboscid moths visit 
these flowers, taking advantage of nectar welling up 
in the tubes, there do not seem to be alternative 
shorter-proboscid insects available for most long- 
proboscid fly flowers to use this resource when their 
primary pollinators are not available. Instead, those 
long-proboscid fly flowers that are self-incompatible 
simply fail to reproduce or reproduce poorly in cer- 
tain seasons at certain sites (Goldblatt & Manning, 
99). 

Fail-safe mechanisms for self-pollination appear 
to be quite common among species pollinated by 


mT 
Ne 


long-proboscid flies. We know now that of the ten 
species of Lapeirousia with flowers adapted for pol- 
lination by long-proboscid flies, four at least are 
self-compatible and autogamous and two are self- 


Representative examples of flower form in species pollinated by the long-proboscid flies of the Prosoeca 


Romulea hantamensis. 
P. magenteum. —F. P. 


ure 4. 
а раж pollination guild. —A. 
Pelargonium ericifolium. —E. 


—B. Lapeirousia oreogen a. —C. Hespe 
crassicaule. —G. Lape 


rantha oligantha. — 


irousia pyramidalis. —H. L. dolomitica. 


ш L. violacea. —Ј. L. silenoides. —К. pude metelerkampiae. —L. Tritonia marlothii. 


162 Annals of the 
Missouri Botanical Garden 


Representative examples of flower form in species pollinated by the long-proboscid flies Moegistorhynchus 
longirostris and Philoliche gulosa. —A. Ixia paniculata. —B. Geissorhiza ехзсара. —C. Pelargonium longicaule. —D. 
P. appendiculatum. —E. P. moniliforme. —F. Babiana tubulosa. —G. Lapeirousia anceps. 


163 


Volume 87, Number 2 


2000 


Goldblatt & Manning 


Long-Proboscid Fly Pollination 


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Annals of the 


164 


Missouri Botanical Garden 


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Long-Proboscid Fly Pollination 


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Annals of the 


166 


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Long-Proboscid Fly Pollination 


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168 


Annals of the 
Missouri Botanical Garden 


proboscis base). —B. Hola crispa (dorsal ed —C. 


Lapeirousia fabricii (dorsal thorax). —D. Pelar, 
pendiculatum dua thorax). 
(ventral abdomen) 


um ap- 
—E. Chania. разый 


incompatible (Goldblatt et al., 1995, and unpub- 
lished data). At least one species of Hesperantha 
(Н. latifolia) and the [ма and Sparaxis species 
known to be pollinated by long-proboscid flies are 
facultatively autogamous, whereas in Gladiolus self- 
incompatibility appears to be frequent (Goldblatt et 
al., 1998a, in prep.). Some populations of Gladiolus 
species with long-proboscid fly flowers have set no 
capsules for three years for which we have obser- 
vations when their pollinators were absent. Al- 


though in Pelargonium self-compatibility appears 
to be the rule (Struck, 1997), autogamy cannot nor- 
mally take place because of mechanical protandry. 
There is therefore no barrier to geitonogamous pol- 
lination in this genus in which inflorescences pro- 
duce numerous flowers contemporaneously but pol- 
lination must be mediated by an external agent. 
Proboscis length is typically shorter than the flo- 
ral tube, a feature that long-proboscid fly pollina- 
tion shares with sphinx moth pollination. This en- 
sures that a visiting insect must push its body into 
flowers so that its proboscis will extend as deeply 
as possible into the tube and reach the nectar res- 
ervoir. Orchid genera are one notable exception. In 
deceptive flowers of Disa amoena and the Disa dra- 


and pollinaria are attached to the upper third of the 
proboscis. Other exceptions are Aristea spiralis and 
species of Brunsvigia and Nerine that are pollinated 
by long-proboscid flies; in these taxa floral tubes 
are less than 10 mm long. These flies, however, 
have bodies large and probosces long 
enough that they will consistently brush against an- 
thers and accumulate deposits of pollen on their 
ventral thoraces and abdomens. Nectar produced 
by such short-tubed flowers is obviously accessible 
о a range 


enough 


of other insects, and we have observed 
effective pollination by bees in Aristea spiralis. We 
have not recorded visits to any other of these short- 
tubed species by insects other than long-proboscid 
flies. The considerable distance of the anthers from 
the mouth of the tube also makes it unlikely that 
even large bees would normally contact anthers 
when foraging for nectar in such flowers because 
their bodies are so much shorter than the filaments. 


LoNc-PnoBoscip FLY POLLINATION IN THE 
HIMALAYAS 


Attention was first drawn to long-proboscid fly 
pollination in Asia by Fletcher and Son (1931), who 
riefly described visits to (and inferring pollination 
of) Roscoea purpurea (Zingiberaceae) by the tabanid 
fly Chorizoneura longirostris (proboscis 50—60 mm 
long). Subsequently, Dierl (1968) described in de- 
tail the behavior of this fly in the course of its visits 
to flowers (presumably foraging for nectar) on Ros- 
coea. The Himalayan region is floristically rich and 
includes several plant species with long-tubed flow- 
ers that may also be pollinated by this fly, e.g., 
other species of Roscoea and Rhododendron spp. 
(Ericaceae). This pollination guild awaits more ex- 
tensive investigation. Involving different fly genera 
and plant species, the Himalayan system presum- 


Volume 87, Number 2 
2000 


Goldblatt & Manning 
Long-Proboscid Fly Pollination 


169 


ably evolved оо of the long-proboscid 
fly guilds of southern Afric 


TAXONOMIC DISTRIBUTION OF LONG-PROBOSCID 
FLY POLLINATION 


According to our survey, at least 83 species have 
been shown to be pollinated solely by long-probos- 
cid flies. Based on their nearly identical floral pre- 
sentation, it seems reasonable to infer that at least 
90 more species also have this pollination strategy. 
Even if this figure is grossly underestimated, there 
seems no doubt that long-proboscid fly pollination 
is of relatively minor importance in the flora of 
southern Africa, which comprises over 20,500 spe- 
cies of flowering plants (Goldblatt, 1997). Never- 
theless, long-proboscid fly pollination assumes 
more than marginal importance in at least two fam- 
ilies in southern Africa, Geraniaceae and Iridaceae. 
Precise figures are not available for Geraniaceae, 
but Struck (1977) has estimated that 25% of the 
southern African species of Pelargonium are pol- 
linated by long-proboscid flies, although the exact 
definition of the system that he entertained is not 
clear. In Iridaceae we have more accurate figures. 
Some 105 of the approximately 1025 species in 
southern Africa appear to have this pollination sys- 
tem exclusively and one or two more (e.g., Hesper- 
antha coccinea—S. D. Johnson, pers. comm.; Ar- 
istea spiralis, Watsonia borbonica—Table 2) may 
have a combined long-proboscid fly and other in- 
sect pollination. Thus, about 10% of southern Af- 
rican Iridaceae have adopted this pollination strat- 

gy. This is substantially more than the estimated 
i species (6.3%) of the southern African Iridaceae 
that are predicted on the basis of floral morphology 
to be pollinated by sunbirds (Goldblatt et al., 
1999 

бош the Iridaceae, long-proboscid fly polli- 
nation appears to be most well developed in Lap- 
eirousia, in which 10 species, or 3046 of the total 
in temperate southern Africa, have flowers polli- 
nated by long-proboscid flies. The system is also 
well developed in Gladiolus (Goldblatt & Manning, 
1999; Manning & Goldblatt, 1999), in which 27 
species are considered to have flowers adapted for 
this particular pollination strategy (1796 of the 
southern African species), and long-proboscid fly 
pollination is inferred for between 10 and 20% of 
the southern African species of five other large gen- 
era, Babiana, Geissorhiza, Hesperantha, Ixia, and 
Tritonia. 

Long-proboscid fly pollination assumes more 
modest importance in Ericaceae (Rebelo et al., 
1985) and Orchidaceae, although prominent in 


Brownleea and Disa, and we have yet to assess its 
significance in Amaryllidaceae and Lamiaceae. The 
strategy seems likely in Acanthaceae, but it has yet 
to be confirmed in that family. In the remaining 
families in which it occurs, long-proboscid fly pol- 
lination is decidedly rare and is evidently confined 
to just one or a few species. Curiously, evolution of 
the system seems highly labile in several genera of 
the Iridaceae and in Disa (Orchidaceae). In Disa 
and Gladiolus it has evolved repeatedly in different 
lineages (Johnson et al., 1998; Goldblatt & Man- 
ning, 1999), an estimated nine times in the latter 
genus. 


CONSERVATION 


Long-proboscid fly pollination poses important 
concerns for conservation. Plants pollinated by a 
single insect, or no more than two over their entire 
range, clearly are at more significant risk than those 
that are pollinated by several different insects 
Bond, 1994). Long-proboscid flies may be regard- 
ed as keystone species, for several plant species 
depend on particular flies for their pollination and 
sexual reproduction. Conservation of plants with 
these specialist pollinators must involve conserva- 
tion of their pollinators, an undertaking fraught with 
unusual difficulties. Both Nemestrinidae and Ta- 
banidae have complex life cycles. Tabanidae have 
carnivorous, aquatic larvae that require wetland 
habitats for their larval development, which may 
take place some distance from sites of the plant 
species on which the adults feed. Female tabanids 
also require a blood meal during their adult phase 
before egg-laying can proceed. This makes the 
presence of suitable host mammals essential. Ne- 
mestrinidae have an equally complex life history. 
Although no details of the life cycles of long-pro- 
boscid flies are known, all members of the Nemes- 
trinidae so far studied have parasitic larvae, often 
on locusts. Obviously, large, relatively undisturbed 
sites with a diversity of habitats are necessary for 
the completion of the life cycles of nemestrinids 
and tabanids 


Literature Cited 
Baker, H. G. & I. Baker. 1983. Floral nectar sugar con- 
es ba red to pollinator type. Pp. 117-141 in 
: & R. J. Little (editors), Handbook of Ex- 
memeni Pollination Biology. Scientific and Academic 
Editions, New York. 
& 


he predictive value of nectar 
chemistry to ihe тєп of pollinator types. Israel 
66. 


994. Do mutualisms matter? Assessing the 
саа va pollinator and disperser disruption on plant 


extinction. Philos. Trans., Ser. В 344: 83—90. 


170 Annals of the 
Missouri Botanical Garden 
Dierl, W. 1968. Zur Nahrungsaufnahme von Chorizoneura А РМ in the fynbos of South Africa. Pp. 


longirostris Патона) (Diptera: Tabanidae). Khumbu 
bn. Forse nternehmens Nepal ae 
76-81. iniversititaverlas Wagner, Innsbruck— 


с 
Faegr L. van der Pijl. 1979. The Principles of 
Pollination Eu. 3rd ed. Pergamon Press, ew York. 
к Р. i 
282-309 in D. J. Futuyma & M. Slatkin 
volution. — Sunderland, Massachusetts. 
Fletcher, T . B. & S. K. Son. а veterinary ento- 
mology = кт Part XIV. Indian J. Veterin. Sci. Anim 
Hush. 1: -199. 
Futuyma, D. | б oo Biology. Sinauer, Sun- 
derland, Massachuse 
Goldblatt, P. 1990. Phylogeny and classification of Iri- 
СЕ. ш Missouri Bot. Gard. 77: 607—627. 
. An overview of the systematics, phylogeny 
and dee of t southern African Iridaceae. Contr. 
Bolus Herb. 13: 1-74. 
1997. Flo ristic diversity in the Cape Flora of 
South Africa. Biodiversity & Conservation 6: 359-377. 
— & Р. Bernhardt. 1990. Pollination biology of Niv- 
enia (Iridaceae) and the presence il кы self- 
compatibility. Israel J. Bot. 39: 93— 
—— & . C. Manning. didis Ph ieu and specia- 
n Lapeirousia subgenus Lapeirousia (lridaceae: 
Ixioideae). Ann. Missouri Bot. Gard. 83: 346—361 
1998. Gladiolus in Southern Africa. 
dii Press, Cape cis 
— 1 


e long-proboscid fly polli- 
nation pde in ue (Iridaceae). Ann. Missouri 
Gard. 86: 758-774 

- Bernhardt. 1995. Pollination bi- 
dar of геа subgenus Lapeirousia (Iridaceae) in 
southern Africa: Floral divergence and adaptation for 
long-tongued fly pollination. Ann. Missouri Bot. Gard. 
82: 517-534. 


& 1997. Notes on the polli- 
dion of Gladiolus brevifolius { (Iridaceae) by bees (An- 
thophoridae) and bee mimicking flies (Pilar: Ac- 
roceridae). J. Kansas Entomol. Soc. 70: 297-3 

998a. Adaptive radiation 
of bee-pollinated ннн species (Iridaceae) in south- 
ern Africa. Ann. Missouri Bot. Gard. 85: 492-517 
, P. Bernhardt & & I. c Manning. 1998b. Pollina- 
tion of petaloid geophytes by monkey beetles (Scara- 
baeidae: Rutelinae: Hopliini) in southern Africa. Ann. 
Missouri Bot. Gard. 85: 215-230. 
, J. C. Man ning & P. Bernhardt. 1999a. Evidence 
of bird pollination in the Iridaceae of southern Africa. 
Adansonia ser. 3, 21: 25—40. 
Grant, V. & K. A. Grant. 1965. Flower Pollination in the 
Phlox Family. Columbia Univ. Press, New k. 
Gregory, D. P. 1963-1964. Hawkmoth pollination in the 
genus Oenothera (Onagraceae). Aliso 5: 357-384, 385- 
419. 


Johnson, S. D. & W. J. Bond. 1994. Red flowers and 


137-148 Arianoutsou & R. Grooves (editors), 
Plant dual Interactions in Mediterranean-Type Eco- 
Academic Press, Dordrecht. 
. 1995. Long proboscid fly pol- 
lination of two orchids in the Cape Drakensberg moun- 
tains, South Africa. Pl. Syst. Evol. 195: 169-175. 
& 1997. Long- tongued fly pollination 
and evolution of floral spur length in the Disa draconis 
mplex. Evolution 51: 45-53. 
. P. Linder & K. S. Steiner. 1998. Phylogeny 
and radiation of pollination systems in Disa (Orchida- 
. J. Bot. 85: 402-411. 
. & P. | 


Tu Cupid comes in 
many gui s: The o-humble fly and : ao 
guild in Tn оо Veld -i КД 50- 

& 996. eca ges ueyi (Dip- 
tera: Nemestrinidae) olli din. guild i in southern Af- 
rica: Long-tongued flies and their tubular flowers. Ann. 
Missouri | Саг d. 83: 67-86. 

1 


e Moegistorhynchus longi- 
rostris (Diptera: Nemestrinidae) pollination guild: Long- 

tubed flowers and a specialized long-proboscid fly-pol- 
lination system in uibem Africa. Pl. Syst. Evol. 206: 
51-69. 


— —— а P. D. J. Winter. 1999. Two new spe- 
cies of Gladiolus (inidapese: Ixioideae) from South Af- 
rica and notes on long- de fly pollination in the 
genus. Bothalia 29: 217-2 

Marloth, R. 1908. Some aie on entomophilous 

flowers. S. African J. Sci. 5: 110-113 

er & J. van 


oo in southern Natal, South Africa. Pl. Sys t 
Evol. 218: 99-112. 

Rebelo, A. G., W. R. Siegfried & E. G. H. Oliver. 1985. 
eyo of Hee ысы in the south-western Cape 

can £r x : 270-280. 

Root, T B. Th niche оет pattern of the 
blue-gray ae Ecol. Monogr. 37: 317-3 

Struck, M. 1997. Floral diveipence and ие in 
the genus Pelargon nium (Geraniaceae) in southern Af- 

cal and evolutionary considerations. Pl. 
Syst. Evol. 208: 71-97. 

к 1. 1973. А new evolutionary law. Evol. The- 


1-30. 
Vogel, s. 1954. 71 ла als Elemente 
der РА ша. 1: 1–338 
Whitehead, У. B., J. H. Giliamce & x С. Rebelo. 1987. 
Insect pollination i in the Cape Flora. Pp. 52-82 in A. 
G. Rebelo (editor), А Preliminary Synthesis of Pollina- 
tion Biology in the Cape Flora. CSIR, Pretoria. 
yk, B.-E. van. ar sugar composition in south- 
ern African Papilionoideae (Fabaceae). Biochem. Syst. 
& Ecol. 21: 271-277. 
ui S. Whitehead, H. F. Glan, D. S. Hardy, ~ » 


van Jaa vel G. F. Smith. 1993. Nectar sugar c 
pos гені in the subfamily Alooideae ао, 
Biochem. Syst. & Ecol. 21. 249-253. 


Alexei А. Oskolski? and 
Porter P. Lowry IP? 


WOOD ANATOMY OF 
MACKINLAYA AND 
APIOPETALUM 
(ARALIACEAE) AND ITS 
SYSTEMATIC IMPLICATIONS! 


ABSTRACT 


Wood anatomy was examined in both species of Apiopetalum (trees endemic to New Caledonia) and i in two species 
of Mackinlaya iube from Queensland, Australia), using light and s 
phylogenetic relationships and taxonomic 
intervessel pits, ranging from 3 to 6 wm; 
Apiopetalum only) axial parenchyma; non-septate fibe 
deposits in the vessels. Differences in wood structure idee the ge 

confirm recent molecular sequence data suggesting that yeso ol and Mackinlaya form a monophyletic group. 
Evidence from ia anatom b validates nor is the hypothesis that these two genera are intermediate between 


Araliaceae and Apiac cod offers no clear indication of the group's phylogenetic position. Several wood characters 
(small jp Aie hic k fiber walls, non-septate fibers) suggest a relationship with Myodocarpus, Delarbrea, and 
Pseudoscia No clear synapomorphies were found to support a sister relationship between Apiopetalum and Mack- 


in nlaya and core Apiaceae as previously suggested by Plunkett, nor between them and other Araliaceae. Helical thick- 
enings on the walls of both ray and axial parenchyma, previously reported only once in Trigonia (Trigoniaceae), were 


observed in one sample of M. macrosciadea. 
Key words: 


Apiales, Apiopetalum, Атаћасеае, Mackinlaya, New Caledonia, phylogeny, Queensland, wood anatomy. 


The phylogenetic positions of Apiopetalum Baill. 
and Mackinlaya F. Muell. within Araliaceae have 
been difficult to assess using traditional approaches 
based on morphology. Recent studies using molec- 
ular data have provided new insights into their evo- 
lutionary relationships, but their exact placement 
remains unresolved. Five species of Mackinlaya 
(including Anomopanax) were recognized by Philip- 
son (1979), extending from Queensland, Australia, 
through the Solomon Islands, Bismarck Archipel- 
ago, New Guinea, and Celebes to the Philippines 
(see also Philipson, 1951). Apiopetalum comprises 


two species, both endemic to New Caledonia (Low- 
ry, in prep.). Mackinlaya species are branched, 
sympodial shrubs with simple and palmately lobed 
or palmately compound leaves. Apiopetalum spe- 
cies are small trees, to ca. 6 m in height, and have 
exclusively simple leave 

n most em ше proposed for Аг- 
aliaceae (Bentham, ; Harms Td Vi- 
guier, : M 55 1967; g & Hoo, 
1982; Takhtajan, 1987), Д ааа jia Mackin- 
laya have been placed in a separate tribe Mack- 
inlayeae, which has traditionally been distin- 


! This research was supported by a "NER to A first author from the German Academic Exchange Service (Deutsch- 


er Akademischer Austauschdienst, DAAD). We thank P. 
suggestions on an earlier draft of the um vhs, and G 


oration during fieldwork in New Papa and Australia. We are 
Js hn, Ch. Wai Ко T. Potsch, J.-M. Veillon, Ph. Morat, Т. Jaffré, J. 
ce was provided to the first author by the following: Ordinariat fiir 


provided: E. S. Chavchavadze, H. С. Richter, 


est, B. Hyland, B. Gray, and R. sen. Assi 


Baas and an anonymo 


us referee for valuable comments and 
18 ene Nama and for his collab- 


yla Jen 
Holzbiologie. Universitat io срне МА АГ fiir Holz- und Forstwirtschaft, Hamburg; and the Botanical 


Museum, the V. L. 


Province Sud, a 


Komarov Botanical Institute, St. Petersburg. The authors acknowledge courtesies an in New 
Caledonia by in staffs of IRD (ex ORSTOM), Service de l'Environnement et ae la Gestion des 
the Direction du Développement Rural et de la Péche, Provin 


et Réserves, 


We also pin the Muséum 


National d'Histoire Naturelle in Paris, the CSIRO spun of Plant таласну (Athertdn), the Sydney Botanical Garden, 
uthor 


and the Australian National laa nile in Canberra 
an herbarium 
erbarium “Travel Award, 
Medical Sciences of Washington University, St. Lou 
2 Botanical Museum, V. L. K 


3 Missouri Botanical Garden, P.O. Box 


ork was also Јавина in part 
чш Missouri Botanical Garden, and the Division of Biology and 


s’ fieldwork was supported by NGS grant 5793-96. The 
by NSF Doctoral Ес Improvement Grant 


omarov Botanical Mo of the Russian Academy of Science, Prof. Popov Str. 2, 
197376 St. Ре Russia, oskolski@AO1818.sp а 
, Missouri 63166-0 


mohot 


and Labor- 


t. Lo ‚ U.S.A., 1 
atoire де Phanérogamie, Muséum National d’Histoire Natusile. 16 rue г '75005 Paris, France, | ки fr. 


ANN. Missour!t Вот. 


GARD. 87: 171-182. 2000. 


172 


Annals of the 
Missouri Botanical Garden 


guished on only a few floral characters, primarily 
the presence of valvate, clawed petals. While 
Mackinlayeae have been recognized in nearly all 
classifications since that of Bentham (1867), this is 
more a reflection of sparse information. on Apiope- 
talum and Mackinlaya than any real confidence in 
their relatedness. We do not consider similarities in 
the form and position of the petals as sufficient ev- 
idence to support the hypothesis that Apiopetalum 
and Mackinlaya form a monophyletic assemblage, 
and additional data are required to clarify their po- 
sition within Araliaceae. 

Apiopetalum and Mackinlaya have generally 
been regarded as most closely related to the well- 
defined group comprising Myodocarpus, Delarbrea, 
and Pseudosciadium (tribe Myodocarpeae), which is 
centered in New Caledonia (Lowry, 1986a, b). Al- 
though the monotypic genus Pseudosciadium was 
included in Mackinlayeae by Harms (1894—1897) 
because of its valvate, clawed petals, Baillon (1878, 
1879) originally suggested that it was most closely 
related to Delarbrea and Myodocarpus. Baillon’s 
initial interpretation has been supported by recent 
systematic studies, and data from the fruit structure 
(Lowry, 1986a, b) and wood anatomy (Oskolski et 
al., 1997) indicated that Myodocarpus, Delarbrea, 
and Pseudosciadium form a monophyletic assem- 
blage. Results of recent phylogenetic analyses 
based on matK, rbcL, and ITS sequence data (Plun- 
kett, 1994; Plunkett et al., 1996a, b, 1997, unpub- 
lished data) further support the hypothesis that 
these three genera form a distinct, basally branch- 
ing clade within Apiales. These studies also suggest 
that Apiopetalum and Mackinlaya comprise a sec- 
ond, isolated clade within the order. 

Apiopetalum and especially Mackinlaya resemble 
many Apiaceae in certain features, such as the pres- 
ence of clawed petals, inflexed anthers in bud, a 
bicarpellate gynoecium, and a petiole base that 
orms a dilated sheath extending around the entire 
circumference of the stem (the latter two characters 
occur only in Mackinlaya). On this basis, Philipson 
(1970) and Rodriguez (1957, 1971) regarded Api- 
opetalum and Mackinlaya as possible intermediates 
between Araliaceae and Apiaceae, traditionally re- 
garded as sister groups. In the molecular studies the 
exact position of the Apiopetalum—Mackinlaya clade 
within Apiales varies depending on the gene se- 
quence and the type of analysis. In some trees, this 
clade is sister to Apiaceae (comprising subfamilies 
Apioideae and Saniculoideae, but excluding many 
genera traditionally placed in Hydrocotyloideae), 
whereas in other analyses the two genera are sister 
to the clade comprising the remaining Araliaceae. 


Wood anatomical characters can provide useful 
complementary data for assessing relationships 
among the genera of Araliaceae, as shown by several 
previous studies (Oskolski, 1994, 1995, 1996; Os- 
kolski et al., 1997). Until now, no information on the 
wood anatomy for either Apiopetalum or Mackinlaya 
has been available. The present study surveys the 
wood anatomy of both species of Apiopetalum and 
two of the five species of Mackinlaya. The results 
are examined with regard to hypothesized relation- 
ships between these genera, as well as with Myodo- 
carpus, Delarbrea, and Pseudosciadium, core Arali- 
aceae, and certain woody Apiaceae (Bupleurum, 
Heteromorpha, Steganotaenia, Myrrhidendron, Eryn- 
gium, Gymnophyton, Asteriscum, and Trachymene). 


MATERIALS AND METHODS 


Most wood specimens examined were collected by 
the authors and C. M. Plunkett; one sample of Mack- 
inlaya macrosciadea was provided by B. Hyland 
(CSIRO Division of Plant Industry, Atherton), and 
another was taken from a plant at the Sydney Bo- 
tanic Gardens. Voucher herbarium specimens from 
New Caledonia are deposited at MO, NOU, and P, 
and from Australia at QRS and MO. For the follow- 
ing descriptions, in cases where multiple samples of 
a species were examined and a feature was seen in 
only a portion of the material, the corresponding col- 
lections are indicated in square brackets. Wood sam- 
ples were taken from trunks in Apiopetalum (from a 
primary branch in A. velutinum [3854], and from 
basal portions of stems in Mackinlaya. 

Standard procedures for the study of wood struc- 
ture were employed to prepare sections and mac- 
erations for light-microscopic (LM) studies (Carlqu- 
ist, 1988). Specimens for scanning electron 
microscopy (SEM) were prepared according to Ex- 
ley et al. (1977). Descriptive terminology and mea- 
surements follow Carlquist (1988) and the IAWA 
List of Microscopic Features for Hardwood Identi- 
fication (АМА Committee, 1989), except that for 
the diameter of intervessel pits the vertical dimen- 
sion was recorded because it is a more constant 
feature than the horizontal diameter in taxa with 
opposite and scalariform pitting. 


=. 


RESULTS 


1. APIOPETALUM (FIGS. 1—4, 8-12; TABLES 1, 2). 


Material studied. Apiopetalum glabratum Baill.: NEW 
CALEDONIA, Mé Ori, 830 m, Lowry 3375; 850 m, Lowry 
4798. A. velutinum Baill.: NEW CALEDONIA, Mt. Mou, 
1080 m, Lowry 3854; 1160 m, Lowry 4700. 


Volume 87, Number 2 


Oskolski & Lowry 
Wood Anatomy 


173 


Growth rings absent in A. glabratum [3375] (Fig. 
1) and A. velutinum [4700], and distinctly marked 
by diffuse-in-aggregates and marginal axial paren- 
chyma forming tangential lines and narrow bands 
near their boundaries (Fig. 2) in other specimens. 

Vessels rounded to slightly angular in outline, nar- 
row to moderately wide (tangential diameter (36—)66— 
87(-152) um), mostly in radial multiples of 2 to 4, 
not numerous (11 to 26 per ши? in A. velutinum 
[4700]; and 27 to 50 ла mn? in other samples). Ves- 
sel walls 2-7 um thick. Tyloses not observed. Vessel 
element length (320—620—820(-1140) e Perfora- 
tion plates simple (more than 5096), and scalariform 
with few bars (up to 18 in A. Boii [4708] and 
reticulate (Figs. 9 and 10), rarely double, in + 
oblique end walls. Intervessel pits alternate (Fig. 11), 
rarely opposite to scalariform, 3—5(-6) jum in vertical 
diameter, rounded or oval with lens- to slit-like ap- 
ertures. Vessel-ray and vessel-axial parenchyma pits 
with distinct borders; similar to intervessel pits in size 
and shape (mostly scalariform in A. glabratum 
[4798], or unilaterally compound (horizontally to ver- 
tically elongated pits on the ray cell walls abut 2 to 
5 pits on the vessel walls), with lens- to slit-like ap- 
ertures surrounded by shallow, groove-like wall sculp- 
tures (Fig. 12). Helical thickenings absent. 

Vasicentric and vascular tracheids not observed. 

Fibers libriform, thick- to very thick-walled (5—22 
p.m), non-septate, with few simple to minutely bor- 
dered pits with slit-like apertures in radial walls. 

xial parenchyma scanty in A. glabratum [3375] 

and A. velutinum [4700], somewhat more abundant 
in other specimens, both paratracheal (appears most- 
ly as solitary parenchyma cells in A. glabratum 
[3375] and A. i no: [4700], or incomplete pa- 
renchyma sheaths near vessels in others) and apo- 
tracheal (diffuse in A. glabratum [3375] (Fig. 1) and 
A. velutinum [4700], diffuse-in-aggregates or margin- 
al parenchyma (Fig. 2) in both other specimens). 
Strands composed of (2)3 to 5(7) cells. 

Rays (3)4 to 6(10) per mm, uni- and multiseriate, 


) (up 
to 8 cells wide in A. Sdn ete [4798]. Ray height 
commonly exceeding 1 mm in A. glabratum (up to 
2.9 mm high in A. p [4798]), and usually 
less than 1 mm in A. velutinum. Multiseriate rays 
formed mostly by square and procumbent cells (the 
latter more numerous in А. velutinum), with 1 to 3 
(up to 6) marginal rows of upright cells, and usually 
with sheath cells of square to upright shape. Uni- 
seriate rays composed of upright cells, rarely with 
some solitary square and procumbent cells. Pits on 
tangential walls of ray cells rounded and oval, very 
small (1-2 рт diam.). Radial canals absent. 


Crystals (appearing mostly as a combination of 
few (1 to 3) large prismatic crystals with numerous 
small ones) common in ray cells (predominantly in 
square and upright ones) in A. glabratum [4798] and 
A. velutinum [4700], and in young parts of stem (near 
the pith) in A. velutinum [3854] (Fig. 8), occurring 
rarely in A. glabratum [3375]. Crystals present also 
in non-chambered axial parenchyma cells of A. ve- 
lutinum [3854]. Brown and yellow deposits con- 
tained in a few vessels in both species examined. 


2. MACKINLAYA (FIGS. 5—7, 13—16; TABLES 1, 2). 


oo studied. Mackinlaya саи Hemsl.: AUS- 
ALIA. Queensland: Bellenden Ker, m, Plunkett 
nim Longlands Gap, 1120 m, E ibd Isabella 
Falls, ca. 30 km NW of Cooktown, 180 m, Plunkett 1549. 
M. macrosciadea (F. Muell.) Е. Muell: AUSTRALIA. 
Queensland: without precise rie p. 1100 
15281; Tolga, 800 m, Plunkett 1 | 
Plunkett 1526; cult. in sdi Botanical Garden (NSW 
208: ои C for original collection: Weston et al. 938 
AUSTRALIA. Queensland: Bellenden Ker, Mt. Bartle 
SW). 


Frere; icd at N 


= 


Growth rings absent or + distinct (Fig. 5), 
marked by lines of marginal parenchyma. 

Vessels rounded, very narrow (tangential diam- 
eter 9-31 um in M. macrosciadea [15281] and 20- 
52 um in other samples), not numerous (20 to 44 
per mm? in M. confusa [1512 and 1520], to rather 
numerous (40 to 72 per mm? in M. confusa [1549] 
and M. macrosciadea [1526 and 938], to 70 to 107 
рег mm? in M. macrosciadea [1497 and 15281]. 
solitary and in radial multiples of 2 to 5 (up to 17 
in M. macrosciadea [15281]). Vessel walls 2—5(-8) 
pm thick. Tyloses not observed. Vessel element 
length (270—)520-770(-1024) qum. Perforation 
plates scalariform with few (up to 14) bars and also 
rarely simple in M. confusa (observed in M. confusa 
[1549] only), or mostly simple (Fig. 14) and some- 
times scalariform with few (up to 6) bars (Fig. 13) 
in M. macrosciadea, occasionally reticulate, in 
oblique and horizontal end walls. Intervessel pits 
transitional to alternate (transitional ones more 
common in M. macrosciadea [1497 and 1526], and 
M. confusa [1512]), rarely opposite, 3—6 um in ver- 
tical diameter, rounded or oval with slit- to lens- 
like apertures commonly surrounded by shallow, 
groove-like wall sculptures (Fig. 14). Vessel-ray 
and vessel-axial parenchyma pits with distinct bor- 
ders, similar to intervessel pits in size and shape, 
or unilaterally compound (then horizontally to ver- 
tically elongated pits on the ray cell walls corre- 
sponding to 2 or 3 pits on the vessel walls). Helical 
thickenings absent. 

Vasicentric and vascular tracheids not observed. 


174 


Annals of the 
Missouri Botanical Garden 


й А 


ы 
. = 


TIE | 

Figures 1-4. eun micrographs of сыно, wood, — . glabratum, Lowry 3375, transverse section, axial 

parenchyma scanty paratracheal and diffuse. —2. А. пе adem, ion) ' 3854, transverse section, growth rings distinct, 

axial parenchyma scanty paratracheal pe rather a diffuse-in-aggregates tending to form long tangential lines 

on growth ring boundaries. —3. A. glabratum, Lowry 3375, tangential section, mostly 4—5-seriate rays with sheath 
| 4. А. velutinum, Duy 3854, tangential section, mostly 3—4-seriate rays with sheath cells. Scale bar — 


pum. 


175 


Oskolski & Lowry 
Wood Anatomy 


Volume 87, Number 2 


2000 


- ua t usi == - 
== X ди у. 
OC er = уз 

==--___--______-_______ нв ÉÓÉ(N 


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во ~ a^ dem 
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== - — 


— YI JITIHILIHIBABAIYXI X 


Mae AAC а, 


5281, 


Hyland 1 
paratracheal and diffuse. —6. M. macrosciadea, 


macrosciadea, 


100 шт. 


. M. 


еішіпит, Lowry 3854, radial section, 


1 


Light micrographs of Mackinlaya and Apiopetalum wood. 


transverse section, growth rings absent, axial parenchyma scanty 


Figures 5-8. 
crystals (arrow) in a ray cell in the young part of the stem (near the pith). Scale bar 


section, helical thickenings on the inner walls of the ray cells (arrows). —8. A. 


Missouri Botanical Garden 


Annals of the 


176 


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Volume 87, Number 2 Oskolski & Lowry 177 
2000 Wood Anatomy 
Table 2. Anatomical characters of libriform fibers and ray parenchyma in Apiopetalum and Mackinlaya. 
umber of Number of 
Length Width Height ипіѕегіаіе multiseriate 
of fibers of rays of rays 
1 2 3 
Apiopetalum glabratum Baill., 1029 + 25.4 4.3/6 1.1/2.9 0.8 3.0 
Lowry 3375 (710-1350) (0—2) (2—5) 
A. glabratum Baill., 1267 + 26.4 3.8/8 1.1/1.9 0.7 4.0 
Lowry 4798 (970-1840) (0-2) (2-5) 
Apiopetalum velutinum Baill., 1196 + 26.8 3.8/6 0.8/1.2 1.2 3.6 
Lowry 3854 (920-1510) (0-2) (2-5) 
A. velutinum Baill., 1074 + 23.1 3.3/5 0.6/1.2 0.5 3.6 
Lowry 4700 (810-1390) (0-1) (2-5) 
Mackinlaya confusa Hemsl., 1324 + 28.3 1.3/3 0.8/3.1 10.5 2.4 
Plunkett 1512 (870-1610) (7-15) (1-3) 
M. confusa Hemsl., 1293 + 27.6 1.3/3 1.4/3.6 14.0 2.5 
Plunkett 1520 (870-1620) (12-16) (1-5) 
M. confusa Hemsl., 1076 = 32.2 2.1/5 1.7/3.6 8.6 4.5 
Plunkett 1549 (730-1530) (6-12) (4-6) 
Ma pa zum (F. Muell) F. Muell 1022 + 21.0 2.2/6 1.1/2.6 1.2 4.7 
Plunkett (730-1300) (3-12) (3-6) 
M. macrosciadea (F. Muell) F. Muell 1045 + 22.2 1.6/4 0.9/2.1 10.2 4.2 
Plunkett 1526 (780-1310) (4-14) (2-7) 
М. macrosciadea (F. Muell) Е. Muell, 801 + 15.3 1.5/3 0.7/1.1 13.9 4.7 
Hyland 15281 (620—1040) (6-21) (2-8) 
M. macrosciadea (F. Muell) F. Muell, 722 * 24.0 1.8/4 0.9/1.9 8.7 12.5 
Weston et a (420-1060) (4-11) (10-14) 


Characters: 1, Length of petty fibers (рт): average + 


average/maximum number ell; 3, Height of rays (mm): 


= 


standard error (minimum — maximum); 2, Width of iyw 
average/maximum; 4, Number of uniseriate rays per m 
). 


average (minimum — о. 5, Number of bias tire rays per mm: average (minimum — maximum 


Fibers libriform, thin- to thick-walled (walls 3- 
5(-8) jum thick), non-septate, with rather numerous 
simple to minutely bordered pits with slit-like ap- 
ertures in radial walls. 

Axial parenchyma scanty paratracheal (appear- 
ing as solitary parenchyma cells near vessels), dif- 
fuse, and marginal, sometimes forming interrupted 
tangential lines (Fig. 5) and narrow bands near 
boundaries of the growth rings. Strands composed 
of 3 to 6(7) cells. Helical thickenings rarely present 
(M. macrosciadea [938]) on inner walls of the axial 
parenchyma cells in contact with vessels (Fig. 15). 

Rays numerous ((14)18 to 21(24) per mm), uni- 
seriate and multiseriate of 2 or 3 cells in width in 
M. confusa [1512 and 1520] and wider in other 
samples (up to 6 cells in M. macrosciadea [1497 ]). 
Uniseriate rays more numerous than multiseriate 
ones in all samples except M. macrosciadea [938]. 
Ray height commonly exceeding 1 mm in M. con- 
fusa [1512 and 1549], and in M. macrosciadea 
[1497], and commonly less than 1 mm in other 
samples. Uniseriate rays composed of upright and 
few square cells. Alternation of uniseriate and mul- 


tiseriate portions common in same ray. Uniseriate 
portions of multiseriate rays very long (up to 20 
rows), formed by upright and solitary square cells, 
multiseriate portions usually as wide as uniseriate 
ones, composed of square and a few procumbent 
cells (Fig. 6). Pits on tangential walls of ray cells 
rounded and oval, very small (1-2 jum diam.). He- 
lical thickenings rarely present in M. macrosciadea 
[938] on inner walls of both upright/square and pro- 
cumbent ray cells in contact with vessels (Figs. 7, 
16). Radial canals absent. Crystals not observed. 
Brown and yellow deposits contained in a few to 
many vessels in both species examined, and also 
in cavities of many fibers and parenchyma cells of 
M. macrosciadea [93 


DISCUSSION 

Very little variation was observed in wood struc- 
ture within the Apiopetalum and Mackinlaya spe- 
cies examined. Apiopetalum glabratum differs from 


A. velutinum by higher and wider rays (Table 2). 


Mackinlaya confusa is distinct from M. macroscia- 


178 


Annals of the 
Missouri Botanical Garden 


Figures 9-12. Scanning electron micrographs of Apiopetalum wood. —9. A. velutinum, Lowry 3854, reticulate 


pl 


perforation plate. —10. A. velutinum, Lowry 3854, radial section, note one scalariform and two simple perforation 
ates. —11. А. glabratum, Lowry 3375, alternate intervessel pitting. —1 


D 


2. A. velutinum, Lowry 3854, vessel-ray pitting 
50 um, 1 


with lens-like apertures surrounded by shallow, groove-like wall sculptures. Scale bar in Figures 9-11 — 
‘igure 1: 20 um. 


Figure 12 = 


dea by the predominance of scalariform perforation 
plates with more numerous bars; it also has less 
numerous vessel lumina, but more samples must be 
studied before the importance of this character can 
be interpreted. 

elical thickenings on the walls of both ray and 
axial parenchyma were observed rarely in one sam- 
ple of Mackinlaya macrosciadea [938] (Figs. 7, 15, 
16). This feature generally appears in tracheal el- 
ements (vessel elements, vascular/vasicentric tra- 
cheids, fibers), and has also been reported very 
rarely in axial parenchyma of some Trigoniaceae 
(Heimsch, 1942), Ancistocladaceae (Gottwald & 
Parameswaran, 1968), and Chrysobalanaceae (ter 
Welle, 1975). The presence of helical thickenings 


in ray cells appears to have been reported previ- 


ously only for Trigonia sericea HBK (Heimsch, 
1942: 133). In the sample of Mackinlaya, helical 
thickenings are found in parenchyma cells adjacent 
to the vessels. 

Mackinlaya and Apiopetalum can be clearly dis- 
tinguished from one another on the basis of their 
wood anatomy. Differences occur in several features, 
e numerous vessels, 
thinner fiber walls, narrower and more numerous 


including narrower and mor 


rays, the absence of diffuse-in-aggregates axial pa- 
renchyma, and the absence of crystals in the ray and 
axial parenchyma cells in Mackinlaya (Tables 1-3). 
The very narrow and relatively numerous vessels, 
and the 1- or 2-seriate rays found in Mackinlaya are 
probably correlated with the shrubby habit of the 
species studied and the correspondingly small di- 


Volume 87, Number 2 
2000 


Oskolski & Lowry 
Wood Anatomy 


179 


$ 
NE 


LE 
+ % 
Әг) 
ха | 
е 9 
= 
VN 
is 


42 


Figures Scanning electron oe of M. macrosciadea, Weston et al. 938, wood. —13. Vessel element, 
scalariform perforation plates with few bars. —14. Simple perforation plate; small intervessel pits with bn to lens-like 
apertures surrounded by shallow ete wall sculptures. —15. Helical thickenings in an i parenchyma cell. 
—16. Helical thickenings in a ray cell. Scale bar in Figure 13 = 50 pum; in Figures 14—16 ) jum. 


ameter of their stems. Other shrubby Araliaceae 
such as species of Oplopanax and Astrotricha (Os- 
kolski, 1994, 1996) have mean values of vessel di- 
ameter and frequency, as well as of ray width, similar 
to those observed in Mackinlaya. Thickness of fiber 
walls, the type of axial parenchyma, and the occur- 
rence of crystals in parenchyma cells appear to re- 
flect relationships more than differences in habit or 
habitat (Oskolski, 1994, 1996), and could be of pos- 
sible taxonomic value (Table 3). 

Despite these differences in their wood anatomy, 
Mackinlaya and Apiopetalum also share a numbe 
of significant features. These include the small in- 
tervessel pits (3 m), the occurrence of both 
paratracheal and apotracheal (diffuse and diffuse- 
in-aggregates, the latter occurring only in Apiope- 


e 


talum) axial parenchyma, non-septate fibers, a pre- 


dominance of upright and square cells in the ray 
composition, and brown deposits in the vessels. Al- 
though each of these characters has also been ob- 
served in other genera of Araliaceae and Apiaceae, 
their combined occurrence in Mackinlaya and Api- 
opetalum is notable for the order Apiales (Table 3) 
and supports the suggestion that these two genera 
are closely related. Diffuse and diffuse-in-aggre- 
gates parenchyma is found alone only in species of 
Myodocarpus, Delarbrea, and Pseudosciadium. Co- 
occurrence of both diffuse and paratracheal axial 
parenchyma is known only from three New Cale- 
hefflera 
| other represen- 
tatives of Apiales examined to date have only a 


donian species of the pan-tropical genus Sc 


(Oskolski & Lowry, in prep.). 


single parenchyma type. The co-occurrence of apo- 
tracheal and paratracheal axial parenchyma could 


Annals of the 


180 


Missouri Botanical Garden 


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Volume 87, Number 2 
2000 


Oskolski & Lowry 181 


Wood Anatomy 


thus be regarded as an apomorphy for Mackinlaya 
and Apiopetalum. 

Besides diffuse and/or diffuse-in-aggregates axial 
parenchyma, several other wood-anatomical fea- 
tures are also shared among Mackinlaya, Apiope- 
talum, and the assemblage comprising Myodocar- 
pus, Delarbrea, and Pseudosciadium, such as the 
presence of small intervessel pits. This character 
occurs nowhere else within Araliaceae except in 
the Australian endemic Astrotricha (Table 3), which 
may also be a basally branching lineage of Apiales 
as indicated by recent molecular sequence data 
from ITS (Mitchell & Wagstaff, 1997; G. M. Plun- 
kett, pers. comm.). However, the wood of Astrotri- 
cha differs strongly from that of Mackinlaya and 
Apiopetalum, notably in its axial parenchyma and 
ray types, and the presence of helical thickenings 
on the vessel walls (Oskolski, 1996). Species of 
Mackinlaya, Apiopetalum, Myodocarpus, Delarbrea, 
and Pseudosciadium also form very thick-walled 
non-septate fibers, which are unusual within the 
order. Each of these wood characters may be a syn- 
apomorphy within Apiales (Table 3), supporting the 
hypothesis of monophyly of the alliance that con- 
tains these five genera. They could likewise be 
symplesiomorphic in that they occur independently 
(but not together) in other groups within the order 
(Table 3), suggesting the possibility of parallel evo- 
lution. Resolving this issue is difficult at present 
because we lack sufficient data on wood features 
(especially the size of intervessel pits) for nearly all 
genera of woody Apiaceae. 

These similarities notwithstanding, wood anatomy 
of Mackinlaya and Apiopetalum is nevertheless very 
distinct from that of Myodocarpus, Delarbrea, and 
Pseudosciadium. In addition to the co-occurrence of 
both paratracheal and apotracheal (diffuse and dif- 
fuse-in-aggregates) axial parenchyma, ray types also 
differ. In Mackinlaya and Apiopetalum, rays are het- 
erogeneous, with both upright and square as well as 
procumbent cells; they are homogeneous with exclu- 
sively procumbent cells in the other genera. Fur- 
thermore, Mackinlaya and Apiopetalum have brown 
and yellow deposits in their vessels and lack radial 
canals. Wood anatomy thus supports the inclusion of 
these genera in a monophyletic Mackinlayeae (Table 

), and refutes the inclusion of Pseudosciadium 
(Baillon, 1878, 1879), which is most closely related 
to Myodocarpus and especially Delarbrea (Lowry, 
1986a, b; Oskolski et al., 1997; Plunkett, 1998, un- 
published data). 

Using generally accepted trends in wood evolution 
(Bailey & Tupper, 1918; Frost, 1930a, b, 1931; Ca- 
rlquist, 1988; Baas & Wheeler, 1996) to determine 


character polarity, several features are regarded as 


plesiomorphic for Mackinlaya and Apiopetalum. 
These include predominantly scalariform perforation 
plates and relatively long vessel elements, whereas 
short vessel elements with exclusively simple рег- 
foration plates of woody Apiaceae (Metcalfe 

Chalk, 1950; Rodriguez, 1957; Greguss, 1959; 
Schweingruber, 1990) are regarded as apomorphic. 
Average vessel element lengths are 651—822 um in 
Apiopetalum and 518-770 jum in Mackinlaya (Table 
1), within the range seen in most other Araliaceae, 
which vary from 650 to 900 jum. The lowest reported 
averages аге 366 шт in Oplopanax horridum (J 
Smith.) Ма. and 374 qum in Eleutherococcus sessi- 
liflorus (Rupr. & Maxim.) S. Y. Hu, with the highest 
of 1339 jum reported in Schefflera gabriellae Baill. 
(Oskolski, 1994, 1996; Oskolski & Lowry, in prep.). 
By contrast, vessel elements in Apiopetalum and 
Mackinlaya are distinctly longer than in woody Api- 
aceae, which have average values that are generally 
less than 400 jum. A notable exception is Hetero- 
morpha arborescens (Thunb.) Cham. & Schlecht., 
whose average vessel length reaches 502 шт (Rod- 
riguez, 1957). Wood features characteristic of Mack- 
inlaya and Apiopetalum, such as the occurrence of 
diffuse and diffuse-in-aggregates apotracheal paren- 
chyma types, small intervessel pits, and heteroge- 
neous rays with distinct uniseriate portions com- 
posed of upright and square cells (Kribs’s (1935) ПА 
type), have not been reported among the woody Api- 
aceae examined, including species of Bupleurum, 
Heteromorpha, Steganotaenia, Myrrhidendron, Eryn- 
gium, nophyton, Asteriscum, and Trachymene 
Table 3), among others (Metcalfe & Chalk, 1950; 
Rodriguez, 1957; Greguss, 1959; Schweingruber, 

) 


— 


Wood anatomical features thus confirm that 
Mackinlaya and Apiopetalum are closely related 
and occupy an isolated position within Apiales, as 
proposed by Plunkett (1998). However, wood char- 
acters neither validate nor refute the hypothesis 
that the two genera under study are intermediate 
between Araliaceae and Apiaceae (Philipson, 1970; 
Rodriguez, 1957, 1971), nor do they offer a clear 
indication of the group’s phylogenetic position with- 
in the order. Based on current data, we cannot 
identify any reliable synapomorphies in the wood 
to support a sister relationship between the Apiope- 
talum—Mackinlaya clade and core Apiaceae (ex- 
cluding most members of subfamily Hydrocotylo- 
ideae), as suggested by Plunkett (1998), nor 
between them and core Araliaceae (excluding My- 
odocarpus, Delarbrea, and Pseudosciadium). How- 
ever, the study of wood structure is of limited use 
in assessing relationships between Mackinlaya and 
Apiopetalum and non-woody members of the Mack- 


182 


Annals of the 
Missouri Botanical Garden 


inlaya group, as defined by Plunkett (1998, pers. 
comm.), or between them and nearly all of the other 
genera of Apiaceae, which are likewise herbaceous. 
Several wood characters (small intervessel pits, 
thick fiber walls, non-septate fibers) do suggest a 
relationship between Mackinlaya and Apiopetalum 
and the alliance comprising Myodocarpus, Delar- 
brea, and Pseudosciadium, as previously hypothe- 
sized (Plunkett et al., 1996a, b, 1997). 


Literature Cited 


Baas, P. & E. A. Wheeler. 1996. Parallelism and revers- 
ibility i in xylem evolution. A review. ТАМА J. 17: 351- 
364. 


Bailey, 1. W. & W. W. Tupper. 1918. Size variation in 
tracheary cells. 1. A comparison between the secondary 
xylems of vascular cryptogams, gymnosperms and an- 
giosperms. Proc. Amer. о s rts 54: 149-204. 

Baillon, H. 1878. Rec es nouvelles sur les Aralié 
et sur la famille Ombelliferes e en мее | Адапзота 12: 
125-178. 

. 1879. Ombelliferes. Histoire des Plantes 7: 66— 


Bentham, G. 1867. Araliaceae. In: С. Bentham & J. I 
ooker, Genera Plantarum 1: 931—947. Lovell ides 
Williams & Norgate, London 

— S. 1988. Comparative Wood Anatomy. Spring- 

r-Ver тар, Berlin. 

Exley, R . В., B. A. Меујап & B. С. Butterfield. 19 
technique for obtaining clear cut surfaces on ds sam- 
ples prepared for | scanning electron microscope. J. 
prey: 110: 7 

Frost, Е. Н. 1930a. n sialization i in sec dd nd in 
в I. Origin of vessel. Bot. Gaz 7-94. 

—— ———. 1930b. Specialization in sec ondary ae in di- 
apg II. Evolution of end wall of vessel segment. 

E 89: 1 анага, 

931. 


Specialization in secondary xylem in di- 

dne III. е of lateral wall of vessel 

segment. Bot. Gaz. 90: 88—96. 

Gottwald, H. & N. Parameswaran. Das sekundüre 
ylem und die systematische Stellung der Ancistrocla- 

daceae und Dioncophyllaceae. Bot. Jahrb. Syst. 88: 49— 
69, 


1968. 


Greguss, P. 1959. Holzanatomie der Europüischen Laub- 
hólzer АТ | Sträucher. Akademiai Kiadó, Budapest. 

Harms, H. Araliaceae. In: ngler & К. ai 
(editors), Die у тоем Pflanzenfamilien III. 8: 1-6 

Enge Poe a ui zig. 

Heimsch, C., 942. Comparative anatomy of the sec- 
ondary ia $ the Gruniales and Terebinthales of 
Wettstein with reference to taxonomic grouping. Lilloa 

: 83-198. 

Hutchinson, J. 1967. Araliaceae. /n: The Genera of Flow- 
ering Plants 2: 52-81. ея Univ. Press, London. 
IAWA Committee. 1989 A list of microscopic fea- 
tures hardwood identification. IAWA Bull., n.s., 10: 

219–: 


Kribs, D. A. 1935. Salient lines of structural o d 
tion in the wood rays of dicotyledons. Bot. 
547-557 

Гомту, Р. Р, П. 1986a. A Systematic Study of Three Gen- 


era of Araliaceae Endemic to or а оп New Са!- 
edonia: Delarbrea, Myodoca ы о, 
Ph.D. ае Washington | vire St. Louis. 


—— ———. 1986b. A systematic ud of Delarbrea Vieill. 
(Araliaceae). Allertonia 4: 169— 

Metcalfe, C. R. & L. Chalk. 1950. Anatom of the Di- 
cotyledons, Vol. 2. Cla gia Press, 

Mitchell, A. D. & S. J. Wagstaff. 1997. Phylogenetic re- 
lationships of P. cin species (Araliaceae) in- 


and m 

Oskolski. A. A 1994. Wood Anatomy of Araliaceae. Ko- 
marov Bot. Institute, St. Petersburg. [In Russian.] 

. 1995. Wood anatomy of Schefflera and related 
taxa о € 16: 191-215. 

——— 96 rvey of the wood anatomy of the Ar- 
me s 99-119 1 т L. A. Donaldson, A. P. Singh, 
B. G. e dix & L. J. Whitehouse (editors), Recent 

vances in Wood Anatomy. New Zealand Forest Re- 
Mah "QE. Rotorua 
= Р. P. Lowry II & H. G. Richter. 1997, pl јин 
wood anatom y of Myodocarpus, Delarbrea, ved 
иш лева) Adansonia, Ser. 3, 61-75. 

Philipson, W. R. 1951. Contributions to our eum dge 
of Old World Araliaceae. Bull. British Mus. Nat. Hist., 
Bot. 1: 3-20. 

1970. Constant e variable features of the Ar- 
aliaceae. Pp. 87-100 in N. K. B. Robson, . Cutler 
& M. ия от) Now Research in Plant Anato- 
my. Academic Press, New York » аа, 

979. Araliaceae, part : С. С. С. J. е 
Steenis mnes press Flora eus ‚ Ser. I, 9 
rtinus Nijhoff, Dr. W. Junk Publishers, The 
Hague, Dior London. 

Plunkett, G. M. 1994. A гоне Ар- 
proach to the “Family-Pair Dilemma" іп Apiales an 
Cyperales. Ph. = Dissertation, и Жон State Uni- 
versity, ME 


айай araliad genera from New Cal 
donia ш ee implications for phylogeny and taxon- 
omy in Apiales = and Araliaceae). Amer. J 
Bot. 85 (6, suppl.): 
"b. E Soltis & B's . Soltis. 1996a. Higher level 
relationships of Apiales (Apiaceae and Ра на 
esti on p d veg analysis of rbcL sequences. 
499—515. 


r. J. Bot 


1996b. Evolutionary pat- 
terns in E Inferenc 'es based on matK sequence 
77-495. 


data. Syst. Bot. 21: 


1997. Clarification of the 
family- "pair E between Apiaceae and Arali- 
e based o nd rbcL sequence data. Amer. 
I Bot. 84. 565-58 
Rodriguez, R. L. Systematic anatomical ee on 
Morrhidendron and ps woody Umbellales. Uni 
lif. Publ. Bot S 145-318 
— 1971. The relationships of the Umbellales. In: 
V. H. Heywood pee r), The Biology and Chemistry of 
gene ines Supplement 1, Bot. J. Linn. Soc. 64: 
63-91. Academic Press, London & New York. 
Schweingruber, Е. H. 1990. Anatomie europäischer Hél- 
zer. үү ч European Woods. Verlag Paul Haupt, 
Bern & Stut 
Takhtajan, A. г “1987. Hic Magnoliophytorum. Nau- 
ka, Leningrad. [In je n.| 
Tseng, С. 1982. А new classification scheme 
for the family Майкл. Acta Phytotax. Sin. 20: 125— 


Viguier, sherches anatomiques sur la clas- 


R. 1906. Rec 
sification des Araliacées. Ann. Sci. Nat. Bot. IX. 4: 1– 


Welle, B. J. H. ter. 1975. Spiral thickenings in the axial 
ое of he Acta Bot. Neerl. 24: 


CARLOS SPEGAZZINI 
(1858-1926): TRAVELS AND 
BOTANICAL WORK ON 
VASCULAR PLANTS! 


Liliana Katinas,? Diego С. Gutiérrez,” 
and Silvia S. Torres Robles? 


ABSTRACT 


Carlos Luis сени номе was a lending figure in Argentinian natural history, mainly recognized for his 


is specimens and publicat 
na are summarize 


in Argentinian journals, describing 


rning Spegazzini’s travels and his botanical work focusing on 
tions, is provided. The i ini 

entin The identification of his typic and non-typic m 
plicated by the lack or sparcity of bae information. Spegazzini published ca. 100 


by Spegazzini is igi i ipud m was men at the University of La Plata and Buenos Aires in Argentina, curator 


of the herbariu 


founder of an сом and an institute of mycology i 


of the National Department of Agric aper first head of the herbarium of Museo de La Plata, and 


city. 


Key words: Argentina, idu. plant collections, iniu travels, vascular plants. 


Carlo Luigi Spegazzini, or “Carlos Luis" Spegaz- 
zini (the Spanish translation of his name by which 


he was recognized in Argentina and in the rest of 


the world), was a leading figure in Argentinian nat- 
ural history, for both his mycological and vascular 
plant studies. Spegazzini was one of the most sig- 
nificant explorers of Patagonia in terms of the vol- 
ume of his collections. He also traveled and col- 
lected extensively in almost all of Argentina, from 
northern Salta to southern Tierra del Fuego. 

This botanist, who was attracted by the South 
American biota since he was a young student in 
Italy (Spegazzini, 1884a), is best known for his 
work in mycology, although his vascular plant sys- 
tematics is of equivalent importance. His herbari- 
um of vascular plants, collected by himself and by 
other collectors, reaches 100,000 specimens (Mol- 
fino, 1929), and he described approximately 1000 
new taxa during his career. In the process, Spegaz- 
zini published more than 320 papers, of which ca. 
100 refer to vascular plants. Furthermore, he was 
a teaching professor of botany, zoology, mineralogy, 
geology, and phytopathology, curator of the Minis- 
terio de Agricultura de la Nación herbarium, first 
head of the herbarium of Museo de La Plata, and 


founder of the Arboretum of the Facultad de Cien- 
cias Agronómicas y Forestales in La Plata city. 
Biographies of Carlos Spegazzini considering his 
mycological work have been published previously 
(Scala, 1919; Hauman, 1923; Molfino, 1929, 1951; 
Arambarri & Spinedi, 1996), but details of his trav- 
els and botanical work on vascular plants are still 
undocumented. This information, 
specimens and publications, is provided herein. 


including his 


BIOGRAPHICAL SKETCH 

A detailed biography of Carlos Spegazzini has 
been published by José Molfino (1929), a botanist 
who married one of Spegazzini's daughters. We pre- 
sent here a brief outline of Spegazzini's life, as it 
pertains to his botanical endeavors. 

Carlo Luigi Spegazzini (Fig. 1) was born on 20 
April 1858 at Bairo in Torino, in northern Italy. He 
took courses in the School of Viticulture and Enol- 
ogy in Conegliano, Venice (Italy), where he gradu- 
ated in 1879. There, he met Pier Andrea Saccardo 
1845-1920), professor and founder of the Scuola 
micologica di Padova (Lazzari, 1973), who would 
be a major influence in Spegazzini's scientific ca- 
reer. Soon after graduating, thinking that botani- 


~ 


! We are grateful to Jorge Сгіѕсі, Genoveva Dawson, Susana Freire, Mario Teruggi (Museo de La Plata), Gustavo 


Giberti (Museo Juan Dominguez), Roberto 
Museum) for providing helpful comments on the man 


for the improvement of the manuscript. L. 
no. 5776-9 


Kiesling (Instituto Darwinion), Hugh Пиз, Bob Kowal, Mark iude (Uni- 
versity of Wisconsin-Madison), Giuseppe Manganelli a di Siena), and Frank Horsman (The Natu 
cript. We are indebted to David Boufford (Harvard University} 
for assistance with Spegazzini's infraspecific taxa from p Gray Herbarium Card Index 
. was supported in this research by the National Geographic Society (Grant 
6) and Consejo Nacional Pa Investigaciones Científicas y Técnicas (CONICET). 


and two anonymous rev iewers 


? Departamento Científico de Plantas Vasculares, Museo de La Plata, Paseo del Bosque, 1900 La Plata, Argen- 


tina. 


ANN. Missouni Bor. Савр. 87: 183-202. 2000. 


184 


Annals of the 
Missouri Botanical Garden 


Figure 1. 


Carlo Luigi Spegazzini in a photograph tak- 
en on 20 April 1922, the day of his 65th birthday. 


cal studies in Italy were well covered and com- 
pleted by other Italian scientists, Spegazzini decid- 
ed to move to another land. In December 1879, he 
disembarked in Buenos Aires harbor. It is interest- 
ing to note that the period from 1850 to 1923 was 
characterized by the arrival in Argentina of a large 
number of European naturalists (Ringuelet, 1960; 
Ragonese, 1986). They came mainly from Germany 
(Hermann Burmeister in 1857, Paul Lorentz and 
Georg Hieronymus in 1870, Fritz Kurtz in 1884), 
Italy (Domingo Parodi in 1875, Spegazzini in 1879, 
Augusto Scala in 1909), Russia (Carl Berg in 1873, 
Nikolai Alboff in 1895), Switzerland (Theodor 
Stuckert in 1869), and Belgium (Lucien Hauman 
in 1904). 

The Spegazzini story in Argentina evolves from 
two fortunate circumstances: his unbounded enthu- 
siasm for the natural sciences, and a scientifically 
young country waiting to be discovered. In the year 
after arriving he made his first field trip and pub- 
lished his first new species of vascular plant, Ca- 
bomba australis Speg. (Nymphaeaceae) (Spegazzini, 
1880). In 1885, he was appointed professor in the 
Instituto Agronómico de Santa Catalina (Buenos 


Aires Province), and professor and temporary dean 
in the Facultad de Agronomfa (University of La Pla- 
ta). Two years later, he was designated Head of the 
Department of Botany in Museo de La Plata. At the 
same time, Spegazzini was a student at the School 
of Agriculture, obtaining his degree on 23 Decem- 
ber 1897 (Ragonese, 1986). 

In 1898 Spegazzini was designated curator of the 
Herbarium of the Ministerio de Agricultura de la 
Nación, in Buenos Aires (currently the Herbarium 
of the Instituto de Recursos Biológicos, INTA), and 
in the following year he founded the Arboretum of 
the Facultad de Agronomía in La Plata. In 1900 he 
was appointed Professor in the Facultad de Far- 
macia y Bioquímica, at the University of La Plata. 
He was a leader in the study of agriculture in the 
country. In his research in this field in Argentina, 
he discovered a fungus that attacks grape vines 
(Plasmospora viticola), the bacterium that causes a 
disease of sugar cane (Bacillus sacchari), and fungi 
that attack tobacco (Peronospora nicotianae) and al- 
falfa plants (Uromyces striatus). 

During the years 1912 to 1914 he returned to 
Europe (Spegazzini, 1916a) to work in herbaria and 
also to visit his mentor and friend Pier Andrea Sac- 
cardo. One of his last botanical activities on 1 April 
1925, was the foundation of a scientific journal in 
Argentina, Revista Argentina de Botánica. [Unfor- 
tunately, after Spegazzini's death the journal ceased 
publication.] 

On 1 July 1926, at the age of 68, Carlos Spe- 
gazzini died in his home at La Plata from kidney 
disease. In his will he donated the house to science 
with the condition that it become an institute of 
botany bearing his name (currently /nstituto de Bo- 
tánica Carlos Spegazzini, LPS; Holmgren et al., 
1990). He also donated his library (ca. 6000 books 
and papers), scientific instruments, and personal 
herbaria of fungi and vascular plants (Anonymous, 


1930). 


COLLECTING EXPEDITIONS 


Carlos Spegazzini was an active field worker 
throughout his life. His principal expeditions and 
collecting efforts are outlined in Table 1 and Fig- 
ures 2 and 3. Only the most representative of his 
frequent collecting trips around the cities of La Pla- 
ta and Buenos Aires are cited in Table 1. 

Spegazzini collected and also received speci- 
mens from other collectors (see Table 1) and his 
own children, Etile, Propile, and Rutile Spegazzini. 
His preferred area of collecting was Patagonia, the 
southern tip of Argentina including Mendoza, Ne- 
uquén, Río Negro, Chubut, Santa Cruz, and Tierra 


Моште 87, Митбег 2 
2000 


Katinas et al. 


185 
Carlos Spegazzini 


=. vid 


CHILE 4... LS 
Siá 25 


t Pie de Palo 
"a • 


76 Viliavicencio 


“о 
Acohcagya 


Џ 
i 
: 10 ' 


б ей Grande 


Ma 


p 1 = Jujuy; 2 = Salta; 3 
Juan; 9 = Córdoba; 10 = Mendoza; 1 


del Fuego (Fig. 3). The countries adjacent to Ar- 
gentina, on the other hand, were scarcely visited 
for collecting. 

One of the first remarkable trips Spegazzini made 
was in 1881, two years after his arrival in Argen- 
tina. On December 18 he embarked as a botanist, 
representing the University of Buenos Aires, in the 
expedition of Lieutenant Santiago Bove aboard the 
ship Cabo de Hornos. Spegazzini, as well as a ge- 
ologist, a geographer, and a zoologist, departed from 
Buenos Aires heading for Isla de los Estados and 
Tierra del Fuego, the southernmost part of America. 
After arriving at Punta Arenas, Chile, Spegazzini, 
Bove, and the geologist embarked in a smaller ship 
to sail through the narrow channels of Tierra del 


| f northern and central Argentina with the main localities (mentioned in Table 1) 
and а) sallean vascular plant ‹ collections that contributed to 


= Catamarca; 4 = Tucumán; 5 = C 
1 = Entre Ríos; 12 = Buenos Aires. 


of Spegazzini's 
ers refer to the Argentine 
haco; 6 — Misiones; 7 — La Rioja; 8 — San 


his herbarium. Numb 


Fuego. A storm caused their shipwreck, but Spe- 
gazzini saved some of the plant collections by 
swimming to the coast and burying them in the 
snow to preserve them. While the three survivors 
awaited rescue, Spegazzini contacted the Indians of 
the area and learned the grammar of their language 
(Spegazzini, 1884b). Spegazzini returned to Buenos 
Aires on 27 September 1882. As a result of this 
adventure, Spegazzini collected 1108 specimens 
(mosses, fungi, lichens, and vascular plants). Forty- 
two years later, in January 1924, he returned to 
Cabo de Hornos in Chile where he discovered very 
different conditions. The Native Indian populations 
had almost disappeared, and the fauna and flora 
were reduced due to pressure from the increasing 


186 


Annals of the 
Missouri Botanical Garden 


-.2'о• = 


pà 
12%) 


ae 


mm. p ERO B 
e = 


** «Вю са ^ 


Гаро qu Ки B 
1 


ә 


\ <. [] 
Cabo de Hornos 


- 7 
Sierras 


de Cura Malal 
SU. Sierras de Tandil 
Eu E A 
ЫЧ se 


22: 


V" Sierra de la Ventana 


D Golfo San Matías 


Carmen de Patagones 


S0? 


0 100 200 300 miles 


U 200 
€ Isla de los Estados 


400 km 


60° | 


Figure 3. Map of southern Argentina with the main localities (mentioned in Table 1) of Spegazzini's and other 
m vasc n plant collections a PT ыы his ipid pe Numbers refer to the Argentine pores 12 
Chubu 


— Buenos Aires; Neuquén; 14 = Rio Negro; 


populations of the growing cities there, the burning 
of native trees, and the introduction of exotic spe- 
cies (Molfino, 1929 

Some trips were undertaken by order of either 
the Argentine government or commercial entities. 
For instance, he went to Chaco to install an alcohol 
factory; to Tucumán to study the sugar cane dis- 
ease; to western Argentina to analyze the viticul- 
tural industry; and to northwestern Argentina to 
search for and study plants that produce rubber. In 
all these trips he looked for the opportunity to col- 


; 16 = Santa Cruz; 17 = Tierra del Fueg, 


lect plants because he found that it was usually 
difficult to organize collecting trips that lacked di- 
rect utility. In his own words: 


“Me es disgustoso declarar que he hallado constante- 
mente una cierta indiferencia, por no decir hostilidad, toda 
vez que he querido llevar a cabo algún viaje botánico que 
no tuviera un fin práctico inmediato, así que para prepar- 
arme debidamente al desempefio de mis funciones de bo- 
tánico oficial he tenido nec i iu de aprovec shar toda 
sión económica o indust 
permitiesen desempeñar ies о 
acumulando asf los materiales que ahora forman Ја base 


Volume 87, Number 2 
2000 


Katinas et al. 
Carlos Spegazzini 


187 


Table 1. 


Carlos Spegazzini’s vascular plant-collecting expeditions, with the names of other collectors who provided 


specimens for him. The dates, names, and description of the travels are taken from herbarium labels and from the 


literature cited. * 


= data taken exclusively from herbarium labels. 


Dates 


Collecting expeditions 


Bibliography 


Nov. 1880 


Dec. 1880 


Dec. 1881-Nov. 1882 


1883 
Early 1883 


Mar.-Nov. 1883 


July-Aug. 1883 


Dec. 1889 
1890 
May-June 1890 
1891 

1892 

Jan. 1894 

Mar. 1894 


Sep. 1894 
Jan.-Mar. 1895 


Nov. 1895 


Dec. 1895 
Late 1895—Early 1896 


С. 


Spegazzini and D. Parodi; Argentina. Prov. Buenos А1- 
res: areas surrounding the city of Buenos Aires (e.g., 
Boca del Riachuelo, Palermo, Recoleta, San José de Flo- 
res, Isla Maciel, Puente Alsina, Montes del Tordillo, 
Montes Grandes, Montes del Real Viej 


C. Spegazzini; b. aie Prov. Buenos PUR General Lav- 
gdalen 


e 


e& n0 


E 


ро 


о t 


ррррь o 


ono 


On 


alle, Ma 


. Spegazzini in Bove's expedition; Argentina. Prov. Santa 


Cruz: Río Gallegos, Río Santa Cruz, Salinas, Isla Pavón, 
Monte León, Isla de los Baguales, Isla de los Le 
Misioneros, Cabo Vírgenes. Prov. Tierra del Fuego: estre- 
cho de Magallanes, Isla de los Estados, Punta Porpesse, 
Cabo Negro, Gregory Bay (Bahía San Gregorio), N coast 
of the province, Ushuaia, Cabo Posesión, Cerro de los 
Caracoles, Bahía Sarmiento, Gente Grande Bay, Punta 
negada, Isla Isabel. Chile: Punta Arenas 


ones, 


. Spegazzini; Uruguay. Arroyo de San Juan, dissi 


Spegazzini and A. Onetto; Argentina. Prov. Santa Cruz: 
Río Santa Cruz region 


. von Gülich; Argentina. Prov. Misiones: Río Piray-Guazíá, 


Río Yacan-Guazü, near Piray. Paraguay. Таригисири, Si- 
erra de Amambay 


. Spegazzini; N Argentina. Prov. Chaco. Paraguay. Río 


Aquidabán, Amambay. 
Tonini del Furia; Argentina. Prov. Santa Cruz: area of río 
Santa Cruz, Lago Argentino. 


Mauri; Argentina. Prov. Santa Cruz: Río Chico. 


. Spegazzini; Argentina. Prov. Chaco: Colonia Resistencia, 
Monte Yponá. 


. Spegazzini; Argentina. Prov. Buenos Aires: isla Santiago. 


oyano; Argentina. Prov. Chubut: Río Carrenleufá, Teka 
-choique P Senguer, Lago Fontana, Río Chubut, Lago 


Musters, Gli Galesa 


. Fischer; Argentina. Pain: Chubut: Cabo Raso, Puerto 


Rawson, Península de Valdez. 


. Spegazzini; Argentina. Prov. Buenos Aires: sierras Cura 
Malal. 


Mauri; Argentina. Prov. Tierra del Fuego: Canal Ultima 
Esperanza. 
Tonnellier; Argentina. Prov. Chubut: Trelew, Rawson. 


. Spegazzini; Argentina. Prov. Buenos Aires: San Nicolás. 
dil. 


Spegazzini; Argentina. Prov. Buenos Aires: Tan 
Spegazzini; Argentina. Prov. Salta: La Viña. 

meghino; Argentina. Prov. Santa Cruz: Río Deseado, 
San Julián, Cafiadón 11 de MES Río Salado. 
Berg; Argentina. Prov. Río Neg 


. Spegazzini; Argentina. Prov. S RM Famaillá. 


. Spegazzini; Argentina. Prov. Buenos Aires: Sierra de la 


Ventana in valle de las Vertientes, between е and 
Sierra de la Ventana, Cerro de la Ventan 
Fischer; Argentina. Prov. Chubut. 


. Spegazzini and S. Venturi; Argentina. Prov. Salta: Cuesta 


de Trancas. 


Spegazzini, 1880, 1914b, 


Spegazzini, 1917c 
Spegazzini, 1883b, 1896, 


1897b, 1901c, 1902b; 
Moore, 1983 


Spegazzini, 1917c 
Spegazzini, 1883a 


Spegazzini, 1883a, 1916с 


m е 
1901c, 19025; Del 

Spegazzini, 1917c 

Spegazzini, 1905 


Spegazzini, 1897c, 1901c 


Spegazzini, 1897c; Hos- 
seus, 1915 
* 


* 


* 
Spegazzini, 1905 
* 
* 
Spegazzini, 1897b, 
1901c 1902b 
Spegazzini, 1901c 
Spegazzini, 1895a, 
1923 
Spegazzini, 1897a, 1905 


Spegazzini, 1902b 
Venturi, 1925 


188 Annals of the 
Missouri Botanical Garden 
Table 1. Continued. 
Dates Collecting expeditions Bibliography 


Late 1895—Кеђ. 1896 
Mar. 1896 
Apr. 1896 
Nov. 1896 
Nov. 1896—Mar. 1897 
Late 1896—Early 1897 


Dec. 1896—Mar. 1897 


Jan.-Mar. 1897 
Feb. 1897 
1897 

1897 

Oct. 1897 
Oct.-Nov. 1897 


Nov. 1897-Jan. 1898 
Oct.-Dec. 1897 


Dec. 1897—Feb. 1898 


1898 
Jan. 1898 
Jan. 1898 


Jan.-Apr. 1898 


Feb.-Mar. 1898 
Fall 1898 


C. Ameghino; Argentina. Prov. Santa Cruz: Golfo San Jorge, 
jan Julián 


J. Koslowski; Argentina. Prov. Chubut: Lago Fontana. 

N. Alboff; Argentina. Prov. Tierra del Fuego: San Sebastián. 
C. cm Argentina. Prov. Buenos Aires: Tornquist, Si- 
a de la Ventana, Río de La Plata, Isla Santiago. 

О. Mauri; Argentina. Prov. Neuquén: Río Aluminé-Neu- 


quén. 
E. Fischer; Argentina. Prov. Chubut: Cabo Raso. 


C. Spegazzini; Argentina. Prov. Salta: Cuesta de Arca-Tran- 
cas, Pampa Grande, Quebrada de Guachipas, Amblaio- 
Cachi, Cafayate, Molinos-Cafayate, Nevado de Cachi, La 
Vifia, cuesta de ntonio, [sonza-Tintín, entre Tala- 

pampa y Valle Calchaquí, Runi-Huasi. Prov. Tucumán: 

Montero, Colalao 

. Ameghino; лета Prov. Santa Cruz: Rfo Chico 

(Chonquen-aik, Emelk- aik), Lago huge com 

. Spegazzini; Chile. Prov. Atacama: Ataca 


С» 


. Spegazzini; Argentina. Prov. ls Air 

. Spegazzini; Argentina. Prov. Catamarca, p Córdoba, 
Prov. La Rioja, Prov. Mendoza, Prov. Salta, Prov. San 
Juan 


оо 


t 


R., V. B. (sic); Argentina. Prov. Santa Cruz: 
Río Santa Cruz, Monte León 
J. Valentín; Argentina. Prov. Chubut: Trelew, Cabo Raso, 
Río Chubut. 
O. Mauri; Argentina. Prov. Neuquén: Lago Traful, Nahuel 
uapí. Prov. Santa Cruz: Río Seco, Canal Ultima Esper- 
anza. 


O 


. Ameghino; Argentina. Prov. Santa Cruz: Pan de Azúcar, 
Ho Chico (Emel-kaik, El Paso), Laguna Seca, Río Santa 


Cruz. 

C. Spegazzini; Argentina. Prov. Buenos Aires: from S Buen- 
os Aires N Patagonia, San Blas, Carmen de Patagones, 
La Pantanosa, Punta Rubia, Bahía San Blas, F 
Lomas de Saladero, Isla de Crespo, Salitral Grande, Sali- 
na de Piedras, Salina del Inglés, Tres Cerros, La Verde, 
Barrancoso. Prov. Neuquén: Lago Nahuel Huapí, Lago 
Traful, Laguna Blanca. Prov. Río Negro: areas of Río Col- 
orado and Río Negro, Salina de Piedras, Puerto 
confluence Río я and Rio Neuquén, Choele Choel, 


uevo, 


Lomas Neg 
C. Moyano; Ris entina. Prov. Río Negro. 
C. Spegazzini; Argentina. Prov. Buenos Aires: sierras de 


Tandil 

F. Lahille; Argentina. Prov. Chubut: Península de Valdéz, 
Caleta Porfirio. 

. Ameghino; Argentina. Prov. Santa Cruz: Río Chico (Ku- 
men-aik, Chonkenk-aik, cerro Kmanaich, Emelk-aik, Bo- 
ron-aik, Parr-aik, Sehuen-aik), Lago Viedma (Orr-aik), 
Lago Argentino d Río Sehuén (Parr-aik), Golfo 
San Jorge, Río Santa Cru 

й . Venturi, Argentina. Prov. ГТА Cruz: Río Santa Cruz. 

. Ameghino; Argentina. Prov. Santa Cruz: between San Ju- 
lián and Rio Deseado. 


C 


pw 


Spegazzini, 1897b, 
le 
Del Vitto et ps 1998 
Del Vitto et al., 1998 
Spegazzini, Ти 1905 
Spegazzini, 1902c; Del 
Vitto et al., 1998 
Spegazzini, 1901c, 
1 
Spegazzini, 18974, 
1899b, 19014, 1916c, 


1917a, b, c, d, 1921a, 
1923c, 1925b 


Spegazzini, 1901c, 1902c 


Spegazzini, 1901b, c 
Spegazzini, 1898 
Spegazzini, 1902b 
Spegazzini, 1899a, 


c, 1902b 
Spegazzini, 1901c, 1902a 
Spegazzini, 1901c, 

1902b, c 


e 1 ји 
19012 )2b, c, 
1905, in 1925b 


ж 


Spegazzini, 1901а 
Spegazzini, 1899а, 1901с 


Spegazzini, 1899a, 19012 
1902b, c 


Spegazzini, 1902b 
Spegazzini, 1902b 


Моште 87, Митбег 2 
2000 


Katinas et al. 
Carlos Spegazzini 


189 


Table 1. 


Continued. 


Dates 


Collecting expeditions 


Bibliography 


Nov. 1898-Early 1899 
Nov. 1898-Feb. 1899 


Nov. 1898—Mar. 1899 
Dec. 1898—Маг. 1899 
(Spring?) 


Јап., Aug. 1899 
Apr.-May 1899 

Nov. 1899-Маг. 1900 
Summer 1899 

Dec. 1899 

Late 1899—Early 1900 
Late 1899—Early 1900 
Dec. 1899-Jan. 1900 
Dec. 1899-Mar., June 
Jan.-Feb. 1900 

Feb. 1900 


Feb.—Mar. 1900 


Fall, Summer 1900 
Nov. 1900 

Summer 1900—1901 
Dec. 1900—Feb. 1901 
Jan. 1901 

Jan. 1901 

Jan.-Apr. 1901 


Feb.—Mar. 1901 


Summer 1901 


J. Koslowsky; Argentina. Prov. Chubut: Lago Blanco, central 
area of the province, Paso de los Indios, Río Senguerr, 
Valle del Río Mayo, Río Chubut. 

N. Illín; Argentina. Prov. Chubut: Lago Musters, Choique 
Lauquen, Angostura, between Trelew and Paso de los In- 
dios, Costa de los Manantiales. 

T. Stuckert; Argentina. Prov. Córdoba. 

C. Ameghino; Argentina. Prov. Chubut: Lago Musters. Prov. 
Santa go Argentino (Karr-aike), San Julián-Río 
Deseado, San Jorge, Lago Buenos Aires, Golfo San Jorge. 


A. Tonnellier; Argentina. Prov. Chubut: Trelew, Rawson, Río 


Е. Lahille; Argentina. Prov. Río Negro: Golfo San Matías. 
Prov. Chubut: Caleta Porfirio. 

N. Illín; Argentina. Prov. Río Negro: Bolsón. Prov. Chubut: 
Río Chubut, between Choique-Laven and Lago M 
о Musters, Teka-dique, Carrenleufti, Teka-choique. 

C. Moyano; Argentina. Prov. Chubut: Carrenleuft. 

C. сардаи Argentina. Prov. Buenos Aires: Sierras de 
Cura Malal, Tornquist. 

A. Larguía; Argentina. Prov. Río Negro: Colonia Valcheta. 

F. Basaldáa; Argentina. Prov. Chubut: Trelew, Río Chubut. 

C. Burmeister; Chile. Río Aisén. 

O. Asp; Argentina. Prov. Neuquén: Sierra de Maichol, Valle 


usters, 


Trolope, Río Manzano, Codihué, Pilahuincó, Río Fiero 
Vega del Pino Hachado, Sierras de Sanquil. 

R. Hauthal; Argentina. Prov. Santa Cruz: Cerro de los Bag- 
uales. 

F. Silvestri; Argentina. Prov. Santa Cruz: Río Santa Cruz, 
Lago Argentino. 

М. Шт; Argentina. Prov. Chubut: Río Carrenleufá, between 
Cholila and Colonia 16 de Octubre 

C. Ameghino; Argentina. Prov. Chubut: Col-huapí an 
huapí). Prov. Santa Cruz: Río Chico, between Río Des 
do and San 

R. Hauthal; aA Prov. Santa Cruz: Sierra de los Bag- 

uales. 

Basaldúa; Argentina. Prov. Chubut: Trelew. 

. Burmeister; Argentina. Prov. Chubut: Arroyo Verde. 


рот 


. Fernández; Argentina. Prov. Neuquén: Lago Nahuel 


парї. 
М. Шт; Argentina. Prov. Chubut: Lago Blanco, Corcovado, 
Puerto Rawson, Carrenleufü, Teka Choique, Nafofo-Ca- 
huellu, Cholila, Manantiales. 
. Spegazzini; Argentina. Prov. Buenos Aires: Tandil. 


Claren; Argentina. Prov. Jujuy: Puna of Santa Catalina. 


dA 


. Burmeister; Argentina. Prov. Chubut: Manantial de la 

Subida. Chile. Río Aisén. 

C. Spegazzini; Argentina. Prov. Mendoza: Punta de Vacas, 
road Mendoza- Villavicencio, road Puente del Inca, Lagu- 

Aconcagua, Cerro Leones, 

Las Cuevas, Cafiadón de los Horquillones. 

én: Fortín Roca. 

Hauthal; Argentina. Prov. Chubut: Río = 


па de Los Horcones, valle del 
Lahille; Argentina. Prov. Neuqu 


Spegazzini; Argentina. Prov. Chaco: Ipagua 


pps 


Spegazzini; Argentina. Prov. Salta: Río Peradi 


Spegazzini, 1899а, 1901c 
1902b 

Spegazzini, 1901c, 
1902b 


Spegazzini, 1899b 
Spegazzini, 1902b, с 


Spegazzini, 1901c, 
1902b 
Spegazzini, 1901c, 
1902b 
Spegazzini, 1902a, b, с 
Spegazzini, 1902c 
Spegazzini, 1905 
Spegazzini, 1902b 
Spegazzini, 1902b 
Spegazzini, 1902b, с 
Spegazzini, 1902b 
Spegazzini, 1902b, с 
Spegazzini, 1902b, c 
Spegazzini, 1902b 


Spegazzini, 1902b, c 


* 


* 
Spegazzini, 1902c 
Spegazzini, 1902b, c 


Spegazzini, 1902b 


Spegazzini, 1901а 

ж 

Spegazzini, 1902c 

Spegazzini, 1901d, 
1917a, b, c, 1925a 

Spegazzini, 1902b 

Del Vitto et al., 1998 

ж 


Spegazzini, 1916c 


Annals of the 


Missouri Botanical Garden 


Bibliography 


juy: Sierra de Santa Bár- 
e Calilegua. Prov Salta: Río Pescado, Río 


: Rio San Francisco, 
ta Bárbara, Santa Cornelia, 


Parque Roca. Prov. Mendoza: cerro 


i; Argen tem Prov. Catamarca: Piedra Blanca. 


Tala. Prov. Mendoza: 


v. Salta: Santa 


190 
Table 1. Continued. 
Dates Collecting expeditions 

1904. С. Spegazzini; Argentina. Prov. San Juan. 

Dec. 1904—Mar. 1905 С. Spegazzini; jd na. Pro 
bara, Sierra 

an Francisco (Ledesma), Río Berm 

Dec. 1905—Арг. 1906 C. Spegazzini; Argentina. Prov. Juj 
Perico, Ledesma, Yuto, Río Saucelito. Prov. Salta: Orán, 
Valle de Lerma, Sierra de San 
Sierra de Maíz Gordo, Sierra de Zenta, Río Santa María. 
Prov. Tucumán: Tafí, 
de los Cordobeses. 

Dec. 1905 C. Burmeister; Argentina. Prov. Santa Cruz: Puerto Desea- 

Dec. 1906—Mar. 1907 C. Spegazzini; Argentina. Prov. Misiones: from Santa Ana to 
Barracón and Río San Antonio, Puerto Esperanza (Río 
Paraná), Posadas, Campina de Américo, San Pedro, Fra- 
crán, Yacán-Guazü, Сагира, Arroyo Dorado, Campo de 
las Cuyas, Cerro Pesegueiro, Campo Grande, Cerro Boni- 
to, Río Bossetti = Grande), Río Acaranguay (Campo 
Grande), Loreto, yado. 

Feb. 1907? C. Spegazzin 

Late 1907 P. збиен Argentina. Prov. Buenos Aires: isla Магі 

rcía. 

Dec. 1909 J. Argerich; Argentina. Prov. Buenos Aires: Estación Arger- 
ich, La Fina 

Dec. 1909-Mar. 1910 P. Spegazzini; Argentina. Prov. Catamarca: Ancasti, Ambato, 
Andalgalá, Huillapina, Pomán, Río 
quebrada de los Horcones, Cacheuta. Pro 
Cornelia, Sierra de Santa Bárbara. Prov. шош: А1ра- 
сћил, Сос 

Маг. 1911 Spegazzini; M Prov. Entre Ríos: Ibicuy. 

Nov. 1911 


1912 
Jan.-Feb. 1914 


June, Oct. 1919, Feb. 
1920 


Feb. 1922 
Summer 1923 


C. Spegazzini; Argentina. — Jujuy: Santa Cornelia, Sierra 
n Negro 


de Santa Barbara, Lecher 


C. Spegazzini; Argentina. B San Juan: 


e 


p e 


. Spegazzini; Paragu 


. Spegazzini; Argentina. Prov. Mendoza: Río Blanco del 


Plata. Prov. San Juan: Paso de los Punta 

uay. Asunción, Puerto Sajonia, Hío Par- 
aguay, San Antonio, Pacü-cuá, road San Lorenzo-San 
Antonio. 

Molfino; Argentina. Prov. Misiones: San Javier, Apóstoles. 


й оа Argentina. Prov. Buenos Aires: Isla Marín 
fa 


Сагсї 

Јап. 1924 pegazzini; Argentina. Prov. Tierra del Fuego: Haberton 
(Shamanes), Isla Grande, Lapataia, Ushuaia, Bahfa 
Orange, Puerto Shell. Chile. Cabo de Hornos, Isla Hoste, 
Penfnsula Hardy. 

Jan. 1926 


A. Raffaelli; Argentina. Prov. Río Negro: S Río Negro. Tala- 
f 


gapa, Barrilniyeo, Carilaufquen. 


Kiesling, um 
Spegazzini, 19 
1917a, n 1923c 
Spegazzini, 1914a, c, 
1917c, 1923c, 1925a, 


с 


сч 1909, 1916с, 
1917a, b, с, а, 1923с, 
19255 


Spegazzini, 19175 
ж 


Spegazzini, 19166, 
1917c, 1923a 


Spegazzini, 1917а 


Kiesling, 1994. 

Spegazzini, 1914a 

Spegazzini, 1921а, b, 
192: 

* 

Anonymous, 1925 


Spegazzini, 1924; Moore, 
1983 


Spegazzini, 1926 


del grandioso Herbario Argentino que figura en la Sección 


am annoy 
tain indifference, if not hostility, every tim 
to make a botanical ition devoid of sisti а profit. 
Thus to properly prepare myself to fulfill my official bo- 


tanical duties 1 have had to take advantage of every eco- 
nomic or industrial mission that my knowledge allowed me 
to do more or less satisfactorily, accumulating i in this way 
the materials that now constitute the basis of the great 
Herbario Argentino of the Secc ión Botánica Ministerio de 
Agricultura and the knowledge that 1 have about Argentine 
phytogeography.] 


Моште 87, Митбег 2 
2000 


Katinas et al. 191 


Carlos Spegazzini 


His partners on the field trips described Spegaz- 
zini as a true naturalist, an enthusiastic and рго- 
found connoisseur of the flora and fauna, which he 
described in simple language, making each expe- 
dition an exciting and enjoyable adventure (Molfi- 
no, 1929; Parodi, 1961) 


SPEGAZZINIS SPECIMENS 


As a result of the collecting expeditions made 
by Carlos Spegazzini and by his collaborators, the 
number of vascular plants in Spegazzini's herbar- 
ium reached 100,000 specimens (Molfino, 1929). 
This number is not surprising if one bears in mind 
that he was able to collect 30,000 specimens in a 
single trip (Venturi, 1925). Due to the abundance 
of fungi and vascular plants collected, Spegazzini 
maintained the majority of them as a personal her- 
barium at home. Several European and North 
American institutions were interested in purchas- 
ing these valuable collections after Spegazzini's 
death, but all offers were rejected. Around 1966 
(Kiesling, 1984), the collection of vascular plants 
was transfered from LPS to the Herbarium of Mu- 
seo de La Plata (LP), also in La Plata city. 

Type materials ascribed to Spegazzini are esti- 
mated at ca. 700 specimens in LP. Staff of this 
herbarium (Katinas et al., in prep.) are currently 
developing a catalogue of these specimens. Other 
specimens remain in Museo de Botánica Juan A. 
Domínguez in Buenos Aires (BAF), in the Instituto 
de Recursos Biológicos (INTA) in Buenos Aires 
(BAB), as well as in the Museo Botánico in Córdoba 
(CORD). Other herbaria with Spegazzini vascular 
plant specimens are: BAE, BR, E, H, IAC, K, L, 
MICH, NY, PAC, PAD, S, U, W, and Z (Stafleu & 
Cowan, 1985). 

Two additional Spegazzini collections of interest 
are, or were, the living types of cacti, and the pho- 
tographs of cacti. The collection of living types is 
practically lost, except for one specimen of Cereus 
still growing in his house, now the institute (Kie- 
sling, 1984). His collection of cacti photographs 
that correspond to type and non-type specimens 
(Kiesling, 1984) is deposited in the Herbarium of 
Museo de La Plata (LP). 

Specimen labels are annotated by Spegazzini or 
stamped with a seal by him, with a few references 
to the locality. He used the initials C. S. for the 
plants he collected, and other initials to differen- 
tiate other collectors: C. A. (for Carlos Ameghino), 
N. 1. (for Nicolás Illín), T. Е (for Tonini del Furia). 
Most of the specimens lack the number of the in- 
dividual collector, although they have the number 
of Spegazzinrs herbarium (LPS). For example, the 


type specimen of Oryzopsis bicolor (Vahl) Speg. var. 
media Speg. collected by Carlos Spegazzini in 18 
should be cited: C. Spegazzini s.n. (ex LPS 12517 
in LP). The name of the collector is frequently not 
mentioned, but can usually be determined by ref- 
erence to the year and locality (Table 1). The date 
of collection usually consists of the month and the 
year, or sometimes the year is accompanied by the 
terms aest. (from the Latin aestivus or summer) or 
aut. (autumnus, fall). The word vere, annotated by 
Spegazzini as *Vere 1889," can be seen only in the 
labels of the specimens collected by Carlos Amegh- 
ino in San Julián-Río Deseado. There are two prob- 
able meanings for this Latin term (Stearn, 1996): 
(1) Ver — spring, a neuter generic noun of the third 
declension. Vere corresponds to the ablative sin- 
gular, i.e., "Ђу the spring," “мић the spring," or 
"from the spring"; and (2) Vere (also vero, revera) — 
an adverb meaning “truly, in fact, rightly, exactly." 
Following other annotations of Spegazzini as “Vere 
1894" (Spegazzini, 1895b) for fungi collected in 
October-November (Southern Hemisphere spring), 
it seems that the mosi reasonable interpretation of 
this term is “spring.” 

For some labels of Patagonian specimens, there 
is no citation of the contemporary political prov- 
inces. From 1878 to 1884 the Argentine territory 
south of Río Colorado (ca. 40°S latitude) to Cabo 
de Hornos was established by the government as 
one unit: Gobernación de la Patagonia. Only in 
1884 was this wide territory divided into the cur- 
rent provinces of La Pampa, Neuquén, Río Negro, 
Chubut, Santa Cruz, and Tierra del Fuego. Another 
problem with some of these Patagonian plants is 
with the specimens collected by C. Moyano in Chu- 
but in 1899. As Moyano did not write the corre- 
sponding labels, Spegazzini annotated the labels 
with probable localities. As he explained (Spegaz- 
zini, 1897c): 


“La colección que voy a publicar se hallaba en bas- 
indicarlos de un modo bastante vago y por recu 


Unfortunately, it completely lacked labels, so I had to 
indicate the habitat very vaguely and more or less by 
the recollections of the collector.] 


The type specimens frequently have the original 
Latin descriptions made in Spegazzini’s handwrit- 
ing. Many of these descriptions were written di- 
rectly in the field or during the specimen prepa- 
ration (Fig. 4). It is also very common to find the 


192 Annals of the 
Missouri Botanical Garden 


Wa A PAZ 22277 1 oho. | 2 
Ly fe. Дола у" pred; gla bor A Cow Але A MEC 


PAL UA > 


P WA ey р == ney, 


uA a 
Crea ed, псе AT, S О REOS 
auo њену pe Ра 


oie E ae СА 
ера. s hn, Baw AE pom 
44. А. BE he ТУ, repr Faber Keke 
W paga РЈ ^ Voi di y 
SAK г 
С Me us aper cm 
Б а Ш. СА 05 би сату h terek 
и. p (aa рима ли ИГ за 


Naw = d~ S вм. pr ша Сам 


AJ ei vs А 


Figure 4. Original description of a new variety of vascular plant made in Spegazzini's handwriting. 


botanist's accompanying drawings, characterized by sling, 1984). The citation of more than one speci- 
their precision and simplicity (Fig. 5). Some sets of men and locality in the original taxa descriptions 
the original Spegazzini illustrations are housed in (species, varieties, and forms) is common, and 
BAF, as is the case with some cactus species (Kie- therefore Spegazzini’s type collection is mainly 


~ 


Моште 87, Митбег 2 
2000 


Katinas et al. 193 
Carlos Spegazzini 


Ld 


Figure 5. Accompanying drawing of a plant specimen made by Spegazzini. 


made up of syntypes. Types are housed in a sepa- 
rate collection in LP, but it is probable that some 
types still remain in the general herbarium. The 
search for these specimens is another step in the 
production of the catalogue mentioned above. 


SPEGAZZINIS PUBLICATIONS 


Carlos Spegazzini published 332 papers from 
1878 to 1926, and ca. 100 refer to vascular plants. 
Almost all of them were published in such Argen- 


194 Annals of the 
Missouri Botanical Garden 


Table 2. New taxa in their corresponding families described by Carlos Spegazzini, as cited in Index Kewensis 
(Anonymous, 1997), and the Gray Card Index Database. The number of taxa described in each taxonomic category is 
in parentheses. Spegazzini is the authority of all names except where marked. The designation of comb. illeg., comb. 
nov., comb. superfl., hom. illeg., nom. illeg., nom. inval., nom. nud., and a change in the authority of the name other 
than Spegazzini (marked with *) follows Zuloaga et al. (1994) and Zuloaga & Morrone (1996, 1999a, b 


AIZOACEAE. Species (1): Tetragonia ameghinoi. Total (1). 

ALISMATACEAE. Species (1): Echinodorus patagonicus. Total (1). 

AMARANTHACEAE. Genera (1): Amarantellus. Species (4): Amarantellus argentinus, Amaranthus cristulatus, A. vul- 

us, Amarantus edulis. Infraspecific taxa (1): Blutaparon portulacoides var. commersonii*. Total (6). 

AMARYLLIDACEAE. Species (3): Zephyranthes lilacina, 2. melanopotamica, Z. oxitepala. Infraspecific taxa (2): 
Hippeastrum bagnoldi var. minor, H. hesperius var. pallida. Total (5). 

ANACARDIACEAE. Species (4): Lithrea chichita*, Schinus chichita, S. longifolia (Lindl.) Speg. comb. nov., S. prae- 
cox. Infraspecific taxa (1): Schinus dependens var. patagonica. Total (5). 

APIACEAE. Genera (1): Notiosciadium. Species (13): Asteriscium fimbriatum, Azorella к А. bovei, А. fuegi- 

dio 


ana, А. patagonica, А. plantaginea, Hydrocotyle cryptocarpa, Mulinum lycopo sud М. т onis (Kuntze) Speg. 
comb. nov., M. patagonicum, M. valentinii, Notiosciadium pampicola, Sanicula patag: а. Infraspecifi taxa (4): 
puce patagonica var. compacta, Bowlesia tropaeolifolia var. heterophylla, B. iropaeolifolia var. patagonica, Hy- 


ocotyle araucana var. тоа Тога! (18). 

APOCYNACEAE. Species (5): Aspidosperma chakensis, A. crotalorum, A. horco-kebracho, A. missionum, Rauvolfia 
schuelii. Infraspecific taxa (1): pie dits nuin blanco var. pendula. Тога! (6). 

AQUIFOLIACEAE. Species (1): /lex tucumanensis nom. nud. Total (1). 

ARACEAE. Genera (1): Lilloa. mein (2): Lilloa puki, Staurostigma vermicida. Total (3). 

ARECACEAE. Species (2): Maximiliana argentinensis, M. orenocensis. Total (2). 

ARISTOLOCHIACEAE. Species (2): Aristolochia melanoglossa, A. stuckertii. Total 

ASCLEPIADACEAE. Genera (1): Dicarpophora. Species (4): Astephanus fruticulosus, Dicarpophora mazzuchii, Oxype- 
talum suaveolens hom. illeg., Vincetoxicum bulligerum. Infraspecific taxa (1): Philibertia gilliesii var. pubescens. To- 
al (6) 


ASTERACEAE. Genera (2): Ameghinoa, Strongylomopsis. Species (58): е leucanthus, Ameghinoa pata- 
gonica, Aplopappus (= Нер moyanoi, А. mustersi, A. patagonicus, А. struthionum, А. tehuelches, Aster 
scorzonerifolius, "Baccha aris chubutensis, B. melanopotamica, B. tandilensis, Brachyclados caespitosus (Phil 
comb. nov., B. megalanthus, B. Bann Chuquiraga argentea (Speg.) Speg. comb. nov., Culcitium gilliesii (Hook. 
& Arn.) Speg. comb. nov., C. sessile, Doniophyton argenteum, Erigeron erianthus, E. platensis, E. remyanus*, Floto- 
via stiffioides, Gutierrezia ameghinoi, Haplopappus ameghinoi, Н. illinii, Н. moyanoi, H. mustersii, H. patagonicus, 
H. struthionum, H. tehuelches, Hieracium врше е, Leuceria inis dium L. ыш Mutisia chubutensis, М. 
moyanoi, M. pulchella, Nassauvia ameghino Ju ibut s, N. patagonica, N. pentacaenoides, Perezia megalan- 
tha, P. pampeana, P. patagonica, P. в Psilocarphus o iferus, ier io argentinensis, S. capillarifolius, S 
choiquelahuensis, S. и 5. colu- pn S. diabolicus, S. inutilis, S. julianus, S. mustersii, S. ке 

5, S. sericeo-nitens, le fuegiana, Vernonia oreophila. Infraspecific taxa (37): Baccharis trime 
viscosissima, Chiliotrichum diffusum var. изин oi Chiliotrichum diffusum var. media, Chiliotrichum diffusum 
var. typica, a argentea var. вет о gayanus var. и Е. philippii var. elatior, Е. philip- 


ри var. humilis, Gnaphalium affine var. medium, С. affine var. E rvulum, G. affine var. pusillum, G. purpureum var. 
sphacelatum (Kunth) Speg. comb. nov., (insani ciosa var. integrifolia, Gutierrezia paniculata var. patagonica, 
Hieracium antarcticum f. fuegiensis, Hypochaeris arenaria var. кы (Sch. Bip.) Speg. comb. nov., Н. агеп- 
aria var. integrifolia (Sch. Bip.) E g. comb. nov., H. variegata var. acutibracteata, H. variegata var. glauc 

H. variegata var. nana, H. variegata var. typica, H. variegata var. pinnatifida, Leuceria ibari var. pr di L “bari 
var. glandulosa, L. iban var. sessiliflora, Nassauvia abbreviata var. azorelloides, N. ‘axillaris var. contracta, N. pata- 


gonica var. elatior, N. struthionum var. robusta, Panargyrum abbreviatum var. subspinosa, Perezia Fs dica var. 
intermedia, Senecio desideratus f. elatiuscula, S. linariifolius var. beg m (Phil.) 5 mb. nov., S. miser 
var. tehuelches, S. trifurcatus var. pentadactylus (Phil.) Speg. comb. nov., S. xanthoxylon var. dinum s. Solidago 
linearifolia var. brachypoda. Total (97). 

BEGONIACEAE. Species (1): Begonia argentinensis. Total (1). 

BIGNONIACEAE. Species (1): Tecoma avellanedae. Infraspecific taxa (1): Argylia potentillaefolia var. australis. Total 


(2). 

BORAGINACEAE. Genera > Oxyosmyles, Valentina, Valentiniella. Species (10): Amsinckia patagonica, A. pseudoly- 
copsicoides, Echinospermum patagonicum, Eritrichium diffusum hom. illeg., E. mesembryanthemoides, E. pampean- 
um, Heliotropium Таоа Oxyosmyles viscosissima, Valentina patagonica, Valentiniella patagonica (Speg.) 
Speg. comb. nov. Infraspecific taxa ар, уон в kia angustifolia var. microcarpa. Total (14 


Volume 87, Number 2 Katinas 195 
2000 Carlos ЕЕ 


Table 2. Continued. 


BRASSICACEAE. "ie ao ): Мени оя Delpinophytum. Species (40): Braya cachensis, В. glebaria, B. lycopodioi- 
des, B. patagonica, B. pectinata, B. pycnophylloides, Cardamine argentina, C. callitrichoides, С. patagonica, Delpi- 
noella ше. + ш phytum E bis а = Speg. comb. nov., Descurainia deserticola (Speg.) Spe 
comb. nov., D. glabrescens (Speg.) Speg. comb. nov., D. heterotricha, Draba ameghinoi, D. argentina, D. chubuten- 
sis, D. glebaria Speg. ex O. E. Schulz, D. gom, D. karr-aikensis, D. oligosperma, D. pectinata Speg. ex O. 
E. Schulz, Menonvillea patagonica, Nasturtium pamparum, N. philippianum nom. illeg., N. EU Schizopetalum 
fuegianum, Sisymbrium ameghinoi, S. paa hom. illeg., S. deserticola, S. fuegianum (Speg.) Speg. comb. nov., 
5. eda А glanduliferum, s maclovianum, S. patagonicum, S. perenne hom. illeg., S. sit ы. S. subscan- 

. tehuelches, Thlaspi chionophilum. Infraspecific taxa (21): Braya lycopodioides var. contracta, Cardamine 
eun var. dichondroides, C. tuberosa var. velutina, Descurainia canescens var. patagonica, D. canescens var. pur- 
pureola, Draba argentina var. grandiflora, D. argentina var. latifolia, D. karr-aikensis var. major, D. karr-aikensis 
var. media, D. karr-aikensis var. minor, D. magellanica var. glabrata Gilg. ex Speg., D. magellanica var. subgla- 
brata, Lepidium pubescens var. salinicola, L. pubescens var. typica, Sisymbrium fuegianum var. glabrum, S. fuegian- 
um var. hispidum, S. sagittatum var. commune, S. sagittatum var. "b gale S. sagittatum var. glaucum, 5. 
sagittatum var. normalis, S. sagittatum var. purpurascens. Total (63 

BROMELIACEAE. Species (6): Aechmea iode Puya flora, P. d Tillandsia chlorantha, T. euosma, Vriesea 
argentinensis. Infraspecific taxa (1): Dyckia montevidensis var. tandilensis. Total (7) 

CACTACEAE. Genera (4): Aylostera, Brittonrosea, Maihueniopsis, Parodia. Species (113): Austrocactus dusenii*, 
intertextus, Aylostera = (Speg.) Speg. comb. nov., Brittonrosea albata, B. anfract pA B. ME B. 
onfusa, B. coptonogona, B. crispata, B. Puoi B. gladiata, B. grandicornis, B. hastata, B. heteracantha, 
B. lamellosa, B. lancifera, B. rdi B. multicostata, B. eps B. за а В. оганда, В. violaciflora, 
В. wippermanni, В. zacatecasensis, Cereus dayami, С. dusenii*, С. guelichii, С. кө" С. platygonus hom. 
Шег., С. roiforus C. santiaguensis, C. silvestrii, C. РАЗНЕ Зеба *, C. thelegonoides, Echinocactus Гоин уа 
Е. baldianus, Е. cachensis, Е. caespitosus, Е. catamarcensis hom. "P Е. hic cA E. famatinensis, E. hae 
паат Е. loricatus hom. illeg., E. pampeanus, E. parvulum, E. platensis, Е. pseudominusculus, E. pygmaeus, 

E. saltensis, E. sanjuanensis, E. stellatus, E. stuckertii, Гр. ү ктын кеге ancistrophora, x baldiana, E. 
cachensis, E. о Е. а Е. melanopotamica, Е. ттиапа, Е. mirabilis, Е. molesta, Е. pseudomi- 
nuscula, E. saltensis, E. silvestrii, Е. spegazzinii К. pa ex Speg.*, Frailea bruchi, Е ые Е рете errima, 

mnoca "esum haldianum (Speg.) Speg. comb. nov., G. brachypetalum, G. chubutense (Speg.) Speg. comb. nov., G. 
leptanthum, G. loricatum, G. parvulum (Speg.) Speg. comb. nov., G. stellatum, Leocereus paulensis, Lobivia areope- 
gon Speg. ex Hosseus, L. hyalacantha, L. oreopepon, L. shaferi*, Maihuenia it cium M. valentinii, Maihueniop- 
sis molfinoi, Malacocarpus sanjuanensis, Opuntia anacantha, O. arechavaletae, O. atro-virens, O. bonarensis, O. 

ruchii, O. canina, O. chaquensis, O. cordobensis, O. halophila, O. hypsophila, O. kiska-loro, O. molinensis, O. 
montevidensis, O. pampeana, O. penicilligera, O. prasina, O. retrorsa, O. Wee d 0. bandage 0. 
tuna-blanca, О. utkilio, О. weberi, Parodia brasiliensis, P. microsperma (F. A. C. ber) Speg. comb. nov., P. para- 
guayensis, Pilocereus decern (Salm-Dyck) Speg. comb. nov., Pterocactus 1 Reb utia famatinensis 
(Speg.) Speg. comb. nov., Tephrocactus hickeni (Britton & Rose) done: comb. nov. Infraspecific taxa (35): Cereus 
poles sai var. кеи ы Echinocactus acuatus var. corynodes, Е. acuatus var. depressus, E. acuatus |. erinacea, 
Е. acuatus var. se i, E. acuatus var. tetracantha, E. catamarcensis var. obscura, E. catamarcensis var. pallida, 
E. gibbosus var. мш к E. gibbosus var. chubutensis, E. gibbosus var. platensis, E. gibbosus var. typica, E. 
gibbosus var. Pisani E. mammulosus var. hircina, E. mammulosus var. pampeanus, E. mammulosus var. sub- 
таттшоза, E. mammulosus var. typica, Е. microspermus var. erythranthus, E. microspermus var. Tae: Е. 
platensis var. Бе E. platensis var. parvulus, E. platensis var. quehlianus, E. platensis var. typica, E. py, 
maeus var. phaeodiscus, Echinopsis leucantha var. brasiliensis, Opuntia bruchii 1 macracantha, О. diademata var. 
inermis, О. diademata var. oligacantha, О. diademata var. polyacantha, О. ficus-indica var. decumana, О. fic 
ка var. gymnocarpa*, Tephrocactus ак f. brachyacantha, T. bruchii f. macracantha, T. glomeratus var. iner- 

‚ T. glomeratus var. alis acanthus. Тога! (152). 

CALYCERACEAE. Species m Boopis ameghinoi, B. chubutensis, B. filifolia, B. leptophylla, B. patagonica, B. raf- 
faellii, Gamocarpha ameghinoi, G. caleofuensis, G. patagonica, G. subandina, Nastanthus chubutensis, N. patagoni- 
cus. Infraspecific taxa (3): pan crassifolia var. spinuligera, B. gracilis var. lazulina, Gamocarpha subandina var. 
glaucescens. Total (15). 

CAMPANULACEAE. Species (1): Downingia pusilla*. Total (1). 

CAPPARACEAE. Species (1): Cleome titubans. Total (1) 


196 Annals 


of the 
Missouri Botanical Garden 


Table 2. Continued. 


CARYOPHYLLACEAE. Genera (1): | Species (7): Lychnis argentina, L. chubutensis, L. patagonica, Me- 
landrium chubutens b. nov., M. patagonicum (Speg.) Speg. comb. nov., Philippiella para mui 
Stellaria ти Infraspecific taxa (11): Arenaria serpens var. апа соја“, Arenaria serpens var. micro 
(Phil.) Speg. comb. поу., Arenaria serpens var. palustris (Naudin) Speg. сот} 
(Phil.) Speg. comb. nov., Arenaria serpens f. robusta, 


). nov., Arenaria serpens var. digi на 
I 
potamica, S. a 
ali. 


m 

Arenaria serpens var. serpylloides, Sagina apetala va 

. apetala var. paludosa, Silene inflata var. patagonica, Stellaria media var. apetala, S. е ка уаг. пог- 
is nom. ај Total (19). 

CELASTRACEAE. Species (2): Schaefferia argentinensis, S. uruguayensis. Total (2). 
CHENOPODIACEAE. Species p Atriplex i, A 


ameghino 
A. macrostyla, A. me 
diu 


. argentina, A. espostoi, А. flavescens hom. illeg., A. frigida, 
. platensis, A. ia = nom. nud., A. sagittifolia, A. ко Сћепор 0- 
т ameghinoi, С. antarcticum in (Hodk. f.) Speg. comb. su ‚ C. fuegianum, C. scabricaule, Holm a tweedii 
Moq.) Speg. comb. nov., Spirostachys olivascens, Suaeda и Infraspecific taxa (20): Atriplex sagittifolia 
var. heterophylla, A. "on ын var. rb A. d var. Ыы А. MORA var. bows no 
inval., Blitum rubrum var. hypoleuca, B. rubru 1 (Hook. f.) S opodium am- 
brosioides var. oblance ла, С. ipi Murs var. idee ue C. ambrosioides var. де мк (Willd) ind comb 

v., C. ambrosioides var. a, C. ambrosioides var. typica, C. scabricaule f. megalosperma, C. scabricaule f. 
pusilla, С. scabricaule f. я "Lerchea вр var. brachyphylla, L. fruticosa var. megalosperma, Salicornia 
corticosa var. procumbens, S. corticos 


var. crosperma 


typica nom. inval., S. fruticosa var. doeringii (Lorentz & Niederl.) Speg. 

comb. nov., S. fruticosa var. шош Тога! (38). 

CONVOLVULACEAE. Species (3): Convolvulus platigena, С. platincola, Ipomoea argentinensis. Total (3). 

CYPERACEAE. Minen (3): Carex patagonica, C. subantarctica, Eleocharis funebris. Infraspecific taxa (3): Carex 
darwinii var. ж, Eleocharis acicularis var. lilliputiana, Uncinia phleoides var. brachytricha. Total (6). 

ELATINACEAE. pan (1): Elatine nivalis. Tot m ). 

ERICACEAE. Species (3): Pernettya chubutensis, P. patagonica, P. philippiana. Total (3). 

EUCRYPHIACEAE. Species (1): Eucryphia M. Total (1). 


EUPHORBIACEAE. Genera (1): dente Species (9): Aonikena patagonica, T chacoensis (Morong) Speg. 
comb. nov., Colliguaja patagonica, Croton ventanicolus, Е ia pampeana, E. pseudopeplus, Jatropha кке: нај 
itica, J. guaranitica, Phyllanthus marginivillosus. [йар Бе {аха (1): MATS а. var. obtusifolia 


Total (11 


FABACEAE. Genera (7): Anadenanthera, Cavaraea, Chiovendea, Manganaroa, Pirottantha, Pithecodendron, Ramori- 
noa. Species (93): Acacia etilis, A. nitidifolia, Adesmia ameghinoi, А. арћапатћа, A. canescens (A. Gray) Speg. 
comb. nov., A. graminidea, A. karraikensis, A. leptopoda, А. pampeana, A. patagonica, А. rudolfi, А. salicornioides, 

А. tehuelcha, Anadenanthera falcata, A. ius жон и patagonicum, А. subandinum, pica acon- 

caguensis, А. ameghinoi, А. benthamianus, A. geanus, А. chubutensis, A. ien Mora Speg. co 

олег. А. hurtadensis, A. ale A. пар ин А. meyenianus, А. mo 

А. reichei, А. rhudolphii, А. sanctae-crucis, ee A. subandinus, A. еи А. tehuelches, А. з и 

adisiensis, А. watsonianus, Calliandra чи ан C. grisebachiana (Harms) Speg. comb. nov., C. part 

(Hook. f. & Arn.) Speg. comb. nov., Cassia carnaval, Cavaraea elegans, Chiovendea jin C. hypoleuca, v. Br. 

thrina chacoensis nom. nud., Ho ipiis t patagonica, Manganaroa ale re 

ta, M. furcata (Gillies ex Hook. & A 


oi, A. id d A. philippii 


andens, M. articula- 
Speg. comb. nov., M. martii (Benth.) Speg. sqm nov., M. gain 
(Willd.) p comb. nov., M. cuin (Willd.) Speg. comb. nov., M. paraensis, M. platensis  (Manganaro) Speg. 
comb. nov., M. subsericea, M. velutina, Mimosa ostenii Speg. ex ч M. striata (Benth.) Speg. comb. nov., M. 
tandilensis, shag ВЕ ameghinoi, Р. aphananthum, P. berteroi, P. canescens, P. carnosum (Dusén) Speg. comb. 
nov., P. filipes, P. gi nideum, P. griseum, P. karraikense (Speg.) Speg. ii. nov., P. leptopodum, P. patagoni- 
cum, P. rudolphi, P aurem (Speg.) Speg. comb. nov., P. serrazzianum, P. silvestrii, P. simonsi, P. tehuelches, 
P. triphyllum, P. vallis-pulchrae, P. villosum, Pirottantha modesta, Pithecodendron argentinensis, Prosopis patagoni- 
ca, Pterocarpus valentinii, Ramorinoa girolae, Trifolium argentinense, Vachellia astringens (Gillies ex Hook. 
Arn.) Speg. comb. nov., Vicia platensis, V. sericella, Xerocladia M Infraspecific taxa (64): Acacia adhaerens 
var. parviceps, А. praecox f. armata, A. praecox f. inermis, A. riparia var. argentinensis, Adesmia filipes var. obtusi- 
folia, A. lotoides var. eins A. lotoides var. elata, А. lotoides var. normalis, A. lotoides var. petiolulata, 
lotoides var. typica, A. patagonica var. nana, Anarthrophyllum desideratum var. bergii (Hieron. ) Speg. comb. nov., 
A. desideratum var. morenonis (Kuntze) Speg. comb. A. desideratu 
A. rigidum var. ср 5 Astraga lus 
sanctae var. cruc 
nseggia infolata va 
malis, H. trifoli 


ov., A. var. mustersil, А. desideratum var. typica, 
us moyanoi var. villosula, А. palenae var. grandiflorus, A. ngii var. lejocarpa, A. 
ndra grisebachiana var. carolae, Erythrina crista-galli var. inermis no 

rrima, H. trifoliata var. glandulosa, H. trifoliata var. Te lla, 
ata var. р thyrus cicera var. patagonica, [. mag cus var. glaucescens, L. magellan- 
icus var. oxyphylla, L. pubescens var. glaucescens, L. pubescens var. о а n ни var. normalis, L. 


offm 
Н. ! ils var. nor- 


Volume 87, Number 2 Katinas et al. 197 
2000 Carlos Spegazzini 


Table 2. Continued. 


stipularis var. patagonica, Lonchocarpus neuroscapha var. pubescens, Manganaroa paniculata var. paraguayensis, M. 
velutina var. Кў таны Patagonium lanatum var. axillaris, Р. lanatum var. parvifolia, P. villosum var. acutifolia, 
P. villosum var. glabratum. P. villosum var. typica, Vachellia farnesiana |. armata, У. farnesiana f. brachypoda, У. 
farnesiana М cavenia ма Speg. comb. поу., V. farnesiana f. inermis, V. farnesiana f. micrantha, V. farnesiana 
f. microcarpa, V. farnesiana f. stenocarpa, V. farnesiana f. typica, V. lutea f. aroma (Gillies ex Hook. & Arn.) Speg. 
comb. nov., V. lutea var. aroma, V. lutea f. leptocarpa, V. lutea f. moniliformis (Griseb.) Speg. comb. nov., V. lutea f. 
oocephala, V. lutea f. pachycarpa, V. lutea f. thlipsacantha, Vicia bijuga var. longipes, V. patagonica var. depaupera- 
ta (Clos) Speg. comb. nov., V. sericella var. glabrata, V. vicina var. azurea, V. vicina var. luteiflora, V. vicina var. 
pallidiflora, V. vicina var. tricolor. Total (164). 

FLACOURTIACEAE. Species (1): Banara glandulosa (Desv.) Speg. comb. nov. Total (1). 

FRANKENIACEAE. Species (3): Frankenia chubutensis, F. pampeana, F. patagonica. Infraspecific taxa (3): Franken- 
ia а var. juniperinoides (Hieron.) Speg. comb. nov., F. microphylla var. relaxata, F. microphylla var. typi- 
ca. Tota 

GENTIANACEAE. Species (1): Erythraea ameghinoi. Infraspecific taxa (5): Gentiana magellanica var. darwinii (Gri- 
seb.) Speg. comb. nov., G. magellanica var. typica, G. magellanica f. albiflora, G. magellanica f. cyanescens, G. 
magellanica f. pumila. Тога! (6). 


GERANIACEAE. Species E» Geranium melanopotamicum. Infraspecific taxa (2): Erodium cicutarium var. arenicola 
(Steud.) Speg. comb. nov., Geranium dissectum var. patagonica. Total (3). 

HALOPHYTACEAE. Genera om Halophytum. Species (1): Halophytum ameghinoi (Speg.) Speg. comb. nov. Total (2). 

HYDNORACEAE. Species (2): Prosopanche bonacinae, P. mazzuchii. Infraspecific taxa (2): Prosopanche burmeisteri 
var. bettfreundii, P. burmeisteri var. minor. Total (4). 

IRIDACEAE. Species (3): Cypella elegans, C. oreophila, S Pm patagonicus, Infraspecific taxa (1): Sisyrin- 
chium striatum var. mrs (Phil.) Speg. comb. nov. Total (4). 

JUNCACEAE. Species (1): Luzula patagonica. Total 

JUNCAGINACEAE. Species ms Triglochin monanthos. Total (1). 

LAMIACEAE. Species (2): Scutellaria platensis, Sphacele pampeana. Infraspecific taxa (5): Micromeria darwinii var. 
imbricatifolia, M. darwinii var. pallida, M. darwinii var. pusilla (Phil.) Speg. comb. nov., M. darwinii var. typica 

| 5 al (7). 


~ 
— 
ү 


= 
S 
= 
= 
= 
E 
< 
e 
ч 
~ 
c 
q 
ao 
< ~ 
% 
PE 


LENTIBULARIACEAE. Species (1): Utricularia platensis. Total (1). 

LILIACEAE. Genera (2): Shien, Schickendantziella. Species (7): Brodiaea ameghinoi, B. patagonica hom. 
illeg., Schickendantzia trichosepala, S. pygmaea, Schickendantziella trichosepala (Speg.) Speg. comb. nov., T. eremo- 
phyllum, T. pulchellum. | шеше taxa (2): Brodiaea patagonica var. angustiloba, Triteleia patagonica var. ап- 
gustiloba. Total (11). 

LOASACEAE. Species (1): Loasa patagonica. Infraspecific taxa (2): Blumenbachia silvestris var. leptocarpa, Loasa 

pinnatifida var. gracilis. Total (3). 

LYTHRACEAE. Species (1): Pleurophora patagonica. Total (1) 

MALVACEAE. Genera (1): Lecanophora. Species (13): Abutilon eriocarpum Speg. ex Stuckert nom. nud., A. vidalii 
(Phil.) Speg. comb. nov., Cristaria kuntzei, С. pues (Hieron.) Speg. comb. nov., Hibiscus argentinus hom. illeg., 

5 


(Hieron.) Speg. comb. nov., S. tehuelches, Sphaeralcea australis, S. patagonica (Niederl.) Speg. comb. nov. Infra- 
нд taxa (4): Sphaeralcea patagonica var. argentea, S. patagonica var. cinerascens, S. patagonica var. normalis, 
S. patagonica var. охуодота. Тога! (18). 

MARTYNIACEAE. Species (1): Craniolaria argentina. Total (1). 

MELASTOMATACEAE. Species (1): Comolia platensis. Total (1). 

MISODENDRACEAE. Species (1): Misodendrum patagonicum. Total (1). 

MORACEAE. Species (1): Urostigma quintuplinerve. Infraspecific taxa (1): Brosimum gaudichaudii var. longifolia. 
Total (2). 

MYRTACEAE. Species (4): Calyptranthes lilloi, C. oreophila, Eugenia guili, E. perorebi Parodi ex Speg. & Girola. 
Infraspecific taxa (1): Tepualia stipularis var. philippiana (Griseb.) Speg. comb. nov. Total (5). 

NYMPHAEACEAE. Species (1): Cabomba australis. Total (1). 

ONAGRACEAE. Species (1): Oenothera pygmaea. Infraspecific taxa (3): Oenothera odorata var. glabrescens, O. odor- 
ata var. media, O. odorata var. undulata. Total (4). 

ORCHIDACEAE. Species (11): Chloraea albo-rosea Kraenzl. ex Speg., C. chica Speg. ex Kraenzl., C. cholilens 
Speg. & Kraenzl.*, C. hookeriana Speg. & Kraenzl.*, C. hystrix Speg. & Kraenzl., C. phoenicea, C. УИ 22 
Kraenzl. & Speg., C. praecincta Speg. & Kraenzl., ГРАН И argentinense, Pleurothallis aurantio-lateritia, Res- 
trepia cogniauxiana Speg. & Kraenzl. Total (11). 

OXALIDACEAE. Species (4): Oxalis chubutensis Speg. ex R. Knuth, O. nahuelhuapiensis, O. patagonica, O. steno- 
phylla. Infraspecific taxa (1): Oxalis valdiviensis var. humilis. Total (5). 


198 Annals of the 
Missouri Botanical Garden 


Table 2. Continued. 


PHYTOLACCACEAE. Species (1): Seguieria guaranitica. Total (1). 

PLANTAGINACEAE. Species (4): Plantago carrenleofuensis, P. oxyphylla, P. pulvinata, P. tehuelcha. Infraspecific 
taxa (10): Plantago macrostachys var. subandina, P. maritima var. macrophylla, P. maritima var. pauciflora, P. 
2s var. baht P. myosuros var. latifolia, P. myosuros var. taraxacoides, P. patagonica var. gracilescens, P. 

ca var. minuscula, P. patagonica var. typica nom. inval., P. pauciflora var. taraxacoides. Total (14). 

PLUMBAGINACEAE. Species (1 ): Statice patagonica. Total (1). 

POACEAE. Species (102): Agrostis eremophila, A. Кијшт, A. moyanoi, А. pyrogea, А. santacruzensis, A. tehuelcha, 
Andropogon agrostoides, Aristida pampeana, Atropis Ышш, Rande fuegiana, C. modesta, C. suka, Cor- 
taderia dioica (Spreng.) Speg. comb. nov., Cryptochloris spatacea nom. nud., Deyeuxia Ка НИ D. freticola, D. 
patagonica, Elymus ааа Е. о Е. танак Festuca chubutensis, Е pampeana, Е pyrogea, Е 
shuka, F. ventanicola, Glyceria antarctica, G. na, G. d Lappago oplismenoides, Leptocoryphium 
penicilligerum, Milium juncoides, Mon onc аа Oplisme из oplismenoides (Speg.) Speg. comb. nov., Ory- 
am bicolor (Vahl) Srk comb. nov., O. grisebachii, O. hackelii Andar] Speg. comb. nov., O. lasiantha (Gri- 

eb.) Speg. comb. nov., O. lejocarpa, O. lejopoda, O. napostaensis, O. ovata, O. panicoides (Lam.) Speg. comb. nov., 
0. ле (E. Desv.) Sa comb. nov., O. stipoides (Trin. & Rupr.) Speg. comb. nov., O. tuberculata (E. 
Desv.) Speg. comb. nov., O. uruguayensis (Griseb.) Speg. comb. nov., О. verrucosa (Phil.) Speg. comb. nov., О. 
verruculosa, Panicum bambusoides Speg. ex Arechav. hom. illeg., P. guaraniticum, Poa argentina, P. chubutensis, 
P. erinacea, P. а P. yaganica, Savastana antarctica (Labill.) Speg. comb. nov., a patagonica, 
Stipa ambigua, S. ameghinoi, S. arcaensis, S. arechavaletae, S. argentina, S. argentinensis, S. bavioensis, S. brac 
chaetoides, S. cacheu ud " caespitosa d Speg. яа nov., 5. calchaquia, S. chubutensis, 5 cordo паа 
curamalalensis, S. dasyantha, S. dasycnemis, S. gracilis, S. hypsophila, S. hystericina, S. jujuyensis, S. juncoides, 

5. xt Si ligularis (Griseb ) Spe. uh nov., 5. molfinoi, S. nana, S. nubicola, S. oreophila, S. pampagran- 

ampeana, 5. paramilloensis, S. patagonica, S. perrigida, S. НЕ E 5. psittacorum, 5. psylantha, 

5. puelches, S. sanluisensis, 5 scirpea Speg. emend. Roig, 5. sublaevis, S. tehuelches, S. torquata, S. uruguaycola, 

S. uspallatensis, Triticum fuegianum, T. magellanicum (Desv.) Speg. comb. nov. Infraspecific taxa (75): Agrostis 

airoides var. flacc MT А. moyanoi var. major, А. moyanoi var. plicatifolia, А. moyanoi var. puberigluma, Bromi- 

dium andinus var. scabrivalvus, Bromus andinus var. scabrivalvus, B. coloratus var. vivipara, B. unioloides var. hir- 
suta, B. nisse var. humilis hom. illeg., B. unioloides var. micrantha, B. unioloides var. rupestris, Cortaderia 
quila var. patagonica, Danthonia picta var. patagonica, Deseo) Dumas var. tandilensis (Kuntze) Speg. 
comb. nov., Festuca erecta var. cirrosa, F. gracillima var. brevifolia, F. gracillima var. patagonica, F. myuros var. 
muralis Kunth ex Speg., Hierochloe redolens var. major, Hordeum murinum var. velutina, Oryzopsis bicolor var. ma- 

Jor, O. bicolor var. media, O. bicolor var. minor, O. lejocarpa var. major, O. montevidensis f. brasiliensis, O. montevi- 

densis f. trachycarpa, O. montevidensis f. typica, O. napostaensis var. brachyphylla, O. napostaensis var. brachysper- 

ma, О. napostaensis var. macrophylla, а var. typica, Poa bergii var. chubutensis, P. lanu rj osa Var. 
elata, P. о var. gracillima, Polypogon elongatus var. patagonica, Stipa bavioensis var. minor, S. brachychae- 
var. major, S. brachychaeta var. minor, S. Nan jitosa. var. elata, S. caespitosa var. subelata, S. eaespitosa var. 
subtypica, S. pomo var. typica, S. clarazii var. bulbosa, S. са var. typica nom. inval., culmis var. 
major, S. filiculmis var. minor, S. kimi var. major, 5. humilis var. minor, 5. manicata var. latifolia ( (Hack. & 
Arechav.) Speg. comb. nov, S. manicata var. media, S. manicata var. Fdo nom. inval., 5. megapotamia var. јип- 
coides (Speg.) Speg. comb. nov., S. megapotamia var. - typica nom. inval., 5. papposa var. major, S. papposa var. 
minor, S. plumosa var. gracilis, S. plumosa var. media, S. plumosa var. micrura, S. setigera var. hispidula, S. setig- 
era f. pallida, S. setigera f. purpurascens, S. Hi versicolor, S. speciosa f. major, S. speciosa var. minor, S. 
tenuis var. argentina (Speg.) Speg. comb. nov., 5. tenuis var. typica, S. tenuissima var. oreophila (Speg.) Spe 
mb. n enuissima var. planicola, Triticum fuegianum var. ion icum, T. magellanicum var. nm (J. 
Presl.) Ри comb. nov., 7: magellanicum var. festucoides, T. magellanicum var. glabrivalva, T. magellanicum var. 
asiopoda, Г ше еч var. pubiflora (Steud.) Speg. comb. поу., T. magellanicum var. secunda (J. Presl.) Speg. 
mb. nov. Total (177). 

POLEMONI eer Species (3): Collomia chubutensis, C. patagonica, Gilia patagonica. Infraspecific taxa (4): Gillia 
gracilis subvar. spathulifolia, Navarretia involucrata var. pumila, N. involucrata var. trichophylla, Polemonium an- 
tarcticum f. violascens*. Total (7 

POLYGALACEAE. Species (6): Acanthocladus moyanoi, A. tehuelchum, Polygala moyanoi (Speg.) Speg. comb. nov., 
P. oedipus, P. pamparum, P. tehuelchum (Speg.) Speg. comb. nov. Total (6). 

POLYGONACEAE. Species (3): Coccoloba argentinensis, Епоропит ameghinoi, E. bonaerense. Infraspecific taxa (1): 
Polygonum spectabile var. patagonica. Total (4). 

PORTULACACEAE. Species (8): Calandrinia chubutensis, C. macrocarpa, C. patagonica, Portulaca amilis, P. crypto- 
petala, P. platensis, P. rosae, Talinum paraguayense. Infraspecific taxa (3): Portulaca cryptopetala f. phenopetala, P. 
oleracea var. macrantha*, P. oleracea var. micrantha*. Total (11). 


Volume 87, Number 2 Katinas et al. 199 
Carlos Spegazzini 


Table 2. Continued. 


RANUNCULACEAE. Species (6): Anemone myriophylla, Myosurus patagonicus, Ranunculus bovei, Ranunculus fuegi- 
anus, R. oligocarpus hom. illeg., R. potamogetonoides. Infraspecific taxa (6): Caltha sagittata f. latifolia, C. sagitta- 
ta f. typica, Myosurus aristatus var. brachypoda, M. aristatus var. 14 acilis, Ranunculus bovei var. potamogetonoides 
(Speg.) Speg. comb. nov., R. peduncularis var. alboffiana. Total А 

RHAMNACEAE. Species (4): Discaria andina*, 1). cognata Mier) Speg. comb. illeg., D. foliosa (Hook. & Arn.) 
Speg. comb. illeg., D. Wr oe piu "ific taxa 4): Colletia ferox var. puberula, Condalia lineata var. erythro- 
carpa, C. lineata var. melanocarpa, C. lin var. ie Re otal (8). 

ROSACEAE. Species (10): pao aca A. tehuelcha, Fragaria pampeana, Margyricarpus acanthocarpus, M. 
ameghinoi, M. niederleinii, M. patagonicus, Prunus argentinensis, Tetraglochin ое (Speg.) Speg. comb. 
nov., T. nere (Speg.) Speg dpt nov. Infraspecific taxa (12): Acaena multifida glaberrima, A. pinnatifida 
var. glabrata, A. platyacantha f. elata, A. platyacantha var. parvifolia, A. pana p ica, А. qe duin f. 
villosa, Margyricarpus setosus var. patagonica, Tetraglochin acanthocarpum var. das ‚ T. acanthocarpum v 
lasiocarpa, T. acanthocarpum var. leiocarpum, T. acanthocarpum var. macropoda, T. B аем var. dedi 

22). 


yis. 


nom. inval. Total (2 

RUBIACEAE. Species M Oreopolus patagonicus. Infraspecific taxa (1): Galium aparine var. pseudoaparine (Griseb.) 
Speg. comb. nov. Total (2). 

SAMYDACEAE. Genera (1): Arechavaletaia. Species (1): Arechavaletaia uruguayensis. Total (2). 

SANTALACEAE. Species (2): Acanthosyris pet bae ameghinoi. Infraspecific taxa (4): Arjona patagonica var. 
tandilensis An Speg. comb. nov., Quinchamalium chilense var. gracile (Brongn.) Speg. comb. nov., Q. chilense 

r. majus (Brongn.) Speg. comb. nov., Q. chilense var. d (Berg.) Speg. Xu nov. Total (6 У 

ЗАРОТАСЕАЕ. сые (2): Siderorylon ligustrinum, S. myrtifolium (Mart.) Speg. comb. nov. Total (2). 

SAXIFRAGACEAE. Infraspecific taxa (3): Saxifraga а var. pavonii, S. magellanica var. trilobata, S. trigyna 
var. patagonica. Тога! (3). 

SCROPHULARIACEAE. Infraspecific taxa (2): Calceolaria bellidifolia var. angustifolia, C. lanceolata var. glabrata. 
Тога! (2). 

SIMAROUBACEAE. Species (1): Picrasma palo-amargo. Total (1). 

SOLANACEAE. Genera (3): т, Pantacantha, Saccardophytum. Species (27): Benthamiella acutifolia, B. 
azorelloides, B. longifolia, B. patagonica, B. pd lloides, Fabiana рик Grabowskia ameghinoi (Speg.) 

peg. с megalosperma, стона eghinoi, H. patagonicus, Jaborosa desiderata, J. leptophylla, J. 
oxipetala, Lycium acad абан, L. ameghinoi, L "halophilum, Г. рн, L. repens, Nicotiana acaulis, N. 
ameghinoi, N. deserticola, N. sylvestris Speg. & Comes, Nierembergia patagonica, Pa EI ameghinoi, Saccar- 
dophyton pycnophylloides, Solanum sidifolium, Trechonaetes leucotricha. аре Б taxa (7): Fabiana patagonica 
var. brachyloba, Е patagonica var. foliosa, Е d var. gracilis, F. patagonica var. nana, F. patagonica var. 
37 


typica nom. inval., Nicotiana alpina var. deserticola, №. alpina var. patagonica. Total (37). 
TROPAEOLACEAE. Species (1): Tropaeolum patagonicum. Infraspecific taxa (1): Tropaeolum polyphyllum var. incis- 
um. Total (2) 


VALERIANACEAE. Species (7): Phyllactis carnosa, P. clarionifolia (Phil.) Speg. comb. nov., P. regularis, P. salicar- 
iaefolia (Vahl) Speg. comb. nov., Valeriana bonariensis, V. chubutensis, V. moyanoi. Infraspecific taxa (3): Phyllactis 
macrorhiza var. pumila, P. magellanica var. azorelloides, Plectritis samolifolia var. pusilla. Total (10). 

VERBENACEAE. Genera (1): Mo n Species (15): Lippia darwinii (Benth. & Hook. f.) Speg. comb. nov., Mon- 
opyrena serpyllifolia, Verbena ameghinoi, V. aurantiaca, V. azorelloides, V. carro, V. chubutensis, V. € и 

mulinoides, V. nubigena, V. patagonica, n serpyllifolia (Speg.) Speg. comb. nov., V. silvestrii, V. struthionum 
е иеа. taxa (3): Verbena crithmifolia var. "latiloba, V. flava var. Mj ах ba, У. flava var. loui 
tal (19). 
VIOLACEAE Species (1): Viola argentinensis. Infraspecific taxa (5): Viola maculata f. calliantha, V. maculata var. 
5 | microphylla f. fimbriata, V. microphylla |. macropoda, У. microphylla f. micropoda. Total (6). 

ZYCOPHY ACEAE. Species (1): Larre a ameghinoi. Total (1). 

ORB. Infraspecific taxa (1): Lycopodium scariosum var. patagonicum. Total (1). 

OPHIOGLOSSACEAE. Infraspecific taxa (1): Botrychium lunaria var. antarctica. Total (1). 


Total: 33 genera; 696 species; 386 infraspecific taxa. TOTAL TAXA: 1115. 


шап journals as Anales de la Sociedad Científica — lications: one ranges from 1878 to 1919 (Scala, 
Argentina, Comunicaciones del Museo Nacional de 1919) and the other Кот 1919 to 1926 (Molfino, 
Buenos Aires, Physis, and Revista de la Facultad de 1929). The subjects of Spegazzini’s publications are 


Agronomía de la Universidad Nacional de La Plata. 
There are two papers listing all of Spegazzini's pub- 


diverse: algae, fungi, vascular plants, anthropology, 
zoology, and medicine. His work on vascular plants 


200 


Annals of the 
Missouri Botanical Garden 


deals with agriculture, floristics, morphology, paleo- 
botany, phytochemistry, phytopathology, taxonomy, 
botanical travels, and xylology. 

Although his taxonomic work on vascular plants 
covers numerous families, he had a special interest 
in Cactaceae (Senet, 1926; Kiesling, 1984), Faba- 
ceae, Orchidaceae, and Poaceae. As mentioned by 
Hauman (1923), his publications of new taxa have 
been subject to criticism because many of them 
were based on taxa already described by other bot- 
anists. Table 2 shows a total of more than 1 
names at the rank of genus, species, variety, sub- 
variety, and form listed as Spegazzini new names 
in Index Kewensis (Anonymous, 1997) and the Gray 
Card Index Database. Relevant literature (Zuloaga 
et al., 1994; Zuloaga & Morrone, 1996, 1999a, b 
has been consulted for oe the current sta- 
tus of these names. As a consequence, some of 
them resulted in illegitimate or invalid names, or 
illegitimate, superfluous, or new combinations. A 
more detailed analysis of the vascular plant names 
published by Carlos Spegazzini will be obtained 
with the production of the type specimens catalogue 
(Katinas et al., in prep.). 

Carlos Spegazzini made some local floristic stud- 
ies, mainly within the Province of Buenos Aires, 
such as in La Plata city, and in the mountain ranges 
of Tandil and Ventana. Although he started the first 
floristic study of the flora of the whole Buenos Aires 
Province, he did not finish this work. 

Most of his travel accounts were first written as 
reports for his sponsoring institution, and then usu- 
ally published in contemporary journals. His ac- 
counts of the trips were thorough and amusing, with 
med of details ranging from his salary to the phy- 
aphy of the area. When he died, many pub- 
foiiis remained unfinished, e.g., a new issue of 
Revista Argentina de Botánica, iconographies of the 
family Commelinaceae in Argentina, a monograph 
of the genus Prosopis (Fabaceae), a work on the 
genus Senecio (Asteraceae), a flora of Patagonia, 
and a catalogue of the vascular plants of Argentina 
with their scientific and vernacular names, includ- 
ing the Indian names (Senet, 1926). 

Carlos Spegazzini was a hard worker who pub- 
lished practically every year of his scientific life, 
with 1925 being his most productive (with 20 pa- 
pers), just one year before his death 


м 


THE LEGACY 


Carlos Luis Spegazzini was an influential force 
in the botanical community of Argentina. As a pro- 
fessor he nurtured enthusiasm and dedication 
among his students, extending his influence beyond 


botanical instruction and encouragement. Inspired 
by his knowledge of agriculture, he helped to solve 
many serious pest problems in Argentina. More- 
over, Spegazzini’s fascination with Patagonia led 
him to the creation of one of the most interesting 
and complete collections of vascular plants of this 
part of the world. His herbarium of vascular plants 
constitutes a fundamental witness to the past veg- 
etation of Argentina, a valuable source of infor- 
mation that contributes to portraying the present- 
day loss of biodiversity. His work has been pivotal 
in awakening botanical science in Argentina and a 
general consciousness of the significance of plant 
and fungal systematics. Those in the botanical com- 
munity who knew Carlos Spegazzini emphasized his 
clear judgment, accessibility, and sense of duty. 
The Institute and Arboretum in La Plata, his lec- 
tures, his publications, and prolific herbaria form 
the backbone of the legacy of Carlos Spegazzini, an 
Italian botanist who loved Argentina as his own 
country. 


Literature Cited 


Anonymous. 1925. Movimiento social. Physis (Buenos 
Aires) 7: ой 138. 

0. Inauguración del Museo Spegazzini. Re- 

vista del M M de La Plata 32: 387—394. 

Index Kewensis on CD-Rom, version 2.0 
for indo. Oxford Univ. Press. Oxforc 

Arambarri, A. N. & H. A. Spinedi. 1996. Садое Taiis Spe- 
gazzini: Micólogo. Revista Museo (Argentina) 2: 15-17. 

Del Уто, А., E. M. Регепаш & M. Correa. 1998 

Evolución del conocimiento botánico de la Patagonia 

n Correa (dir.), Flora 


23. La obra botánica del Dr. Carlos Spe- 
gazzini. Physis (Buenos Aires) 6: 303-308. 

Holmgren, P. K., N. H. Holmgren & L. C. Barnett (Edi- 
tors). 1990. Index Herbariorum. je I, The Herbaria 
of the World, 8 ed. Regnum Veg. 

Hosseus, C. C. 1915. Algunas ы де = Казо 

. Physis (Buenos Aires) 1: 534—54 

Kiesling, R. 1984. Recopilación, en edición ciis de 
todos los trabajos o referencias sobre Cactaceas publi- 
cadas por el Dr. Carlos Spegazzini. Librosur edic "lones, 
Quilmes, gres Aires, Argentina. 

1 storia del conocimiento de la flora san- 
juanina. Pp. 8-15 п В. Kiesling (editor), Flora de San 
Juan, bd l. үйнө Mazzini Editores, Buenos Aires, 


rgen 
— с. 1923. Storia della поа парада: Ка. Arti 
rafiche Saturnia 5. A. 5., 
1929. Carlos сы 5и vida y su obra. An- 
ales 506. Ci. Argent. 108: 7-81. 
———. 1951. Semblanza del doctor Carlos Spegazzini. 


n. Ci. Mus. La Plata 2: 1-6. 


Moore. D. M. 1983. Flora of Tierra del Fuego. Antony 

bis son, идее England, and Missouri Botanical 
arden, St. Louis, U.S.A. 

hu L. R. Ciento cincuenta afios de Botánica 


1961 : 
en с pnm Argentina. Bol. Soc. Argent. Bot. 11: 


Volume 87, Number 2 
2000 


Katinas et al. 
Carlos Spegazzini 


Ragonese, A. 1986. Desarrollo de los bu he botánicos 
e investigaciones sobre pastizales naturales en la Re- 
püblica Argentina. Ed. Libro del асраи Buenos 
Aires, Argentina. 

были, E. J. 1960. Bosquejo histórico de los natur- 
alistas extranjeros en Argentina. Boletín del Laboratorio 
de Botánica, Facultad de Agronomía, Universidad Na- 
cional de La Plata 4: 3—67. 

Seala, А. С. 1919. La labor a a | ps IUS 
Spegazzini. Anales Soc. Ci. Arge 

Senet, R. 6. La labor del oed Ceca ^ P 
Diario 2 dap edición del 19 de agosto, Buenos Ai- 
res, Arg 

E uM c. L 1880. Plantae d novae vel cri- 
ticae. cs Soc. Ci. Argent. 10 23. 

3a. Plantae novae E Americae aus- 

tralis. Piin Soc. Ci. Argent. 16: 1-16. 

188 Relazione pp sulle collezioni 
botaniche fatte in Patagonia e nella Terra del Fuoco. 
Pp. 3-16 in S. Bove, tac soe Italia. 

a. Costumbres de 

Soc. Ci. Argent. 17: 221—240. 


8 uo de la Tierra del Fuego. An- 


s Patagones. Anales 


ales Soc. Ci. Argen : 3- 
. 1895a. El dh de là саћа de azúcar en Tu- 
cumán. у Azucarera (Buenos Aires) 16: 5—29. 
——. 189 na nueva enfermedad de las peras. Re- 
vista Fac. Agron. Univ. Nac. La Plata 1: 1—13. 

—— ———. 1896. Plantae per Fuegiam a Carolo те 
аппо `1882 collectae. Anales Mus. Nac. Hist. Nat. 

Buenos pets 5: 39-104. 

Contribución al estudio de la Flora de 

la Sierra 5 la eee oe Fac. Agron. Univ. Nac. 

La Plata 32/33: 591-6. 

ae Patagoniae australis. нан 
Fac. Agron. Univ. Nac. La Plata 30/31: 485-58 

—— ———. 1897c. Primitiae Florae Chubutensis. evista 

Agron. Univ. Nac. La Plata 32/33: 591—6 

7d. Plantae novae nonnullae өнер Aus- 
tralis. | pium Mus. Nac. Buenos Aires 2: 46— 

— 1 Sobre una nueva enfermedad del tabaco. 
Oficina Químico-Agrícola de la provincia de Buenos Ai- 
res 4: : 


Fac. 


18994. Моуа Ae ad Floram Patagonicam. 
ји ми Nec Nac. t. Nat. Buenos Aires 1: 44—59. 
99b. Pla пи novae nonnullae Americae Aus- 
Tini ] yo Mus s Aires 1: 81—90. 
901a. Contribución de de lá Ey = 
Tandil. rh Sessé, Larran a y Renovales, La Pla 
Buenos е Argentina. 
1 с Se platenses. Anales Mus. Nac. 
coded 19: 1—56. 
1901c. Nova Addenda ad Floram Patagonicam. 
I. таер у = Editora de Coni Hermanos, Buenos 
Aires, Argentin 
1901 


wo novae nonnullae Americae Aus- 
жай; Ш. Conan: Mus. Nac. Buenos Aires 1: 312-324, 
— 


902a. Nova Addenda ad Floram m 
II. т. Mus. Nac. Hist. Nat. Buenos Aires 3/4: 13 
308 


. 1902b. Nova Addenda ad Floram Patagonicam. 
II. Anales Soc. Ci. Argent. 30: 13-18, 136-144, 166- 
185, 275-291. 

. 1902с. Nova Addenda ad Floram Patagonicam. 
Ш. Anales Mus. Nac. Hist. Nat. Buenos Aires 7: 135- 
308. 


905. Flora de la provincia de Buenos Aires. 
PRO del krea de Agricultura, Sección Biología 
=e –161. 
Al través ni Misiones. 
Ares. mid Nac. La Plata 5: 9-95 
1 bre се parásitas fanerogámicas 
de la ‘Republica Argentina. Anales Soc. Ci. Argent. 77: 


Revista Fac. 


—— ———. 1914b. Notas у sup sobre plantas bets 
para los ganados. Ana oc. Ci. Argent. 77: 
—— ——. 1914с. La iid ruins. nueva нек: ds 
caucho. Anales Soc. Ci. Argent. 78: 71-73. 
. 1916a. Una nueva especie de Cachiyuyo del 
Рега. Physis (Buenos Aires) 2: 241-244. 
1916b. Aceitunas de manantial (Nostoc pruni- 
forme C. A. Agard var. andicola Speg. n. var.). Physis 
ез ka 2: 282-283 
—— 6c. Algunas Orquidácesa argentinas. Anales 
Mus. б Hist. Nat. Buenos Aires 28: 131-140 
. 1917а. Ramillete de plantas argentinas nuevas 
o interesantes. Physis (Buenos Aires) 3: 37-47. 
———. 1917b. Ramillete de plantas iab inas nuevas 
o interesantes (continuación). Physis (Buenos Aires) 3: 


155-179 


7c. Ramillete de plantas argentinas nuevas 
o interesantes (conclusión). Physis (Buenos Aires) 3 
325-351. 

7d. Espigando en el herbario. Imprenta y 
Casa Editora de Coni Hermanos, Buenos Aires, Argen- 
Ina. 


. 1921a. Plantas nuevas o interesantes. Imprenta 
y Casa Editora de Coni Hermanos, Buenos Aires, Ar- 
gentina. 


1921b. Sobre algunas enfermedades y hongos 

que afectan a las plantas de “agrios” en Paraguay. Im- 

prenta y Casa Editora de Coni Hexe. Buenos Aires, 

Argentina. 

. 1923a. Fitoadémonas. Physis (Buenos Aires) 6: 

325-327. 

. 1923b. |. sobre Alpa-Sandia. Physis (Buenos 
= : 135-13 

————. 1923c. i 'ácieas Argentinas. Bol. Acad. Nac. 

34. 


. Relación de un paseo hasta el Cabo de 
Hoorn. a Acad. Nac. Ci. 27: 26—404. 

—— ———. 1925a. Stipeae platenses novae v. criticae. Re- 

vista Argent Bot. 1: 9-51. 

el Argentinas. Revista Argent. 


Pa patagónico. Revista Argent. 
17. 


К. S. Cowan. 1985. Taxonomic Litera- 
‚ pp. 776-785. Bohn, Scheltema and Hol- 


. Botanical Latin. Timber Press, Port- 


Venturi, S. 1925. Supercherías Spegazzinianas, o palos y 
más palos. Miguel Violetto y Cia., Tucumán, Argentina. 

Zuloaga, F. O. & O. Morrone (Editors). 1996. Catálogo 
de las plantas vasculares de la República Argentina. I. 
Pteridophyta, Gymnospermae y Angiospermae (Mono- 
cotyledoneae). Monogr. Syst. Bot. Missouri Bot. Gard 
60: 1—323. 


——— (Editors). 1999a. Catálogo de las plan- 
tas vasculares de la Repüblica Argentina. II. Acantha- 
ceae-Euphorbiaceae (Dicotyledoneae). Monogr. Syst. 
Bot. Missouri Bot. Gard. 74: 1—621. 


Annals of the 
Missouri Botanical Garden 


—— — (Editors). 1999b. Catálogo de las 
plantas vasculares de la Republica Argentina. II. Fa- 
aceae ). Monogr. Syst. 
Bot. find Bot. Gard. 74: 623-1269. 


, E. G. Nicora, Z. E. Rugolo de Agrasar, O. Mor 
rone, J. e A. M. Cialdella. 1994. Catálogo de 
ч familia Poaceae en la República Argentina. Monogr. 


yst. Bot. Missouri Bot. Gard. 47: 1-178. 


PHENETIC SIMILARITY 
PATTERNS OF DIOECIOUS 
SPECIES OF POA FROM 
ARGENTINA AND 
NEIGHBORING COUNTRIES! 


Liliana Mónica Giussani? 


ABSTRACT 


Dioecious species of Poa L. are mainly American with a total of 41 taxa in South America, 11 in North America, 2 


were establishe 


American species, considered within Poa sect. 


r. ibari, are proposed herein. Bivariate analyses showed other non- ‘linear patterns of variation. These results, together 


with univariate analyses, were used to 


select diagnostic characters. Sexual dimorphism was the majo 


or source of species 


variation, and ranges for pistillate ind. staminate specimens were also calculated. An identification key for all taxa is 


include 


Key words: Poa, dioecious species, numerical taxonomy, morphological variability, synonymy, sexual dimorphism. 


Poa L. is one of the largest genera within Po- 
aceae, including over 200 species according to 
Hartley (1961), Nicora (1978), and Kellogg (1985). 
Additionally, Clayton and Renvoize (1986), Soreng 
1990), Watson and Dallwitz (1992), and Anton and 
Connor (1995) mentioned 400 to 500 taxa within 
oa. 

Poa is distributed worldwide, particularly at high 
altitudes and latitudes in both hemispheres; the 
taxon is largely absent from low areas of tropical 
regions (Hartley, 1961). It is a recognizable, well- 
defined (Clayton & Renvoize, 1986; Nicora & Ки- 
golo de Agrasar, 1987; Soreng, 1990), and mono- 
phyletic genus (Hartley, 1961; Kellogg, 1990). 

Interspecific variation in Poa is due principally 
to quantitative diagnostic characters (Kellogg, 
1990). Discrete morphological variables are mostly 
unreliable, often varying widely with the environ- 
ment, and even within a single plant (Kellogg, 
1985; Rúa, 1996). Species are frequently grouped 
into complexes based on their morphological simi- 
larities. These species complexes further comprise 


different chromosome numbers (Akerberg, 1942; 
Kellogg, 1985). Taxonomists who have studied the 
genus (Bor, 1952; Marsh, 1952; Torres, 1969, 1970; 
Vickery, 1970; Nicora, 1978; Moore, 1983; Kellogg, 
1985; Tateoka, 1985; Edgar, 1986) agree on the 
difficulty in delimiting taxonomic boundaries within 
Poa. Clausen (1961) and Soreng (1990) both con- 
sidered that this variability in chromosome number 
results from polyploidy, introgression as well as 
apomixis. Phenotypic plasticity as stimulated by 
environmental variability also obscures species de- 
limitation within Poa (Vickery, 1970; Giussani & 
Collantes, 1997). 

Breeding systems are highly diverse within Poa. 
Perfect flowers are typical, but dioecism, gynomon- 
oecism, and gynodioecism are also found among 
American species (Anton & Connor, 1995). Apo- 
mixis on exclusively pistillate plants or facultative 
apomixis on perfect flowers are both well docu- 
mented (Connor, 1979, 1981; Kellogg, 1987). 

Evolution of dioecism within Poa could have de- 
rived from gynodioecious species, following species 


| | am grateful to Universidad de Buenos Aires (ОВА), Instituto de Botánica Darwinion (IBODA), and Consejo 


Nacional de Investigaciones Científicas y Técnicas (CONIC 
e Fernando O. Zuloaga for helpful suggestions on the research and critical Eu on the 


dissertation. I acknowledg 


ET), which supported this research as part of my Ph.D. 


manuscript, O. Morrone for suggestions on the manuscript, M. B. Collantes for guidance on the beginning of this project, 


and E. Nicora 


specimen loans. 


also thank two anonymous reviewers for comments and Victoria Hollowell for her сна reading 


and — that improved different aspects of this manuscript. 
2 Instituto de Botanica Darwinion- CONICET, Labardén 200, Casilla de Correo 22, San Isidro (1642), Buenos Aires, 
Mina. 


ANN. Missouni Bor. GARD. 87: 203—233. 2000. 


204 


Annals of the 
Missouri Botanical Garden 


migration from North to South America (Anton & 
Connor, 1995). Dioecism is well represented in the 
Americas: 11 dioecious species are distributed in 
the Northern Hemisphere, while 41 range south- 
ward. These South American species were gener- 
ally treated as Poa subg. Dioicopoa (E. Desv.) J. R. 
Edm., although Soreng (1998a, b) considered them 
within Poa sect. Dioicopoa E. Desv., to conserve 
Poa subg. Poa as monophyletic. In order to main- 
tain a uniform treatment of the genus Poa, the tax- 
onomic delimitation proposed by Soreng (1998a, b) 
and based on a phylogenetic approach will be fol- 
lowed. 

South American dioecious species are mainly 
distributed in Argentina and Chile (Marticorena & 
Quezada, 1985; Zuloaga et al., 1994). Parodi 
(1936) and Rosengurtt et al. (1970) cited three en- 
demic species for Uruguay: P. arechavaletae Parodi, 
P. uruguayensis Parodi, and P. megalantha (Parodi) 
Herter. Smith et al. (1981) and Longhi-Wagner and 
Boldrini (1988) mentioned one endemic species in 
southern Brazil: P. reitzii Swallen. Nicora (1995) 
described one new species from Paraguay, Р. ped- 
ersenii Nicora; and Hitchcock (1927) and Renvoize 
(1998) cited one dioecious, species in Bolivia, Р 
buchtienii Hack. Although it was originally intend- 
ed herein to consider all South American species 
of Poa sect. Dioicopoa, some taxa were not included 
due to the lack of material or difficulties in the 
identification of doubtful taxa, especially with Chi- 
lean material, and endemic species of Uruguay and 

razil. 

Partial taxonomic revisions of some South Amer- 
ican taxa (Parodi, 1936; Torres, 1969, 1970; Ni- 
cora, 1978; Moore, 1983) used quantitative char- 
acters as diagnostic features, although there was 
much overlap in their character ranges among spe- 
cies. Differences in pistillate floret hairiness were 
also utilized for species classification, with stami- 
nate specimens, which are generally glabrous, dis- 
counted (Nicora, 1978). 

The aim of this study is to analyze the pattern of 
morphological variation within Poa sect. Dioicopoa 
based on pistillate and staminate plants. Grouping 
from multivariate analyses was used to look for sim- 
ilarity among taxa and this, along with study of 
types, helped circumscribe species and establish 
synonymy when necessary. Ап identification key is 
presented as the result of numerical analyses. This 
key is based on groups of selected characters that 
are strongly correlated, while unique characters 
were only used when they presented conspicuous 
discontinuities. 


MATERIALS AND METHODS 
SPECIES AND SPECIMENS ANALYZED 


Morphological variation among 34 dioecious spe- 
cies and three varieties was analyzed. A list of spe- 
cies is presented herein as Appendix 1 at the end 
of this manuscript. Species included in this study 
derive from the following regional taxonomic treat- 
ments: Nicora (1978) and Moore (1983) for Pata- 
gonia; Torres (1969, 1970) for Entre Ríos and 
Buenos Aires, Argentina; Parodi (1932, 1937, 
1940, 1950, 1961, 1962) and Zuloaga et al. (1994) 
for Argentina; Hitchcock (1927) and Renvoize 
(1998) for Bolivia; Smith et al. (1981) and Longhi- 
Wagner and Boldrini (1988) for Brazil; Marticorena 
and Quezada (1985) for Chile; and Parodi (1936) 
and Rosengurtt et al. (1970) for Uruguay. 

Three hundred seventy-six exsiccatae out of ap- 
proximately 800 specimens examined by the author 
were analyzed and regarded as operational taxo- 
nomic units (OTUs) for numerical analysis (Appen- 

dix 2). The number of specimens considered per 
species varies according to their representation in 
herbaria. Some endemic species are represented by 
only a few specimens, while other widespread spe- 
cies are represented by 15 or more. Types of all 
taxa were seen, and most of these were included in 
the numerical analysis. Only types of Poa bonar- 
iensis (Lam.) Kunth, Р. iridifolia Hauman, P. lan- 
uginosa Poir., P. prichardii Rendle, and P. stuckertii 
(Hack.) Parodi were not recorded. An attempt was 
made to cover the full range of morphological var- 
iation used by earlier authors to discriminate taxa, 
and to reflect distributional range for all species 
considered. Voucher specimens are deposited in 
the following herbaria: BAA, BAB, CORD, LP, 
LPB, and SI. 


MORPHOLOGICAL VARIABLES 


Forty-four morphological characters, including 
anatomical and epidermal leaf characters, were 
measured for this numerical analysis. Of these, 29 
are quantitative multistate characters and 15 are 
discrete variables (see Appendix 3). Most of them 
were considered diagnostic in previous taxonomic 
studies (Torres, 1969, 1970; Nicora, 1978; Moore, 
1983). Others were added to increase the character 
database. Vegetative characters were measured on 
the penultimate leaf of a sterile innovation. Leaf 
anatomical characters were sampled on a cross sec- 
tion from the blade midregion. Epidermal charac- 
ters represent the average of 10 measurements from 
the abaxial epidermis of the leaf blade midregion. 
Epidermal and anatomical characters follow the 


Volume 87, Number 2 
200 


Giussani 
Phenetic Patterns in Poa Sect. Dioicopoa 


character descriptions from Ellis (1976, 1979). Re- 
productive characters were measured on the tallest 
fertile culm. 


ASSESSMENT OF THE TAXONOMIC COMPLEXES AND 
SPECIES 


Phenetic similarities were examined by principal 
component analysis 
me represents distances between major groups 

ore accurately than any clustering method 
(Sneath & Sokal, 1973). A correlation coefficient is 
suggested for mixed data with predominantly quan- 
titative multistate characters (Crisci & López Аг- 
mengol, 1983). Numerical analysis was performed 
on a standardized character matrix. Data were 
transformed by the Pearson-moment correlation co- 
efficient on a similarity matrix, which was used to 
obtain the principal components. The basic data 
matrix is deposited in the Facultad de Ciencias Ex- 
actas y Naturales, Universidad de Buenos Aires, 
Argentina (Giussani, 1997: appendix B), and is also 
available from the author upon request. 

A analysis was repeated several times in order 
to detect different morphological patterns within the 
species studied. Toward this, different steps of mul- 
tivariate analysis were followed: 

(1) A single species or similar species groups 
were recognized by means of PCA. 

ach species or species group was charac- 
terized by the contribution of each character to the 


is ordination tech- 


first five principal components. 

(3) Distinctive species or species groups were 
separated and re-analyzed to recognize another var- 
iation pattern. 

(4) Species that were not initially delimited were 
re-analyzed until all taxonomic entities assorted 
into species complexes or were recognized as in- 
dependent species. 

Invariant characters were removed before PCA 
was performed. PCA distortion was measured due 
to the relatively low variability seen in the first five 
principal components. For this purpose, a Euclid- 
ean distance matrix deriving from the first five PCA 
axes was compared with a similarity matrix by a 
cophenetic correlation coefficient (Sneath & Sokal, 
1973). Two dissimilarity matrixes were obtained 
based on two indexes: taxonomic distance coeffi- 
cient and Manhattan distance, with гү, and ry the 
cophenetic correlation values, ий Жы Of 
these, only the highest scored value is presented. 

Minimum spanning tree or MST procedure (Gow- 
er & Ross, 1969; Rohlf, 1992) was additionally 
used to examine the similarity relationships among 
OTUs. This distance tree was imposed on the PCA 


plot with the OTUs linked by lines representing 
their minimum total distance (Cliffor tephen- 
son, 1975) 5 was performed on dissimilarity 
matrixes derived from taxonomic distance coeffi- 
cient or Manhattan distance. 


DISCRIMINATION AMONG TAXONOMIC ENTITIES 


Taxonomic groups were first defined by PCA and 
then considered as a priori groupings for discrim- 
inant analysis or DA (Sneath & Sokal, 1973; Affifi 
& Clark, 1984). DA was performed to identify lev- 
els of certainty within previously recognized groups 
as well as to select diagnostic characters based on 
standardized coefficients of the canonical variables. 

For discriminant analysis, only those characters 
that contributed most to the variability of the first 
five components of the PCA (r > 0.5) and that had 
the least correlation among each other (г < 0.5) 
were included. The only discrete characters ana- 
ed by DA were those of diagnostic value for pre- 
vious classifications (Torres, 1969, 1970; Nicora, 
1978; Moore, 1983), but quantitative multistate 
characters were preferred. 

The empirical method (Affifi & Clark, 1984) was 
used to estimate goodness of fit of the classification 
procedure. Thus, the discriminant function was ap- 
plied to the same samples used for deriving it, and 
the proportion of individuals correctly classified for 
each group was computed. 


e 


UNIVARIATE AND BIVARIATE ANALYSES. A KEY FOR 
IDENTIFICATION OF SPECIES AND TAXONOMIC 
COMPLEXES 


Mean and standard deviations for quantitative 
characters, as well as mode for discrete variables, 
were calculated for each species and species com- 
plex: these were plotted to reveal discontinuities 
and the pattern of variation among taxonomic en- 
tities (Sokal & Rohlf, 1969). Variation of a single 
character was also analyzed to consider dimor- 
phism between pistillate and staminate plants. 
Thus, averages of pistillate and staminate plants 
were calculated. Bivariate analysis was used to de- 
tect morphological patterns based on different char- 
acter combinations. After numerical analyses were 
completed, diagnostic characters were selected, 
and a key to identify species or taxonomic com- 
plexes was срок. constructed. 

umerical ana made using either 
NTSYS-pc (Rohlf, 1992) o or т STATGRAPHICS (Sta- 
tistical Graphics System by Statistical Graphics 
Corporation, 1992). 


206 


Annals of the 
Missouri Botanical Garden 


1.6 Kr ila а 
ad zu Poa bergii complex 
| Elg 
1.07 
Poa dolichophylla complex 
ё 
П 0.3]: 


-0.37 
-0.9 T T T 
-1.6 -1.0 -0.4 I 0.3 0.9 
Figure 1. First set of multivariate analyses. Plot of 376 individuals on the first two principal components. Two 


groups of species are distinguished: t 


мн dolichophylla complex (DO, IR, PC, ST), and the P. bergii complex (BE 
BA). Each label corresponds to the species name, with the 


first two letters abbreviated as in Appendix 1 (oistillate 


individuala i in all capital letters, and ie individuals in lowercase letters); the third T or t letter indicates the 


type species = a non-grouped specimen. 


RESULTS 


Taxonomic groups were selected based on the 
distribution of OTUs in the first five axes of PCA, 
since distortion of these axes was less than 10%. 


FIRST SET OF MULTIVARIATE ANALYSES 


The Poa dolichophylla Complex, the Poa ber- 
gii Complex, and Poa schizantha. Ordination 
of all 376 OTUs on the first five axes, utilizing 44 
characters, would account for 52.4% of total vari- 
ability. Although this is a small percentage of total 
variability, PCA distortion was low: ги = 0.92 

he first three principal components showed two 
groups of similar species. Axes I and II show the 
Poa dolichophylla complex, ыш, Р. dolicho- 
phylla Hack., Р. pilcomayensis va magrosto- 
idea Hack., P. stuckertii, and P. Я (Fig. 1). 
These principally grouped along the negative edge 
of axis I. From analysis of character loadings, they 
are characterized by long leaves, blades, and 
sheaths; plant stature over 50 cm tall; broad blades; 
long and broad panicles; numerous vascular bun- 
dles with sclerenchyma girders on both abaxial and 
adaxial epidermis; short and truncate ligules; 
groups of sclerenchyma cells extending on abaxial 


and adaxial epidermis at blade margin; small sto- 
mata; and short spikelets, glumes, florets, and lod- 
icules. 

The second species group was defined by axes 
I, II, and Ш and comprises three taxa: P. bergii 
Hieron. var. bergii, P. barrosiana Parodi, and Р 
schizantha Parodi. Specimens from these species 
clustered at the positive end of these axes (Fig. 1). 
They also present long leaves, blades, and sheaths, 
tall plants, and long panicles. However, these spe- 
cies are distinguished from the previous group by 
longer ligules, thicker blades, larger stomata, lon- 
ger spikelets, glumes, and florets, additional nerv- 
ing on the first glume, as well as numerous vascular 
bundles with sclerenchyma girders only on abaxial 
epidermis. 

The second group was analyzed separately. PCA 
was performed on a subset of s based on 
39 morphological variables. Poa bergii var. bergii, 
P. barrosiana, and P. schizantha share some attri- 
butes that were revealed as invariant characters be- 
fore PCA was performed: long rhizomes (HAB 2, 
see Appendix 3), no viviparous florets (FLOv 0), 
glabrous lemmas between nerves (HAIbet 0), no 
vascular bundles with only adaxial sclerenchyma 
girders (SCLad 0), and numerous silico-suberose 


Volume 87, Number 2 
2000 


Giussani 207 
Phenetic Patterns in Poa Sect. Dioicopoa 


“—~<— ~— 


SC 


"= ~ - 


= 
| 


Poa bergii complex 


КЕ Pistillate 


re 2. First set of multivariate analyses. Plot of 33 individuals on the first three PCA axes: Роа schizantha (SC) 
and pistillate and staminate individuals of the P. bergii complex are in different shades of gray. See Figure 1 caption 


for explanation of labe 


paired cells (SISU 2). These first five components 
accounted for 5596 of total variation; meanwhile, 
distortion of PCA was low: r, — 0.93. Specimens 
of P. schizantha were distinguished from those of 
the Poa bergii complex: P. bergii var. bergii and P. 
barrosiana; sexual dimorphism within the P. bergii 
complex was also supported. Figure 2 shows OTU 
distribution on the first three principal components. 
Poa schizantha falls on the negative end of axis I, 
and toward the positive ends of axes II and III. It 
is clearly defined by a smaller plant size; smaller, 
narrower, and thinner leaves; smaller fertile struc- 
tures; long and rigid callus hairs; and convolute 
blades. It also has smaller stomata and bulliform 
cells only slightly or not differentiated from other 
epidermal cells. А clear dimorphism between pis- 
tillate and staminate plants of the P. bergii complex 
was also distinguished. Pistillate specimens asso- 
ciate with the positive end of axes I and III: they 
are larger plants, with more hairy florets than sta- 
minate specimens. 


iscrimination among taxonomic entities. The 
Poa dolichophylla complex, the P. bergii complex 
(P. bergii and P. barrosiana), P. schizantha, and а 
remaining group of miscellaneous species com- 
prised the four a priori groups for an initial dis- 
criminant analysis. Standardized coefficient values 
for the first three canonical variables were used to 


characterize groups. Species within the P. dolicho- 
phylla complex have wide leaves and blade margins 
with a group of sclerenchyma cells extending on 
the adaxial and abaxial epidermis; the Р. bergii 
complex has long ligules, sheaths, lemmas, stomata, 
as well as numerous glume nerves. Poa schizantha 
is characterized by intermediate ligule, lemma, and 
stomata lengths, as well as narrower blades and 
thinner long cells. These groups appeared appro- 
priately classified by discriminant functions. 
Eighty-eight percent of specimens of the P. doli- 
chophylla complex, 8596 of the P. bergii complex, 
and 100% of P. schizantha correctly assorted to the 
a priori groups. 


SECOND SET OF MULTIVARIATE ANALYSES 


After subtracting specimens of the Poa dolicho- 
phylla complex, the P. bergii complex, and P. schi- 
zantha from the original data matrix, the 302 re- 
maining specimens were re-analyzed with the 
original 44 morphological and anatomical charac- 
ters. The first five PCA axes accounted for 49% of 
total variability; in spite of this percentage, distor- 
tion was low at r, — 0.89. 

OTU distribution on the first three axes of PCA 
discriminates three groups of species (Fig. 3): 
GROUP A, constituted by P. bonariensis, P. buch- 
tienit, P. calchaquiensis Hack., P. lanigera Nees, P. 


Annals of the 
Missouri Botanical Garden 


, та ` 
ME \ 
, m 
~ 


r 


Poa ligularis 


L 


es 3. Second set of multivariate analyses. Plot of 302 individuals on the first three principal components 


1807 
showing Group A (Poa bonariensis, P. buc 
senii, P. pilcomayensis, P. montevidensis 


of la 


patagonica var. neuquina Nicora, P. pedersenii, P. 
pilcomayensis Hack., montevidensis Arechav., 
and P. resinulosa Nees ex Steud; GROUP B, con- 
sisting only of specimens of P. ligularis Nees ex 
Steud.; and GROUP C, which includes P. alope- 
curus (Gaudich.) Kunth, P. boelckei Nicora, P. po- 
gonantha (Franch.) Parodi, P. prichardii, P. shuka 
(Speg.) Parodi, P. superbiens (Steud.) Hauman & 
Parodi, and P. tristigmatica E. Desv. ex Gay. 

Analysis of character correlation with the first 
three PCA axes revealed that species of Group A, 
placed on the positive end of axis I and the negative 
end of axis III, are characterized by long blades; 
short, truncate ligules; and smaller spikelets, 
glumes, florets, and lodicules. Group B, on the pos- 
itive end of axes I and III but the negative end of 
axis П, also has shorter spikelets and florets, but is 
distinguished by its longer ligules, filiform leaves, 
smaller stomata, as well as smaller plants and in- 
florescences. Species of GROUP C aligned on the 
negative end of axis I but the positive end of axis 
II. These species are distinguished by their larger 
spikelets, glumes, and florets; wider, thicker blades; 
additional vascular bundles with sclerenchyma 
girders on both abaxial and adaxial epidermis; and 
longer stomata. 


agonica var. neuquina, P. peder- 
р opecurus, P. 


boelckei, P. pogonantha, P. prichardii, P. is P. superbiens, and P. tristigmatica). See Figure 1 caption for explanation 
f labels. 


Discrimination among taxonomic entities. Groups 
A, B, C, and all remaining species were the four a 
priori groups used for the discriminant analysis 
(DA). Specimens assorting to Group A and B sep- 
arated along the first canonical variable. They are 
characterized by long blades, long hairs along lem- 
ma nerves, and sharp leaf blade apices. Specimens 
of Group C are distinguished by longer glumes. Poa 
ligularis, separated by the second canonical vari- 
able, is discriminated by a long ligule and numer- 
ous vascular bundles with sclerenchyma girders 
only on the abaxial epidermis. Group A is also 
characterized by wide blades associated with the 
second variable. All groups were assigned by the 
classification procedure with the percentages of 
correct classification being 86%, 90%, and 80% for 
groups A, B, and C, respectively. 


The Poa bonariensis Complex, Poa lanigera, 
Poa pilcomayensis var. pilcomayensis, and the 
Poa resinulosa Complex. 
within Group A include Poa bonariensis, P. buch- 
пепи, P. calchaquiensis, P. lanigera, P. monteviden- 
sis, P. patagonica var. neuquina, P. pedersenii, P. 
pilcomayensis var. pilcomayensis, and P. resinulosa. 
Their phenetic relationships were investigated by 


Species assorted 


Volume 87, Number 2 Giussani 209 
2000 Phenetic Patterns in Poa Sect. Dioicopoa 
Poa resinulosa complex ~. рых 
ш n: IM p 
, Poa bonariensis complex 
БЕЈ | `... й 
mum 
e 4. Second set of multivariate analyses. Plot of 84 individuals of Group A on the first three PCA axes showing 
the Poa resinulosa complex (BT, CL, PE, RE), the P. bonariensis complex (BN, MO, PN), P. lanigera (LG), and P. 
pilcomayensis (PI). See Figure 1 caption for explanation of labels. 


PCA. A data matrix was based on 84 OTUs and 43 
variables. Only 1 character out of the 44 was in- 
variant: the absence of viviparous florets (FLOv 0, 
see Appendix 3). The first five components ac- 
counted for 5396 of total variability. PCA distortion 
was low, being гм = 

A plot of the first ilios principal components 
showed four distinct groups of species (Fig. 4). The 
P. resinulosa complex consists of P. buchtienii, P. 
calchaquiensis, P. pedersenii, and P. resinulosa. А 
P. bonariensis complex contains P. bonariensis, P. 
patagonica var. neuquina, and Р. montevidensis. 
Two other groups each consist of two independent 
taxa, P. pilcomayensis var. pilcomayensis and P. lan- 
igera. The P. resinulosa complex, located at the 
negative end of axis I and the positive end of axis 
III, comprises specimens of relatively small stature, 
with small stomata, narrow blades, as well as short- 
er spikelets, glumes, and florets. The P. bonariensis 
complex distributed to the positive end of axes I 
and III. This complex includes rhizomatous, larger 
plants, with longer stomata, wider blades, as we 
as larger spikelets, glumes, and florets. Vascular 
bundles with sclerenchyma girders are more nu- 
merous in blade cross section. Poa pilcomayensis 
var. pilcomayensis and P. lanigera both fell at the 
negative end of axis III. They share flat, narrow 
blades. However, P. lanigera is distinguished along 
the first principal component by its longer glumes, 


florets, and spikelets than are seen for P. pilcomay- 
ensis var. pilcomayensis. 

Specimens within the P. resinulosa complex, in- 
cluding P. buchtienii, P. calchaquiensis, P. pedersen- 
ii, and P. resinulosa, were separately re-analyzed. 
PCA revealed a sexual dimorphism among speci- 
mens with staminate individuals presenting smaller 
fertile structures and less hairy florets than pistil- 
The first five PCA axes accounted for 
54% of the whole variance, and the cophenetic cor- 
relation coefficient showed a low PCA distortion, ry 


late ones. 


Specimens of the P. bonariensis complex were 
also separately analyzed. Principal differences 
among OTUs were again due to sexual dimorphism. 
This P. bonariensis group distributed on PCA axes 
I, II, and IV and was also sustained on a minimum 
spanning tree (Fig. 5). Staminate specimens, in 
contrast with pistillate specimens, again show 
smaller spikelets, glumes, and florets, as well as 
less hairiness on these florets. 


Discrimination among taxonomic entities. А 
priori groups for DA corresponded to those ob- 
tained from PCA: (1) P. bonariensis complex; (2) P. 
resinulosa complex; (3) P. pilcomayensis var. pilco- 
mayensis; and (4) P. lanigera. Two groups were sep- 
arated along the first two canonical variables. The 
P. resinulosa complex is discriminated by a rela- 


210 


Annals of the 
Missouri Botanical Garden 


Poa bonariensis complex 


Pistillate 


Figure 5. Second set of multivariate analyses. Plot of 25 individuals of the Poa bonariensis complex (BN, MO, PN) 
distributed on the I, Ш, and IV principal components. MST is superimposed on the PCA plot (lines). Dimorphism 
between pistillate and staminate specimens is the main source of variation among specimens of the complex. See Figure 


1 caption for explanation of labels. 


tively long ligule and subconvolute blades. The Р 
bonariensis complex is characterized by large pan- 
icles and glumes, wide paleas, numerous silico-su- 
berose paired cells, and vascular bundles with scle- 


aligned along the third canonical variable. Poa lan- 
igera has broader paleas and hairier lemma nerves 
than Р. pilcomayensis. Discriminant classification of 
the a priori groups showed a good fit: 9296 of spec- 
imens for the P. resinulosa complex, 90% for the P. 
bonariensis complex, 9296 for P. pilcomayensis, and 
75% for P. lanigera were correctly assigned. Some 
misidentified specimens of the P. resinulosa com- 
plex associated with the Р bonariensis complex. 
Others of P. pilcomayensis were related to P lani- 
gera. Some specimens of P lanigera intermixed 
with the P. bonariensis complex. 


The Poa alopecurus, Poa pogonantha, and 
Poa tristigmatica Complexes. PCA ordination 
of specimens of Group C revealed two species 
groupings. The Poa pogonantha complex consists 
of P. pogonantha and P. prichardii, whereas speci- 
mens of P. alopecurus, P. boelckei, P. shuka, P. su- 
perbiens, and P. tristigmatica fall into the second 


group. OTU distribution (Fig. 6) is based on PCA 
axes I, II, and IV. Analysis of character loadings 
revealed that specimens within the P. pogonantha 
complex are characterized by viviparous florets and 
long panicles, but few have sclerenchyma girders 
associated with vascular bundles. A second species 
grouping features larger plants, longer ligules, as 
well as longer glumes, florets, and lodicules. The 
first five PCA axes explained only 4796 of total var- 
iation. Nevertheless, its distortion remained low, ry 


Specimens of P. alopecurus, P. boelckei, P. shuka, 
P. superbiens, and P. tristigmatica were individually 
analyzed. Ordination along the first three principal 
component axes revealed two groups of species 
(Fig. 7): a P. tristigmatica complex with two spe- 
cies, P. boelckei and P. tristigmatica; and a P. alo- 
pecurus complex including P. alopecurus, P. shuka, 
and P. superbiens. The P. tristigmatica complex as- 
sociated with the positive end of the first component 
axis due to smaller plants, leaves, and fertile struc- 
tures. The Р. alopecurus group principally located 
at the opposite side of axis I, associated with larger 
plants, leaves, and fertile structures. Pistillate and 
staminate specimens of P. boelckei and P. tristig- 


Volume 87, Number 2 
2000 


Giussan 211 


Phenetic Patterns in Poa Sect. Dioicopoa 


e 6. Second set of multivariate analyses. Plot of 76 individuals of Group C (Poa alopecurus, P. Á— P. 


igure 
pogonantha, P. prichardii, P. shuka, P. superi 


iens, and P. tristigmatica) оп 


the I, II, and ТУ РСА axes. The P. p 


onantha 


complex (PG, PR) is clearly distinguished from the remaining species. See Figure 1 caption for nsi ad A of labels. 


matica differentiated on axis II (Fig. 7). Pistillate 
plants present wider glumes and florets, longer lod- 
icules, and thicker blades than staminate speci- 
mens. PCA distortion was low, at r, — 0.92, al- 
though the first five axes accounted for just 4996 of 
total variability. 

When based only on specimens of the P. pogon- 
antha complex, PCA revealed morphological dif- 
ferences between pistillate and staminate individ- 
uals. Pistillate specimens have hairy calluses, hairy 
lemma nerves, and vascular bundles with scleren- 
chyma girders on both abaxial and adaxial epider- 
mis. Staminate specimens are distinguished by hav- 
ing more nerves on the first glume and 
well-differentiated bulliform cells. The first five 
axes accounted for 5796 of total variability with 
PCA distortion being low, гг = 0.92. Only three 
variables, navicular apices (АРГ 1, see Appendix 

) no vascular bundles with few sclerenchyma 
cells (SCLin 0), and group of sclerenchyma cells at 
blade margin not extending on abaxial and adaxial 
epidermis (CAP 1) were invariant and removed 
from the matrix before performing P 


Discrimination among taxonomic entities. DA 
was performed to discriminate relevant groups 
emerging from the second ordination set. А priori 


groups were defined as: (1) the P. tristigmatica com- 
plex (P boelckei and P. tristigmatica); (2) the P. 
pogonantha complex (P pogonantha and P pri- 
chardii); and (3) the P. alopecurus complex (P. al- 
opecurus, P. shuka, and P. superbiens). Characters 
that best reflect specific differences were selected 
following standardized coefficients of canonical var- 


ized by broad glumes, paleas, and blades. The P. 
pogonantha complex is defined by long spikelets in 
association with viviparous florets, and long blades. 
Finally, the Р alopecurus complex has more nodes 
on the panicle along the principal axis, as well as 
more nerves on the first glume. These groups clas- 
sified correctly: 91% were related to the P. tristig- 
matica complex; 8296 to the P. pogonantha com- 
plex; and 96% to the P. alopecurus complex. 


THIRD SET OF MULTIVARIATE ANALYSES 


Specimens of species previously analyzed were 
removed from the data matrix. The 121 remaining 
OTUs were then considered for this subsequent 
study. PCA was performed; the first five compo- 
nents accounted for 48% of total variability. PCA 
distortion was low, r, = 0.91. A group was differ- 
entiated along the I, II, and V axes, including P. 


Annals of the 
Missouri Botanical Garden 


~ 
N 


Poa tristigmatica™ 
complex 


к? 
ви ES 


EM y 


Џ 
\ 


== 


Е... 


| 
Em 


Poa alopecurus complex 


a 


Figure T. 
of specie 
for аи of labels. 


bergii var. chubutensis Speg., P. boecheri Parodi, P. 
hubbardiana Parodi, P. lanuginosa, and P. pata- 
gonica Phil. var. patagonica. According to their 
character loadings, these species have longer 
leaves and inflorescences, wider and thicker 
blades, and larger stomata than the remaining spe- 
cies. 


Poa hubbardiana and the Poa lanuginosa 
Complex. The previously defined group (Роа 
bergii var. chubutensis, P. boecheri, P. hubbardiana, 
P. lanuginosa, and P. patagonica var. patagonica) 
was further analyzed by РСА. Figure 8 is based on 


the first three principal components. Two groups of 


species appear, with sexual dimorphism being the 
main source of variation within these groups. Caes- 
pitose P. hubbardiana separated along the third 
component and placed on its negative edge. This 
species is differentiated because it has a short lig- 
ule, small stomata, and long hairs lying between 
the nerves of the lemma. А P. lanuginosa complex 
(Р. bergii var. chubutensis, P. boecheri, P. lanugi- 
nosa, and P. patagonica var. patagonica) principal- 
ly distributed along the positive edge of axis III. 
Examining their character loadings, this group is 
defined by long ligules and small florets. Axes I 
and II showed a clear dimorphism between pistil- 


Second set of multivariate analyses. Plot of 52 individuals on the first three PCA axes showing two groups 
s: the Poa alopecurus complex (AL, SK, SU) and the P. tristigmatica complex (BK, TR). See Figure 1 caption 


late and staminate specimens of P. hubbariana as 
well as the P. lanuginosa complex. Pistillate plants 
are more robust, with longer spikelets, glumes, and 
florets, with these florets being distinctly hairy. 

e first five components accounted for just 5296 
of total variability. However, the cophenetic corre- 
lation coefficient derived from PCA was high (r, — 
0.86), showing little distortion. 


rimination ng entities. Species 
of the P ae ae complex, P. hubbardiana, and 
the remaining specimens were used as a priori 
groups for DA. The first two canonical variables 
clearly discriminated these groups. The P. lanugi- 
nosa. complex presents long panicles and large sto- 
mata. Poa hubbardiana has long florets, brief lig- 
ules, and sharp blade apices. Classification of the 
groups showed a few mismatches. Nevertheless, 
91% of specimens of the P. lanuginosa comple 
were correctly classified. Meanwhile, 7896 of ex- 
pected specimens were associated with P. hubbar- 
iana 


FOURTH SET OF MULTIVARIATE ANALYSES 


Poa holciformis, Poa huecu, and Poa indiges- 
ta. After discrimination and removal of Poa hub- 


Volume 87, Number 2 
2000 


Giussani 213 
Phenetic Patterns in Poa Sect. Dioicopoa 


, 


Pistillate 


Poa hubbardiana 


igure 8. Third set of multivariate analyses. Plot of 43 individuals on the first three PCA axes: sexual dimorphism 
is clearly defined in Poa hubbardiana (HB) and the P. lanuginosa complex (BO, BU, LA, PA). See Figure 1 caption 


for explanation of labels. 


bardiana and the Р lanuginosa complex, a new 
ordination analysis was performed on the 78 re- 
maining specimens based on 43 morphological var- 
iables. One invariant character was excluded before 
performing PCA: groups of sclerenchyma cells at 
blade margins do not extend on abaxial and adaxial 
epidermis (CAP 1, see Appendix 3). The first five 
PCA axes explained 49% of total variability, with 
PCA distortion being low, r, — 0.91. 

OTU distribution on the first three axes showed 
a group of three caespitose species, P. holciformis 
Presl, P. huecu Parodi, and P. indigesta Parodi (Fig. 
9). Specimens for those species principally asso- 
ciated with the positive end of axis I but the neg- 
ative end of axis III. They are distinguished by their 
relatively long leaves and inflorescence, and rela- 
tively wide, thick blades. Spikelets, glumes, and 
florets are smaller than the same structures on the 
remaining species. Pistillate and staminate florets 
are glabrous. 

Three species were independently analyzed: P. 
holciformis, P. huecu, and P. indigesta. These are 
recognized as single discrete entities on the first 
three РСА axes. Роа indigesta is the largest in size 
among the three taxa, and also has the longest, wid- 
est, and thickest leaves. Poa holciformis is differ- 
entiated from P. huecu by its longer spikelets, 


glumes, and florets, and wider blades. The first five 
components analyzed accounted for 59% of total 
variability, but their distortion was very low, with 
ги = 0 


Discrimination among taxonomic entities. These 
previous three species were considered the a priori 
groups to discriminant analysis. Standardized co- 
efficients of canonical variables revealed Р. indi- 
gesta to present long blades, abundant cuticular 
prickles, and broad panicles. Poa holciformis is 
characterized by long lemmas and ligules. In con- 
trast, P. huecu, at the opposite site, has shorter lem- 
mas and more nerves on the first glume. Classifi- 
cation of specimens based on discriminant 
functions showed a good fit to data. One hundred 
percent of the specimens of a priori groups were 
included in P. huecu and P. indigesta, respectively. 
Eighty-three percent were correctly assigned to P. 
holciformis, but some specimens here associated 


with P. huecu 


FIFTH SET OF MULTIVARIATE ANALYSES 


The Роа denudata and Роа rigidifolia Com- 
plexes. Upon the removal of Р. holciformis, P. 
huecu, and P indigesta, a new PCA involving the 
54 remaining specimens and using 41 morpholog- 


214 


Annals of the 
Missouri Botanical Garden 


Роа holciformis, Р. huecu, P. indigesta 


ure 9. Fourth set of multivariate analyses. Plot of 78 individuals on the first three PCA a 


axes showing Poa 


ler P. huecu, and P. indigesta, clearly separated from the rest of species. See Figure 1 caption for explanation 
of la 


ical variables was conducted. Three characters 
were excluded as invariant for the whole group. Ap- 
ice blades are all navicular (API 1, see Appendix 
3), viviparous florets are absent (FLOv 0), and all 
groups of sclerenchyma cells at blade margins do 
not extend on abaxial and adaxial epidermis (CAP 
1). The first five principal components accounted 
for 50% of variance among specimens, yielding a 
low distortion for the analysis of r, = 0.91 
Distribution of OTUs is represented on the first 
three principal components (Fig. 10). Specimens 
are gathered into two groups. The Poa rigidifolia 
complex (as defined by Giussani et al., con- 
sists of P. dusenii Hack., P. ibari Phil., and P. rig- 
idifolia Steud. The P. denudata complex comprises 
P. denudata Steud. and P. nahuelhuapiensis Nicora. 
The P. rigidifolia complex principally associated 
with the negative end of axis I. This complex is 
distinguished by longer spikelets, glumes, and flo- 
rets than seen in the P. denudata complex. Con- 
versely, P. denudata and P. nahuelhuapiensis have 
longer leaves and inflorescences, wider blades, and 
more sclerenchyma girders on both sides of the vas- 
cular bundles. Minimum spanning tree or MST 
technique confirmed species groupings. This tech- 
nique was performed because two specimens, the 
types of P. dusenii and P. rigidifolia, appeared iso- 
lated from their respective groups. Analysis of min- 
imum distances among these OTUs showed rela- 


tionship of the isolated types to the Р. rigidifolia 
complex (Fig. 10) 

pecimen ordination for only the Р. denudata 
complex, P denudata and Р. nahuelhuapiensis, 
showed a clear dimorphism among pistillate and 
staminate individuals along the first three principal 
components. Pistillate specimens are characterized 
by hairy florets and longer leaves, inflorescences, 
and florets than staminate plants. 


Discrimination among  taxonomic entities. 

Specimens of both the P. rigidifolia and the P. 
denudata complexes were considered as a priori 
groups for DA and were characterized by analysis 
of standardized coefficients. The P. denudata com- 
plex presents long blades, large stomata, and broad 
panicles. Meanwhile, the P rigidifolia complex 
presents long lemmas. Classification percentages 
showed a good fit between groups: 9096 were in- 
cluded in the P. denudata complex, and 94% сог- 
rectly classified into the P. rigidifolia complex. 


UNIVARIATE AND BIVARIATE ANALYSIS 


Previous analyses showed a phenetic pattern 
among dioecious species based on linear relation- 
ships, and it was possible to recognize complexes 
of similar species, as well as independent species. 

Bivariate analysis revealed the correlation be- 
tween different pairs of morphological and anatom- 


Volume 87, Number 2 
2000 


Giussani 215 


Phenetic Patterns in Poa Sect. Dioicopoa 


Figure 10. 


Fifth set of multivariate analyses. Plot of 54 individuals on the first three PCA axes and minimum total 


distances between individuals obtained from the minimum spanning tree represented by lines. The Poa rigidifolia 
complex is represented on black labels (DU, IB, RI), and the P. denudata complex is on gray labels (DE, NA). See 


Figure 1 caption for explanation of labels. 


ical characters. Pairwise combinations were drawn 
for fertile characters (GLUle, GLUw, LEMle, 
LEMw, PALle, PALw, LODle, and LODw) as well 
as paired combinations of characters for relative 
plant sizes (LEAle, BLAle, SHEle, HEIG, PANle, 
PANn, PANw, BLAw, SCL2). All analyses showed 
linear relationships. Fertile characters are contin- 
uously distributed along the whole range of varia- 
tion. In contrast, those characters for plant size pre- 
sent a conspicuous discontinuity between two main 
groups (Fig. 11). One of these main groups contains 
plants of large size including the P. dolichophylla 
complex (P. stuckertii, P. iridifolia, P. pilcomayensis 
var. calamagrostoidea, and P. dolichophylla), the P. 
bergii complex (P. barrosiana and P. bergii var. ber- 
git), the P. bonariensis complex (P. bonariensis, P. 
montevidensis, and P. patagonica var. neuquina), 
and Р. indigesta. Smaller plants distinguish the sec- 
ond group and include all remaining species. 
Another series of pairwise character combination 
showed a non-linear pattern of variation. Four tax- 
onomic groupings are shown in Figure 12 and are 
based on the relation between ligule length and leaf 
length. One group includes species with short lig- 
ules but long leaves; a second species group, by 
contrast, has short ligules and short leaves. A third 
group combines species with both long ligules and 


long leaves. Finally, a fourth group consists of spe- 
cies with long ligules but short leaves. A similar 
grouping pattern was observed from the relation- 
ship between ligule length and any other character 
related to plant size (LEAle, BLAle, SHEle, HEIG, 
PANle, PANn). Another important character com- 
bination involves leaf length and stomata size (Fig. 
13). Here, groups of species assembled according 
to: (1) small stomata but long leaves, (2) large sto- 
mata and long leaves, (3) large stomata but short 
leaves, and (4) short leaves but stomata of medium 
size. 

Univariate analysis considered the average (for 
quantitative variables) or mode (for discrete vari- 
ables) for all complexes and individual species, as 
well as pistillate and staminate specimens, sepa- 
rately. Sexual dimorphism was noted on fertile 
characters related to spikelet, glume, and floret, as 
well as floret hairiness. Table 1 shows a list of se- 
lected characters representing the range of varia- 
tion among complexes and species, as well as sex- 
ual dimorphism within entities. 


DISCUSSION 


Taxonomic problems within Poa were accurately 


expressed by Bor (1952: 7-8), and his words re- 


Annals of the 
Missouri Botanical Garden 


54 
meum mom Se HONO up t арн e m ~ 
-77 e ~ 
A ћ 
49 и“ • e e. \ 
2 I 
/ e , 
44 { ! 
ө e e, 
39 о E 
-~ т оъ ч чу > === - 
LET 75 А\ 
- , 
„ р 
29 ра у 
-47 A , 
„т П 
2 і 
24 Г a ^^ | 
Н А ^ A і 
= я А A! 
Е Gre 
P4 
‘ at 
14 „ aA А A „ 
Га 
24 
9 М А A yen 
у 2 
а Бане 
4 + + ~ 
10 20 30 40 50 60 70 


e Large size д Small size Plant height (cm) 


54 = 
at `~ -2-170 
‘A [А ett о ` 
+ 
aa} | | / „—--77 
` | D 
-~ ` А ) N о "m 
Е ~ vussa? 
= 34 Pub: -—Ó 
t "А ML irc У 
Ё / 4 , ® ' 
2 | / e | 9 
р \ a 
% 24 LE № P4 ө % ө / 
& ЈА у z 4 
$ А ~ ^^ e. / 
= ~ ® 2 7 
Sa А, ice Pd • „= 
~ — 
14 У А `i a us 
^_ of | + 
`o..." 19 “7 
аи 
4 
2 10 12 
< > < ETAT 
АСЗ E = а length (mm) 
О >LIG/>LEAF >LIH/<LE AF 
55 
777 7« 
50 e^ И аи ~ 
/ ) » 
Ir J pd а Е \ 
45 Џ T Ра m у 
4 ГА 
/ / 
40 : и и п 2^ 
= Ц vut 
g 35 Е -7 
јат 
-— ~ 
30 " à о 
{ i А ~ кее ы ад ; 
Џ ~ 2 
25 A n 2 AA' „7 e e, 
Н | a А | P d i 
= 20 i А А П 4 e ' 
‘А А АЈ ni 
7] 4 
15 N (e? Fu e ө, 
2 ~ Јо - 
10 Э А P2 Ы === 
ead 24 
5 
0,037 0,042 0,047 0,052 


0,032 
Ф => > Stom ata length (mm) 
@>STOM/<LEAF A medium STOM/<LEAF 
hol | (top). Linear corre- 


Figures 11-13.  Bivariate analyses showing different patterns of morphological variation. —1 
— 12 (middle). Non-linear correlation pattern: ligule length vs. leaf length 
gth. 


ns pattern: plant height vs. leaf length. 
(bottom). Non-linear correlation pattern: stomata length vs. leaf len 


Volume 87, Number 2 
2000 


Giussani 217 
Phenetic Patterns in Poa Sect. Dioicopoa 


main relevant, as emphasized by more recent au- 


thors (Vickery, 1970; Soreng, 1990): 


“The systematic treatment of the species ... is one 
of the most bewildering and difficult of taxonomic stud- 
е гес- 


separate species in such groups, but Е of 
more or less variable ит must be и 


This study of Poa sect. Dioicopoa utilized mul- 
tivariate taxonomic techniques, as well as univari- 
ate and bivariate analyses, to better understand the 
morphological variation pattern—a different meth- 
odology from traditional taxonomic treatments. 
Analyses of both pistillate and staminate specimens 
denote complexes of species of great similarity, but 
it was also possible to clearly recognize some valid 
species. A multivariate key for identification among 
taxonomic entities of Poa sect. Dioicopoa was pro- 
duced (see key herein). 

Identification of varieties for three species ana- 
lyzed here could not be supported. Thus, Poa bergii 
and P. bergii var. chubutensis, P. patagonica and P. 
patagonica var. neuquina, and P. pilcomayensis and 
P. pilcomayensis var. calamagrostoidea did not 
group together by similarity and were either in- 
cluded in different species complexes or main- 
tained as independent entities. Poa bergii was 
grouped with P. barrosiana in a P. bergii complex, 
while its variety P. bergii var. chubutensis associated 
differently within the P. lanuginosa complex. Poa 
patagonica was more similar to species of the P. 
lanuginosa complex than to its variety P. patagon- 
ica var. neuquina, which clustered within the P. 
bonariensis complex. Finally, P. pilcomayensis was 
recognized as independent, while its variety P. pil- 
comayensis var. calamagrostoidea related to a P. 
dolichophylla complex. 

Poa schizantha, a particular case within Poa 
sect. Dioicopoa, has been found only twice, and 
appears restricted to the dunes of Monte Hermoso, 
Argentina (Parodi, 1940). It is recognized by pre- 
sent results as a single species, related to the P. 
bergii complex. The three species (P. barrosiana, P. 
bergii, and P. schizantha) were found in similar en- 
vironments and geographic areas. However, P. schi- 
zantha is clearly distinguished by a particular bi- 
lobed lemma, 
different anatomical blade structure; among other 
characters, it lacks а well-developed midrib and 
bulliform cells. These features could originate by 
hybridization or odd mutation. Possibly, its off- 


an interrupted panicle, and a 


spring had low fertility ы жуа the case, it 
has never been collected a 

Plant stature of less сена versus greater than 50 
cm was a significant criterion by which to discrim- 
inate species within Poa sect. Dioicopoa. Plant 
height correlates with characters such as leaf blade 
and sheath length, panicle length, and number of 
nodes per panicle (Fig. 11). Other characters, of no 
linear variation, also address species discontinu- 
ities. Ligule length divides Poa sect. Dioicopoa into 
those species with short ligules (less than 2(-3) 
mm) and those with long ligules (more than (3—)4 
mm). 

Plant habit did not show, despite traditional 
treatments (Parodi, 1936; Torres, 1969, 1970; Ni- 
cora, 1978), a clear pattern of variation, nor any 
character correlation that could clarify the taxo- 
nomic structure within Poa sect. Dioicopoa. Some 
species could be differentiated only by the pres- 
ence or absence of a rhizome. For example, species 
of the P. resinulosa complex, P. jr edm and P. 
resinulosa, are caespitose, while uchtienii and 

> calchaquiensis are rhizomatous. Parodi (1936) 
suggested that the rhizome is systematically valu- 
able, dividing Poa sect. Dioicopoa by rhizomatous 
and caespitose habits. Later, Hunziker (1978) o 
served that crossings of rhizomatous and и M 
species of Poa produced fertile offspring, although 
the pattern of inheritance depended on the species 
that were crossed. Therefore, it remains preferable 
to group otherwise similar rhizomatous and caes- 
pitose species in the same taxonomic complex. 

Sexual dimorphism was the principal source of 
variation within species and taxonomic complexes 
of Poa sect. Dioicopoa. Thus, it was necessary to 
establish ranges of variation for reproductive char- 
acters in pistillate and staminate specimens sepa- 
rately. Sizes of spikelets, glumes, and florets, as 
well as hairiness of florets, indicated major differ- 
ences between pistillate and staminate e plante Tra- 
ditionally, hairiness of the pistillate was 
ae a useful И character (Torres, 

1970; Nicora, 1978). Since staminate florets are 
usually glabrous, it was not possible to investigate 
interspecific limits based on similar characters from 
staminate specimens. This study showed a great 
variation in hairiness of pistillate florets within, and 
among, analyzed groups. Therefore, these charac- 
ters are less informative for interspecific variability. 
Callus hairiness on pistillate florets could be a syn- 


variation. The absence of hairs on pistillate florets 
occurs only in P. huecu, P. holciformis, and Р. 
digesta. Both pistillate and staminate саа іп 


218 


Annals of the 
Missouri Botanical Garden 


Table 1. 


Morphological variation of species of Poa sect. Dioicopoa characterized by vegetative and reproductive 
characters. Median and standard deviation between par 
ma 


entheses for quantitative characters. Mode, and minimum and 


m values are between parentheses for variable discrete characters. Pi corresponds to number of pistillate 
specimens analyzed per species; St to number of staminate specimens. Character abbreviations as in Appendix 3. 


Leaf Ligule Plant Panicle _ Nodes/ Blade Stomata 
length length height length Panicle width length 
Taxa (LEAle) (LIGle) (HEIG) (РАМе) — (PANn) (BLAw) (STOM) 
Poa alopecurus 24 6 34 7.2 10 1.3 .042 
(Pi = 22, St = 8) (+ 9.86) (+ 2.2 (+ 16. ш (+ 2.28) (6-15) (+ 0.35) (= 0.0055) 
P. bergii 48.3 10.4 56. 15.9 15 1.3 0.049 
(Pi = 16, St = 11) (+ 14.92) (+ 3.48) =. | E (= 5.1) (11-20) (+ 0.44) (= 0.0051) 
P. bonariensis 42.7 1.1 А 16.3 13 1.3 0.047 
i= 14, St = 11) (+ 13.42) (+061) (+ p (+ 4.13) (10-18) (+ 0.29) (= 0.0044) 
P. denudata 16.7 24 28.9 6.5 11 0.8 041 
Pi = 8, St = 8) (+ 7.00 (+ 1.61) (+ 6.29) (+ 1.81) (9-14) (+ 0.18) (= 0.055) 
P. dolichophylla complex 49.2 1.2 69.3 20.4 17 3.4 0.034 
(Pi = 23, St = 18) (+ 18.57) (+ 0.66) (+ 19.56) (+ 5.85) (11-20) (+ 0.98) (= 0.0028) 
P. holciformis 16.2 5.7 9.2 1.6 0 1.3 0.04. 
(Pi = 8, St = 4) (+ 7.68) (+ 2.35) (+ 1431) (+ 3.3) (7-14) (+ 0.28) (= 0.0044) 
P. hubbardiana 30.9 1.1 37.3 9.1 11 0.036 
(Pi = 4, St = 5) (+ 11.62) (+ 0.41) (+ 13.51) (+ 2.57) (8-13) (= 0.21) (+ 0.0051) 
P. huecu 18.8 5 34.5 8.7 12 1.1 0.040 
(Pi = 6, St = 3) (+ 8.32) (+ 2.2) (+ 11.94 (+ 1.02) (10-16) (+ 0.17) (= 0.0049) 
P. та гема 40.8 7.9 57.6 18.5 15 1.5 0.048 
(Pi = 2, St = 1) (33.5—48.0) (6.3-8.7) (40—71.7) (11.5-29) (15-20) (1.11-1.76) (0.042-0.046) 
P. lanigera 24.3 0.7 36.2 9.2 13 1.5 0.040 
і = 5, 5) (= 10.56) (= 0.28) (+ 1348) (+ 2.87) (10-18) (= 0.46) (= 0.0039) 
Р. Іапиріпоѕа 25 7.8 38 10.3 13 0.9 0.048 
(Pi = 20, St = 14) (+ 10.21) (+ 3.19) (+ 15.44) (+ 4.11) (9-18) (+ 0.22) (= 0.0048) 
P. ligular 22.6 7.4 29.8 7.3 12 0.7 0.038 
(Pi = 12, St = 9) (+ 14.78) (+ 3.01) (+ 12.34) (+ 3.22) (7-20) (+ 0.31) (+ 0.0037) 
P. pilcomayensis 21.3 0.8 33.3 9.3 8 0.037 
(Pi = 5, St = 5) (+ 8.25) (+ 0.41) (+ 10.43) (+ 4.26) (8-14) (+ 0.31) (+ 0.003) 
P. pogonantha 16.5 2 37.7 7 8 0.043 
(Pi = 14, St = 9) (+ 6.58) (+ 1.08) (+ 16.37) (+ 2.32 (6-12) (= 0.30) (+ 0.0049) 
P. resinulosa complex 18.8 1.4 32.3 7.9 12 .79 0.036 
Pi = 19, ) (+ 12.39) (+ 1.01) (+ 17.71) (+ 4.23) (8-16) (+ 0.22) (+ 0.0042) 
P. rigidifolia 9.7 4.7 17.2 4.3 9 7 0.037 
i = 23, 15) (+ 6.17) (+ 1.95) (+ 868) (+ 1.79) (6-14) (= 0.17) (+ 0.0056) 
P. schizantha 8.1 7.3 38.7 20.5 13 8 0.038 
(Pi = 5, St = 1) (+ 6.45) (= 2.31) (= 6.94) (+ 6.57) (9-13) (= 0.26) (+ 0.0028) 
P. tristigmatica 4 3.6 26 6.5 0 1.6 048 
(Pi = 14, St = 9) (+ 5.57) (+ 2.15) (+ 10.65) (+ 1.78) (7-12) (+ 0.38) (+ 0.0048) 


these three species are completely glabrous. This 
absence of hairs in these taxa could represent a 
loss with respect to the evolution of the entire ge- 
nus, as suggested by Kellogg (1990) for other spe- 
cies of Poa. Pistillate anthoecia of P. hubbardiana 
are remarkably hairy, and clearly differentiated 
from other species. Its woolly hairs on the floret 
callus are numerous, long, and folded, while hairs 
on and between palea and lemma nerves are curled, 
and twice or three times longer than those seen in 


other species. 


Out of the 34 species analyzed, only 8 were dis- 


criminated by numerical taxonomic methods: Poa 
holciformis, P. hubbardiana, P. huecu, P. indigesta, 
P. lanigera, P. ligularis, P. pilcomayensis, and P. 
schizantha. The remaining species showed congru- 
ent morphologies, aggregating into species com- 
plexes by phenetic similarities. Any hierarchical 
assignment to taxonomic varieties was not consis- 
tent with current species alignments. The validity 
of species assorting into complexes remains contro- 
versial. For classification purposes, a species com- 
prises a taxonomic grouping of pistillate and sta- 
minate specimens of sympatric populations, sharing 


Volume 87, Number 2 


iussani 
Phenetic Patterns in Poa Sect. Dioicopoa 


2000 
Table 1. Extended. 
Glume length Lemma length Hairs on callus Hairs on nerves 
(GLUle) (LEMle) (HAlIcal) (HAInrv) 
Pistillate Staminate Pistillate Staminate Pistillate | Staminate Pistillate ^ Staminate 
6.8 6 7.5 T 4 4 2 l 
(+ 1.16) (+ 1.27) (+ 0.98) (+ 1.42) (0—4) (0—4) (0—3) (0—3) 
5 8 6 4 0 3 0 
(+ 1.25) (+1.00) (+ 0.99) (+ 0.95) (04) (0-3) (0-3) (0-1) 
3 3.2 5.3 4 4 4 3 
(+ 0.62) (+ 0.48) (+ 0.74) (+ 0.25) (0-4) (2-3) (0-2) 
4.3 6 3 .5 4 0 2 0 
(+ 0.73) (+ 0.39) (+ 0.48) (+ 0.33) (0-4) (0-4) (0-2) 
E! 2.9 4.1 6 4 0 2 0 
(+ 0.72) (+ 0.78) (+ 0.68) (+ 0.53) (0-4) (0-4) (2-3) (0-1) 
4.7 "i 5.4 4.7 0 0 0 
(+ 0.69) (+ 0.72) (+ 0.77) (+ 0.86) 
5.8 4.2 6.6 5 4 4 3 0 
(+ 1.31) (+ 0.51) (+ 1.35) (+ 0.23) (0-4) (0-3) 
4 2.8 И | l 0 0 0 
(+ 0.67) (+ 0.70) (+ 0.63) (+ 0.68) 
3.7 2.5 4.5 3 0 0 0 0 
„2 ‚2 B .8 4 0 3 
(+ 0.37) (+ 0.43) (+ 0.53) (+ 0.42) (0—4) (2—3) (0-2) 
1 3.8 5.8 4.7 4 4 3 0 
(+ 1.03) (+ 0.71) (+ 0.99) (+ 0.61) (0-4) (0-4) (2-3) (0-2) 
6 2.7 4.8 3.4 4 0 0 
(+ 0.67) (+ 0.35) (+ 0.83) (+ 0.24) (0-4) (2-3) 
9 2.5 4.1 3.4 4 4 0 
(+ 0.37) (+ 0.51) (+ 0.51) (+ 0.46) (0-4) (0-1) 
5.6 7 2 0 2 0 
(+0.79) (+ 1.28) (+ 1.12) (+ 0.59) (0-4) (0-3) (1-2) (0-2) 
3 A 3.3 4 0 0 
(+ 0.71) (+ 0.42) (+ 0.79) (+ 0.43) (0—4) (0—4) (0—3) (0—2) 
5.1 9 9 5.6 4 0 0 
(+ 0.95) (+ 0.82) (+ 0.79) (+ 0.85) (0-4) (0-3) (2-3) (0-2) 
5.7 4.1 6.6 2 0 0 0 
(+1.32) (+ 1.51) (0-2) 
5 4.5 6.7 5.4 2 0 2 0 
(+ 0.86) (+ 0.69) (+ 0.97) (+ 0.83) (0-4) (0-4) (1-2) (0-1) 


similar variation pattern for several characters, not 
a single one. In general, morphological variation 
presents clear discontinuities identifying species. 


Moreover, the predominant variation pattern of a 


species is clearly associated with sexual dimor- 


phism. Morphological variation within only pistil- 


late plants or staminate specimens, correlated with 


environmental factors, appears significant only at 
the infraspecific level. By these criteria, sympatric 
species of Poa sect. Dioicopoa within the same tax- 
onomic complex are synonymized herein. One new 
variety of P. rigidifolia is further proposed based 
on sexual dimorphism associated with environmen- 


tal discontinuities. Precise ranges of morphological 
and anatomical variation are established herein and 
also correspond to Giussani (1997). 


TAXONOMIC TREATMENT 


Poa alopecurus (Gaudich. ex Mirb.) Kunth, Re- 


vis. Gramin. 1: 116. 1829. Arundo alopecurus 
Gaudich. ex Mirb., Ann. Sci. Nat., Bot. 5: 100: 
1825. TYPE: Falkland Islands, East Falkland: 
Port Louis, 14 Feb.-28 Apr. 1820, С. Gaudi- 
chaud s.n. (holotype, P not seen, isotype, US 
78849 [fragment ex P] not seen). 


220 


Annals of the 
Missouri Botanical Garden 


Poa shuka Speg) Parodi, Revista Argent. Agron. 20(4): 
180. 1953. Festuca shuka 5рев.. Anales Mus. Nac. 
His te Buenos Aires 5: 95. 1896. TYPE: Argen- 
ae Tierra del Fuego е Is. Ж yee Sur: Us- 
huaia, Isla de los Estados, Port Vancouver, Дека 

ау, Spegazzini s.n. (holotype, LPS 14322 
een; 18 icky pe. LP. 

Poa ара Po (Steud.) Hauman & Parodi, Physis (Buenos 
Aires) 9: 344. 1929. Aira superbiens Steud., Syn. Pl. 
uae 1: | ) i 
Sandy Point, Dec., W. Lechler 1194 (holotype, P not 
seen; isotype, BAA fragment!, US 2695872 ex Hb. 
Cosson P not seen, US 76311 ex W not seen). 


This species inhabits the southern portion of Pa- 
tagonia, covering western and southern regions of 
Santa Cruz, western areas of Tierra del Fuego e 
Islas del Atlántico Sur, Argentina, as well as 
boundary zones of XII Región, Chile. 


Poa bergii Hieron., Bol. Acad. Nac. Ci. 3: 374. 
1879. TYPE: Argentina. Río Negro: boca de 
Río Negro, C. Berg 205 (holotype, CORD!). 


Poa barrosiana Parodi, Physis (Buenos Aires) 11: 134. 

32. E: Argentina. Buenos Aires: Miramar, 31 

Jan. 1930, L. R. Parodi 9820 (holotype, BAA!; iso- 
type, US 89694. not seen). 


Poa bergii inhabits costal dunes of Buenos Aires 
and Río Negro provinces, Argentina. It may extend 
to similar habitats in Uruguay and Brazil. 


Poa bonariensis (Lam.) Kunth, Revis. Gramin. 1: 

15. 1829. Festuca bonariensis Lam., Tabl. 

Encycl. 1: 192. 1791. TYPE: Argentina. E. 

Bonaria Circa Monte-Video, inter rupes et 

maritimas, 1767, E. Commerson s.n. (holotype, 

P not seen; US 2875384 ex P not seen; iso- 
type, BAA fragment!). 


Poa pnr Arechav., Anales Mus. Nac. Montevi- 
o 1: 479. 1897. чы Uruguay. Montevideo: en 
а hümedos ‚ Arechavaleta 5101 (lecto- 
type, designated by Parodi (1936), MVM not seen; 

isolectotype, fragment LP!, type photo, LP!). 
Poa patagonica Phil. var. neuquina Nicora, e 
1(18): 107. 1977. TYPE: Argentina. Neuquén: De- 
partamento Lácar, San Martín de los Andes, fi Ruiz 

Leal 20315 (holotype, BAA!). 


Distributed in the northeastern Buenos Aires 
province, this species reaches its southern limit at 
the system of Tandilia and Sierras Australes. It ex- 
tends to the north and east, into southern Entre 
Ríos, Uruguay, and Brazil. One of its synonyms, P. 
patagonica var. neuquina, was found in a Ciprés 
forest near San Martín de Los Andes, Neuquén, 
disjunct from its main area of distribution. 


Poa denudata Steud., Syn. Pl. Glumac. 1: 259. 
1854. TYPE: Chile. Valdivia, W. Lechler 578 
(holotype, LE not seen; pistillate fragment, 
BAA!; isotype, US not seen). 


Poa nahuelhuapiensis Nicora, Hickenia 1(18): 106. 1977. 
TYPE: Argentina. Neuquén: Departamento Los La- 
gos, Península Quetrihué, 1 Nov. 1949, О. Boelcke 
& J. H. Hunziker 3458 (holotype, BAA!). 


Poa denudata grows in western Neuquén, Rfo 
Negro, and Chubut, Argentina, and along the Chi- 
lean boundary in Regions VIII to X (Toledo & Za- 
pater, 1991), near lake margins, among rocks, or in 
grasslands below elevations of 1 


Poa lanuginosa Poir., Encycl. 5: 91. 1804. TYPE: 
Uruguay. Montevideo: E. Commerson s.n. (ho- 
lotype, P not seen; fragment BAA!, US 88769 
fragment ex P not seen). 


Poa bergii Hieron. var. chubutensis Speg., Revista Fac. 
Agron. Univ. Nac. La E un 3: 628. 1897. d 
Argentina. Chubut: artamento Florentin 
Ameghino, Cabo Raso, C. Spegazzini 938 (DE. 
LP not seen; fragment BAA 

Poa boecheri Parodi, Revista 7 Артоп. 28: 100. 
1961 [1962]. TYPE: Argentina. Mendoza: Departa- 
ved San Rafael, Valle del Atuel, El Sosneado, 35? 

. 5, 4 Oct. 1955, T. W. Bücher, J. P. Hjerting & K. 
Rahn 801 (holotype, BAA!; isotype, C not seen). 
Poa patagouea Phil., Anales Univ. Chile 94: 168. 1896. 
Chile. pe Ultima Esperanza, Lago 
Pinto, m dic. 1877, H. Баг s.n. (holotype, SGO not 
seen; me BAA!, US 88748 fragment ex SGO 
en). 


not s 


= 


Роа lanuginosa is a widespread species, distrib- 
uted in Argentina from 35°S to the steppes in Tierra 
del Fuego. It grows in highly degraded soils, and 
on dunes and sandy soils in Patagonia. 


Poa pogonantha (Franch.) Parodi, Revista Ar- 
gent. Agron. 20: 180. 1953. Festuca pogon- 
antha Franch., Miss. Sci. Cape Horn, Bot. 5: 
87. 1889. TYPE: Argentina. Patagonia, Port 
Eden, 24 Jan. 1879, Savatier 1844 (holotype, 
P not seen, fragment ex P, BAA!; isotype, US 
s.n. fragment and photo ex P not seen). 


Poa prichardii Rendle, J. Bot. 42: = 1904. ТҮРЕ: Аг- 
gentina. зе Cruz: Lago Argentino, Monte Buenos 
Aires, 1900-1, H. Prichard s.n. (holotype, BM not 
seen; ae ВАА!). 


This species occurs in humid forests of Río Ne- 
gro, Chubut, Santa Cruz, and Tierra del Fuego, Ar- 
gentina (Nicora, 1978), and in the XII Region of 
Chile (Toledo & Zapater, 1991). This area is in- 
cluded within the Subantarctic phytogeographic 
province (Cabrera & Willink, 1973; Cabrera, 
1994) 


Volume 87, Number 2 
2000 


Giussani 221 


Phenetic Patterns in Poa Sect. Dioicopoa 


Poa rigidifolia Steud. var. rigidifolia, Syn. Pl. 
Glumac. 1: 260. 1854. TYPE: Argentina. Ti- 
erra del Fuego e Is. del Atlántico Sur: Islas 
Malvinas, Is. Soledad, Pto. William, Sep. 
1850, W. Lechler s.n. (holotype, P not seen; 
fragment BAA!, US 88734 fragment not seen). 


Due to the results obtained in this research, the 
P. rigidifolia complex 1s considered a single taxo- 
nomic species. Two varieties are proposed in agree- 
ment with the previous study of Giussani et al. 
(1996). Pistillate individuals of each variety showed 
a different pattern of variation to that present in 
staminate specimens, although correlation between 
morphology and environment was high for both pis- 
tillate and staminate plants. Poa rigidifolia var. rig- 
idifolia includes P. spiciformis (Steud.) Hauman & 
Parodi (= P. poecila Phil., Giussani, 1993), previ- 
ously synonymized by Giussani and Collantes 
(1997). It is distinguished by the woolly hairs on 
the floret callus but an absence of hairs between 
the palea and lemma nerves on pistillate florets, 
thinner blades, longer sheaths, ligules, and blades, 
and smaller stomata than pistillate specimens of Р. 
rigidifolia var. ibari. 


Poa rigidifolia var. ibari (Phil.) Giussani, comb. 
et stat. nov. Basionym: Poa ibari Phil., Anales 
Univ. Chile 94: 170. 1896. TYPE: Chile. De- 
partamento Ultima Esperanza, Lago Pinto, Jan. 
1844, H. Ibar s.n. (holotype, SGO not seen; 
isotype, BAA!) 


Poa dusenii Hack., in Dusén, Ark. Bot. 7(2): 8. 1908. 
TYPE: Argentina. Santa Cruz: Mazaredo portum, 
47°41'S, Jan. 1905, Dusén 5318 (holotype, W not 
seen; fragment BAA!; isotypes, US 89702 not seen, 
US 1161178 not seen). 


Poa rigidifolia var. ibari is characterized by not 
having woolly callus hairs on the floret callus but 
having them on, and between, principal palea and 
lemma nerves of pistillate florets. 

Further treatment of morphological and environ- 
mental variation within P. rigidifolia and a discus- 
sion of character assessment can be found in Gius- 
sani and Collantes (1997). Poa rigidifolia inhabits 
steppes of Chubut, Santa Cruz, and Tierra del Fue- 
go, and similar environments in neighboring Chile. 
Poa rigidifolia var. rigidifolia is present in sub- 
humid austral areas, whereas P. rigidifolia var. ibari 
grows in more xeric environments (Giussani et al., 


1996; Giussani & Collantes, 1997). 


Poa tristigmatica E. Desv., in Gay, Fl. Chil. 6: 
419. 1853. SYNTYPES: Chile. Magallanes, 
Cordillera de Talcarengue, Feb. 1831, Gay 49 
(P not seen; fragment, BAA!, US 88717 frag- 
ment not seen); Bahía Duclos, Estrecho de Ma- 
gallanes, Commerson s.n. (P not seen). 


Poa boelckei Nicora, qp Bes Е bei TYPE: 
Argentina. Neuqué ar, Chapelco, 
encima del e apis "1800-1870 n m, т Fe 1. 1974, М. 
N. Correa et al. 5926 (holotype, ВАВ!). 


Poa tristigmatica is found in Chile and in Ar- 
gentina from Mendoza to Tierra del Fuego. Its dis- 
tributional area falls within the Altoandina phyto- 
geographic province (Cabrera & Willink, 1973; 
Cabrera, 1994), ranging from 1600 to 2000 m in 
Neuquén, and at 500 m, or higher, at the highest- 
latitude areas of Tierra del Fuego. Poa boelckei was 
collected only in Cerro Chapelco at approximately 


For the remaining P. dolichophylla and P. resi- 
nulosa complexes, species show allopatric distri- 
bution, and a study of population variability and 
their ecological relationship is required to resolve 
taxonomic problems. In addition, types of older 
named taxa, particularly those of Philippi, need to 
be studied. For this reason, synonymy has not yet 
been established. 

However, the P. dolichophylla complex is well 
segregated from other species within Poa sect. 
Dioicopoa. Species distributions are clearly distinct 
biogeographically. Poa dolichophylla and P. pilco- 
mayensis var. calamagrostoidea inhabit Sistema de 
Cumbres Calchaqufes-Aconquija-Famatina іп 
northwestern Argentina (González Bonorino, 1958). 
Poa stuckertii is restricted to central Argentina, on 
hills of Córdoba and San Luis, reaching 1500 m. 
This taxon is associated with lowland humid soils. 
Poa iridifolia occurs in the Sierras Australes and 
Tandilia, in Buenos Aires, Argentina (González 
Bonorino, 1958). 

Species of the Р. resinulosa complex are mor- 
phologically similar but inhabit disjunct areas. Poa 
resinulosa grows in central Argentina, between 30? 
and 45°S, in steppes, on stony, dry soils (Nicora, 
1978), and in the Sierras de Tandil and Ventana, 
in Buenos Aires. Poa pedersenii was only collected 
in Yhu, Paraguay. Poa calchaquiensis grows in the 
Argentinian Puna (the southern extreme of the Pe- 
ruvian-Bolivian Altiplano) in Jujuy, Salta, Tucu- 
mán, and La Rioja and could also be spread over 
Catamarca. Poa buchtienii is distributed on the AI- 
tiplano and the Cordillera Oriental of Bolivia. 

Morphological variation of both complexes must 
be addressed by populational and biological studies 


222 


Annals of the 
Missouri Botanical Garden 


to better resolve the degree of similarities and tax- 
onomic boundaries among these species complexes 
within Poa sect. Dioicopoa. 


MULTIVARIATE KEY: CLASSIFICATION OF THE DIOECIOUS 
SPECIES OF ARGENTINA 


ры ен (ог taxa were selected according 
sults i 


a 
suites of correlated characters, increasing the accuracy of 
identification 
Thus, this а није key is not a conventional key. It 
e 


d maximum values indicate the largest range ob- 


ve 

То identify dioecious species of Poa, it is desirable to 
have in hand both pistillate as and staminate (St) spec- 
imens of the population examin hen only a pistillate 
specimen is available, сесије species from Poa 
subg. Andinae Nicora cannot be excluded. 


Бам 


1. Plants 45-80 ст tall (64 (31–136)); leaves 


40 cm long (28.2 (8—61)); = 
em long (18.4 (5.5-39)); ae 13-25 
em long (18.4 (6—37)); nodes per panicle, 
13-18 (16 (10–20)) __________________ 
15-55 ст tall (32.4 (4.3-75)); 
leaves 5-35 em long (20.5 (3.5—54.5)); 
blades 5-20 ст lo (12.8 (1.3-37)); 
sheaths 4—12 cm long (7.6 (1.7-22)); pan- 
icles 5-13 ст long (8.6 (1.7-29)); nodes 
per panicle, 9-13 (11 vole 0)) _________ 
Ligules truncate to obtuse, 0.6-2 mm long 
(1.2 (0.4—4.5)); panicle 2 5-6 ст wide (4 
©. 8–10)); abaxial sclerenchyma girders, 1— 
3 (5) 


a 


2(1). 


2'.  Ligules acuminate, 6-12 mm long (6.6 
(4.5-19.6)); panicle 1.5-3 cm wide (2.5 
(1-4.5)); abaxial sclerenchyma girders: 3-7 


3(2). d caespitosa, iridaceous, laterally com- 
essed with coriaceous and wide 
an panicles. at maturity. Leaf blades flat 


and carinate, mm wide (6.6 (1.8— 


13)). Glume I, 1-nerved; pistillate (Pi) 
and staminate (St) glu 2.5-4 m 
pss 0 < тт wide 


mas 3-4.5 mm long (4 5-5). 0. 7-0.9 
mm wide (0.8 (0.6—1.1)). EA and sta- 
minate paleas 2.5-3.5 mm long (3 (2- 
3.8), 0.45-0.65 mm wide (0.5 (0.35— 
0.8)). Keel and marginal nerve hairs of 
pistillate florets, when present, less than 0.5 
mm long. Sclerenchyma girders at both sides 


of each vascular bundle: 15-35 (23 (6—45)); 


= 
~ 
~ 
м 


5(4). 


5'. 


sclerenchyma cell groups at blade margin, 
extending on abaxial and adaxial epidermis. 


Stomata infrequent on abaxial epide 
0.03-0.037 mm pus (0.034 (0.027— 
0.042) _____ a dolichophylla complex 


Plants d not t laterally y compressed, 
with herbaceous 


8 mm wide 
; quss Pi: 


(4 (3.7—4.5)); Pi and St, 0.85-1.1 mm 
wide (1 (0.8—1.3)). Paleas, Pi: 3.4—4.2 mm 
long (3.8 (3-4.6)), St: P 6 mm long 
(3.4 (3—4)); Pi T St: 0.7 mm wide 
(0.65 (0.5-0.9)). Keel he ен! чи пегуе 
hairs of pistillate e more than 0.5 mm 
long. Sclerenchyma girders at both sides of 
each vascular bundle. 6-12 (9 (5—15)); scle- 
renchyma cell groups at blade margin, not 
extending on abaxial and adaxial epidermis. 
Stomata infrequent on adaxial epidermis. 
Stomata frequent on adaxial epidermis, 


0.042-0.050 mm long (0.047 (0. 037- 


0.054)) Poa bonariensis 


Plants with long and deep rhizomes. Pistil- 
late and staminate spikelets 5- to 6-fl 


-9 mm long ion (3.6- 


145 mm wide (1 (0.6-1.7)). Lem- 


5.9 mm pedea (4.2 (3-6. "yy I 
0.8-1.3 mm wide (0.96 (0.5-1.8)) ............- 
Plants caespitous. Pistillate and staminate 
spikelets 3" flowered; Pi: 4.5-6 
4 mm); Pi and § 
G ae I, Served, Pi 3.5-4 mm pen 
St: 0.7-0.8 bini 
(St: гы тт desi уна Pi: ра m 
long, St: 3 mm long; Pi and St: 0.9-1. 1 


m wide. Restric ted to Zapala, Neu- 

én, а — - 
Pistillate and staminate spikelets 5.5-8.: 
mm wide 


(0.67—1.23)), St: 0.7-0.9 mm wide (0.82 
(0.6—1.15)). Lemmas, Pi: 5-7 mm long (5.8 
(4.2-7.5)), St: 4–5.5 mm long (4.7 (3.9- 
5.8)); Pi and St: 0.9-1.2 mm wide (1.2 
(0.7-2.2)). Paleas, Pi: 4-5 mm long (4.2 
(3.3-5.1)), St: 3-4 mm long (3.6 (3–5.2)); 
Pi and St: 0.8—0.9 mm wide (0.83 (0.5— 
1.4)). Plants of sandy soils of semiarid Pa- 
tagonia and Cuyo, Argentina " 
Pistillate and staminate spikelets 8-1 1 mm 


Poa indigesta 


Poa lanuginosa 


Volume 87, Number 2 
000 


Giussani 
Phenetic Patterns in Poa Sect. Dioicopoa 


223 


long (9 (6-12)), 3.5-8 mm wide (6 (3-10)). 
Glume I, 3-5 (7)-nerved, Pi: 6.5-9 mm 


(1.1 (0.7-1.8)), St: 0.6-1.1 mm wide (0.9 
(0.6—1.7)). esa of coastal dunes of Buenos 
Aires, Агре oa bergii 

601).  Ligules ac Pam 3-8 mm long (5.7 (1– 
16)) 


6'. Ligules truncate to obtuse, 0.5-2 mm long 
(1.5 (0.2—5.8)). [Only P. denudata and P. po- 
ое с a present lig- 
ules 2 to mm long |. one in 


- 
~ 
D 
EE 
f 
= 
n 
а Б 
B" de 
S 
5 
oe 
"m 
= 
2 
Bur 
Ф 
Ф 
"о 
ч 
=й 
N 
e 
3 
Ф 
Ф 
= 
р 
=. 
Oy 
Ф 
„л 


ст long (20.5 (13-29)). Leaf blades sub- 
convolute; 0.14—0.21 mm thick (0.17 
9. 12-0. 24). Bulliform cells not differenti- 


| ue ма n Hermoso and Bahía Blanca, 
Arge Poa schizantha 
T. Plants caespitous or long-rhizomatous. Pan- 


РА 


0.25 


© 
3 
а. 
= 
БРЕ 
= 

= 8 

T 

Е) 
B 
= 
E. 
6 
| 


Шо, уюы Клен. по! ей pis- 


brous or scarcely pubescent 


Be 
КӘ] 
az 
р 
= 
т 
~ 
w 
e 
un 
B, 
5 
= 
n 
= 
= 
[e] 
m 
5: 
м 
= 
=. 


wide (0.7 (0.4—1.1)). Lemmas, Pi and St: 
3. mm long (4.4 (3—6.4)); Pi and St: 
0.8-1.1 mm wide (1 (0.6-1.3)). Paleas, Pi 
and St: 2.3-4 mm long (3.3 (1.84. e Pi 
and St: 0.55-0.8 mm wide (0.65 (0.4— 
0.9 


8. Plants caespitous or long-rhizomatous. 


nd 
St: 0.7-1.2 mm wid (0. 9 (0.5—1.4)). Lem- 
mas, Pi and St: 5-7 mm long (6 (4—8.5)); 
Pi and St: 1-1. тт .2 (0.7-2.2)). 
Paleas, Pi and St: 3-6 mm long (4.5 (2.6– 
6.7)); Pi and St: 0.7-1.2 mm wide (0.8 


(0.5-1.5)) 10 


ae 
59 


gules 3-6 mm long (5 (3—9.3)). Pistillate 
and staminate florets glabrous. Steppes on 
plains and mountains from San Juan to Neu- 


9". 


10(8). 


1110 


1200). 


— 


quén, between 1000 and 3000 m, Argentina 


tagonia and central regions of Argentina — 


Leaf blades 2-4 mm wide (2.8 (1.6—5)). 
Florets frequently viviparous. Glume I, Pi: 

—8 mm long (6.2 (3.7—9)), St: 4.5-6.5 
mm long (5.2 (3.6–8)). Lemmas, Pi: 6-8 
mm long (7.1 (5.5-9.2)), St: 4.7-7.5 mm 


mm wide (0.9 (0.5—1. 5). Plants at Pagon 
ian Andes mountains; also widespread on 
steppes of Santa Cruz and Tierra del Fuego, 
Argentina naa e a 
Leaf blades 1-2.5 mm wide (1.6 (0, 8-3)). 

: 4-6 


wide (0.75 (0.5—1.4)). Plants of Patagonian 
steppe and semiarid areas of Cuyo and cen- 
tral Argentina | 
Ligules 2-5.5 mm long (3.6 (1.2-9)). 
Plants 18-38 cm tall (26 (15—62)); leaves 
8-18 cm long (14 (5.5—30)); leaf blades 5- 
10 ст long (8.3 (3-15)), 0.25-0.40 mm 


long (6 (2.5—16)). Stomata 0.044—0. 053 
mm long (0.048 (0.042—0.062)). Glume I, 
Pi: 4.5-6.2 mm long (5.5 (3.7-7)). St: 4— 
5.3 mm long (4.5 (3.6–5.6)). Lemmas, Pi: 


ian Altoandina biogeogaphic province, Ar- 
1 


һиеси 


ae Poa ligularis 


gentina and Chile _________ oa tristigmatica 


Ligules 4–8 mm long (6 (1.5—11)). Plants 
25-50 ст tall | (34 (9-73)); leaves 15-35 
ст long (24 (4.5—52)); leaf blades 8-20 
em long (13.4 (2.2-30)), 0.20-0.28 mm 
thick (0.25 (0.17—0.36)); sheaths 6-14 ст 
long oe (2.5—22)). Stomata 0.036–0.046 

ng (0.042 (0.032—0. Pus Chime I, 


mm long (7 (5—9)). Paleas, Pi and St: 4.5- 
6 mm long (5.2 (4.2-6.7)). Southeast Pata- 


onia and Chile |... oa alopecurus 
). 


5 

Leaf blades 2-3 mm wide (2.6 (1.8—4 
istillate and staminate florets glabrous. 
teppes and mountains from San Juan to 
Mendoza, between 1900 and 3900 m, Argen- 


Leaf blades 1.2-2 mm wide (1.6 (0.8—3)). 
Pistillate Da callus with woolly, long hairs; 

marginal nerves, and intercostal epi- 
делије zones glabrous or with hairs 0.5 mm 


224 Annals of the 
Missouri Botanical Garden 


or longer; staminate florets glabrous ог Pi and St: 3.5-5 me long (4.1 (2.6—5.3)); 
зен pubescent. Steppes of semiarid Pa- Pi and St: 0.7-0.8 mm wide (0.75 (0.5— 
pn 13 1) Non 
13(12). Plants long-rhizomatous, 20-50 cm tall (38 16(15). Plants. caespitous o ог r long- rhizomatous, Leaf 
(13-61)); panicles 7-14 ст long (10.3 blades 1-2 mm wide (1.6 (0.9—3.2)). Sto- 
(4.5-19.8)); panicle nodes: 11-15 (13 (9- mata 0.032-0.040 mm long (0.037 
18)). Leaves 15-35 cm long (25 (7.2-47)); (0.027—0.0496)). | wo Pi: 4-6.5 mm 
leaf blades 7-27 cm long (17 (4.7—35)) and long (5.6 (3.3-7.2)), St: 4-5 mm long (4.6 
.23-0.34. mm thick (0.28 (0.18—0.41)); (3.4—5.4)). Glume I, Pi: 2.5-3.5 mm long 
sheaths 4—14 em long (8.25 (2.5-20)); lig- (3 (1.8—4.8)), St: 2-3 mm long (2.5 (1.5— 
ules 5-11 mm long (7.8 (3.3—16.4)). Sto- 3.3); Pi and St: 0.4—0.7 mm wide (0.54 
a 0.044—0.054 mm long (0.048 (0.33-1)). Lemmas, Pi: 3-5 mm long (4.1 
(0.040—0.060)); keel and marginal nerves of (2.8–5.4)), St: 2.8-3.5 mm long (3.4 (2.8— 
pistillate florets with hairs longer than 0.5 4.1)). Paleas, Pi and St: 2.5-3.5 mm long 
mm long, intercostal epidermis zones gla- О rd Y REPRISE КИНИ 
brous. Widespread on semiarid Patagonia, 16’. Plants caespitous. Leaf blades 2-4 mm 
and coasts of Argentina, Brazil, ie diam wide (3.1 (2—4.4)). Stomata 0.037-0.044 
a lanuginosa mm is ҮЙҮ, (0.036–0.046)). Spikelets, 
13’. — Plants har 8-25 ст ull. (17.6 (6– Pi: 6.4-7 mm long (6.8 mm (6.3-7.2)), S 
42); panicles 3-7 em p (4.4 (1.7-9)); 4.5-6 mm long (5.6 (4.8—6.3)). Glume I, 
d aeri 7-11 (8 (6–14)). Leaves 5- Рі: 3.5-4.5 mm long (4.2 (3.8-4.6)), St: 
18 cm long (10.2 (3.5-36.5)); leaf blades 2.8-3.5 mm long (3.2 (2.8-3.8)); Pi and 
3-8 ст long (5.6 (1.3-24)), 0.15-0.23 St: 0.5-0.8 mm wide (0.67 (0.40-0.9)). 
mm thick (0.19 (0.11—0.28)); sheaths 2-7 Lemmas, Pi: 4.8-5.8 mm long (5.3 (4.7— 
em long (4.6 (1.7-14.8)); ligules 3-7 mm 6)). St: 3.5-4 mm long (3.8 (3.2—4.1)). Pa- 
long (4.7 (1.1–10.3)). Stomata 0.033- leas, Pi: 3.2—4 m m long (3.6 (3.1-4.1)), 
0.044. mm long (0.038 (0. 030-0. 054)); St: 2.8-3.5 mm long (32 2 nd 3.8) ___ 
keel and marginal nerves of pistillateflorets — — ||| —— Poa lanigera 
with hairs shorter than mm | m" 17(15). Plants ава оца. ИИ ‘ences usually not 
д epidermis zones p or gla longer than leaf length. Ligules obtuse, 0.6— 
brous. Southern Patagonia, Argentin 1 mm long (1.1 (0.5—1.9)). Stomata scarce 
14(13). Callus of пи а eg "d woolly, on abaxial epidermis. 0.230.040 mm 
ong hairs, intercostal epidermis zones of long (0.036 (0.027-0.045)). Leaf blades with 


lemma and palea generally glabrous. On pis- 
tillate specimens: sheaths 3.5-6.5 cm 


long, blades 0.8-0.1 1 mm thick, and sto- 


apex acuminate to acute. Florets not vivipa- 
rous. Pistillate floret hairy, cottonlike; callus 


: with woolly, long hairs; keel, marginal 
mata 0.029—0.038 mm long; on staminate E 
aser · spikelets 5.4-7.8 ^ idu nerves, and pa in epidermis zones pem 
"Le PORC ___ pecu rues và hairs over 1 mm long. Staminate florets gla- 


leaf blades 1-1.5 mm long. Sarine га 

calities of Santa KS = Tierra del Fu 

Argentina _____ rigidifolia var. О 
14'. Callus of abi ded glabrous, interc 

tal epidermis zones of lemma and palea usu- 

ally pubescent. On pistillate specimens: 

sheaths 1.8—4.8 cm long, blades 0.12- 


brous or with a few long, woolly hairs on cal- 
lus. Grasslands above 1500 m in Córdoba 
and San Luis hills, Argentina — 
— m Po oa hubbardiana 
17'. Plants кни bullorcscime 'es longer 
than the leaf length. Ligules acuminate, 1— 


0.16 mm thick; stomata 0.034—0.042 4 mm long (2.2 (0.5—5.8)). Stomata frequent 
mm long; on staminate specimens: spikelets on the abaxial epidermis, 0.035-0.045 
4.7-6.5 mm long and leaf blades 0.8-1.3 mm long (0.042 (0.034—0. 055). Leaf blades 
mm long. Xeric localities of Santa Cruz and with apex navicular to obtuse. Florets usually 
Chubut, Argentina __ Poa rigidifolia var. ibari viviparous. Pistillate floret callus without 
15(6). Florets not viviparous. Glume I, Pi: 2.5- hairs, or with short or long, woolly hairs; 
4.5 mm g (3.4 (1.84.8), St: 2-3.5 keel, marginal nerves, and intercostal epi- 
mm long (2.7 (1.5-3.8)); Pi and St: 0.45- dermis zones glabrous, or with hairs shorter 
.8 mm wide (0.58 (0.33-1)). Lemmas, E than 0.5 mm. Staminate florets glabrous or 
3-5.5 mm long (4.5 (2.8—6)), St: 3—4 m scarcely pubesc pod acad Andes 
long (3.5 (2.8—4.1)); Pi and St: 0.6-1 mm mountains, up to 1000 m ________________ 
wide (0.78 (0.54—1.2)). Paleas, Pi and St: 18(16). Plants caespitous or fol вве Leaf 
2.8-3.8 mm long (3.1 (1.8—4.1)); Pi and blades conduplicate; 0.15-0.24 mm thick 
$1: 0.4—0.7 mm wide (0.54 (0.3-1) ___ 16 (0.20 © 17-0.48)). Glume I, 1-пегуеа. 
15'.  Florets usually viviparous [not in P. hubbar- Lem Pi and St: 0.7-0.9 mm wide 
diana, infrequently in P. wire Glume Е (0. 65-1. 2)). Paleas, Pi and St: 0.45- 
I, Pi: 4-7 mm long (5.4 (3.3-7.5)), St: 0.7 mm wide uibs )4—-1)) —————— — 
3.5-6.5 mm long (4.5 (2.0-7.5); Pi and . | | = Poa rine nues 
St: 0.7-1.1 mm wide (0.82 (0.52—1.3)). 18'. Plants caespitus Leaf blades flat; 0. 
Lemmas, Pi: 4.5-8 mm long (6.3 (5—8.8)), 0.17 mm thick s 15 (0.13-0. А lume 
St: 4—6.5 mm long (5.1 (4–7)); Pi and St: I, нан (1). Lemmas, Pi and St: 0.6- 


0.9-1.3 mm wide (1.1 (0.72-1.5)). Paleas, 0.7 mm wide (0.65 (0.54—0.86)). Paleas, Pi 


Volume 87, Number 2 Giussani 225 
2000 Phenetic Patterns in Poa Sect. Dioicopoa 
and St: 0.35-0.45 mm wide (0.4 (0.3— el complejo Poa rigidifolia asociada al efecto del pas- 
0.7) ____ Poa pilcomayensis var. pilcomayensis toreo ovino y al ambiente. Revista Chilena Hist. Nat 
19(17). Florets usually viviparous. Spikelets, Pi and 70: 421-434 
St: 7-20 mm long (14 (5.6–28.6)). Glume ‚ A. J. Martínez & M. B. Collantes. 1996. Mor- 
I, Рі: 5-6.8 mm long (6 (4.3–7.1)), St: 4— pholo iral variation M Бан with environment in four 
6.5 mm long (5.6 (3.9–7.5)); Pi and St: јен dg dan; gonian Poa species: The Poa rigidifolia 
0.7-1 mm wide (0.90 (0.64—1 .3)). ea mplex. C №. J. Вог. 74: 762-772 
Pi: 5.5-8 mm (7 (5-8.8)), St: 5-6.5 m p: y Белове Е. 1958. Сарпшо І, Orografía. In: La 
(6 (4.8–6.9)). Pistillate floret callus with Argentina Suma de Geograffa. Tomo Ш. Ed. Francisco 
short or long, woolly hairs ___ Poa pogonantha de rear Ediciones Peuser, Buenos dinis. 
19'. Viviparous florets absent, occasionally pre- Gower, J. C. & G. J. S. Ross. 1969. Minimum spanning 


sent. Spikelets, Pi and St: 5-7 mm long 
(6 (5–7.7)). Glume I, Pi: 3.5-5 mm long 
(4.3 (3.3—5.3)), St: 3-4 mm long (3.6 (3- 
4.1)); Pi and St: 0.6-0.8 mm wide (0.72 
(0.51—0.83)). Lemmas, Pi: 4.5-5.5 mm 
long (5.3 (4.3—5.8)), St: 4-5 mm long (4.5 
(4—5.2)). ir Md GT callus with н! nd 
woolly hai Poa denudata 


Literature Cited 


Affifi, A. A. & V. Clark. 1984. ed Multi- 
variate Analysis. Wadsworth, California, U.S 
Akerberg, E. 1942. y ырай dieti in эз жайнай 
and из hybri ан alpina. Hereditas 28: 1–126. 
доп, A. M. & H. E. Connor. "199 Floral biology and 
reproduction in Pao (Poeae: Сипае, Austral. J. 
Bot. 43: 
Bor, B L. 1952. The ig Hg L. 
. Hist. Soc. 50: 787-8 
З nus A. L. 1994. Сону ОРТА Argentinas 
Fascfculo I. Enciclopedia Argentina de Agricultura y 
Jardinería, П. Ed. Acme, Buenos Aires. 
& A. Willink. 3 peus de América 


in India. J. Bombay 


Clausen, J. 1961. е, faite by apomixis in 
olyploid Poas. Eup a 10 

p W. D. & S. А, н. g е пега id 

m. Grasses of the World. Kew Bull. Aes Ser. - 


Clifford, H. T. & W. Stephenson. 1975. An Introduction 
to Numerical а Academic Press, New Yor 
Connor, H. E. 1979. Breeding b pm in the grasses: A 

survey. New 2 nd J. Bot. 17: 547—574. 
19 Evolution of reproductive systems in the 
T dn. Missouri Bot. Gard. 68: 48—74. 
Crisci, J. V. & M. F. López о 1983. Introducción 
a la teoría y práctica de la taxonomía numérica. Pro- 
grama Regional de sida e Científico y Tecnológico. 
OEA, Washington, D.C. Monografía, Serie Biología 26. 
Edgar, : pee Poa L. in New Zealand. New Zealand J. 
Bot. 
Ellis, ^ т 0 А procedure for erc compar- 
oaceae. I. The leaf-blade as 


A pro ocedure for standardizing compara- 
tive leaf арй in the Poaceae. II. The е 
seen in surface view. Bothalia 12: 641-671. 

Giussani, L. М. 1993. Sobre la validez de Роа breviculmis 

pe y Poa spicaeformis (Steud.) Hauman et Parodi. Bol. 

‚. Argent. Bot. 29(3—4): 249. 

1997. Variabilidad Morfológica y Biogeográfica 
de PA Especies Argentinas del Subgénero Dioicopoa 
(Poaceae). Tesis Doctoral, Facultad de M Exactas 
y Naturales, Universidad de Buenos Aire 

M. B. Collantes. 1997. Variación fenotípica en 


pidermis as 


trees and вй! cluster analysis. Applied Sta- 
tistics 18: 54—64. 

Hartley, W. 1961. Studies on the origin, evolution, and 
distribution of the Gramineae 4: The genus Poa. Aus- 
tral. J. Bot. 9: 152-161. 

Hitchcock, A. 5. 1927. Poa L. In The Grasses from Ec- 
uador, Peru and Bolivia. Contr. U.S. Natl. Herb. 24(8): 


326-333. 

Hunziker, J. H. 1978. Cytogenetics and о of some 
species of Poa (Gramineae). Pp. 144—148 M. E. 
Drets, N. Brum-Zorrilla & G. A. Folle (editors), Actas 

t 


del Ш Congreso Latinoamericano de Genética. Monte- 


eo. 

Kellogg, E. A. 1985. A biosystematic study of the Poa 
secunda Па J. Arnold Arbor. 66: 201–242. 

. Apomixis in the Poa secunda complex. 

Anan р [p 74: 1431-1437. 

990. Ontogenetic studies of florets in Poa (Gra- 


ets Alfons and heterochrony. Evolution 44: 
1978-1989. 
Longhi-Wagne I. Boldrini. 1988. Ров 


A. R. 
Schultz (editor), Flog Ilustrada de Río "ias do Sul. 
Porto Alegre, Río Grande do Sul. 

Marsh, V. L. 1952. A taxonomic revision of the genus 
Poa of United States and southern Canada. Amer. Midl. 
Naturalist 47: 202—250. 

Marticorena, C. & M. Quezada. 1985. ме ме d la flora 

-2) 1 
1983. Flora of Tierra del d pees 
IM Shropshire, and Missouri Botanical Garden, St. 


uis. 
Nicora, E. G. 1978. Poa. In: M. ea, Gramineae, 
Flora Patni. Colección Ciéntfüca del ТАТА. 8(3): 

138—2( 


Роа pedersenii Nicora, una nueva especie 
para la ora у: Paraguay. СапдоПеа 50: 544—546. 

о де Артазаг. 1987. Los géneros de 
на i yeh n Austral. Ed. Hemisferio Sur, 


uenos 

Parodi, L. R. 1 932. Algunas gramíneas argentinas nuevas 
o críticas. Physis 11: 129-138. 

————. 19 Contribución al conocimiento de las es- 
pecies del género Poa de la flora uruguaya. Revista 
Argent. Agron. 3: 133-152. 

1937. Gramíneas n nuevas o críticas. 
Notas Mus. La Plata, Bot. 2: 13-16. 

1940. Una nueva especie de Poa para la Flora 
Argenta, Notas Mus. La Plata, Bot. 31: 325-330. 

————. 1950. Las дрес. ah лек para el ganado еп 
la Repdblica Argentina. Revista Argent. Agron. 17: 
163- ar 


. Gramíneas argentinas nuevas o críticas. 
enu. ч Артоп. 28: 100—105. 
Gramíneas argentinas nuevas o críticas 
IV. би redo Agron. 29: 12-22. 


226 


Annals of the 
Missouri Botanical Garden 


Renvoize, S. А. 1998. Gramíneas de Bolivia. The Royal 
Botanic Gardens, Kew 

Rohlf, F. J. 1992. NTSYS- -рс Numerical Taxonomy and 
i a Analysis System. Applied Biostatistics, 
New 


Rosengnti, B., B. Arrillaga de Maffei & P. Izaguirre de 
Artucio. 1970. Gramíneas Uruguayas. Universidad de 
id 


"0. 

e genus Poa in Patagonia (Argen- 

tina): A phenetic = is species of Poa subgenus 

Poa. Bot. J. Linn 29-24]. 

i hae & R. M. Klein. А 
Parte b Сга. In R. Reitz ш Flora Ша 
Catarinense. Itajaf, 

Sneath, Р. H. A. & R. R. Sokal. 1973. Numerical Тах- 
onomy. The Principles and Practice of Numerical Clas- 
sification. Freeman, San Francisco. 

Sokal, R. R. & F. J. Rohlf. 1969. Biometry. The Princi- 
ples and Practice of ша in Biological Research. 
H. dear pud Fra 

Soreng, R. J. 990. Chloroplast- DNA phylogenetics and 
Ca in a reticulating group: Study in Poa (Po- 

aceae). Amer. J. Bot. 77: 1383-1400 

1998a. An 5. classification for dcn in 
North America, and o notes on sections, speci 
and subspecies of Роа, Puccinellia. and Dissanthelium 
(Poaceae). Novon 8: 187-202. 

———. 1 . Erratum for an i 1 
for Роа in North America, and minal notes on sections, 

species, and subspecies of Puccinellia, and Dis- 
santhelium (Poaceae). Novo 326. 
gr eese bres с 1992. Statgraphics 
e. STSC Inc., Rockville, Maryland. 

и. T 1985. Chromosome numbers and their taxo- 
nomic implications in the genus Poa of Japan. Bot. 
Mag. (Tokyo) 98: 413—437. 

Toledo, O. X. & E. A 


A 


1991. ND. ded General 
I. ан. La Familia 
: A. Burkart (editor), Flora 
Ilustrada de Entre Ríos i o Colección Cientí- 
fica del I.N.T.A. Bueno 3 
1970. Poa: In A. L. Cabrera, Flora de la Prov- 
тела de Buenos Aires. Colección Científica del I.N.T.A. 
4(2): 102-125 
Vickery, W. 19 70. А taxonomic study of the genus Роа 
ve in Australia. Contr. New South Wales Natl. Herb. 4: 


145-169. 
Watson, L. & M. J. Dallwitz. 1992. The Grass rod of 
the World. C A B International, — iy 
Zuloaga, F. O., E. G. Nicora, olo de i. [0] 
Могтопе i Ciadella. 1994. Catálogo 
de la Familia Poaceae en la Repüblica Argentina. Mon- 
ogr. Syst. Bot. Missouri Bot. Gard. 47: 1-178. 


APPENDIX 1. 


List of species. Letters in parentheses correspond to the 
abbreviation of the species name used in the phenetic 
lysis. 


Poa alopecurus n Kunth (AL) 
P. barrosiana Parodi (BA) 
Р. bergii кады ( 
bergii var. chubutensis Speg. (BU) 
€ (BO) 


b (BK) 
P. bonariensis Lam.) Kunth (BN) 


P. buchtienii Hack. (BT) 

P. ое Hack. (CL) 
P. denudata 5 DE 

dolichophylla Hack. (DO) 

P. dusenii Hack. (DU) 

P. holciformis J. Presl (HO) 

P. hubbardiana Parodi (HB) 


P ibari Phil. (IB 


P. patagonica var. neuquina Nicora (PN) 

P. pedersenii Nicora (PE) 

P. pilcomayensis Hack. (PI) 

P. pilcomayensis var. calamagrostoidea Hack. (PC) 
P. pogonantha (Franch.) Parodi (PG) 

р prichardii Rendle (PR) 

P. resinulosa Nees ex Steud. (RE) 

P rigidifolia Steud. (RI) 


ST) 
Р. superbiens (Steud.) Hauman & Parodi (SU) 
(TR) 


P. tristigmatica E. Desv. ex Gay (TR 


APPENDIX 2. 


ecimens listed here were used as OTUs in the phe- 
netic analysis. They are sorted alphabetically by species 


Malvinas represents a similar location to the Falkland Is- 
lands on specimen labels. 


Poa alopecurus 


ARGENTINA. Santa Cruz: Depto. Güer Aike, Entre 
Glencross у Pta. Alta, 51°46’S, 71743", 1976, Latour et 
al. 1004 (ВАВ) [AL, Pi and St]; Barranca al norte de 28 
de Noviembre, 51735'5, 72?11'W, 13 Jan. 1976, Latour et 
al. 382 (ВАВ) [SK, Pi]; ruta 3, a 8 km al У de Río 
Gallegos, 51°37'S, 69°25'W, 5 Nov. 1977, Roig & Méndez 
2361 (ВАВ) [SK, Pi and St]; Estancia Cerro Castillo, La- 
guna Figueroa, 51?28'S, 72°27'W, 12 Jan. 1977, Latour 
et al. 1367 a? ISU, St.]; Lago Ar репне. Puerto Ban 
dera, 50°18'5, 72747", Correa et al. 3032 (BAA) [SU, 
Pi]; Estancia Пи Viscachas, Cerro Las Viscachas, 
50°40'S, 72°01'W, 25 Jan. 1977, Arroyo et al. 2428 (BAB) 
[SU, Ру; Cerro sin nombre, 50°46'S, 72°08'W, 29 Jan. 
1977, Arroyo et al. 2643 (BAB) [SU, Pi]. Tierra del Fue- 
go e Isla del Atlántico Sur: Islas Malvinas, Isla Sole- 
dad, 52°00’S, 59*00'W, Gaudichaud С. s.n. (Arundo alo- 
ecurus Gaudich. ex Mirb., syntype, BAA, fragment) [AL, 
Pi]; Islas Malvinas, 52?00'S, 59?00'W, Hooker s.n. (Fes- 
tuca antarctica var. у Hook. f., isotype, BAA fragment) 
[AL, Pi and St]; Ushuaia, Is. de los Estados, Port Vancou- 
ver, 54^49'S, 64°20'W, Spegazzini s.n. (Festuca shuka 
Speg., isotype, LP) [SK, Pi]; Canal a Beagle, Isla de los 
Gaviotines, 54°52'5, 68°18'W, Vervoorst 161 (BAA) [AL, 


Volume 87, Number 2 
2000 


Giussani 227 
Phenetic Patterns in Poa Sect. Dioicopoa 


St]; Islas de los Estados, Bahía Liberty, 54^50'S, 64725" W, 
2 Nov. 1971, Dudley et al. 1301 (BAB) [AL, St]; Islas de 
los Estados, Puerto Vancouver, 54°49'S, 64°20'W, 16 Jan. 
1934, Castellanos 12868 (BAB) [AL, St]; * los Es- 
tados, Puerto Cook, 54^50'S, 64°20'W, 4 4, Cas- 
tellanos 12829 (BAB) [AL, St]; Is. de ie Па ed Pto. 
Cook, 54?49'S, 64°20'Ұ, 4 Jan. 1934, Castellanos 12829b 
(51) [SK, BE Bahia Thetis, Región del Río del Fuego, 
54^12'S, 67?23'W, Маг. 1902, Holmberg & Calcagnini 
3604 EA [AL, Pi]; Ea. Harberton, Campo Rancho Tam- 
bo, shore of Bahía Cambaceres, 54°52'S, 67?16' W, 8 Jan. 
1968, Moore 1373 € [AL, Pi]; Bahía Agüirre, de 
Español, E end beyond settlement Ostoic, 
65°58'W, 19 Feb. 1968, "Moor re 1875 (BAB) [AL, St]; гы 
molino, 54?50'S, 67?53'W, 21 Dec. 1932, Castellanos 
7579 (ВАВ) [AL, St]. CHILE. XII Región: Sandy Point, 
53°10'S, 70°54'W, № Lechler 1194 (Ата superbiens 
Steud., isotype, BAA fragment) [SU, Pi]; Islote Pto. Luisa, 
54756'5, те W, Roig et al. 2838 (ВА А.) [AL, у Sierra 
Baguales, umbre, Co. sin nombre, 50°42'S, 
72°22'W, js ro 1975, Boelcke et al. 682 (BAB) [AL, Pi 
and St]; Punto Bella Vista, 51°30’S, 73?15'W, 5 Dec 
1979, Roig et al. 5097 (BAB) [AL, Pi]; Estancia бано 
Castillo, Lago Sofía, 51*32'S, 72737' W, 14 Jan. 1977, La- 
tour et al. 1493 (ВАВ) [SK, Pt]; Laguna Figueroa, 
51?28'S, 72°27'W, 12 Jan. 1977, Latour et al. 1367 (ВАВ) 
[SU, St.]; Sección Tres Pasos, Hotel, 51725'5, 72?29'W, 
14 lus. 1977, Latour et al. 1600 (ВАВ) [SU, Рај; Latour 
et al. 1574 (BAB) [SU, Pi]. 


Poa bergii 


ARGENTINA. Buenos Aires: Bahía Blanca, Punta 
Alta, 38°53'5, 62°04’W, Feb. 1902, Spegazzini et al. 2381 
(BAB) [BE, Pi]; Partido General Dorrego, Monte Hermoso 
38"59'5, 61?17' W, 30 Oct. 1985, Villamil 3393 (SI) [BE, 
Pi and St], 31 Oct. 1985, Villamil 3413 (SI) [BE, Pi], 9 
Nov. 1986, Villamil 4291 (SI) [BE, Pi]; Camino a Monte 
Hermoso, Balneario Sauce Grande, 38°59’S, 61?17'W, 31 
Oct. 1986, Villamil 3432 (SI) [BE, Pi], 30 Oct. 1965, Ca- 
brera et al. 17061 (LP) [BE, Pi], Cabrera et al. 17063 (LP) 
[BE, St], Cabrera et al. 17065 1/2 (LP) [BE, St], 24 Nov. 
1963, Fabris & Schwabe 4821 (LP) [BE, Pi]; Partido Pe- 
llegrini, Pellegrini, 36°16'5, 63°10'W, 28 Nov. 1940, Ca- 
brera 6948 (LP), [BE, Pi]; Partido Coronel Rosales, Pe- 
huen-Co, 39?00'S, 61?33'W, 19 Nov. 1962, Cabrera & 
Fabris 14914 (LP) [BE, P Partido Villarino, Médanos, 
38°50'S, 62°42'W, 6 Nov. 1940, Cabrera 6673 (LP) [BE, 
Рај; Partido General Madar Pinamar, 37?07'S 
56°51'№, 11 jen 1950, Cabrera 10707 (LP) [BE, Pil 
Partido Tres Arroyos, Claromecó, 38°52'S, 60704", 8 
Nov. 1986, ш 4211 (SI) [BE, Рај; Partido P bm 
Alvarado, Miramar, 38?16'S, 57?52'W, 31 Jan. 1930, Pa- 
rodi 9820 (holotype, BAA) [BA, Pi]. 11 Nov. 1939, Ca- 
brera 5555 (BAA) [BA, St]. дыла 1922, Наитап 5762 
(ВАА) [ВА, +: Partido Lobe Quequén, 38°32'S, 

58°42'W, Jan. 1930, Cabrera 1317 (SI) [BA, St], Feb. 
1925, Parodi 6398 (BAA) [BA, St], 7 Nov. 1943, Boelcke 
s.n. (SI) [BA, Pi and St]; Partido General Pueyrredón, Mar 
del Plata, 38°00’S, 57?34'W, 9 Dec. 1947, Boelcke 2838 
(BAA) [BA, Pi], Jan. 1923, Barros 4898 (BAA) [BA, Pi], 
9 Dec. 1947, Boelcke 2837 (BAA) [BA, St]. Río Negro: 
Adolfo Alsina, médanos en la el Hío Negro, 
41°03'S, 62?48' W, Berg 205 (holotype, CORD) [BE, Pi]. 
URUGUAY. Colonia: Riachuelo, 34?25'S, 57?52'W, 12 
Oct. 1936, Cabrera 3872 (BAA) [BE, Pi]. 


Poa bonariensis 


ARGENTINA. Buenos Aires: Río Viedma, Sep. 1934, 
Ruiguelet 409 (BAA) [BN, Pi and St]; Partido Avellaneda, 
Avellaneda, 34?40'S, 58?23'W, 14 Oct. 22, Parodi 
4747 (BAA) [BN, Pi and ый Partido Dolores, Laguna Se- 
vigné, 36°12'5, 57?44'W, 11 Oct. 1962, Cano 1 (BAA) 
[ВМ, St]; Partido La au La Plata, en el Jardín Zooló- 
gico, 34?55'S, 57°57'W, Nov. 1962, Torres 1023 (LP) [MO, 
St]; Partido Cañuelas, Ruta 3, km 64, entre Brandsen у 
Monte, 35?03'S, 58°46’W, 13 Nov. 1962, Cano-Cámara З 
(SI) [MO, Pi]. Entre Ríos: Depto. Gualeguaychú, E. 
Carbó, 33?09'S, 59°14'%, 23 Oct. 1949, Burkart 18132 
(SI) [BN, Рај; Paranacito, 33?42'S, 59°01’W, Oct. 1949, 

agonese & Crovetto 25 (BAB) [BN, Pi and St]; Médanos, 
33°25'S, 59°04’ W, 2 Dec. 1930, Parodi 9445 (BAA) [BN, 
Pi and St]; Delta inferior, Arroyo Martínez, 33752'S, 
58*40' W, Oct. 1944, Burkart 15044 (SI) (МО, St], Burkart 
15043 (SI) [MO, St]; Depto. aleguay, La Verde, 

33°09'S, 59°20'W, 21 Oct. 1949, Burkart 18068 (SI) [BN, 
Pi]; Depto. La Paz, Arroyo Feliciano, 30°44'S, пи 
17 Oct. 1980, Mufioz 1353 (SI) [MO, Pi and St]. N 
quén: Depto. Lácar, San Martín de Los Andes, 40°10’ s. 
71°21'W, 30 Jan. 1959, Ruiz Leal 20315 (holotype, BAA) 
[PN, Pi], 6 Jan. 1938, Giacobbi 12961 (BAA) [PN, Pi], 22 
Jan. 1966, Eskuche 591 (BAA) [PN, Pi]; Cerro al Norte 
de San Martín de los Andes, 40?10'S, 71?21'W, 10 Dec. 
1946, Dawson 1287 1/2 (BAA) [PN, Pi]. URUGUAY. 
Montevideo: Montevideo, 35?00'S, 56°05'W, en parajes 
hümedos, Nov., Arechavaleta 5101 (isotype, LP fragment) 
[MO, Pi and St]; Fray Bentos, 33?10'S, 58°15'W, 5 Oct. 
1934, Meyer 1037 (BAA) [MO, St]. Soriano: 
Grande, Paso de Los Loros, 19 Nov. 1937, Rosengurtt 
2294 (BAA) [MO, St]. 


Poa denudata 


ARGENTINA. Neuquén: Depto. Picunches, Pino 
Hachado, Orilla afluente Arroyo Haichol, 38°40’S, 
70*54' W, 21 Jan. 1963, Valla et al. 3024 (BAA) [DE, E 
and St]; Depto. Los Lagos, Villa Puerto Manzano, 40°49’ 

А °37'W, 8 Dec. 1963, Diem 3190 (BAA) [DE, Pi and si 
rro O'Connor, 40°49’S, 71°37'W, 25 Nov. 1963, Die 
3009 (BAA) [DE, Pi and St]; Isla Victoria, 40° 54'S, 
71°34'W, 11 Nov. 1946, Perrone 15212 (BAA) [NA, St]; 
epto. Minas, o Epu-Lauquen, Arroyo Pincheira, 
36^50'S, 71°03'W, 18 ie 1964, Boelcke et al. 11003 (SI) 
[DE, Pi]; Península Quetrihué, 40°52’S, 71°38’W, 1 Jan. 
1949, Boelcke & Hunziker 3458 (holotype, BAA) [NA, Pi 
and St]; Puerto San Patricio, 40%52'5, 71?38'W, 7 Nov. 
1940, Diem 285 (BAA) [NA, St], 30 Nov. 1942, Diem 415 


iérre $, 71°20’W, 28 Oct. 

Hunziker 3418 (BAA) [DE, Pi and St]. CHILE. X Regién: 
Valdivia, 39°49'S, 73°14’ W, Lechler W. 578 (BAA, frag- 
ment) [DE, Pi] 


Poa dolichophylla complex 


Poa dolichophylla: ARGENTINA. La Rioja: Depto. 
Famatina, Sierra Famatina, Mina El Oro, 28'55'5, 
67°31'W, 6 Feb. 1956, Calderón 1115 (BAA) [DO. Ру, 7 
Feb. 1956, Calderón 1168 (BAA) [DO, Pi]. Tucumán: 
Depto. оро La Banderita, ruta 65, 27°20’S, 
66°00'W, 14 . 1966, Boelcke et al. 5505 (BAA) [DO, 
Pi and pi Dade Taff, Bajo de Anfama, 27°40'S, 
65°34'W, 6 Aug. 1906, Lillo 5066 (isotype, BAA) [DO, 
Pi]; Valle Calchaquí, Peñas Azules, 26°35'S, 65°40'W, 27 


228 


Annals of the 
Missouri Botanical Garden 


Jan. 1933, eer Poen (BAA) [DO, Pi[; Cerro El Ne- 
grito, 26°37'S, 6 9 Jan. 1964, Giusti et al. 3863 
(BAA) [DO, Pi]; je Puerta, 29 Jan. 1933, Parodi 10891 
(LP) [DO, d € Calchaquí, Peñas Azules, 26°35’S, 
65°40'W, 27 1933, Parodi 10927 (BAA) [DO, Pi and 
St]. Р iridifolia; ARGENTINA. Buenos Aires: Partido 
Balcarce, Quinta Sagenave, 37°52'S, 58°15'W, 12-25 Oct. 
1943, Hunziker 3930 (BAA) [IR, St]; Partido General 
Pueyrredán, Mar del Plata, Sierra La Brava, 37°50'S, 
57°55'\, 4 Dec. 1930, Hicken 40 (SI) ПВ, Pi]; id 
Saavedra, Sierra de Curamalal, La Gruta, 37°5 
62°21'W, 12 Nov. 1938, Cala eo (SI) ПК, St]; en 
tido Tandil, Sierra de las Animas, 37°29'$, 59718", 2 
Nov. 1940, Cabrera 6810 (SI) [IR, a Tandil, 37?20'S, 
59°08' W, 20 Nov. 1929, Hunziker 8971 (BAA) ПВ, St], 15 
Nov. 1953, Burkart 19263 (SI) [IR, Pi]; Sierras de Tandil, 
37°20'S, 59°08’W, 1 Nov. 1919, Hicken 21 (SI) ПВ, St]; 
Partido Tornquist, Sierra de la Ventana, Abra de la Ven- 
tana, 38?08'S, 62°00’\, 7 Nov. 1938, Cabrera 4687 (SI 
ПК, Ру; Cerro de la Ventana, 38°08’S, 61°47'W, 7 Oct. 
1939, Cabrera 5336 (SI) [IR, Pi], 9 Nov. 1938, Cabrera 
4746 (SI) [IR, St]; Cerro Napostá, Estancia Laurina, 
38°08'S, 61747", 17 Nov. 1972, Gómez et al. 11719 
(BAA) [IR, Pi]. P. ir din var. calamagrostidea: AR- 
GENTINA. Tuc o. Chicligasta, Estancia Las 
Pavas, Puerto El "Ва а 27°19" 56 65°55'W, Маг. 1924, Ven- 
turi 3077 (SI) [PC, St], Venturi 3074 (SI) [PC, Pi]; Depto. 
Famaillá, Quebrada del Río Colorado, 27^10'S, 65°22'W, 
26 Aug. 1939, Meyer 14980 (BAA) [PC, Pi id St]; Que- 
brada de Lules, 26"56'5, 65?21'W, 5 Nov. 1929, Venturi 
9066 (LP) [PC, St]; b Monteros, Quebr ada de los 
Sosa, 27°09'S, 65°34’ W, Dec. 1960, Burkart 22098 (BAA) 
[PC, Ру; Depto. Taff, Bajo de Anfama, 27°46'S, 65°34’ W, 
6 Sep. 1906, Lillo 5064 oe z fragment) [РС, St]; 
Puerta de San Javier, 26°46'S, 65°21'W, Dec. 1923, Ven- 
turi 2540 (SI, LP) [PC, Pi and si; ы. j Playa 
del río de la Hoyada, 26"34'5, 66724", 23 Nov. 1921, 
Scheiter 1855 (BAA) [PC, Pi]. P. чискепи: ARGENTINA. 

órdoba: Depto. Pocho, Camino a los Gigantes, Cerro 
Blanco, próximo al Río Juspe, 31°27'S, 64738", 5 Dec. 
1958, Nicora 6642 (BAA) [ST, Pi]; Depto. San Alberto, 
Pampa de Achala, 31*36'S, 64°45'W, 10 Nov. 1925, Hick- 
en 16565 (BAA) [ST, St], 1—4 Dec. 1926, Parodi 7582 
(BAA) [ST, Pi], Parodi 7621 (BAA) [ST, Pi and St]; La 
Posta, 31°36’S, 64?45' W, 7 Dec. 1958, Burkart 20901 (SI) 
[ST, St], 9 Dec. 1995, Giussani stm2 (SI) [ST, St], Giussani 
pne (SI) [ST, St]. San Luis: Depto. Chacabuco, Sierra 
Comechingones, 32°54'S, 65704", 15 Nov. 1925, Caste- 
llanos s.n. (BAA) [ST, Pi]; Depto. Pringles, La Carolina, 

'S, 66°06'W, 8 Nov. 1940, Burkart 10787 (BAA) 

[ST, Pi]. 


— 


Poa holciformis 


ARGENTINA. Mendoza: Depto. Las Heras, Las Cue- 
vas, Refugio Militar General Lamadrid, 32° 40'S, vum W, 
10 Jan. 1963, Boelcke et al. 9720 (BAB, SI) [HO, Pi ii 
St]; Depto. San Carlos, Los Paramillos, camino a Lagu 
Diamante, 34?10'S, 69°35'W, 23 Jan. 1989, Gómez- om 
345 (SI) [HO, Pi]; 6 km W Refugio Militar General Al- 
varado, 34°10’S, 69?45'W, 17 Jan. 1963, Boelcke et al. 
9985 (BAB) [HO, Pi]. Neuquén: Depto. Chos Malal Ca- 
jon del Arroyo del cruce, 36°43'5, 70?23' W, 27 Jan. 1964, 
Boelcke et al. 11265 (ВАВ) [HO, Pi]; Vegas del у 
camino а Riscos Bayos, 36*54'S, 70720", 24 Jan 
Boelcke et al. 11157 (BAA) [HO, Pi and St]; Risco ы 


i 
confluencia Arroyo Olletas con el Arroyo Curileuvü, 


36?57'S, 70?20'W, 25 Jan. 1964, Boelcke et al. 11175 
(BAA, ВАВ) [HO, Pi]; Depto. Minas, Sierra de Cochicó, 
36°21'S, 70734", 29 Jan. 1970, Boelcke et al. 14087 
(BAA) [HO, St]; Nacimiento de la cordillera del Viento, 
36°46'S, 70?31'W, 2 Feb. 1964, Boelcke et al. 11540 
(BAB) [HO, Pi and St]. CHILE. In Cordillera chilensibus, 
Haenke s.n. (isotype, BAA fragment) [HO, Pi]. 


Poa hubbardiana 


ARGENTINA. Córdoba: Depto. San Alberto, Sierra de 
Achala, al bajar de la Pampa de Achala, 31°36’S, 
64745", 1-4 Dec. 1926, Parodi 7501 (holotype, BAA) 
[HB, Pil; Sierra de Achala, 31?36'S, 64^45'W, 15 Nov. 
1878, Hieronymus s.n. (ВАА) [HB, Pi and St]; Pampa de 
Achala, La Posta, 31736"5, 64°45'W, 9 Dec. 1995, Giussa- 
ni s.n. (SI) [HB, St]. San Luis: Depto. Chacabuco, Sierra 
de Comechingones, 32°54'5, 65°04’ W, 23 Nov. 1925, Ca- 
stellanos s.n. (BAA) [HB, St]; Depto. La Capital, Ciudad, 
33°18'S, 66°22'W, Parodi 2615 ee [HB, St]; Depto. 

Pringles L Carolina, 32°49'S, )6'W, 8 Nov. 1940, 
Burka 10792 ( (SI) [HB, Pi and E Burkart 10781 (SI) 
HB, Pi]. 


Poa huecu 


ARGENTINA. Mendoza: Depto. Las Heras, Puente 
del Inca, Valle de los Horcones, 32°50'S , 69°55'\, 12 
Jan. 1963, Boelcke et al. 9818 (BAB) [HU, Sij. Neuquén: 
Depto. Chos Malal, Cajón Grande, Cordillera del Viento, 
36°58'5, 70*30' W, 25 Jan. 1935, Ragonese А. 284A (ho- 
lotype, BAA) [HU, Pi]; Cajón de en o, Cordillera del 
Viento, 36°58'S, 70°30'W, 25 Jan. 193 agonese 284 
(BAA) [HU, St]; Cajón inferior del еке Turbio llamado 
localmente Arroyo Domuyo, 36744'5, 70?23'W, 28 Jan. 
1964, Boelcke et т 11323 (BAA) (HU, Pi]; Depto. Minas, 

una Varvarco C у та curso infe- 
rior, 36°17'S, 70°30" W, 29 Jan. 1910, Boelcke et al. 14061 
(BAB) [HU, St]; Arroyo Enfermera, extremo sur, 36?23'S, 
70°37'W, 28 Jan. 1970, Boelcke e а та. 14036 (ВАВ) bot 
Pi]; Baños Calientes, Río Varvarco, 36°42'S, 70°37'W, З 
Jan. 1964, Boelcke et al. 11424 (ВАВ) [HU, Рі]; Abs 
Zapala, Zapala, 38°55’S, а Ніскеп 16564 (51 
[HU, Рі]; El Sauce, sd $, 70?00' W, 11 Dec. 1952, Ca- 
brera 11175 (LP) [HU, Р 


Poa indigesta 


ARGENTINA. Мере aria Zapala, Zapala, 
38°55'S, 70°04’W, 7 Dec. 1946, Dawson С. 1227 (holo- 
type, BAA) x Pi and 3 e Feb. 1920, Parodi 2721 
(BAA) [IN, Pi]. 


Poa lanigera 


ARGENTINA. Buenos Aires: Partido Pehuajó, Pe- 
ч 35°48'$, 61°52'W, 15 Oct. 1950, Burkart 18465 
SI) [LG, St]; Partido Tigre, FNGBM, 34°25’S, 
58°35'W, 13 Oct. 1946, Lanfranchi 535 ( (SI) [LG, St]. En- 
tre Ríos: Depto. Federación, Federación, 31°01'S, 
57°53'W, 23 Sep. 1961, Burkart 22437 (BAA) [LG, Pi and 

St]; Salto Grande, 31?13'S, 57?56' W, fi Oct. 1950, Hun- 
ziker 4431 (SI) [LG, Pi]; Depto. Uruguay, Concepción del 
Uruguay, 32°29'S, 58°14’ W, 17 Oct. 1949, Burkart 17969 
(BAA) [LG, Pi and St]. PARAGUAY. Alto Paraná: Santa 
Teresa, 24°43’S, 54°21'W, 4 Aug. 1945, Bertoni 1706 
(BAA, SI) [LG, Pi] URUGUAY. Montevideo: Sellow s.n. 
(isotype, BAA) [LG, Pi and St]. 


Dh 


Volume 87, Number 2 
2000 


Giussani 229 


Phenetic Patterns in Poa Sect. Dioicopoa 


Poa lanuginosa 


ARGENTINA. Buenos Aires: Partido Dorrego, Monte 
Hermoso, 38°59'S, 61?17'W, 30 Oct. 1965, Cabrera et al. 
17065 (LP) [LA, Pi and St]; Partido General Madariaga, 
Pinamar, 37°07'S, 56°51'%, +" Oct. 1973, Zardini 215 
(LP) [LA, Pi]; at Junin, Médano Grande, 34?27'S, 
LER W, 17 Oct. 1940, Cabrera 6503 (LP) vus St]; ic 
tido Tornquist, Sierra de la Ventana, estancia El Car 
38°09'S, 61?55'W, 6 Oct. 1939, Cabrera 5309 (LP) [L А, 
St]. Chubut: Depto. Río Sengüerr, Estancia La Laurita, 
44^44'S, 70^15'W, 18 Jan. 1949, ан 3849 (BAA) 
[PA, Pij Río Mayo, Estación Zootecnia Mallín, El Tacho, 
S, 70°16'W, 4 Feb. 1954, Ad 3545 (BAA) 
. Cabo Raso, 
938 alis eA BAA fra 
ment) © к ко pe 1922, ае 3447 (ВАА) [PA, PI. 
Mendoza: Depto. San Rafael, Valle del Atuel, El Sos- 
neado, 35?05'S, 69*34' W, 4 Oct. 1955, Bócher et al. 801 
(holotype, BAA) [BO, Pi and St]. Neuquén: Depto. n 
Lagos, Nahuel Huapi, Estancia Fortín Chacabu 
T. 71909", З Nov. 1949, Boelcke & Hunziker 3492 

BAA) [LA, Pi]: e a Мир sd ríos Pichi- 
E uén y Neuq Yeg 36735'S, 
70*48' W, а el ie үг > ns LA. Pi а St]; а 
5 km de Las Ovejas, camino a la laguna Epu-Lauquén, 
36°57 S, 70°45’ W. 14 Jan. 1964, Bo nee et al. 10772 
(BAA) [LA, Pi]. Rio Nears : Depto. Avellaneda, ruta 22, 
50 km antes de Choel choel, 30°10" S. 65°05’ W, Bacigalu- 

о М. & Nicora Е. e 1/2 por [BO, Pi]; Depto. 
or rquinco, Río Chico, 41742'S, 70°27'W, 9 Nov. 1949, 
Soriano 3751 (BAA) Io Pi and St Depto. Avellaneda, 
ruta 6 km antes | General Conesa, 39?45'S, 
65"26' W, 1 Nov. 1972, Bacigalupo & Nicora 11676 (BAA) 
[LA, Ру; Isla Choel Choel, pen del Río Negro, 39?16'S, 
65^39' W, 7 Nov. 1972, Bacigalupo & Nicora 11654 (BAA) 
[LA, Pi and St]; Choel Fw 39*10'S, 65°05’ W, Bacigalu- 
po & Nicora 11651 (BAA) [BO, St]; Depto. Pichi Mahuida, 

a 20 km S de Río Colorado, 39°03'5, 64?20'W, 7 Nov. 
1965, Correa & Nicora 3168 (BAA) SN Pi and St]. San- 
ta Cruz: Depto. Magallanes, San Julián, 5 km al sur por 
ruta З, Bajo Salado, 49°19’S, 67?42'W, 22 Nov. 1963, 
T et al. 2683 (BAB) [PA, Pi]; Depto. Corpen Aike, 

E co, Establecimiento Las Vegas, 50°07’S, 68°37'W, 
No. 922, Dauber 165 (BAA) [PA, Pi and St]; Orilla del 
Río Br ruta 3, 49°49'S, chile 3 Dec. 1971, Boel- 
Depto. КЕ Puerto 
n °45'5, 6 
Ya 1965. С ea & Nicora oca dx s Depto. 
Lago Argentino, o, Campos сн Bilbao 
50°38" S, 72°54’ W, 3 Feb. 
[PA, St]; шт Río his :0, "Cañadón León, 4874. 
70°16' W, Маг. 1952, Cittadini 18 (BAA) [PA, St], Citta- 
dini 21 (BAA) КН, Ру. Tierra del Fuego e Islas del 
Atlántico Sur: Depto. Rí i 

W de Río yw le. 53°41'% 
Moore 1493 (BAA) [РА, Pi]. CHILE. XII Regi 
Pinto, cerca del origen ia río, 52°00'S, 72°24’ Ww т ы 
877, Ibar s.n. (holotype, BAA fragment) [PA, St]. 


Poa ligularis 


ARGENTINA. vw me Partido Bahía owns 
Bahía Blanca, 38°44’S, 62°16'W, Hanslow 552 (typ 
BAA ТЕ [LL St]. Clara 34 (P. denudata var “ж 
Ball, BAA fragment) [LI, S 
~ мили Мадгуп, Laguna d. a, А 
Oct. 1990, Вегі: ег & Beeskow 1132 (SI) (LI, St], 30 Oct. 


1992, Bertiller 3150 (SI) [LL Pi]; Bose Sarasa a 40 
m de Puerto Madryn, 42°37'S, 6 ct. 1995, 
Bertiller 3395 (SI) (LI, St. Вен E (SI) Hi s 
epto. Rawson, Trelew, 4. р W, 16 Oct. 
Soriano 1867 (SI) [LI, Pi]. La E Dept о. Chica nts 
Cerro Los Guanacos, Oct. 1960, Cano 1119 (BAB) (LI. 
St]; е Utracán, Entre Atreucó y Quehué, 37°05’ 
64°11'W, 10 Jan. 1995, Ragonese hu асти 18205 
(BAB) [11, Pi]. Mendoza: Depto 
Obligación, 20 Nov. 1943, Covas 2097 (SD "m Pi] Nia 
uén: Dept o. Eon Bes. pog ores de l 
Neuquén, m 58' S, 68703", J 
[LL Pi], Ке 2257 (BAA) їп, "bi and St]. Rí 
Depto. Avellaneda, ruta 22, 50 km antes de Choel Chocl. 
39°10 'S, 65°05'W, 6 Nov. 1972, Bacigalupo & Nicora 
11646 (SI) [LI, Pi]; DP Торе ruta 251 a 13 km al 
de Genera о Соно. 40°01'S, 


i 
Piccinini & Leguizamón 1610 (BAA) [LL Pi]; ruta За 8 
km sur de San Antonio, 40?45'S, 64°56’W, Correa & Ni- 
cora 3193 (BAA) [LI, Pi and St]. Santa Cruz: Depto. 
Deseado, Camino a Cabo Blanco, Tellier, entre Estancia 
El Chara y Tellier, 47?26'S, 66°02'W, Correa et al. 3362 
BAA) [LI, Pi and St]. 


~ 


Poa pilcomayensis 


ARGENTINA. Entre Rios: Depto. Concordia, Parque 
Rivadavia, 31723'S, 58°01’\, 22 Sep. 1961, Burkart 
22456 (SI) [PL Pi and St]; Depto. Gualeguay, La Verde, 
21 Nov. 1949, Burkart 18064 n [РІ, Pi and St]; Islas 
Lechiguanas, Delta medio frente а Ramallo, 33?29'S, 
6001", 30 Dec. 1941, Burkart 12874 (SI) [PI, St]; Dep- 
to. Victoria, Isla del Francés frente a Rosario, 32°57'S 
60*39' W, 15 Dec. 1937, Burkart 8854 (SI) [PI, Pi and Stl, 
Burkart 8817 (SI) [PL Pi]. 


Poa pogonantha 


ARGENTINA. Patagonia: Port Eden, 24 Jan. 1879, 
Savatier 1844 (Festuca pogonantha Franch., holotype, 
BAA fragment) [PG, Pi]. Chubut: Depto. Futaleuft, 
Parque Nacional Los Alerces, Laguna Cisne, 42°30'S, 
dod W, 16 Dec. 1962, Roquero 5389 (BAA) [PG, Pi]; 
. Río Sengüerr, Lago Fontana, Estancia La Pepita, 
44°55 S. 70°58’ W, 29 Jan. 1960, Soriano 5662 (BAA) 
[РЕ, Pi]; Lago Fontana, Lote 15, 44?56'S, 71°30’\, Mar- 
tinoli & Boggiano 15080 (BAA) [PR, Pi and Si} tus. ш 
& Boggiano 15079 (BAA) [PR, St], 11 Feb. 
tellanos 9968 (BAA) [PR, Pi], Martinoli ^ ы 
15077 Fei [PR, 5]; Martinoli et Boggiano 15078 


(ВАА) [PR, St]; Lago Fontana, Estancia La Pepita, 
i 5, vod W, 29 Jan. 1960, Soriano 2618 (BAA) 
Pi]; Depto. Tehuelches, Lago Vintter, 43?58'S, 


is "d W, 2 Feb. 1989, Nicora 9473 b (SI) [PG, St]; Lago 
Vin laya arenosa, 43°58'5, 71733", 7 Feb. 1989, 
Nicora 9537 (SI) [PG, Pi]. Río Negro: Depto. Bariloche, 

arque Nacional Nahuel Huapi, Ventisquero Frías, 
41°11'S, 71?49'W, 13 Jan. 1952, Boelcke & Correa 5500 
(ВАВ) [PG, Pi and St]; Laguna Frías, 41°04’S, 71?49' W, 
8 Jan. 1952, Boelcke & Correa 5373 (BAB) [PG, Pi], Boel- 
cke & Correa 5380 (BAA) [PG, Pi]; Lago Roca, Arroyo 
Apoco, 41°23'S, 71°47' W, 26 Jan. 1952, Boelcke & Correa 
6045 1/2 (BAA) [PG, St]. Tierra del Fuego e Islas del 
Atlántico Sur: Depto. Ushuaia, Glaciar Martiales, 
54^45'S, 68°18'W, 9 Dec. 1962, Correa et al. 5365 (BAA) 
[PG, Pi]. CHILE. ХИ Región: Lago Azul, 51°27’S, 


230 


Annals of the 
Missouri Botanical Garden 


73^18'W, 10 Jan. 1977, Moore & Pisano 1575 (BAB) [PG, 
Pi]; Islas Wollaston, Caleta Lientur, 55?44'S, 67°19'W, 23 
Jan. 1980, Pisano 5112 (SI) [PG, St]; Estancia Cerro Cas- 
tillo, Cerro Solitario, 51?20'S, 72°37'W, 18 Jan. 1977, La- 
tour et al. 1739 (ВАВ) [PG, Pi]; Península W пад а 
cadura Río Serrano, 51?25'S, 73?04'W, 23 Jan. 

Moore & Pisano 1905 (BAB) [PG, Pi], Moore 367 (SI) т 
Pi]. 


Poa resinulosa complex 


Poa buchtienii: BOLIVIA. La Paz: Manco Kapac, Lago 
Titicaca, Isla del Sol, 16?03'S, 69?10'W, 26 Jan. 1986, 
Liberman 1289 (28) [BT, St]; Murillo, La Paz-Calacoto, 

acia el n o Illimani, 16?40'S, 67°45'\, 19 
Jan. 1983. Beck 9076 (LPB) [BT, Pi]. Oruro: Avaroa en- 
tre Challapata у Tolapalca, 19*20'S, 66°50'W, Feb. 1979, 
Ceballos et al. 236 (SI) [BT, Pi]; La Paz, 18°36’S, 
66*55'W, Feb. 1911, Buchtien O. 2467 (syntype, BAA) 
[BT, St]; Poopo, a 4.5 km al norte de La Paz, sobre la ruta 
hacia Oruro, 16?20'S, 68?05' W, 6 Mar. 1993, Peterson et 
al. 12714 (LPB) [BT, Pi and St]. Potosí: Surchichas, Po- 
1081, а 12 km al NW de Salo, 20°55’S, 66?18' W, 21 Mar. 
1992, Peterson & Annable 11823 (LPB) p 9 and eh 
Tarija: Avilez Cerca de Cobres, 21°38’S, ; 
Јап. 1986, iei Ne (LPB) [BT, Pi and B : B 
quiensis: ARGENTINA. Jujuy: Depto. DR. Manuel Bel- 
grano, Camino de n de Alto Lozano a Tiraxi, 24?05'S, 
65740", 3 Nov. 1974, Correa et al. 6106 (SI) [CL, St]; 
Depto. Humahuaca, Mina Aguilar, 23?12'S, 65741", 23 
Feb. 1972, Ruthsatz 14577 (BAA) [CL, Pi]; Abra entre 
Iruya e Iturbe, 22°58’S, 65?21'W, 25 Jan. 19 die Ruthsatz 
14589 (BAA) [CL, St]; Depto. er Bárbara, El Fuerte, 
22°07'S, 65?26'W, 18 Feb. 1972, Ca s et al. 22236 
(BAA) [CL, Pi]; Minn adig Cerro Tuzgle, 23"26'5, 
66°30'W, 3 Маг. 1967, Werner 76 (LP) [CL, Pi]; Depto. 
Tumbaya, Volcán, ‘Abe га del Paraguay, 23°55’S, 65°27'W, 
Feb. 1927, Venturi 4905 (SI) [CL, St]; Depto. Yay), Yavi 
Chico, 22°06'S, 65°28'W, 7 Маг. 1940, Meyer 14926 
(BAA) [CL, St]; Quebrada de Roquero, 22°18'S, 65°35’ W, 
20 Feb. 1963, Cabrera 15369 (SI) а ni ami Depto. 
Guachipas, Pampa Grande, 25°52’S, 6 r. 1900, 
Holmberg 2616 (BAA) [CL, Pi]. поени D Taff, 


Cumbres Calchaqufes, 26°35’ 5, 65°40'W, 29 Jan. 1907, 
Lillo 5065 A р [CL, St]. Р ни PARA- 
GUAY. Caaguaz 25*01'S, 55°58'W, 19 Sep. 1988, 


Pedersen 15049 e SI) [PE, Pi и ST. P. resinu- 
losa: ARGENTINA. Buenos Aires: Part General 
Pueyrredón, Estancia La Brava, Sierra de Valdes, 37°54'S, 
58°00'W, 18 Nov. 1977, Boelcke et al. 791 (SI) [RE, Pi]; 
— Tornquist, Estancia Chica y Estancia ddp Cha- 
co, Cerro La Vieja, 38*06'S, 62°14’ W, 17 Nov. 1981, Vi- 
Шат 2073 (SI) [RE, Pi S Su; Se ла | Mamim- PA 
Cerro del Potrero, 38%03'5 56'W, 18 Nov. 1981, V 
Пати 2110 (SI) [RE, St], cmd 2054 (SI) [RE, Pi ind 
t]. Chubut: Depto. Futaleufti, Estancia Sufiica, 43*03'S 
1°04'W, Lahitte s.n. (BAA) [RE, Pi]; Parque Nacional 
Los Alerces, Río Percey, Carre Ceballos, 42°55'S, 
71°20'W, Lahitte & Roquero 192 (ВАА) [RE, St]; Depto. 
Languifieo, Río Tecka, Pampa Chica, 43?28'S, 70°51'W, 
8 Nov., Skottsberg s.n. (P. decolorata Pilg., isotype, BAA 
fragment) [RE, St]. Córdoba: Depto. Río Seco, Саћа Cruz 
m de Villa de María, 29°54’S, 63*44'W, 7 Nov. 
1949, Hunziker 8003 (SI) [RE, St]. Mendoza: Mendoza, 
Gillies s.n. (isotype, BAA, fragment) [RE, Pi]. — nin 
Depto. Huiliches, Parque Nacional Lanin, a Lago Trom 
39*34'S, 71°32'W, Lahitte et al. 606 (BAA) IRE, Pi ind 


St]; San Martín de los Andes, camino entre Lolog y Ma- 
muil-Malal, 39°40'S, 71722", Neumeyer 31 (ВАА) [RE, 
Pi and 51]; Depto. Los Lagos, Estancia Fortín Chacabuco 
41*02'S, 71°15’ he е 3210 1/2 (BAA) i E ind 
St]. Santa Fe: Depto. Vera, Calchaquí, 29°54’ 8'W, 
7 June 1965, Alonso 866 (SI) [RE, Pi]. 


Poa rigidifolia 
age aie Santa Cruz: Depto. Спег Aike, Estan- 


a Stag River, meseta Latorre, Cerro Punta Gruesa, 
51-328, 72°05'W, 25 Jan. 1978, Roig et al. 2950 (ВАВ) 
le Pi]; Estancia Punta Alta, 51743'S, 71°58’W, 24 Jan. 

976, Latour et al. 536 (ВАВ) [RI, Pi]; Estancia Punta 
Lov 51744'5, 68*56' W, 1976, Nicora 828 (BAB) [RI, 
Ру; 42 km oeste de Estancia Punta del Monte, cruce a 
Sección Magán, 51732'S, 71735'W, 1977, Roig & Méndez 
2494 (BAB) [RI. Pi and St]; entre Estancia Pose y 
Laguna Condor, 51?48'S, 71°42'W, 14 Dec. 1976, Latour 
et al. 1057 (BAB) [RI, St]; Estancia и, гша 293, 
51727'5, 72°15'W, 1976, Latour et al. 1198 (BAB) [RI, 
Pi]; Punta Loyola, 51750"5, 69*11'W, 6 Dec. 1976, Latour 
: al. 906 (ВАВ) [RI, St]; Bajo La Leona, 51°31'S, 
6946", 11 Nov. 1977, Roig & Méndez 2437 (BAB) [RI, 
St]; Río Gallegos, entre Estancia Maragata y Las Buitrer- 
аз, 51^45'S, 70^10'W, 10 Dec. 1975, Arroyo et al. 410 
(BAB) [RI, St]; Estancia Guakenken Aike, 51?27'S, 
69°48'W, 14 Nov. 1977, Roig & Méndez 2485 (ВАВ) [RI, 
Pi and St]. Tierra del Fuego e Islas del Atlántico Sur: 
Depto. Río Grande, Estancia El Salvador, 53?39'S, 
68°35'W, 20 Nov. 1971, Boelcke et al. 15093 (ВАВ) [RI, 
Ру; 51 km N de San Sebastián, Estancia Cullen, 52°53’S, 
68°30'W, 21 Nov. 1971, Boelcke et al. 15156 (BAA) [RI, 
Ру; Estancia Secunda Argentina, Jan. 1933, Castellanos 
7597 (BAA) [RI, Pi]; А е William, 
52°00'S, 59°00'W, Sep. 1850, Lechler s.n. (P. rigidifolia 
ери holotype, жа пау Pi]. Da re 
estate, Mar. 6, Philippi s.n. (P. poecila Phil., 
ODE ВАА шы. [RI, Рі; Sandy Point, 53°10 'S, 
70°54'W, Oct. 1852, Lechler 10686 (Aira spiciformis 
Steud., holotype, BAA apap [RI, Pi]. 


Poa rigidifolia var. ibari 


ARGENTINA. Chubut: 
44°39'S, 66°10' W, 19 Oct. 


Depto. Ameghino, Lochiel, 
1946, Soriano 1915 (B AA) 


Mes г. E Chico, Chimen Aike, 51744 
Sleumer 790 (BAA) [IB, Pi]; pu de la 

nien. Ghies, a Punta Alta, 51?27'S, 72°06'W, 5 
Feb. 1978, Ambrosetti & ucl 3747 SN Ме. Pi]; 
Sección San D 51°24” 34'W, 19 Jan. 1978, 
Roig et al. 2756 (BAA) ES E and ed Estancia ee 
51°50'S, 69*11'W, 8 Dec. 1976, Latour et al. 877 (BAB) 
1]; Bajo La Leona, А 69°46'W, 1977, Roig & 
Méndez 2436 (BAB) ПВ, St]; Estancia Los Vascos, 
5'S, 70°52'W, 21 Jan. 1978, Roig et Men 2877 (BAB) 
i Ю km al NW 
go Argentino, 50° 3 "5, 71*40' W, 
v. 1963, Du. et "pe 2888 (BAA) [DU, Pi and St]; 
que Пе: Patagonia orientalis ad Mazaredo ропшт, 
47905'5, 66°42'W, Jan. . Dusén 5318 (P. dusenii 
Hack., holotype, BAA fragment) [DU, Pi]; Camino а 


Volume 87, Number 2 
2000 


iussani 231 
Phenetic Patterns in Poa Sect. Dioicopoa 


rto Deseado, 47?48'S, 66?12' W, 18 Nov. 1963, Correa 
et ны 2590 (ВАА) [DU, Pi and St]; Depto. Río с 
rnador Gregores, Hotel Las 


I into, 
72°24’ W, Jan. 1844, Ibar s.n. (Р ibari Phil., isotype, ВАА 
т [IB, Pi]. 
Poa schizantha 


ARGENTINA. Buenos Aires: Partido Bahía Blanc 
Bahía Blanca, 3844 5, 62"16' W. Nov. 1941, Zafjanella 


M ‚е 
las dunas maríti 38°59'5 '\ 8 Nov. 0 Pa- 
rodi 13672 (holotype, BAA, SI) [SC, Pi], Parodi 13673 
(BAA) [SC, St], Parodi 13675 (BAB) [SC, Pi], Cabrera 


6752 (BAA) [SC, Pi], Jan. 1941, Zaffanella 14402 (BAA) 


с, 


Poa tristigmatica 


ARGENTINA. Chubut: Departamento Río Sengiierr, 
Pampa de ge Estancia La Media Luna, 45°35'S, 

ei Villamil 2236 (SI) TR. St]; Lien 
5 37, Martinoli & и ne 
(BAA) |ТЕ, Pi]; Depo Tehuelches, Gober 


uqué epto. 
pelco, = е filo, 40°09’S, 71°20'%, 23 Feb. 1974, Correa 
et al. 5926 (P. boelckei Nicora, holotype, ВАВ) [BK, Pi], 
Correa et = 5928 (BAB) [BK, Pi]; Cordón del Cerro Сћа- 
pelco, Portezuelo Trahunco, Faldeo S, 40?09'S, 71720", 
13 Feb. 1978, Gentili 695 (BAB) [BK, St]; Cerro Chapel- 
co, 40?09'S, T Feb. 1961, León & Calderón 963 
(BAA) [BK, Pi], 26 1966, Eskuche 603-2 (BAA) [BK, 
St]. Eskuche 603-1 UPS [BK, St], Eskuche 599-3 
[BK, Pi], 12 Jan. 1961, León & Calderón 903 (BAA) [ВК, 
Pil; nolan Minas, Paso del Macho, 36°26'S, 70°46’ W, 26 
Jan. 1970, Boelcke et bs 13931 (ВАА) [TR, Pi], Boelcke 
et К 13926 (B. ) [T nd Laguna oF Campos, 
Cajón Benítez, paso a Vieja, 36°17'5, 70741", 1 
Nov. 1970, Boelcke et a 1428) (BAA) [TR, Pil сш 


Е 964 
(BAB) [TR, St]; Cajón del Portillo, 
70°36’ W, 31 Jan. 1970, Boelcke et al 14168 (BAA) [TR, 
Pi and St]; Cordillera del Viento cruzada de Tricao Маја! 
al Cajón de Butaló, en vertiente, 36°58'S, 70*30'W, З Nov. 
1964, Boelcke et al. 11567 (BAA) ipd St]; Depto. Ñor- 


ín, ei 37°49'S, 71?06'W, 3 . 1930, Hirsch- 
horn 23 (BAA) [TR, Pi and St]. Rio Negro: Depto. Na- 
1 Huapi, Cerro Catedral, 41°05’S, 71945, Feb. 1954, 


1831, Сау 49 (Р. a ia E. Desv., in Gay, syntype, 
Pi]. 


BAA fragment) [TR, 
APPENDIX 3. 
orphological variables used in numerical analys 
their ге айыда | in ане ig codification, and brief 
explanations of character variation 
VEGETATIVE VARIABLES 
l. e habit (HAB). е ог short-rhizomatous 


toloniferous (3). Plants 


long- rhizomatous as P. lanuginosa. Caespitose plants 


sometimes develop aiti as in P. alopecurus or long 

es as in P. tristigma 
2. Leaf pedes (LEAle, ст). This i is a variable character 
within Poa 


with sheath and leaf blade length, plant height, pan- 
icle length, and number of nodes on principal panicle 
axis. The highest value, 93 cm, was measured on Р 
не [о the minimum value, 3.5 ст, согте- 
onded to P. rigidifolia. 
3. Sheath length (SHEle, cm). The highest value found, 
cm, correspon nded to 5 bergii and P. stuckertii, 
ib the minimum value, 1.7 cm, was measured on 
P. rigidifolia. 
4. Leaf blade length (BLAle, cm). The highest value 
found, 61 cm, corresponded to P. dolichophylla, n 
the minimum value, 1.3 cm, was measured on Р ri 


idifolia. 
5. Ligule length (LIGle, mm). This character ео а 
conspi cuous discontinuity, dividing Poa sect. Dioi- 
d species and Tien liguled 
as measured in 
n P. pedersenii. 
6. Lra shape (LIGsp). лаје (1); truncate to ођ- 
tuse (2). Long ligules are usually acuminate (P. ligu- 


7. Leaf blade apex (API). Navicular or ae (1); sharp 
or acuminate (2). Poa sect. Dioicopoa is characterized 
by a navicular or obtuse leaf blade apex as in Р. lani- 
gera. Sharp or acuminate apices also occur within 
species such as P. hubbariana and P. ber, 


BLADE CROSS SECTIONS 


8. Blade ош пе (OUTIi). Flat (1); conduplicate (2); 1 соп- 
volute ог subconvolute (3). Species of the Р do 
ae Puro are [rh onim by E 

schizantha nts volute to 
ts id Blade, rad pati. ah the ab- 
sence of bulliform cells; blades in other species vary 
from conduplicate to subconvolute. 

9. Blade width, measured on the adaxial epidermis be- 
tween blade margin and midrib (BLAw, mm). This 
measure generally correlates with the number of vas- 
cular bundles with sclerenchyma girders on both ad- 
axial and abaxial epidermis. The highest value was 
recorded in P. iridifolia, 6.8 mm, and the minimum 
value was found in P. ligularis, 0.37 mm. 

10. Blade maximal thickness between abaxial and adaxial 
epidermis layers (BLAt, mm). This character fre- 
quently correlates with и length. The maximum 
а 0.55 pee was seen in P. Re с the тіп- 

value, 0.1 n P. pilcom 

11. Bulliform call dedita ent BULeo). Not mi little. dif- 
ferentiated (1); inflated, and well developed (2). Poa 


drib as in P. holciformis. Poa schizan- 

bulliform cells, 
while these are generally little differentiated in P. po- 
gonantha. 

12. Shape of sclerenchyma at blade margin (CAP). 

tending on abaxial or adaxial epidermis (1); crescent- 

shaped cap, with sclerenchyma briefly extending on 


232 


Annals of the 
Missouri Botanical Garden 


abaxial and adaxial epidermis (2) Ld of 

states as in Ellis, 1976). Poa sect. Dioicopoa gener- 

ally presents a group p of sclerenchyma cells at "lade 

bod seen in cross section. Terminal sclerenchy- 

are ommonly о to pointed like т 

P rigidifelio, with species of t du ophylla 
complex presenting a crescent- shape 


рй» complex (45), whereas minimum 
only 1 or 2 were found in P. rigidifolia, Р ligularis, 
and the P. 


girder only on abaxial epidermis (SCLab). These are 
commonly few (1 or 2) in the Р dolichophylla com- 
plex, but are more numerous (5 to 8) in P. bergii. 


t. Dioicopoa, they are often present (3 to 5) 
in the P dolichophylla complex 

16. Number of vascular bundles saith only a few scleren- 
chyma cells surrounding vee m n This 
character is variable in species of Poa . Dioico- 
poa, si e few даны ла od 
to this 


ABAXIAL BLADE EPIDERMIS 


17. Stomata length (STOM, mm). This character, usually 
indicative for different ploidy levels, presents three 
ranges of vari Ра 


phylla complex, Minas thier: species [us inter- 
ediate values. 

18. Thickness of the long cells (CELt, mm). This varies 

between 0.016 mm in Р. ligularis and 0.033 mm in 


P. lanuginosa. 

19. Abaxial epidermal prickles (PRIC). Absent to infre- 
quent (1); frequent to numerous (2). Prickles are gen- 
erally ш; distributed on ү epidermis, al- 
though they are infrequent in the Р о 
сотрјех, Р. bergii, P. КЫ айал, P lanigera, P. p 
comayensis, and P. tristigmatica. 

20. Silico- süberose cell pairs on intercostal epidermis 


(SISU 


condition in Poa sect. Dioicopoa, though they are in- 
frequent in P. lanigera and P. pilcomayensis. 


FERTILE VARIABLES 


21. Plant height, i from the longest culm of fer- 


tile plants (HEIG, с 


corded in the P. dolichophylla jae (136 с hi: the 
smallest plant, in P. rigidifolia (6 cm 
22. Number | culm nodes (CULM). A vertes between 
n small plants and 4 to 5 in taller plants. 
23. sign lai (PANle, cm). The longest panicle (37 
was recorded in P. dolichophylla; the shortest (1.7 
cm) was in Р. rigidifolia. 
24. Panic le width, measured on widest panicle (PANw, 
cm). Panicles are contracted within Poa sect. Dioi- 
copoa, with the widest recorded in P. bonariensis (2— 


26. 


21. 


Ме 
20 
~ 
© 


w 


— 


МАЈ 


w 
D 


. First glume 


5 cm). The Poa dolichophylla complex резе» broad 
panicles (5—8 cm), which are usually expanded 


| Number of nodes on principal panicle axis (PANn). 


to only 6 in Р. alopecurus, P. pogonantha, and P. rig 
a 

Len of terminal, well-developed spikelet of a 
branch panicle (ЗРПе, mm). Well-developed spikelets 
were selected from the upper half of a panicle, par- 
eb from terminal spikelets of a branch. Spikelet 
length and correlated measures do not usually va 
within the same panicle. The highest value was found 
in P. pogonantha (28 mm), : 


with the shortest among species of the Р dolicho- 
phylla De. (2.8 mm). 

Width of terminal well-developed spikelet of a branch 
panicle (SPIw, mm). This character correlates with 
flower number per spikelet. The broadest spikelet was 
recorded in P. bergii (10 mm). 


~ 


. Number of florets per spikelet (FLOn?). Spikelets in 


a sect. Dioicopoa generally have 3 to 5 flowers. 
d in P. schizantha, and 11 
in P. bergii. The minimum e Pid (2). is com- 
monly present among species 


| Viviparous florets (FLOv). in (0); present (1). Vi- 


re 
season, sa as P. alopecurus, P pogonantha, and P. 
tristigma 


. First iue length (GLUle, mm). This correlates “ш 


lemma and palea length as well as 


i e specimens of P bergii (10.3 
2 ae (8.9 mm). The : ripa glumes 
were recorded in a staminate specimen of P. dolicho- 
phylla (1.38 mm). 
DB sured from principal nerve to 
1m). This varies from 0.33 mm in Р. 
: in P. 
in P h 
Ratio of first glume length/sec a glume length (eu 
G2). Equal to 1 (0); less than 1 (1); m 1 (2). 


Having the first glume shorter than Tena sec ка glume 


glume length/lemma length (G2/LE). 
to 1 (0); less than 1 (1); more than 1 (2). The 
se econd glume is generally shorter than the lower lem- 
in Poa sect. Dioicopoa. 

Number of nerves on the first glume (GLUn). Glumes 
in Poa sect. Dioicopoa generally present a principal 
nerve and two marginal nerves (sometimes absent). 
Poa bergii pec two additional marginal nerves (in 
total, 3 to 5 o 7). 


. Prickles on he, rac ichilla (RACH). Absent to infrequent 


YT 


(0); frequent to numerous prickles (1). Poa sect. Dioi- 
copoa generally present these prickles on rachilla and 
panicle axes. 

Lemma length (LEMle, mm). Dimorphism between 
pistillate and staminate plants impacts lemma lengths 
within species of Poa sect. Dioicopoa. Species dis- 
crimination is considered among specimens of the 


Volume 87, Number 2 
000 


Giussani 233 
Phenetic Patterns in Poa Sect. Dioicopoa 


Qo 
24 


o 
© 


~ 
Кој 


= 
м 


B 
— 


mm); the minimum value in species of the 
P. resinulosa complex (0.54 mm). 

. Palea length (РА е, mm). The shortest palea was re- 
corded on a staminate specimen of the P. resinulosa 
complex (1.75 mm); the longest on a staminate spec- 
imen of P. alopecurus (6.72 mm). 

. Palea width between nerves (PALw, mm). Dimorphism 
between pistillate and staminate plants is not always 
evident in palea width. The maximum value was re- 
corded in P. bergii (1.8 mm); the minimum value cor- 
responds to species of P. pilcomayensis (0.30 mm). 

. Lodicule length (LODle, mm). Only entire and well- 
developed lodicules were considered for recording 
data. The longest lodicules were measured on P. alo- 
pecurus, P. bergii, P. indigest lanuginosa, P. po- 
pot P. rigidifolia, and Р tristigmatica (1.2—1.7 
mm long). The shortest lodicules were recorded on P. 
bonariensis and Р. ligularis (0.25—0.33 mm long). 

. Lodicule width, including lobes (LODw, mm). Species 
variation within Poa sect. Dioicopoa ranges between 
0.2 mm and 0.7 mm wide, being more than 1 mm 


~ 


= in a few specimens of P. huecu and P. tristig- 
atica. 


: Hairs on the callus of the first floret (HAIcal). Absent 


(0); rigid and short, less than 1/2 of the floret (1); 
rigid and long, more than 1/2 of the floret (2); woolly 
and short, less than 1/2 of the floret (3); woolly and 
long, more than io of the floret (4). Hairiness is a 

hic character between pistillate and staminate 


florets. Pistillate foots usually have a hairy callus in 
ct. Dioico, nd pistillate florets of only P. 
indigesta, P. huecu, P. holciformis are complete] 


js they can also present a few rigid or woolly, long 
r short hairs. 


3. Hanes along lemma nerves lie gs Absent (0); 


scabrous (1); уа less than 0.5 m ; hairs more 
than 0. 5 mm (3). Pistillate florets of Poa sect. Dioi- 
copoa are characterized by the presence of hae on 
principal and marginal nerves that are 0.5 mm long 

lanigera, P. ligularis, P. 
lanuginosa, and P. hubbardiana present hairs longer 
han 0.5 mm, these being conspicuous and abundant 
in P. hubbardiana. Staminate floret nerves are typi- 
cally glabrous 


. Pubescence between principal and secondary lemma 


nerves (HAlbet). Absent (0); scabrous (1); hairs less 


rigidifolia, Tr [^ than 0.5 mm long well as 
abundant and longer in P. hubbardiana. 


STUDIES IN ANNONACEAE 
XXXVI. THE DUGUETIA 
ALLIANCE: WHERE THE 
WAYS PART! 


Lars W. Chatrou,? Jifke Koek-Noorman,? 
and Paul J. M. Maas? 


ABSTRACT 


Results of a cladistic analysis of morphological and anatomical data of the Duguetia alliance (Annonaceae) are 


the past distinction of Pachypodanthium from Duguetia appears to be absent. Characters used to resolve relationships 


the tw 


between and within 


o clades are difficult to polarize by outgroup comparison. It is de 


monstrated that the critical 


reassessment of classical morphological characters, and the search for new ones, may well advance phylogenetic res- 


olution within Annonaceae. 


ords: Annonaceae, cladistics, Duckeanthus, Duguetia, Fusaea, Letestudoxa, morphology, Pachypodanthium, 


Pseudartabotrys. 


The classification of the Annonaceae presents 
workers on this family with a Herculean challenge. 
Early classifications of the family, as in Hooker and 
Thomson (1855), emphasize identification and only 
incidentally reflect phylogeny. This also applies to 
a more recent classification (Hutchinson, 1964). 
However, at higher, tribal levels, these two classi- 
fications have little in common. То date, the clas- 
sification by Fries (1959) resolves most subgroups. 
Exclusively on the basis of inflorescence and floral 
characters, he distinguished two subfamilies, three 
tribes, and 14 informal genus groupings. The com- 
position of many of these genus groups has been 
amended after phenetic analyses of flower and fruit 
morphology (van Heusden, 1992; van Setten & 
Koek-Noorman, 1992; Koek-Noorman et al., 1997), 
and phylogenetic analyses based on gross morpho- 
logical and palynological data (Doyle & Le Thomas, 

996, 1997) 


One of Fries's genus groups is the Duguetia al- 
liance, comprising West African and tropical Amer- 
ican genera. The composition of this alliance re- 
mained untouched to date except for the exclusion 
of Malmea and the inclusion of the monotypic ge- 
nus Pseudartabotrys (van Setten & Koek-Noorman, 


1992). Characteristic features of the genera belong- 
ing to this alliance include valvate sepals, imbri- 
cate petals, one basal ovule, and the presence of a 
rudimentary aril. Most distinctive for the alliance 
is the presence of pseudosyncarpous fruits. These 
are aggregates of astipitate carpels, which become 
fused with one another and/or adnate to the recep- 
tacle. Genera usually considered to fit into the Du- 
guetia alliance are the Neotropical genera Duck- 
eanthus (1 sp.), Duguetia (95 spp.), and Fusaea (2 
spp.), together with Letestudoxa (3 spp.), Расћуро- 
danthium (4 spp.), and Pseudartabotrys (1 sp.) from 
West Africa (van Setten & Koek-Noorman, 1992; 
Le Thomas et al, 1994; Koek-Noorman et al., 
Van Heusden (1992) dissentingly placed 
Duguetia and Pachypodanthium in one informa 
group, separate from Duckeanthus, Fusaea, Letes- 
tudoxa, and Pseudartabotrys (plus Afroguatteria, 
Enicosanthellum, and Disepalum) in another. 
us, the majority opinion on the circumscrip- 
tion of the Duguetia alliance seems to prevail. Yet 
closer examination reveals some problems. Recent 
cladistic analyses (Doyle & Le Thomas, 1994, 
1995, 1996, 1997; Doyle et al., 2000) array Du- 
guetia, Pachypodanthium, Letestudoxa, Fusaea, 


The authors thank Jan van Veldhuizen, Jos van der Maesen, and Jan Wieringa of the National Herbarium of the 
arira Wageningen Agricultural University branch, for РИМА herbarium на ud сне апа рћо- 
Rain a, Jan Maas echni 


r for information o on Ann 
ч rt and Peter уап и Welzen је: hos. ind diseussion during the 


stance, Hendrik 


onymous reviewer for critical comments manuscript. 


Financial support was provided by the Netherlands а for Scientific ae (NWO; uem no. . 805. 40.201) 
d. 


to the first author, which is gratefully acknowledge 


? National Herbarium of the Netherlands, He University branch, Heidelberglaan 2, 3584 CS Utrecht, the Neth- 


erlands. 


ANN. Missouni Bor. GARD. 87: 234—245. 2000. 


Volume 87, Number 2 
2000 


Chatrou et al. 
Duguetia Alliance 


and Duckeanthus in one clade, linked to the clade 
of the "xylopioids." Pseudartabotrys was not in- 
cluded in these analyses. 

Pseudosyncarpy, which otherwise only occurs in 
the Annona group, is an obvious synapomorphy for 
this “Duguetia clade." During pseudosyncarpous 
fruit development the postgenital aggregating of the 
carpels can occur through two processes, viz. the 
lateral fusion of carpel walls, and the inclusion of 
the very basal parts of the carpels into the fruiting 
receptacle. The former case has been extensively 
documented for the genera Annona and Rollinia by 
Впесћје-Маск (1994). The fusion of carpels starts 
with dovetailing of the epidermal cells of adjacent 
carpels, and ends with complete fusion. The fruit- 
ing receptacle does not contribute to the aggregat- 
ing of the fruit. А similar fruit development has 
been described for Fusaea (Chatrou & He, 1999). 
The inclusion of the very basal parts of the carpels 
into the fruiting receptacle (see Svoma, 1998) orig- 
inates from acropetal development of the receptacle 
after flowering has been completed. This type of 
aggregating of the fruit is present in all species of 
Duguetia, whereas the degree of lateral fusion of 
the carpels varies from completely free to complete- 
ly fused among the species of this genus. Thus, 
both origins of pseudosyncarpy occur within the 
Duguetia alliance and can be traced when closely 
inspecting fruit morphology. Pseudosyncarpy 
should therefore be considered as a non-homolo- 
gous similarity. However, the fruit type can be in- 
corporated into analyses in a more straightforward 
way by unravelling it ontogenetically (Patterson, 
1982). In this analysis we will consider the differ- 
ential origins of pseudosyncarpy, separating it into 
two morphological characters (see Data and Anal- 


A family-wide phenetic analysis based on flower 
and fruit morphology resulted in inclusion of Duck- 
eanthus, Letestudoxa, Pseudartabotrys, Fusaea, and 
Pachypodanthium in one cluster (Koek-Noorman et 
al., 1997). Duguetia appeared in another cluster, 
together with Guatteria and the Annona group. Nev- 
ertheless, the overall similarity between all genera, 
including Duguetia, was perceived so strongly by 
the authors that the signal appearing from the 
phenogram was ignored, and all genera were 
grouped together in a tentative scheme of genus 
groups. In the same paper, Koek-Noorman et al. 
(1997) concluded that some of the principal char- 
acters used by Fries (1959) for the distinction of 
genus groups, viz. sepal and petal aestivation, bare- 
ly contribute to their phenetic clustering. 

The following paradox thus emerges: a genus 
group, or clade, has long been recognized intui- 


tively but is weakly supported by morphological ev- 
idence. Schatz and Le Thomas (1993) stated that 
confusing phylogenetic pattems based on macro- 
morphological character distribution within Annon- 
aceae have been clarified during the past two de- 
cades by new palynological and karyological 
evidence. In spite of its general validity, this state- 
ment cannot be applied to the Duguetia alliance. 
Karyological evidence is too scattered to be un- 
equivocal (Doyle & Le Thomas, 1996). Palynolog- 
ical data reveal too many autapomorphies among 
the genera of this alliance to be illuminating. Based 
wholly on palynological data, Walker (1971) even 
erected the informal Fusaea subfamily, accommo- 
dating Fusaea and Duckeanthus, but placed Du- 
guetia in another subfamily. Walker's data were re- 
interpreted by Le Thomas (1980-1981) and Le 
Thomas et al. (1994). However, Le Thomas et al. 
(1994) did not clarify the phylogeny of the Dugue- 
tia alliance with pollen ultrastructural data, but 
conversely discussed the implications of their re- 
sulting phylogenies for the evolution of pollen mor- 
phology. 

Doyle and Le Thomas (1996) stated that given 
the high level of morphological homoplasy in An- 
nonaceae, only molecular analysis might be able to 
resolve higher-level relationships. The Duguetia al- 
liance was addressed by van Zuilen (1996) with her 
cladistic analysis of trnL-F sequences, combined 
with morphological characters, favoring the inclu- 
sion of Duguetia, Fusaea, Pachypodanthium, and 
Pseudartabotrys as one clade. Duckeanthus and Le- 
testudoxa were not included in her analysis. 

Except for most seed characters, many of the 
morphological characters used in the above-men- 
tioned analyses still are conventional characters in 
a Friesian vein (e.g., Fries, 1934, 1959), which 
have been subject to little recent critical revision. 
Moreover, regarding the reticulate nature of char- 
acter expression in Annonaceae, the taxonomic lev- 
el at which a phylogenetic analysis is performed 
determines the character choice. Contrasting with 
a family-wide phylogenetic analysis, an analysis at 
the tribal or genus group level requires different 
data matrices informative only for the particular 
group examined, as was elegantly shown by John- 
son and Murray (1995) in their analysis of the tribe 
Bocageeae. 

In this paper we address the phylogeny of the 
Duguetia alliance sensu Koek-Noorman et al. 
(1997) and Le Thomas et al. (1994), by conducting 
a cladistic analysis based on leaf, flower, fruit, and 
seed characters, many of which have not been used 
in cladistic analyses of Annonaceae before now. We 
provide the rationale for the recent submersion of 


236 Annals of the 
Missouri Botanical Garden 


Table 1. Data matrix with taxa, characters, and character states used. The species abbreviations, as used in Figure 


2, are in parentheses. *: African species of Duguetia. ?: character state unknown. 


1 2 

123456789012345678901234 

Annona sericea Dunal (Annser) 000000000000000000100001 
Duckeanthus grandiflorus R. E. Fr. (Рисрта) 000010200001001020110011 
Duguetia argentea (R. E. Fr.) R. E. Fr. (Dugarg) 020001000020000000000110 
uguetia asterotricha (Diels) R. E. Fr. (Dugast) 010000000120020200001110 
Duguetia barteri (Benth.) Chatrou* Dugbar) 011001002120000101000110 
Duguetia confinis ( ee & Diels) ето Dugcon) 011001002120000101000110 
Duguetia dilabens Chatrou & Repetur* Dugdil) 011001000020000101000110 
ANGE S alee (A. s -Hil.) end & Hook.f. Dugfur) 020001000020200100000110 
uguetia inconspicua Sagot Duginc) 010000000020000101000110 
Duguetia eie e A. St.-Hil. Duglan) 020001000020200100000110 
Duguetia neglecta Sandw. Dugneg) 010000001010000000000110 
Duguetia quitarensis Benth. Dugqui) 020001000020000100000110 
Duguetia riberensis Aristeg. ex Maas & Boon Dugrib) 011001001020000200000110 
Duguetia riparia Ни ак ег Dugrip) 010000000120000000000110 
Duguetia spixiana Dugspi) 020001000021020000000110 
pee staudtii а: & Diels) Chatrou* Dugsta) 011001000020000101000110 
Duguetia uniflora (DC. ex ad Mart. Duguni 010000000020000110000110 
Fusaea ое (Aubl.) Safi Fuslon) 000110110121011020110001 
usaea peru а В. E. Fr. Fusper) 000110110121011020110001 
Letestudoxa bella Pellegr. Letbel) 100100110011121120100011 
Let xa glabrifolia Chatrou & Repetur tgla) 100100100011?21?20100011 
Letestudoxa lanuginosa Le Thomas (Letlan) 100100100011221?2010???? 
Pseudartabotrys letestui Pellegr. (Pselet) 100100000011101120101001 


1—habit; 0 = tree/treelet, 1 = liana 

2—trichomes; 0 = simple, 1 = bip os = lepidote (+ stellate) 

3—leaf shape asymmetric; 0 = no, 

4—secondary veins on upper side h- leaf: 0 = = flat to нЕ 1 = impressed 
5—весопдагу veins joining to form marginal vein; 0 = no, 1 = у 
6—mesophyll type; 0 = dorsiventral, 1 = isobilateral 


7—histology of primary vein; 0 = phloem and/or ` sclerenchyma surrounding xylem body, 1 = phloem and/or scle- 
le 


renchyma surrounding and intruding xylem body, 2 = phloem only abaxially accompanying xylem 
8—curly trichomes; 0 = absent, 1 = present 
9— inflorescence position: terminal on ee axillary shoot; 0 = never, 1 = ee 2 = always (ordered) 
10— inflorescence: abnormal displacement of prophyll; 0 = i ent, is presen 
11—bracts; 0 = non-cucullate, scale-like, 1 = non-cucullate, foliaceous, 2 - = cucullate 
12—pedicel abruptly and distinctly widening into flowering rec о 0 = по, 1 = yes 
13—flower color (in vivo); 0 = white to cream, 1 = yellow to ora 


ge, 2 = pink to red 
14—fusion sepals; 0 = free or basally connate, i — suben usb connate, rupturing longitudinally, 2 
connate, rupturing irregularly 
5 


о, 1 = yes 
16—stamen color (in vivo); О = white-cream-yellow, 1 = pink to red 
17—stamens sclerified; 0 = no, 1 = only on peru E 2 — on both sides 
18— position of anther thecae; 0 = 6 1 = Іа 
19—style; 0 = absent, 1 = presen 
20—styles coherent by sie arem papillae: 0 = no, 1 = yes 
21— sepals persistent in fruit; 0 = 
22— basal carpels fused with fruiting Node 0- пена 1 = yes 
23—fruiting receptacle protruding between carpels; yes 
24—direction of aril fibers; О = toward distal pum P seed, 1 = toward proximal епа of seed 


entirely 


Pachypodanthium into the synonymy of Duguetia аз well as for most Annonaceae (Schatz & Le 


nthium will uS be referred to as “Afri- 
can species of Duguetia." А biogeographical ques- DATA AND ANALYSES 


(Chatrou. 1998). Species formerly known as Pachy- Thomas, 1993). 
po 


tion comprises whether the break-up of West The data matrix includes 23 taxa and 24 char- 
Gondwana was a vicariance event for this subgroup acters (Table 1). All species of Duckeanthus, Fu- 


Volume 87, Number 2 
2000 


Chatrou et al. 
Duguetia Alliance 


237 


saea, Letestudoxa, and Pseudartabotrys are includ- 
ed in the analysis. (For vouchers, see Appendix 1.) 
All 4 African species, and 11 Neotropical species 
of Duguetia are selected out of ca. 95 that consti- 
tute the genus. Annona sericea is included as out- 
group taxon for two reasons. First, Annona is in- 
cluded in a clade that is directly linked to the 
pseudosyncarpous clade on the basis of sequence 
data (van Zuilen, 1996). Furthermore, of the genera 
that appear close to the pseudosyncarps in van Zu- 
ilen’s analysis, Annona is the only genus with pseu- 
dosyncarpous fruits, and (often) with seeds provid- 
ed with a rudimentary aril. Therefore, the scoring 
of characters 22—24 for the outgroup is enabled. 
The character set has been designed to incorporate 
independently evolving morphological and anatom- 
ical data from different plant parts. It comprises 17 
binary characters and 7 three-state characters. Of 
the latter characters, only character 9 is quantita- 
tively ordered, scoring for the relative abundance 
of the particular inflorescence position within a 
species. Although our objectives pertain to genera, 
the characters are scored at the species level. This 
allows the scoring of several characters with incon- 
sistent character states within a genus. Characters 
3, 6, 8, 9, 10, 11, 12, 13, 14, 16, and 18 are het- 
erogeneous within a particular genus, while homo- 
geneous within another. Duguetia possesses multi- 
ple states for all of these characters except for 
trichome character 8, while one of the other, non- 
monotypic genera is scored uniformly. Scoring at 
the species level also 
question of the relationship between the Neotropi- 
cal and African species of Duguetia. Bootstrap val- 
ues of a previous phylogenetic analysis of Duguetia 
inspire little confidence in the clades found, nor in 
most of the sections of Duguetia as recognized by 
Fries (van Zuilen et al., 1 
choice of the 11 species for our analysis is such 
that they display the phenetic variation within the 
genus well. We decided to include Letestudoxa lan- 
uginosa in the analysis, despite the fact that its 
fruits are unknown, and therefore characters 21—24 
could not be scored. Inclusion may possibly allow 
more insight into the evolution of flower color (char- 
acter 13), as L. lanuginosa has its flower color in 
common with only two species of Duguetia (D. fur- 
furacea and D. lanceolata). The missing values for 
characters 21–24 do not present any problem for 
the analysis, as they are simply treated as uninfor- 
mative 


permits us to address the 


erefore, our 


Some of the characters in our analysis are 
straightforward morphological (or anatomical) ones. 
ther characters have hardly been documented, or 


are ate for the first time, with the following 
explanation 

ne 7: histology of the primary vein. De- 
tails hereon can be found in van Setten and Koek- 
Noorman (1986). In their survey of leaf anatomy of 
Annonaceae, Duckeanthus has not been taken into 
account. We sectioned leaf parts of D. grandiflorus 
according to the same methods as described in van 
Setten and Koek-Noorman (1986). The histology of 
the primary vein of Duckeanthus shows a pattern 
that is hitherto unknown in Annonaceae. The phlo- 
em only abaxially accompanies the xylem (Fig. 1A). 
This pattern is an autapomorphy of Duckeanthus, 
and we scored it as a separate character state. 

Character 8: curly trichomes. Curly trichomes 
have been described for Fusaea (Chatrou & He, 
1999) and Letestudoxa (Chatrou, 1998). In both 
genera these trichomes occur on the lower side of 
the leaves, on the petioles, and on the young twigs. 
In Letestudoxa they occur on the outer side of the 
calyx as well. Besides curly trichomes, normal 
straight trichomes occur as well in both genera. 

Character 9: position of inflorescence. Two Af- 
rican species of Duguetia exclusively have terminal 
inflorescences on reduced axillary leafy shoots. Le 
Thomas (1969) described them as axillary for D. 

arteri. Two Neotropical species of Duguetia (D. 
neglecta and D. riberensis) exhibit the same position 
of the inflorescence, though not in all cases. Hence, 
the latter two species have been scored as 1. The 
position of the inflorescences in other Duguetia 
species is terminal on leafy twigs, and never on 
reduced axillary leafy shoots. 

Character 10: inflorescence, abnormal displace- 
ment of prophyll. This phenomenon has been de- 
scribed for inflorescences of Fusaea (Chatrou & 
He, 1999), and is also present in four species of 
Duguetia. Normally subsequent fertile prophylls al- 
ternate at angles of 180°. In Fusaea, D. asterotri- 
cha, D. barteri, D. confinis, and D. riparia the pro- 
phylls alternate at angles of ca. 90° only. 

Character 11: shape of bracts. Cucullate bracts 
have been documented for Fusaea by Chatrou and 
He (1999), and have been found in all species of 
Duguetia, except for D. neglecta, which has folia- 
ceous bracts. 

Character 17: sclerified stamens. Van Heusden 
(1992) mentioned the occurrence of indurate (more 
or less lignified) stamens in Duckeanthus, Fusaea, 
Letestudoxa, and Pseudartabotrys. We made medial 
cross sections for at least 10 stamens per species, 
staining with Astra-blue and Safranin, to check for 
the occurrence of sclerenchyma. Two basic patterns 
were found: (1) sclerenchyma is either absent; or 
(2) sclerenchyma is present on both the entire inner 


238 Annals of the 
Missouri Botanical Garden 


Sclerenchyma 
ИЩ Phicem 
Xylem 

A Parenchyma 


Figure 1. —A. ра drawing а transverse section through primary vein of Duckeanthus grandiflorus. В, С. 
Cross sections through stamens. —B. Fusaea longifolia, showing sclerified tissue (dotted region) and extrorse locules. 
—C. Duguetia staudtii, rus latrorse booa nd no те D, Е. s transversely sectioned through raphe, 
showing direction of aril fibers. —D. Directed toward proximal епа о E aas longifolia. —E. Directed toward 
distal end of seed: Duguetia confinis. Scale bars: A-C = 0.1 mm; D, E = 1 mm. 


Volume 87, Number 2 
2000 


Chatrou et al. 239 


Duguetia Alliance 


side, as well as on the outer side between the the- 
cae (Fig. 1B, C). Only Duguetia uniflora showed an 
intermediate pattern, with sclerenchyma found only 
on the inner side of the stamen, and absent between 
the thecae. 

Characters 19 and 20: styles. For Annonaceae, 
the presence of a style is often difficult to judge. 
Fusaea has clearly distinct ovaries, styles, and stig- 
mas. Its transition between ovary and style is in- 
dicated by a constriction, and by differences in epi- 
dermal outgrowths, and in shape in transverse 
section. The stigma can be discerned from the style 
by differences in color and epidermal outgrowths 
(Chatrou & He, 1999). The same pattern occurs in 
Duckeanthus, Letestudoxa, and Pseudartabotrys. As 
in Fusaea, the styles of Duckeanthus interlock by 
means of papillae. In those species of Duguetia ex- 
amined, a clear distinction can be seen between 
the ovary and the apical part of the carpel, but 
subsequent transitions are absent. Therefore a style 
is considered to be absent in these genera, in spite 
of their presumed presence according to van Heus- 
den (1992) and Doyle and Le Thomas (1996). 

Characters 22 and 23: fruit type. Among fruits 
of different species of Duguetia, different degrees 
of fusion of the carpels occur. However, in all spe- 
cies the fruiting receptacle protrudes between the 
carpels by acropetal growth (Svoma, pers. comm. 
1996). In fruits with a low degree of carpellary fu- 
sion this is very noticeable, especially when dried. 
Here, the surface of the receptacle shows shallow 
concavities in which the carpels are loosely posi- 
tioned. Yet even in fruits with a high degree of car- 
pellary fusion (e.g., Duguetia furfuracea, D. barteri) 
the protrusion of the receptacle between the carpels 
is discernible. Fruits of Duckeanthus and Letestu- 
doxa have free, stipeless carpels, attached to the 
fruiting receptacle in shallow concavities, resulting 
in a functional syncarp (Schatz & Le Thomas, 
1993) similar to those of Duguetia. An important 
difference between fruits of Duckeanthus and Le- 
testudoxa on the one hand, and those of Duguetia 
on the other, is the position of the basal sterile car- 
pels. In the latter genus, these basal carpels insep- 
arably coalesce with the fruiting receptacle and 
constitute a proximal collar on the fruiting recep- 
tacle. In Duckeanthus and Letestudoxa, the basal, 
sterile carpels contribute to the functional syncarp 
and readily detach from the fruiting receptacle. 

Character 24: direction of aril fibers. Arillate 
seeds are found in all species of the Duguetia al- 
liance. The aril is considered to be rudimentary 
(van Setten & Koek-Noorman, 1992) as it covers 
considerably smaller parts of the seed than seen in 
species of the tribe Bocageeae. Among the Dugue- 


tia alliance, the aril develops from the base of the 
testa, distinguished from the other parts of the testa 
by its closely packed, long parallel cells (Garwood, 
1995). We found that these rudimentary arils fur- 
ther assort into two types. The first type has the 
long, parallel cells directed toward the distal end 
of the seed, while in the second type they are di- 
rected proximally (Fig. 

he data were analyzed using PAUP version 
3.1.1 (Swofford, 1993). Heuristic searches for most 
parsimonious trees were performed by random step- 
wise addition with 100 repetitions, the Tree-Bisec- 
tion-Reconnection (TBR) branch swapping algo- 
rithm, and the MULPARS and STEEPEST 
DESCENT options in effect. Only minimal trees 
were retained, and zero-length branches were bro- 
ken down with the COLLAPSE option. The use of 
either the DELTRAN or the ACCTRAN optimiza- 
tion criterion produced identical tree topologies. 
Bootstrapping was performed with the TBR swap- 
ping algorithm, simple addition sequence, and 250 
repetitions. The relative robustness of the clades 
was assessed additionally by performing a decay 
analysis (Bremer, 1988; Donoghue et al., 1992) for 
all clades of the strict consensus tree. Character 
evolution was analyzed using MacClade 3.04 (Mad- 
dison & Maddison, 1992). 


RESULTS AND DISCUSSION 


Parsimony analysis of the data matrix in Table 1 
resulted in 63 shortest trees of 49 steps. All 63 
trees belong to one island of trees, with each tree 
connected to every other tree in the island through 
a series of trees, and each one differing from the 
next by a single rearrangement of branches (Mad- 
dison, 1991). The strict consensus tree has a con- 
sistency index (CT) of 0.65 and a retention index 
(RI) of 0.85 (Fig. 2). Bootstrap values are indicated 
above the nodes for each clade of the consensus 
tree that is maintained after bootstrap analysis. 
Bootstrap values = 50 are given. Bootstrap values 
= 70 are considered to be high. Our consensus tree 
satisfactorily meets the conditions under which 
bootstrap values = 70 correspond to a probability 
of = 9546 that the corresponding clade accurately 
reflects the true phylogeny (Hillis & Bull, 1993). 
Only the condition of internodal change of = 2096 
is not fully met: the basal nodes with bootstrap val- 
ues of 95 and 88 both have an internodal change 
of 2596 of the characters. Decay values are indi- 
cated below the nodes. We were unable to realize 
a decay analysis in which trees of three steps longer 
were retained. The large number of trees resulting 


240 


Annals of the 
Missouri Botanical Garden 


with the first three letters of both g 
the nodes. Decay values are indicated below the nodes. Amer. 


Annser Amer. 


Dugarg Amer. 


66 [ —- Dugfur Amer. 


d1 


60 


d Г Duglan Amer. 


Dugqui Amer. 


Dugspi Amer. 
[ Dugast Amer. 


d1 


mE c Dugrip Amer. 


95 


Duginc Amer. 


Dugneg Amer. 


Dugrib Amer. 


87 г Dugbar Afr. 


d1 


da LL Dugcon Afr. 


Dugdil Afr. 


Dugsta Afr. 


Duguni Amer. 


| Ducgra Amer. 


d»2 


77 


d2 


65 


98 Fuslon Amer. 
d>2 Fusper Amer. 


Letbel Afr. 


d2 


Letgla Afr. 


Letlan Afr. 


Pselet Afr. 


Figure 2. Strict consensus tree of 63 most parsimonious trees after analysis g. Taxa are рея 


taxon 


eneric name and epithet (see also Table D. e е are indica 


and Afr. indicate American or African ина of 


Volume 87, Number 2 
2000 


Chatrou et al. 
Duguetia Alliance 


Table 2. Synapomorphies for the combined Duguetia-Fusaea clade, and for the Duguetia clade and the Fusaea 
clade separately. The number of the character as described in Table 1 is included in parentheses. 


Duguetia-Fusaea 
clade Duguetia clade 


Fusaea clade 


No synapomorphies (19) style absent 


(22) basal carpels fused with fruiting 
receptacle 


(23) fruiting receptacle protruding be- 


els 


(12) pedicel abruptly and distinctly widening into 
flowering receptacle 

(15) petals velutinous 

(17) stamens sclerified on both sides 


tween carp 
(24) aril fibers directed toward distal 


end of see 


from this search exceeded the maximum number of 
trees that PAUP can retain. 

The high amount of synapomorphy, as expressed 
by the high RI, the high bootstrap values, and the 
highest decay values for (1) the clade formed by all 
species of Duguetia (Duguetia clade), and (2) the 
clade formed by Duckeanthus, Fusaea, Letestudoxa, 
and Pseudartabotrys (Fusaea clade), arouse high 
шр in these clades. These results disap- 

ve placement of all six genera into one clade or 
Mane: (Doyle & Le Thomas, 1994, 1996; Koek- 
Noorman et al., 1997; Le Thomas et al., 1994), and 
support the distinction made among them by van 
Heusden (1992). The inclusion of Afroguatteria, 
Disepalum, and Enicosanthellum into the Fusaea 
group by van Heusden (1992), however, is contra- 
dicted by strong evidence, both from general mor- 
phology as well as from molecular evidence (Doyle 
& Le Thomas, 1994, 1996; Koek-Noorman et al., 
1997; Doyle et al., 2000 

African species of obs form a relatively dis- 
tinct clade with Duguetia riberensis within Dugue- 
tia. Continued recognition of Pachypodanthium 
would have rendered Duguetia paraphyletic, and 
consequently Pachypodanthium species recently 

ave been transferred to Duguetia (Chatrou, 1998). 
In the past Pachypodanthium has been considered 
to be different from Duguetia primarily on the basis 
of wood anatomy and palynology. Vander Wyk and 
Canright (1956) pointed out a difference in vessel 
density, being low for Pachypodanthium and high 
for Duguetia. Increased sampling has rendered the 
argument untenable (Ter Welle, pers. comm. 1997) 
Pollen grains of the African species of Duguetia 
possess an extremely reduced exine consisting of 
only spinules. Le Thomas et al. (1994) interpreted 
these spinules as homologous with verrucae in the 
Neotropical species, which also show varying exine 
reductions. Both pollen and wood indicate that Du- 
guetia, despite its uniform appearance and its pre- 
sumable monophyly (van Zuilen, 1996), remains a 


variable genus. Such is also demonstrated by our 
data matrix, with by far the largest part of the ho- 
moplasy deriving from Duguetia. The type of tri- 
chomes (character 2), asymmetric leaves (character 
3), inflorescence position (character 9), and position 
of the thecae (character 18) represent features in 
our data matrix for which the African species seem 
anomalous. These aberrant character states are also 
encountered in a small subset of the Neotropical 
species of Duguetia. This reticulate nature of char- 
acter expression, which was also found in a study 
of Duguetia leaf anatomy (Bakker & Visser, 1994), 
requires broader sampling of species; this is under 
way in a forthcoming analysis of Duguetia (Koek- 
Noorman & Maas, in prep.). 

What are the character states that identify the 
combined Duguetia-Fusaea clade, the Duguetia 
clade, and the Fusaea clade? We traced all char- 
acter states at the ingroup node, as well as at the 
internal nodes basal to the Duguetia clade and the 
Fusaea clade, respectively. The Duguetia-Fusaea 
clade is only characterized by traits that appear 
extensively within the Annonaceae, either by par- 
allelism or by mosaic retention. Examples of these 
features include attributes such as trees, simple tri- 
chomes, symmetric leaves, free or basally connate 
sepals, and white to cream flower color. Thus, the 
Duguetia-Fusaea clade is not characterized by any 
synapomorphies (Table 2). The only character state 
that comes close to being synapomorphic is cucul- 
late bracts, present in all Duguetia (except D. neg- 
lecta) and in Fusaea. However, it is one step more 
parsimonious to assume parallel evolution in Du- 


clade are characterized by four and three synapo- 
morphies, respectively (Table 2 

To resolve relationships within the Duguetia-Fu- 
saea clade we added some characters to the matrix 
that are novel, and which do not appear in the ma- 
jority of other Annonaceae. This novelty does not 


242 


Annals of the 
Missouri Botanical Garden 


imply the change of one character state to another 
character state, and consequently these characters 
are difficult to polarize by outgroup comparison. 
This affects inflorescence character 10, implying a 
sympodial development of the inflorescence. The 
lack of any fertile inflorescence bracts in the tribe 
Bocageeae, and the presence of lower bracts that 
do not produce axillary buds in most Xylopia and 
many Guatteria, illustrate the paucity of sympodi- 
ally developing inflorescences in Annonaceae. 
Character 20 for stylar papillae can only be scored 
for those few annonaceous species in which the car- 
pels are provided with a style. Receptacular char- 
acters 22 and 23 require the presence of pseudo- 
syncarpous fruits, which outside the group under 
study are only present in Anonidium and in the 
Annona group. Arillate character 24 can only be 
scored for a small group of genera where the seeds 
possess a rudimentary aril (van Setten & Koek- 
Noorman, 

Considering this, outgroup comparison is diffi- 
cult to use to polarize the set of characters at hand. 
Possible outgroup taxa may be selected from pre- 
vious phylogenetic analyses. Doyle and Le Thomas 
(1994) found Toussaintia and the xylopioids con- 
nected basally to the pseudosyncarps, all of them 
together forming a monophyletic group. Van Zuilen 
(1996), based on limited sampling, however, found 
the Duguetia-Fusaea clade attached to a clade 
comprising Uvaria, Uvariopsis, Isolona, Monodora, 
Annona, and Rollinia. All these genera suffer from 
the comparative lack of the above-mentioned char- 
acters. The only exceptions are genera from the An- 
nona group, which share sympodially developing 
inflorescences, pseudosyncarpous fruits, and rudi- 
mentary arils with the Duguetia alliance. 

The character set as herein designed cannot es- 
tablish the monophyly of the combined Duguetia- 
Fusaea clade, and cannot yield a corroborated po- 
sitioning of the clade within the Annonaceae, as 
the selected characters obscure the choice of out- 
group taxa. For the monophyly of the combined Du- 

uetia-Fusaea clade we rely on Doyle and Le 
Thomas (1996) and Le Thomas et al. (1994). 

Our study, as well as the cladistic analysis of the 
tribe Bocageeae (Johnson & Murray, 1995), shows 
that the search for new morphological characters 
with little generality, can be illuminative for anal- 
yses at low taxonomic level. The combined Dugue- 
tia-Fusaea clade (the *pseudosyncarps") shows по 
synapomorphies or unique combinations of char- 
acters. The challenge for future research will be to 
find synapomorphies that circumscribe more inclu- 
sive, monophyletic groups, to find new characters 
with more generality. Otherwise, it will be difficult 


to bring the phylogeny of Annonaceae beyond float- 
ing groups of genera with rather well resolved in- 
ternal relationships. 

Considering the geographical distibution of the 
taxa (Fig. 2), the pattern is straightforward, deriving 
from the break-up of Gondwana. Both the Duguetia 
clade and the Fusaea clade apparently existed be- 
fore this event. Within the Fusaea clade, closer 
phylogenetic relationships correspond with geo- 
graphical proximity. Fusaea and Duckeanthus 
evolved as the Neotropical representatives, and Le- 
testudoxa and Pseudartabotrys as the African ones. 
We cannot address whether the biogeographic iso- 
lation of the African species of Duguetia corre- 
sponds with their constituting a separate clade 
within Duguetia, or whether some of the African 
species have closer sister-group relationships with 
Neotropical species. Analyses including more spe- 
cies of this genus, which will have to resolve this, 
are forthcoming. 


Literature Cited 


Bakker, M. E. & W. J. Visser. 1994. Studies in Annon- 
aceae XIX. пи anatomy of Duguetia St. Hil. (Annon- 
aceae). Bot. Jahrb. Syst. 116: 83-111. 

Bremer, K. 1988. The limits of amino acid sequence data 
in angiosperm phylogenetic reconstruction. Evolution 
42: 795—803. 

Впесћје-Маск, М. Н. 1994. isla zur Histogenese der 
Blüten und Früchte pseudosynkarper Annonaceen-Ar- 
ten. Deutsche Hochschulschriften 1028. Hünsel-Hoh- 

ch. 


hanging с сенн Stud- 
ies in Neotropical "me West African Annonaceae. Ph.D. 
Dissertation, Utrecht University, Ur ins 

P. He. 1999, Studies in A eae XXXIII. 
A revision of Fusaea (Baill.) Saff. Ванта 52: 181- 
203. 


Donoghue, M. J., R. G. Olmstead, J. F. Smith & J. D. 
Palmer. 1992. ренина. Mais of i 
based on rbcL sequences. Ann. Missouri Bot. Gard. 
333-345. 

Doyle, J. А. & A. Le Thomas. 1994. Cladistic e 
and pollen evolution in Annonaceae. Acta Bot. Gallic 


141: 149-170. 


& 995. Evolution of pollen characters 
and relationships of African n саней 
of a cladistic analysis. Pp. ~ in A. Le Thom 
m de Palyn ologie af 
International po 
la Formation et les Exchanges Géologiqu s. Orleans. 
Phylogenetic analysis “ 
сһагас!ег evolution in Annonaceae. Bull. 
Hist. Nat nos 18: 279-334. 

& - а E palynology for 

phylogeny of о Ехрепте bd removal of 
pollen characters. Pl. Syst. Evol. 206: 9. 
, P. Bygrave & A. Le Thomas. 2600. eu ИИ 
of rer: data for pollen evolution in Annonaceae. 
Pp. 259-284 in M. M. Harley, C. M. Morton & S. Black- 
more (editors), Pollen and Spores: Morphology and Bi- 
ology. Royal Botanic Gardens, Kew. 


Volume 87, Number 2 
2000 


Chatrou et al. 
Duguetia Alliance 


243 


Fries, R. Е. 1934. Revision der Апеп einiger Anona- 
ceen- m III. Acta Horti Berg. 12(1): 1-220. 

. 1959. Annonaceae. Pp. | прјег 
K. Prantl (editors), Die већа Pfanzenfamilien, 
2nd ed., 17аП. Duncker & Humblot, Berlin. 

Garwood, N. C. 1995. Studies ac 
phology and ecology of seedlings, fruits and seeds of 
selected Panamanian species. Bot. Jahrb. Syst. 117: 1– 


Имде, E .C.H 1992. Flowers of Annonaceae: 
Morphology, алайы, and evolution. Blumea 
uppl. 7: 1-218. 

Hillis, D. M. & J. J. Bull. 1993. An empirical test of 
bootstrapping as a method for assessing confidence in 
phylogenetic fei Syst. Biol. 42: 182-192. 

rs ‚ J. D. & T. Thomson. 1855. Flora Indica 1: 86– 

153. W. cae London 

Hutchinson, J The бане era of € Plants 
Sess die Ww 1. P свеце Press, Oxfor 

Johnson, D. M. & N. A. Murray. 1995. ynop sis of the 
tribe Bocageeae о un revisions of Cardi- 
opetalum, Froesiodendron, Trigynaea, Bocagea, and 

ittoni 19. 


K. van Setten & C. M. van Zuilen. 
1997. — in ро XXVI. Flower and fruit 
morpho logy in Annonaceae. Their contribution to pat- 
terns in aln dae Bot. Jahrb. Syst. 119: 213- 
230. 


Le Thomas, A. Annonacées. Pp. 1-371 in A. Au- 
p eres cin du и Vol. 16. Muséum па- 

nal d’histoire naturelle, Pari 

1981. а characters of the 

#90 grains of African Аппопасеае and their signifi- 

cance for the phylogeny of the primitive angiosperms 

Pollen & = 22: 267-342, 23: 5— 

‚ B. Lugardon & J. A. Doyle. 1 Pollen ultra- 
structure and relationships of Fusaea ee Safford 
and Duguetia A. Saint-Hilaire (Annonaceae). Rev. Pa- 
laeobot. Palynol. 83: 

Maddison, D. R. 1991. The Ни and importance of 
multiple islands of most-parsimonious trees. Syst. Zool. 
40: 315-328. 

Maddison, W. P. & D. R. Maddison. 1992. MacClade, 


version 3.04. P Associates, Sunderland, Massa- 


usetts. 

Рено С. 1982. Morphological characters and ho- 
mology. Pp. 21-74 in К. A. Joysey & riday 
(editors), Problems of Phylogenetic Reconstruction. The 
Systematics Association Special Volume no. 21. Aca- 
demic Press, London and New York. 

Schatz, G. E. & A. Le Thomas. 1993 dine. d A 
primitive dicot family with an ancient center rica— 
South America. Pp. 86—104 in P. Goldblatt (edili Bi- 
ological к Between Africa and South Amer- 

v. Press, New Haven. 

n & J. Koek-Noorman. 1986. Studies in 

Пике VL A leaf anatomical survey of genera of 

Annonaceae in the Neotropics. Bot. Jahrb. Syst. 108: 

7-50. 


—. 1992. Fruits and seeds of Annona- 
ceae. Morphology and its significance for classification 
and M lorc in Annonaceae XVII. Bib- 


lioth. 142: 1- om Schweizerbart’sche Verlags- 
менам и: 
Зуота, Е. 1998 


Fruit hon diversity in Annonaceae. 
Annonaceae Newslett. 12: 68— 


Swofford, D. L. 1993. РАЏР: Ранке Analysis Us- 


ing Parsimony, version 3.1.1. Computer program dis- 
tributed by the Illinois Natural History Survey, Cham- 
paign, Illinois. 

Vander Wyk, R. W. & J. E. Canright. 1956. The anatomy 
e relationships of the Annonaceae. Trop. Woods 104: 


Valker J. W. 1971. Pollen morphology, phytogeography, 
phylogeny of the Annonaceae. Contr. Gray Herb. 
= P 131. 
Zuilen, Ae M. van. 1996. Patterns and Affinities in the 
Duguetia Alliance (Annonaceae). Molecular and Mor 
ied Studies. Ph.D. Dissertation, Utrecht аи 
sity, Utrecht. 


, J. Koek-Noorman & Р. J. M. Maas. 1995. А 
phylogenetic analysis of Duguetia Mi rupe based 
on morphological data. Pl. Syst. Evol. 194: —188. 


Appendix 1. Voucher specimens for taxa studied. 
Annona sericea Dunal 

FRENCH GUIANA. Saint-Maurice-Region de Saint 
Laurent, Piste d'Apatou, 8 Feb. 1990 (fr), Cremers & Hoff 
11256 (P, U, US). GUYANA. East Berbice-Core vs 

ear Thompson's farm (Timehri), 

Maas et al. 5932 (NY, U, WU). U 
bice: vicinity of Mabura Hill, trail from Mabura Hill-Lin- 
den road to Demerara Landing, 26 Aug. 1988 (fl buds, ба 
bes et al. 7144 (K, P, U, WU). SURINAM. Marowi 

ong road near Mongotapoe, 10 June 1954 (fl buds, vin), 
rne 6133 (NY, U). VENEZUELA. Bolívar: Street 
Alcabala Casa Blanca towards Isla Anacoco, 28 July 1981 


5. 
5 
ч 
S 
ч 
= 
-- 
$ 
LI 


Rio Negro, 13 Oct. 1932 (fl), Ducke 23904 B, K, 'RB, 

US); right bank of Rio Negro, Ilha Tamanduá ner ha 

Marajó), near Сагарапа, 18 Oct. 1987 (fl), M. 

6772 (МУ, U), Maas et al. 6778 (NY, U); Пћа P Flores, 

Rio Negro, 17 Feb. 1959 (fr), Rodrigues (S). 

Duguetia argentea (R. E. Fr. 2: E. Fr. 
RAZIL. Amazonas: km 124 of Manaus-Porto Velho 

Hwy., 25 Mar. 1974 (fr), Tice et al. P20915 (G, 

INPA, U); ws e A чы Rio Negro, 17 Oct. 1987 (fl), 

IA. 


Maas et al. 6764 (NY, U). COLOMB azonas-Vau- 
pés: R па cd Jine Gojé, arri as Piraparaná 
and Río Popeyaká, Cafio Unguyá —1 


1952 (fl, fr), Garcta-Barriga E (COL, US). 
ZUELA. Amazonas: arlos de Río Negro, 21 
17 Apr. 1981 (yfr), Delascio С. et al. чи (VEN); Isla 
Sebástian, Río Casiquiare, between Boca and Chapazón, 
alt. 120 ue 31 Jan. 1980 (fl, fr). pow & Clark 8939 
MO, U, VEN). 
Duguetia meds (Diels) R. 11 Fr. 

AZI zonas: km 118 of Manaus—Caracaraf 
Hwy., Mar. 1976 (fl), D. ai & Damião 767 (INPA); 
Reserva Florestal Ducke, alt. 80 m, 16 Jan. 1990 (fr), 
Gentry & Revilla 69142 (U); ibidem, 18 Jan. 1990 (fl), 
Gentry & Nelson 69219 (U); Manaus, 2 km from Taru- 
mazinho, 18 Nov. 1975 (8), О. P. Monteiro INPA53548 
INPA); km 70 of Мапаиз—ПасоаНага Hwy., 31 Мау 1994 
(8), Webber 1477 (HUAM). PERU. Loreto: Mishuyacu, 
near Iquitos, Oct.-Nov. 1929 (fl), Klug 86 (F, NY, US). 
Duguetia barteri (Benth.) Chatrou 

N. Centre-Sud: bank of the Nyong River, 

40 km SE of Yaoundé, alt. 550 m, 9 Nov. 1961 (fr), Bre- 
teler 2013 (BR, K, Р, WAG). Littoral: right bank of Опет 


— 


~ 


244 


Annals of the 
Missouri Botanical Garden 


River, near "opp with pepe River, 6 km SW of 


‚ 4 Apr. 1965 (fl), Leeuwenberg 5377 


; Lombé, ew ss 6 July 
1976 (fr), McKey & Gartlan 139 (Е). GABON. Ogooué- 
Ivindo: Ipassa, 10 km 5 of Makokou, small island in 


Ivindo River, alt. = bs 27 ed nie (ir), "Hladik 1835C 
bee Ipassa, 10 km ako vindo River, alt. 500 
, 14 Mar. 1975 ra 'Hladik 2641 (P). NIGERIA. Ondo: 
Ma rbara Rise Ibaji-Ojoku Reserve, 11 Mar. 1934 (fr), 
Taylor 13 (FHO). 
uguetia confinis (Engl. & Diels) Chatrou 
CAMEROON. Cen tre-Sud: ca. 16 km from Kribi, 


Zenker Кеч (В, BR, E, G, GO 
Z). GABON. Estuaire: near Libreville, Apr. 1897 (fl), 
oyen-Ogooué: SW of Lambaréné, near 
anga, Conoco drilling site, alt. 20 m, 3 Feb. 1991 

(fr), McPherson 15195 (MO). Nyanga: 10 km on Maambi 
iver, ug. 2 (fl), Wieringa ~ van de Poll 1360 
о е: Rabi, Shell camp, alt. 
1994 (fl, fr), Wein & van Nek 3290 


Klaine s.n. (P). 


(WAG). Og 
50 m, 26 Nov 


(W. 
Duguetia dilabens Chatrou & Repetur 
GABO werten forest reserve of Kienké Kribi, 
Ebolowa km 16, 5 Jan. 1968 (fr), Bamps 1679 (BR). Lit- 
toral: Lombé, Tissongo, Е: Aug. 1976 (fr), МсКеу & Gar- 
tlan 194 (K). Ngounié: new road from Mouila to Yeno, 5 
km on either side of Kembele village, alt. 500 m, 20 July 
1986 (fl, fr), Thomas & Wilks 6510 (MO, P, WAG) 
Duguetia furfuracea (A. St.-Hil.) Benth. & Hook.f. 
BOLIVIA. Santa Cruz: Prov. Velasco, Serrania de 
uanchaca, alt. 800 m, 3—4 Dec. 1987 (fl, fr), Thomas et 
al. 5578 (U). BRAZIL. Bahia: 15-20 km from Andarai, 
along the road to Itaeté which branches E off the road to 
Mucugé, alt. 500—600 m, 13 Feb. 1977 (fl, fr), Harley et 
al. 18634 (CEPEC, F, IPA, K, MO, NY, P. U, US). Dis- 
trito Federal: 20 km S of Brasília, on dd to Belo Ho- 
rizonte, alt. 000 m, 26 Aug. 1964 (fl), Irwin & Sod- 
erstrom 5572 (NY, S, SP, TEX). Goiás: Mun. Mineiros, 14 
km E of turn-off for Mineiros, 1 Feb. 1986 (fl, fr), An- 
e & pes 1635 (GB, U). Mato Grosso: Mun. 
mpo Grande, road from Campo Grande to Rochedo, 12 
July aor (8, fr), "Hatschbach & Guimarães 21837 (S, UC). 
as Gerais: Serra do Espinhaço, 6 km N of Gouvêia 
on =. to Diamantina, alt. 1250 m, 10 Apr. 1973 (fl, = 
W. nderson et al. 8585 (F, MO, NY, RB, U, UB 
São Paulo: Fazenda Hollambra, 35 km оси аѕ, 
alt. 600 т, 25 Feb. 1976 (fl), Shepherd & ‘Gibbs 11246 
(K, MG, NY). PARAGUAY. Amambay: 14 km S of Bella 
Vista, alt. 250 m, 25 Mar. 1983 (fl), Simonis et al. 197 
(AAU, F, G, U). 
Duguetia inconspicua Sagot 
RAZIL. Amapá: Rio Araguari, camp 13, 9 Oct. 1961 
(fr), J. M. Pires et al. 51633 (NY). Pará: basin of Rio 
Trombetas, 3 km up Rio Mapueira from Cachoeira Por- 
teira, 30 May 1974 (fr), Campbell et al. P22301 (NY, U). 
FRENCH GUIANA. Mt. Bellevue de l'Inini, alt. 700 m 
17 Aug. 1985 (fr), de Granville et al. 7580 (B, CAY, P, U). 
GUYANA. Seballi Compartment, ca. 3 km S of Mabura, 


F, K, LZ, M 
Marowijne River, alt. 430-520 m, 31 Dec. 1954 (fr), Cow- 
an & Lindeman 39044 (NY, S, U, US). 
Duguetia lanceolata A. St.-Hil. 

BRAZIL. Minas Gerais: Lagoa Santa, 8 Маг. 1865 (fl, 
fr), Warming s.n. (C, F, K, NY, P, S). Parana: Sengés, 29 


June 1910 (fl buds), Dusén 9939 (GH, NY, S). Santa Ca- 
tarina: Vargem Grande, Lauro Müller, alt. 350 m, 24 Oct. 
1958 (fl), Reitz & Klein 7483 (B, K, S). Sào Paulo: Mun. 
Brotas, Pepira-Mirim, Experimental Station of Mogi-Gua- 
cu, Arboretum, alt. 500 m, 22 Sep. 1992 (fl), Maas et al. 
8043 (LZ, U, UEC, ULM, WU). 

Duguetia neglecta Sandw. 

YANA. Base of Mt. Makarapan, near rapids of Mak- 
arapan Creek, 15 Sep. 1988 (fr), Maas et al. 7433 (B, 
BBS, Е, MO, МУ, U, VEN, WIS); Mabura Hill Nature 
Reserve, 25 Aug. 1990 (fl, fr), Polak et al. 28 (U); Lab- 
bakabra Creek, Tiger Creek, Essequibo River, 26 Aug. 
1937 (fl), Sandwith 1214 (G, K, NY). SURINAM. Area of 
Kabalebo Dam project, along road between km 29 and 30, 
1 Sep. 1980 (fl, fr), Lindeman, Górts-van Куп et al. 59 (F, 
K, NY, U 
Duguetia quitarensis Ben 

BOLIVIA. Pando: Río Madeira, 12 km above Ађипа, 
20 July 1968 (yfr), Prance et al. 6213 (INPA, MG). BRA- 
IL. Amazonas: Mun. São Paulo de Olivença, near Pal- 
mares, 11 Sep. to 26 Oct. 1936 (fl), Krukoff 8260 (^, BM, 
Е, С, К, LE, а а МУ, S, 0). Para: Rio Cuminá- 
Mirim, 13 Dec. 1906 (fl), Ducke Hallie (BM, G, MG). 
COLOMBIA. Meta: Sierra de la Macarena, Сайо Ciervo, 
alt. 600 m, 12 Jan. 1950 (fr), Philipson e et ge 2084 (COL, 
S, US). ECUADOR. Napo: La Joya de los Sachas, Parque 
Nacional de Yasuní, alt. 230 m, 8-15 July 1993 (fl), Dik 
54 (MO). GUYANA. Kanuku Mts., Puwib River, alt. 
100 m, 13 Feb. 1985 (fl, fr), Jansen-Jacobs e n 184 (K, 
U, WIS). PERU. Huánuco: Pachitea, W o rto Inca, 
alt. 250—300 m, 14 Sep. 1982 (fr), ви ста (MO. U). 
Loreto: Prov. Maynas, Río Momon, 0—5 km from conflu- 


a О m, 2 Nov. 
(U, USM). EE 
E of San Fernando de Atabapo, N ban 
of Río а alt. 95 т, 4 May 1979 (fr), Davidse et al. 
17183 (MO 
Duguetia = Aristeg. ex Maas & Boo 
VENEZUELA. Apure: Distr. San Fernando, mouth 2 
Río Arauca at its intersection with Río Orinoco, alt. 3 
14-15 May 1977 (fl), Davidse & González one (MO, 0), 
Bolivar: Puerto Ordaz, ed Félix, Apr. iii irc 
guieta 5308 (HBG, U, . Guárico: margin 
Orituco, 5 km of им a 1963 (fl), о © 
Tamayo 5087 (HBG, VEN). 
Duguetia riparia Huber 
BOLIVIA. Pando: S bank of Rio Abunda, between 
Cachoeiras Tres S and Fortaleza, 3-16 km above mouth, 
18 July 1968 (fl buds), Prance et al. 6132 (INPA, NY). 
ZIL. Amazonas: Reserva Forestal Ducke, km 26 of 
Manaus-Itacoatiara Hwy., 12 Oct. 1995 (fl), Miralha, 
08 (INPA, 0, Para: Belém, Reserva Мо- 
cambo, 8 Nov. 1995 (fl, fr), Maas et al. 8360 (INPA, K, 
LZ, MG, MO, NY, U, ULM, WIS). COLOMBIA. Caqueta: 
Quebrada El Engafio, 26 Nov. 1991 (fl), Duivenvoorden et 
al. 16694 (0). FRENCH GUIANA. Oyapock River, 
Grande Roche, Saut Cafesoca, 31 July 1969 (fl), Oldeman 
T.425 (CAY, P, U). SURINAM. S of Juliana top, 13 km N 
of Lucie River, alt. 350 m, 9 Aug. 1963 (fl), Irwin et al. 
54639 (B, F, G, M, NY, P, US) 
Duguetia spixiana Mart. 
BOLIVIA. Beni: Prov. Yacuma, SE of San Borja, alt 
350 m, 14 Sep. 1988 (fl), Beck 16749 (LPB, U). BRAZIL. 
Acre: Mun. Senador Guiomard, km 33 of BR 317, 11 


Volume 87, Number 2 
2000 


Chatrou et al. 245 


Duguetia Alliance 


Oct. 1980 (fl), Cid & Nelson 2834 (MG, NY, U). Ama- 
zonas: Mun. Sao Paulo de Olivenga, near Palmares, basin 
of Rio oe 11 Sep.-26 Oct. us P h Krukoff 
8402 (А, Е, С, К, LE, MICH, PS. 0). 
COL MR Аана Quebrada ree 7 yw N of 
Leticia, near Río — е 4 Јап. 1971 id сз кан 
(МО). ECUADOR. Napo: 4 km N of Coca, alt. 
15 Sep. 1986 e» Palacios & mm 1312 о, U). ERU. 
oreto: 7 km of Iquitos, 30 in ind 72 (fl, fr), Croat 
Е. (AAU, E F, MO, NA, NY). Ucayali: rw Coronel 
rtillo, Carretera Vigna ca. 8 km osque von 
броја alt. 250 т, 2 Nov. 1984 (fl, fr), Maas et al. 
6183 (MO, U, USM. WIS). 
Duguetia staudtii (Engl. & Diels) Chatrou 

N. Littoral: Douala-Edea Reserve, Tisson- 
go Study Area, Transect B, June 1976 (fr), Waterman & 
McKey 879 (U). CENTRAL AFRICAN REPUBLIC. San- 
gha Economique Prefecture, Ndakan, Gorilla study area, 
alt. 350 m, 1 Apr. 1988 (fr), Harris & Fay 416 (MO, P). 
CONGO-BRAZZAVILLE. Sangha: W slope of Mt. Na- 
bema, alt. 900 m, 15 Nov. 1991 (fr), Thomas et al. 
8877 (MO). GABON. Woleu-Ntem: Bitam, 20 Dec. 1933 
(yfr), Le Testu 9424 m BR). IVORY ee Néy: 61 

N of Sassandra, W of Niapidou, alt. 
1959 (fl, fr), oeque 2579 (BR, uc: p 
Bong: Bong range, 15 Aug. 1962 (fl, fr), n 1176 
(B, BR, K). Nimba: Nimba Mts., alt. 500 m, 28 1962 
(fl), Voorhoeve 891 (WAG). NIGERIA. с ‘Oban, 9 
Mar. 1959 (fl), Talbot 1494 (BM, Z). 

Duguetia uniflora (DC. ex Dunal) Mart 

AIL. Amazonas: Hio [сапа, 18 Nov. 1945 (fl), 
Fróes 21407 (F, IAN, K, NY, US); Igarapé Тагита-Аси, 
30 km NW of Manaus, 8 Oct. 1990 (fl), Miralha, Maas et 
al. 232 (U, ULM). Roraima: Rio Branco, at mouth of Rio 
Itapera, 8 June 1989 (fr), bu et al. 20424 (U). VENE- 
ZUELA. Amazonas: Tamatama, Upper Río Orinoco, alt. 
125 m, 13 July 1942 (iD. n Williams 15859 (A, F, G, 
NY, RB, 5 ). 
Fusaea са (Aubl.) Saff. 

Beni: km 13 of Riberalta-Guayaramerin 
road, 19 Nov. 1989 (fl), Daly et al. 6260 (MO, U). BRA- 
ZIL. Amapá: Mun. Oia km SSE of Oiapoque, 
1 Dec. 1984 (8), Mori et al. 17100 (NY, U). Amazonas: 
km 155 of Manaus-ltacoatiara Road, 16 Dec. 1974 (fr), 
Gentry & Ramos 13345 (MO, U). Rondônia: Angustura, 
is Machado de ^ Dec. 1931 (fr), Krukoff 1537 (A, 

. MICH, MO, . Roraima: Serra da Lua, 21 
d 1969 (fl we du et al. 9367 (NY, U). COLOM- 

. Antioquia: Mun. Caucasia, road to Nechí, 14 km 

fran Caucasia—Planeta Pica Road, Hacienda La Cande- 

leria, alt. 50 m, 24 Nov. 1986 (fl), Zarucchi & Cárdenas 

4250 (COL, MO, Ат, U). Santander: Puerto Parra-Cam- 

. 1979 (fl, fr), Renterta Arriaga et al. 

; Vaupés: vicinity of Мий, 20 May 

1976 (fr), Zeke 1607 (COL, GH, K). FRENCH e 
A. Les Eaux Claires, near Crique Tortue, alt. 2 


10 Feb. 1993 (fl). Maas et al. 8066 (U). GUYANA. тай 


ее Essequibo: Kuyuwini landing, Kuyuwini 
River, alt. 200 m, 2 Feb. 1991 (fl), Jansen-Jacobs et al. 
2313 (U). PERU. Loreto: Jenaro Herrera, ПАР, Arbore- 
tum, alt. 120 m, 25 Oct. 1994 (fl buds, m apre et 


al. 2 (AMAZ, U, USM, WU). Madre de s: Parque 
у del Maná, 14 Oct. 1986 (fl buds) / "oster et al. 
11824 (0). 

nc. peruviana R. E. Fr. 

PERU. Amazonas: Río Santiago, 2 km from Caterpiza, 
19 Nov. ee (fr), Huashikes 1326 (МО). Huánuco: W 
Sira Mts., 26 km 5 Бај: по Inca, Ра Biologic 
Field edis alt. 2 ‚ 1 Feb. 1 T ce & 
Vasquez 11-1293 on at 260-350 m, 20 199. 
Rainer 244 (О, WU). Loreto: Río Маро, =. 5 


о Camp, alt. 140 m, 3 No 


et al. 29779 (F, 
Dec. 1988 (fl, fr), Vasquez & Jaramillo 11378 (MO, U, 
SM). 


Letestudoxa bella Pellegr. 

CAMEROON. Centre-Sud: near Kom Valley, 25 km E 
of confluent of Ntem River and Akom River, near Ebolo- 
wa, 5 Mar. 1970 (fl, fr), Letouzey 10097 (BR, P). GABON. 

aut-Ogooué: 70 km SSW of Moanda, alt. 730 m, 13 
Oct. 1970 (fl buds), Breteler 6877 (WAG); 42 km SE of 
Lambaréné, 3 Oct. 1968 (fl), Breteler 5805 (WAG). 
Ngounié: Moucongo, 19 Oct. 1926 (fl), Le Testu 6336 
(BR, P). Ogooué-Maritime: Doudou L 
Doussala, 27 Aug. 1985 (1), Reitsma 1432 (МАС); 
3 km S of platform Rabi 13, near old incat camp, alt. 5 
m, 28 Sep. 1994 (fl), Wieringa & Nzabi 2797 (WAC). 
Letestudoxa glabrifolia Chatrou & Repetur 

oleu-Ntem: ca. 10 km on Tchimbélé—As- 
sok road, alt. 630 m, 14 Sep. 1994 (fl buds), Breteler 
12858 (U, WAG); Tchimbélé, on lake border near dam, 
alt. 530 m, 26 Dec. 1989 (fr), Wieringa 293 (WAG); Cris- 
tal Mts., 10 km on Tchimbélé-Kinguélé road, alt. 570 m, 
24 Jan. 1983 (fr), de Wilde 198 (WAG 
Letestudoxa lanuginosa Le Thomas 

BON. Woleu-Ntem: Nkout, 13 Oct. 1933 (fl), Le 

Testu 9320 (BR, MO, P); Oyem, 7 May 1934 (fl), Le Testu 


Peeudlartabotrys eo Pellegr. 

: E part of presidential reserve Won- 
ga- Wongué, ca. 100 ы S of Libreville, alt. 100 m, 3 Mar. 
1983 (fr), de Wilde et al. 890 (WAG). Ogooué-Maritime: 

abi-Kounga, E of Rabi, 29 Oct. 1991 (fl), Breteler & 
Jongkind 10211 (WAG); near Rabi, Shell Oil Company’s 
amp, 24 Nov. 1991 (fl), McPherson 15564 (BR); Rabi, 
1.5 km along pipeline to Echira, alt. 40 m, 24 Nov. 1994 
(8, fr), Wieringa & van Nek 3273 (WAG); 22 km along a 
track leading in W direction into the Doudou Mts., alt. 
150 m, 3 Dec. 1986 (fl), de Wilde et al. 9136 (P, WAG); 
Rabi, Shell-Gabon, just E of the airstrip, alt. 80 m, 22 
Jan. 1993 (fl, fr), de Wilde & van der Maesen 10888 
(WAG 


MOLECULAR SYSTEMATICS 
OF THE CHINESE YINSHANIA 
(BRASSICACEAE): EVIDENCE 
FROM PLASTID AND 
NUCLEAR ITS DNA 
SEQUENCE DATA! 


Marcus Koch? and Ihsan A. Al-Shehbaz? 


ABSTRACT 


Species of the Chinese endemic genera Yinshania, Hilliella, and Cochleariella were originally placed in or closely 


associated with Cochlearia. A previous prelimin 


ary molecular study m 


ainly on European Cochlearia and detailed 


morphological studies by us showed that this complex was not affined to Cochlearia s. str. Depending on the authority 


consulted, the number of taxa recognized in this 


ested by ne 


wit earia, as was su 


complex ranged from 


illiella pi Cochleariella The second lineage зө the diploid taxa from Yinshania. Ho 


e lineage combines exclusively the highly ote 


arly all previous authors. 


ords: шаш. “СоеМеапейа, Hilliella, molecular systematics, reticulate evolution, Yinshania. 


Many authors follow Schulz (1936) and Schultze- 
Motel (1986) in dividing Cochlearia into the sec- 
tions Pseudosempervivum Boiss., Glaucocochleria O. 
= Eucochlearia Prantl), and 
ulz. As shown by Koch 

999a), иц this sectional classification is 
highly artificial. Section Cochlearia is widely dis- 
tributed in Europe and the circumpolar region, 
whereas section Glaucocochlearia, which was 
raised to the generic rank by Pobedimova (1968), 
is restricted to southwestern Europe. The latter sec- 
tion consists of C. glastifolia L. and C. megalos- 
perma (Maire) Vogt, as well as C. aragonensis Coste 
& Soulié, which was only recently included (Koch 
et al., 1996), although considered to be distantly 
related to the other two species (Koch et al., 

a). Section Cochlearia consists of a species 
complex that demonstrates highly polymorphic 
chromosome numbers, and diverse ecological ad- 
aptation and geographic distributions. Morphologi- 
cal differences between phylogenetically sister taxa 
are often weak and poorly defined (Koch et al., 
1996). Both sections Cochlearia and Glaucococh- 
learia are closely related to the genus Jonopsidium 


et al. 


Rchb. (Koch et al., 1999a). Section Pseudosemper- 
vivum, which is centered in the Middle East and 
clearly unrelated to Cochlearia, is most closely re- 
lated to Masmenia F. K. Mey. and Noccaea Moench 
(Koch et al., 1999a), both of which were segregated 
by Meyer (1973, 1979, 1991) from Thlaspi L. 5.1. 
The family of Brassicaceae is divided into sev- 
eral tribes and subtribes. Most of them are highly 
artificial, such as tribe Arabideae (Koch et al., 
1999b) or Lepidieae (Zunk et al., 1996). Following 
classical tribal concepts, Cochlearia sect. Pse 
sempervivum, sect. Cochlearia, and sect. Glauco- 
cochlearia are members of tribe Lepidieae. Species 
originally assigned by Schulz (1923) to section Hil- 
liella (Yinshania, Hilliella, and Cochleariella) were 
excluded from Cochlearia by Pobedimova (1970), 
who did not assign them to any genus. However, 
these species have recently been placed in three 
Chinese endemic genera, Yinshania Y. C 
Z. Zhao, Cochleariella Y. H. Zhang & Vogt, and 
Hilliella (0. Е. Schulz) У. H. Zhang, each of which 
was assigned to a different subtribe. The genus Yin- 
shania (Ma & Zhao, 1979) was placed in subtribe 


' We are grateful to Zhang Yu-hua for providing some of the samples. The curators of A, B, ВМ, E, GH, HAST, 


IBSC, K, KUN, LE, M 


2 Institute of Botan 


MO, NAS, NY, Р, PE, TAI, TI, TNS, US, W, and WU are thanked for the loan of specimens. 
ny, University for Agricultural Science, Gregor-Mendel-Str. 33, A-1180 Vienna, Austria 


* Missouri Botanical Garden, P.O. Box 299, St. Louis, Missouri 63166-0299, U.S. 
ANN. Missouni Вот. GARD. 87: 246-272. 2000. 


Volume 87, Number 2 
2000 


Koch & Al-Shehbaz 
Chinese Yinshania 


247 


Descurainiinae, tribe Sisymbrieae. The genus 
Cochleariopsis (Zhang, 1985), renamed as Coch- 
leariella (Zhang & Cai, 1989), was placed in sub- 
tribe Cochleariinae, tribe Lepidieae, along with Hil- 
liella s. str. (Zhang, 1986). Although a few studies 
on taxonomy, evolution, and origin of these genera 
(Zhang, 1987; Zhang & Xu, 1990; Zhang, 1993) 
have been made, nothing was said about their sys- 
tematic position in relation to the remaining Asian 
and European taxa of Cochlearia. Zhang’s (1987) 
division of Yinshania (excluding Hilliella) into two 
sections and two series and Zhao’s (1992) classifi- 
cation of Yinshania (including Hilliella) into two 
sections and six series, were shown by Al-Shehbaz 
et al. (1998) to be highly artificial. In fact, one of 
the species assigned by Zhang (1987) to Hilliella 
and by Zhao (1992) to Yinshania was placed by Al- 
Shehbaz and Yang (1998) in the synonymy of Car- 
damine fragariifolia O. E. Schulz 
n the basis of a comprehensive morphological 

survey of Yinshania, Hilliella, and Cochleariella, 
Al-Shehbaz et al. (1998) reduced the latter two to 
synonymy of Yinshania, and concluded that there 
is no need for infrageneric subdivisions that do not 
reflect the phylogenetic relationships of this small 
genus of 13 species 

They also ра that morphological сћаг- 
acters previously used in the delimitation of species 
(e.g., density of papillae on the fruit valves, fruit 
shape, and seed number per fruit) are highly vari- 
able among and within different populations of the 
same species. Furthermore, differences in the com- 
pression of fruit (terete vs. latiseptate or angusti- 
septate) that were emphasized heavily by earlier 
authors (e.g., Schulz, 1936) in the delineation of 
genera were not found to be taxonomically useful 
in the Yinshania complex. As shown by Koch et al. 
(1999a), the placement of heavy emphasis on fruit 
compression has led to the artificial integration of 
several taxa into Cochlearia sect. Pseudosempervi- 
vum instead of Thlaspi s.l. In fact, terete and var- 
iously flattened fruits occur in numerous genera of 
the Brassicaceae, and in many cases this aspect of 
fruit morphology is taxonomically insignificant. 

Morphological convergence and parallelism are 
widespread in the Brassicaceae (Dvorák, 1971; 
Meyer, 1973; Avetesian, 1983; Endress, 1992), and 
the dependence on such characters to construct 
phylogenies often leads to erroneous conclusions 
(Sytsma, 1990; Meyer, 1991). Recent molecular 
analyses (e.g., Warwick et al., 1992; Price et al., 
1994; Mummenhoff & Koch, 1994; Zunk et al., 
1996; Mummenhoff et al., 1997; Koch et al., 1998a, 
b; Koch et al., 1999a, b) have made significant con- 


tributions to a better understanding of the classifi- 
cation, generic delimitation, and phylogenetic re- 
lationships in the Brassicaceae. A preliminary 
study (Koch et al., 1999a) utilizing ITS nrDNA and 
cp trnL intron sequence data of four species of the 
Yinshania complex (including Hilliella and Coch- 
leariella) clearly showed that the complex is unre- 
lated to Cochlearia. In this analysis it has been 
shown that ITS and trnL intron sequence data pro- 
vide sufficient sequence variation to distinguish 
significantly between Yinshania and Cochleariella/ 
Hilliella accessions with both data sets. In order to 
gain a better insight of the phylogenetic relation- 
ships within this Chinese complex, we examined 
sequence variation of the internal spacer regions 
(ITS1 and ITS2) of nrDNA (Baldwin et al., 1995; 
Campbell et al., 1995) and of the cp trnL intron 
(Bóhle et al., 1994; Gielly & Taberlet, 1994; van 
Ham et al., 1994; Koch et al., 1999a), and com- 
pared the derived molecular phylogenies with tra- 
ditional concepts based on morphological data. 
This approach provided us with the opportunity to 
characterize species lineages and to analyze incon- 
gruencies between different data sets in order to 
test hypotheses of gene flow over lineages and chlo- 
roplast capture. 


MATERIALS AND METHODS 
PLANT MATERIAL 


Leaf material for DNA extraction was obtained 
from herbarium specimens (Table 1), most of which 
were provided and determined by Zhang Yu-hua 
(Institute of Materia Medica, Zhejiang Academy of 
Medicine, Hangzhou, People's Republic of China). 
We did not examine vouchers to verify Zhang's de- 
terminations. The samples represent a broad spec- 
trum of species of Yinshania, Hilliella, and Coch- 
leariella. Cardamine flexuosa With. and Rorippa 
palustris (L.) Besser served as the outgroups. The 
DNA sequences for the outgroups were obtained 
from Franzke et al. (1998). 


DNA EXTRACTION, PCR-AMPLIFICATION, AND 
SEQUENCING 


The total DNA for the outgroups was isolated 
from leaf tissues following the CTAB (cethyltriam- 
moniumbromide) method of Doyle and Doyle 
(1987), as modified by Mummenhoff and Koch 
(1994). DNA extraction from herbarium material 
was performed in a mini preparation in Eppendorf 
reaction tubes from 50-ug dried tissue. Tissue was 


ground with sand and prewarmed 2X CTAB-buffer. 


Annals of the 


248 


Missouri Botanical Garden 


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249 


Koch 4 Al-Shehbaz 
Chinese Yinshania 


Volume 87, Number 2 


2000 


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250 


Annals of th 
Missouri Botanical Garden 


Organic extraction and DNA isolation from herbar- 
ium specimens followed Koch et al. ( | 

Double-stranded DNA of the complete ITS ге- 
gion, including the 5.85 rDNA gene, was amplified 
by 30 cycles of symmetric PCR using ITS primers 
initially designed by White et al. (1990) and mod- 
ified by Mummenhoff et al. (1997). The 18F primer 
(5'-ССААССАСААСТССТААСААСС-3') is lo- 
cated at the 3’-end of the 18S rDNA gene, and 
primer 25R (5'-TCCTCCGCTTAT ТСАТАТСС-3') 
is located at the 5'-епа of the 25S rDNA. It has 
been reported that PCR selection of rDNA para- 
logues has occurred (Buckler et al., 1997). How- 
ever, PCR selection might have only been impor- 
tant in high G+C content sequences (Buckler et 
al., 1997). Sequences from Yinshania and Hilliella 
(Koch et al., 1999a) are comparable in G+C con- 
tent to sequences from Gossypium, in which PCR 
selection was probably weak (Wendel et al., 19954). 
The resulting amplification product included 1751, 
5.85 rDNA, and ITS2. Only those PCR products 
were cloned into the pGEM-T-Easy cloning vector 
(PROMEGA) that showed a single band on ethi- 
dium bromide stained agarose gels. Two cloned ITS 
regions from two independent PCR reactions were 
sequenced (forward and reverse) with both ampli- 
fication primers and two universal primers located 
in the flanking sites of the pGEM-T-Easy vector (t7- 
forward: 5'-gtaacgatttaggtgacactatcg-3, m13-re- 
verse: 5'-agcggataacaatttcacacagga-3). This means 
that every single clone was sequenced four times 
to avoid sequence errors. The trnL (UAA) intron 
was amplified and sequenced by using the univer- 
sal primer B49318 (5'-CGAAATCGGTAGACGCT- 
АСС-3'’) located at the 3'-end of the trnL(UAA)5'- 
exon and A49855 (5'-GGGGATAGAGGGACTTG- 
ААС-3') located at the 5'-end of the trnL(UAA)3'- 
exon (Taberlet et al., 1991). The PCR profile used 
to amplify the trnL intron followed the following 
profile: hot start with 5 min. at 94?C, and 35 cycles 
of amplification (1 min. 94?C, 45 min. 50?C, 45 
min. 72°C), final elongation step for 10 min. 72°C, 
and storage at 4°C. DNAs were cycle-sequenced 
using the Taq DyeDeoxy Terminator Cycle Se- 
quencing Kit (ABI Applied Biosystems, Inc.). Prod- 
ucts of the cycle sequencing reactions were run on 
an ABI 377XL automated sequencer (ABI Applied 
Biosystems, Inc.). Material from accession numbers 
22-26 (Table 1) was only used for sequencing the 
trnL intron sequence, because amplification of the 
ITS regions failed totally. 


DATA ANALYSIS 


ITS data. Boundaries of the ITS regions and 
coding sequences were determined by comparison 


to those of Sinapis alba L. (Rathgeber & Capesius, 
1989) and other Brassicaceae (Mummenhoff et al., 
1997; Koch et al., 1999а). DNA sequences were 
aligned by hand. Parsimony analyses were рег- 
formed with unordered Fitch parsimony and 
weighted parsimony with a transition : transversion 
weighting of 1.0:1.08 using PAUP version 3.1 
(Swofford, 1993). The BRANCH-AND-BOUND al- 
gorithm was used to find maximally parsimonious 
trees. Bootstrap analysis (Felsenstein, 1985) was 
performed using 1000 replicates and the HEURIS- 
TIC search algorithm with the MULPARS option. 
We combined the GAPMODE=MISSING option 
with the coding of the gaps as additional presence/ 
absence characters (Downie & Katz-Downie, 1996). 
This option decreases the number of equally par- 
simonious trees because of the redundancy result- 
ing from having two sets of scored characters for 
the same indel events (Wojciechowski et al., 1993). 
Evolutionary data are most often presented as a 
phylogenetic tree, the underlying assumption being 
that evolution is a branching process. However, em- 
pirical data is rarely ideal and often supports sev- 
eral trees instead of one unique tree. Hence, it 
makes sense to consider tree reconstruction meth- 
ods that produce a tree if the given data heavily 
favor one tree over all others. Otherwise, methods 
that produce a more general graph that indicates 
different possible phylogenies are useful (Huson, 
1998). One such method is the Split Decomposition 
introduced by Bandelt and Dress (1992) and its 
variations. In order to visualize conflicting phylo- 
genetic signals, we analyzed all ingroup ITS se- 
quences (see Fig. 6), using the software gres 
SplitsTree version 1.0.3. (Huson & Wetzel, 1995 
shareware ftp://ftp.uni-bielefeld. eel). 
trnL intron data. DNA sequences were aligned 
by hand. Parsimony analysis was performed with 
PAUP (version 3.1; for options, see ITS data). Gaps 
were treated as additional unweighted binary char- 
acters. These gaps were coded using strict criteria: 
gaps must occur at the same position and have the 
same aligned length to be treated as homologous, 
and no splitting of one gap in two or more char- 
acters was performed (Koch et al., 1999a). Boot- 
strap analysis was performed as described above. 


TRIBAL RELATIONSHIPS 


To estimate the tribal affinity of Yinshania, we 
derived an ITS phylogeny with sequences from 
Capsella rubella Reut. (Koch et al., 1999b), Ara- 
bidopsis thaliana (L.) Heynh. (GenBank U43224), 
Yinshania acutangula (O. E. Schulz) Y. H. Zhang, 


Volume 87, Number 2 


Koch & Al-Shehbaz 
Chinese Yinshania 


251 


Barbarea vulgaris R. Br. (EMBO X98632), Carda- 
mine flexuosa With. (Franzke et al., 1998), Thlaspi 
arvense L. (Koch et al., 1999a), Cochlearia aestu- 
aria (Lloyd) Heywood (Koch et al., 1999a), Brassica 
oleracea L. (GenBank AF039994/AF040038), and 
Sinapis alba (EMBO X66325). DNA sequences 
were aligned by hand, and the alignment is shown 
in Figure 4. Alignment positions 109-160 were re- 
moved from subsequent analysis. Parsimony anal- 
yses were performed with unordered Fitch parsi- 
mony using PAUP version 3.1 (Swofford, 1993). 
The BRANCH-AND-BOUND algorithm was used 
to find maximally parsimonious trees with the GAP- 
MODE=NEWSTATE option. Bootstrap analysis 
(Felsenstein, 1985) was performed using 1000 rep- 
licates and the HEURISTIC search algorithm em- 
ployed with the MULPARS option. A decay anal- 
ysis (Bremer, 1988) was performed in addition to 
the bootstrap approach, in order to assess the con- 
fidence that could be placed in the monophyly of 
clades. Decay indices (DI) were estimated accord- 
ing to Baum et al. (1994). 


RESULTS 
ITS DATA 


The total length of the alignment with accession 
numbers 1–21 and outgroups Cardamine flexuosa 
and Rorippa palustris is 464 bp, with 283 and 181 
nucleotides in ITS] and ITS2 spacer regions, re- 
spectively. The alignment required 36 (7.8%) gap 
positions, including the outgroups, and is shown in 
Figure 1. Sequence data were submitted to Gen- 
Bank with accession numbers AF100793- 
AF100852 (Table 1). One third of these gaps is 
located between positions 117 and 133 in the ITS1. 
This region was completely excluded from subse- 
quent analysis. Additionally, we excluded nucleo- 
tide position 465—467 and 473-484 from subse- 

uent data analysis because of an ambiguous 
alignment (Fig. 1). Gaps from nucleotide positions 
11-14 and 293-294 were treated as one single gap, 
respectively. The number of introduced gaps is 
comparable to a phylogenetic analysis of Thlaspi 
s.l. with 4.8% of the sites (Mummenhoff et al., 
1997). In Krigia and outgroups, Kim and Jansen 
(1994) had to introduce gaps in 3.9% of the sites. 
Total lengths of ITS1 and ITS2 are nearly identical 
among the taxa surveyed, and vary between 447 bp 
(Hilliella alatipes (Hand.-Mazz.) Y. H. Zhang & H. 
i var. micrantha Y. H. Zhang [= Yinshania 
rivulorum (Dunn) Al-Shehbaz et al.], accession no. 
10) and 457 bp (Cardamine flexuosa and Rorippa 
palustris). 
Phylogenetic analysis using Fitch parsimony, in- 


cluding the additional 0/1 matrix for the gap posi- 
tion, resulted in 24 most parsimonious trees (MPTs) 
with a length of 538 and a consistency index (CI) 
of 71.4% (66.9% if autapomorphies are excluded). 
Of the 264 variable nucleotide positions, 166 in- 
formative positions were in the ITS] region (in- 
cluding 48 autapomorphies) and 98 in the ITS2 
region (including 25 autapomorphies). Four out of 
36 gap positions within the total sequence align- 
ment are unique to a particular sequence (Rorippa 
palustris 2 gaps, acc. no. 3, and acc. по. 14). A 
calculation of the transition/transversion ratio for 
the МРТ» revealed a ratio of 1.00: 1.08. Therefore, 
we used a weighted parsimony approach with a 
character state weighting of 1.00: 1.08 (transition: 
transversion), which resulted in one MPT that is 
also represented among the 24 MPTs from the Fitch 
parsimony with a consistency index of 67.796 (CI 
57.9% if autapomorphies were excluded). We pre- 
sent the MPT from the weighted parsimony ap- 
proach to demonstrate relative branch length (Fig. 
2). Bootstrap values are provided from 1000 repli- 
cates using the weighted parsimony approach. For 
most taxa we identified only one ITS sequence type 
within a single specimen. For Hilliella lichuanensis 
Y. H. Zhang (acc. no. 13), Cochleariella zhejian- 
gensis (Y. H. Zhang) Y. H. Zhang & R. Vogt (acc. 
no. 21), Yinshania acutangula (acc. no. 5), Y. henryi 
(Oliv.) Y. H. Zhang (acc. no. 7), and Y. furcatopilosa 
(K. C. Kuan) Y. H. Zhang (acc. no. 8), we found 
two very similar ITS sequences among the two 
clones sequenced. In the case of accession num- 
bers 13, 5, 7, and 8, ITS1 and ITS2 regions differed 
by only a single site mutation. In the case of C. 
zhejiangensis (acc. no. 21), the two ITS types from 
the same individual differed by 15 mutations, 
which might indicate that two different ITS loci 
were cloned and sequenced. Both sequences clus- 
tered within the same clade. In Hilliella fumarioi- 
des (Dunn) Y. H. Zhang & H. W. Li (acc. no. 17), 
we detected two ITS sequences from a single in- 
dividual that clustered in different clades (Figs. 2, 
6). Both sequences differed by 76 mutations. The 
ITS data clearly support a separation of two clades 
consisting of Yinshania sensu Zhang (1987) and 
Hilliella/Cochleariella. Different accessions from 
one taxon (sensu Al-Shehbaz et al., 1998, refer to 
fig. 2) grouped at different positions in the case of 
C. zhejiangensis (acc. no. 20, not related to acc. nos. 
18, 19, 21 

Using the taxonomic concept of Al-Shehbaz et 
al. (1998) and merging Hilliella changhuaensis Y. 
H. Zhang, H. guangdongensis Y. H. Zhang, and H. 
lichuanensis (acc. nos. 11, 12, 13, respectively) into 
H. lichuanensis, only accession numbers 11 and 13 


Annals of the 


252 


Missouri Botanical Garden 


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Koch & Al-Shehbaz 
Chinese Yinshania 


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255 


Моште 87, Митбег 2 


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Koch & Al-Shehbaz 
Chinese Yinshania 


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Koch 4 Al-Shehbaz 
Chinese Yinshania 


Volume 87, Number 2 


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Volume 87, Number 2 
2000 


Koch & Al-Shehbaz 
Chinese Yinshania 


259 


grouped together by ITS data. Accession number 
12 (H. guangdongensis) is unrelated to these ac- 
cessions based on ITS data. Similarly, H. warburgii 
(O. E. Schulz) Y. H. Zhang & H. W. Li (acc. nos. 
18, 19), Cochleariella zhejiangensis (acc. nos. 20, 
21), and H. fumarioides (acc. no. 17) merge into Y. 
fumarioides (Dunn) Y. Z. Zhao, as proposed by AI- 
Shehbaz et al. (1998). ITS sequences from Y. fu- 
marioides sensu Al-Shehbaz et al. (1998) are found 
in three different positions among the Hilliella/ 
Cochleariella clade (Fig. 2 


trnL DATA 


The alignment of 514 bp is interspersed with 
seven gaps as shown in Figure 3. Seq 
were submitted to GenBank with accession 
bers AF100853 through AF100881 (Table 1). "The 
lengths of trnL intron sequences range from 311 bp 
in Hilliella rivulorum to 514 bp in Cardamine flex- 
uosa. Of the 38 variable nucleotide positions in the 
alignment, 12 sites are autapomorphic. In addition, 
one of the seven gaps is autapomorphic. Phyloge- 
MPTs with a length 
of 53 steps and a consistency index of 92.5% 
(90.0% if autapomorphies were excluded). One 
most parsimonious tree is shown to demonstrate rel- 
ative branch length. The strict consensus tree could 
be generated easily by drawing branches that are 
indicated in Figure 2 by heavy bars with zero 
length. Bootstrap values are given from 1000 rep- 
licates. 

crimination of a Yinshania clade from a Coch- 
leariella/Hilliella clade is not as obvious with trnL 
intron data as compared to ITS sequence data (Fig. 
2). In the шп], tree the branch setting of the Yin- 
shania clade as a sister group to the Hilliella/Coch- 
leariella clade is not highly supported, and boot- 
strap ~~ for this branching point is less than 
50% (Fig. 2). Removing additional gap characters 
from the matrix parsimony analysis resulted in one 
MPT with the Yinshania clade as a sister group to 
Н. hunanensis (acc. no. 16), H. guandongensis (acc. 


uence data 


netic analysis resulted in two 


no. 12), C. zhejiangensis (acc. no. 20), and H. ala- 

es var. micrantha (acc. no. 10). Remaining Hil- 
liella/Cochleariella taxa appeared in this analysis 

or trnL data excluding gap information as a sister 
group to these two clades. Nonetheless, integration 
of Yins а qianningensis Y. ang into the 
HilliellalCochleariella clade is 0 оа for trnL 
data, in contrast to its segregation by ITS. Esti- 
mation of decay indices (DI) using the trnL intron 
matrix without gap information revealed a high val- 
ue, DI = 3-, for the branch setting of the Yin- 
shania clade. Different accessions of C. zhejiangen- 
sis (acc. nos. 20, 21) did not group closely together; 
this has also been documented for the ITS data 
(Figs. 2, 6). Hilliella changhuaensis, H. guangdon- 
gensis, and H. lichuanensis do not group together 
in the cpDNA-based tree as proposed by the mor- 
phology-based concept combining them in H. li- 
chuanensis (Al-Shehbaz et al., 1998); the same dis- 
cordance holds for H. warburgii, C. zhejiangensis, 
and H. fumarioides. Based on the trnL sequence 
data, they are not combined in one single clade that 
could be named as H. fumarioides as proposed in 
the revison of Al-Shehbaz et al. (1998). 


ITS VERSUS TRNL INTRON DATA 


Both phylogenetic trees (ITS vs. trnL data) from 
Figure 2 show some congruencies (all following ar- 
guments are also true when comparing the strict 
consensus trees from Fitch parsimony, which could 
be easily deduced by drawing branches indicated 
by heavy bars with zero length): 

(1) Hilliella guangdongensis [sensu Zhang]. 
which has been merged in H. lichuanensis sensu 
Al-Shehbaz et al. (1998), is separated from remain- 
ing H. lichuanensis sensu Al-Shehbaz et al. (1998), 
and it is more closely related to H. hunanensis, a 
taxon that was recognized by Al-Shehbaz et al. 


998). 
(2) Hilliella warburgit and Cochleariella zhejian- 
are not integrated into H. fumarioides sensu 


Al-Shehbaz et к] (1998), but (3) most accessions 


Comparison of the ITS-derived ж аы with those from the trnL sequence data from Yinshania, ла 


5 
leariella, and Hilliella, as well as outgroups 
enumeration in brackets follows Table 1. For f ITS 


values are provided from 1000 replicates and show 


amine an 

and the trnL tree the nomenclature sen 

. In between both trees the taxonomic treatment of pe taxa E nint to iy Shehbaz et 

weighted ај арргоасћ) (то 
да 


orippa from subtribe Arabideae siege ви Accessio 
g (1985, 1986. 


— 


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ches if greater than 50%. Distance scale indicating 


branch lengths is given below. ITS designation (a) and (b) of accession nos. 5, 7, 8, 13, 17, and 21 indicate different 
] 


ITS types from a single individual. 


Annals of the 


260 


Missouri Botanical Garden 


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Koch 4 Al-Shehbaz 
Chinese Yinshania 


Volume 87, Number 2 


2000 


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2000 


Koch 4 Al-Shehbaz 
Chinese Yinshania 


263 


of H. warburgii and C. zhejiangensis formed one 
single cluster with no separation of H. warburgii 
versus C. zhejiangensis. This is in agreement with 
previous concepts combining both taxa, C. zhejian- 
gensis and H. warburgii, in one single taxon. These 
congruent ITS and trnL intron findings demonstrate 
that both У lichuanensis and Y. fumarioides species 
complexes, treated as well-defined taxa according 
to the morphological revision of Al-Shehbaz et al. 
(1998), do not form monophyletic groups by molec- 
ular evidences. 

However, some incongruencies among the two 
molecular data sets could be detected: (1) Yinshan- 
ia qianningensis grouped either inside the Hilliella/ 
Cochleariella clade (trnL intron data) or into Yin- 
shania s. str. (ITS data); (2) Н. rupicola (D. С. 
Zhang & J. Z. Shao) Y. H. Zhang also clustered 
into two different subgroups within Hilliella/Coch- 
leariella. 


TRIBAL RELATIONSHIPS 


The alignment of ITS1—5.8SrDNA-ITS2 is 644bp 
in length (Fig. 4). Within the ITS1 region (bp 1– 
301) there are 143 variable nucleotide positions 
(including 62 autapomorphies). The ITS2 region 
(position 456—644) contains 85 variable nucleotide 
positions (including 57 autapomorphies). The 5.8S 
rDNA gene, located between both spacer regions 
(bp 302—455), contains 11 variable nucleotide po- 
sitions (including 6 autapomorphies). Because of an 
ambiguous alignment, nucleotide positions 107— 
153 and 460—484 were removed from the original 
data matrix (Fig. 4), resulting in a final data matrix 
of 412 bp. 

Fitch parsimony analysis resulted in one MPT 
with a length of 288 steps and a consistency index 
of 79.5% (66.5% if autapomorphies are excluded). 
The phylogenetic tree (Fig. 5) showed closer rela- 
tionships of Yinshania to taxa from tribe Arabideae 
sensu Janchen (1942) (Arabidopsis (DC.) Heynh., 
Barbarea В. Br., and Cardamine L. were used to 
represent tribe Arabideae). However, Cochlearia 
(including species loosely affined to the Yinshania 
complex) and Capsella rubella were placed by 
Janchen in the tribe Lepidieae, where they had 
been put by Hayek (1911) and Schulz (1936). Our 
ITS data do not support this latter placement. A 
molecular analysis of Arabidopsis, Arabis L., and 
their relatives shows a close relationship of Cap- 
sella rubella to Arabidopsis thaliana (Koch et al., 
1999b). Also demonstrated is the polyphyly of Ar- 
abis and Arabidopsis, indicating that tribal struc- 
tures within Brassicaceae are highly artificial. The 
ITS phylogeny does suggest that Chinese Yinshania 


and related taxa are closer to Cardamine and Bar- 
barea from tribe Arabideae than to genera like 
Thlaspi or Cochlearia s. str. of the tribe Lepidieae. 


CYTOLOGY 


Little is known about the cytology of Yinshania. 
Zhang (1995, and pers. comm.) counted 2n = 12 
for Y. qianningensis Y. H. Zhang [= Y. acutangula 
sensu Al-Shehbaz et al., 1998], Y henryi (Oliv.) Y. 
H. Zhang, and Y. furcatopilosa (K. C. Kuan) Y. H. 
Zhang, 2n = 42 for Hilliella yixianensis Y. 
Zhang, H. paradoxa (Hance) Y. H. Zhang & H. W. 
Li, and H. changhuaensis [= Y. lichuanensis sensu 
Al-Shehbaz et al., 1998], and 2n = 44 for H. 
shuangpaiensis Z. Y. Li [= Y. rupicola sensu А]- 
Shehbaz et al., 1998]. These data correspond to the 
ITS-derived phylogeny, in which the diploid Y 
qianningensis, Y. henryi, and У furcatopilosa, to- 
gether with Y. acutangula, are separated from the 
polyploid Hilliella. Within polyploid Hilliella, the 
Hilliella taxa with 2n — 42 are combined. Hilliella 
shuangpaiensis (represented in this study by acc. 
no. 4, Fig. 2) with 2n — 44 did not group closely 
to the known 2n = 42 taxa (represented in this 
study by acc. nos. 2, 11, and 15, Fig. 2). However, 
any conclusions based on cytology must be only 
preliminary. It remains possible that the Hilliella/ 
Cochleariella group could also be represented by 
polyploid taxa with 2n = 42 and 44. Base chro- 
mosome number for this hexaploid group is x = 7, 
instead of x = 6 as in the Yinshania group. А few 
taxa within the Hilliella/Cochleariella clade may be 
aneuploids (2n — 44) that derived from 2n — 42. 


DISTRIBUTION 


Some interesting features emerge when topolo- 
gies of the phylogenetic trees are compared to the 
geographic distribution of Yinshania s.l. Geograph- 
ic distribution of ITS sequence types from taxa of 
the four main clades within Hilliella/Cochleariella 
do not follow their phylogenetic relationships (Fig. 
7), and they are randomly mixed in Southeast China 
in the provinces of Guangdong, Jiangxi, Zhejang, 
Anhui, Hubei, and eastern Sichuan. However, they 
are separated geographically and phylogenetically 
from the Yinshania clade from Hunan, Xizang, and 
Sichuan. Taxa from the Yinshania clade extend the 
distribution to the southwest. А mixed distribution 
of DNA types also holds for the trnL data. No geo- 
graphic structuring of plastome types could be ob- 
served among taxa from the Hilliella/Cochleariella 
clade (Fig. 8). Based on trnL intron data Y. qian- 
ningensis (acc. no. 1) from Sichuan integrates into 
the Hilliella/Cochleariella clade. Geographically, 


Annals of the 


264 


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Koch & Al-Shehbaz 
Chinese Yinshania 


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Volume 87, Number 2 Koch & Al-Shehbaz 
2000 


267 
Chinese Yinshania 
0.1 substitutions/site » 
93 Capsella 
72 Arabidopsis 
97 Yinshania Arabideae 
= 
96 Вагђагеа 
100 | 
— Cardamine З 
Thlaspi 
Lepidieae 
Cochlearia 
100 Brassica . 
Brassiceae 
Sinapis 


Figure 5. Phylogenetic relationships of cruciferous taxa from different tribes. Genetic distances were calculated 
using the program TREECON (van de Peer & de Wachter, 1994) under the Kimura model (Kimura, 19 


80). Gaps were 
not taken into account, and the neighbor joining algorithm was used to calculate genetic distances. The robustness of 
the tree was tested by 1000 bootstrap replicates. 


Yinshania clade 


Hilliella/Cochleariella clade 


Figure 6. Split Decomposition of the ITS sequence data set analyzing the ingroup taxa. The DRAW-EQUAL-EDGES 
option was used to draw the network, and thus distances are not drawn to scale. 


268 


Annals of the 
Missouri Botanical Garden 


Sa 


- Pacific Ocean z 
600 km 
95° 100° 105° С? 115° 120° 
1 [ | \ \ 
Figure 7. Distribution of ITS types among Yinshania and Hilliella/Cochleariella accessions under study in China. 


Taxon enumeration follows Table 1. The Hilliella-Cochleariella clade is separated into four ITS types indicated by 
circles and boxes (gray or white) well suppported by high bootstrap values in Figure 2. Accessions marked by open 


circles and boxes are two well-separated subgroups, which are comb 
. Phylogenetic relationships are shown schematically in the upper left box and follow Figure 2. For the Yinshania 


data 


— 


ined to a single group in Figure 8 (plastid шта, 


clade phylogenetic relationships are shown to demonstrate the position of Yinshania qianningensis [У. acutangula sensu 


Al-Shehbaz] (асс. по. 1) (refer to Fig. 2). 


this accession lies close to a similar plastome type 
(acc. no. 15) from the Hilliella/Cochleariella clade 
(Fig. 8). Upon comparison of the ITS and trnL in- 
tron-derived phylogenies, taxa that showed differ- 
ent positions in the phylogenetic analysis (e.g., H. 
rupicola, acc. no. 14; H. fumarioides, acc. no. 17: 
one additional ITS sequence copy) were located at 
the center of the Hilliella/Cochleariella distribu- 
tional area. We interpret these results as a first bio- 
geographical documentation of reticulation within 
polyploids. Hilliella fumarioides (acc. no. 17) pos- 
sessed a trnL intron type similar to samples from 
surrounding areas. The two ITS types found in that 
particular individual were also present in adjacent 
regions and from different species. From Hilliella 
rupicola (acc. no. 14) we could isolate an ITS DNA 
type, which is also present in adjacent regions in 
accession numbers 10, 16, 17, and 20. trnL intron 
DNA type from accession number 14 also corre- 


sponds to trnL intron DNA types from 2, 3, 11, 13, 
17, and 20 


MORPHOLOGICAL VARIATION AND TAXONOMIC 
CONSIDERATIONS 


s in numerous other cases in the Brassicaceae, 
morphological differentiation among and within so- 
called genera and taxa of the Yinshania s.l. com- 
plex does not provide ample characters to draw 
sharp and uncontroversial generic boundaries. 
Flower morphology in the entire complex is of no 
predictive diagnostic value, even for the separation 
of species (Al-Shehbaz et al., 1998). Only leaves 
and fruits offer characters useful for the separation 
of species, and all taxa appear to be not well de- 
fined. As shown by Al-Shehbaz et al. (1998), seed 
sculpture and number per locule, cotyledonary po- 
sition, development of the fruit septum, type (if any) 


Volume 87, Number 2 
2000 


Koch & Al-Shehbaz 
Chinese Yinshania 


269 


trnL ^ © Yinsha 
Hilliella - 
n Cochleari 
ls Pacific Ocean z 
600 km 
959 1009 1059 Cd 115? 1209 
[ L 1 | | 


Figure 8. Distribution of trnL intron types among Yinshania and Hilliella/Cochleariella accessions under study in 


China. 


Taxon enumeration follows Table 1. The Hilliella- Cochleariella clade is separated 


into three plastid trnL types 


indicated by circles and boxes (gray or white) well suppported by high bootstrap values in Figure 2. Phylogenetic 
relationships are shown schematically in the upper left box and follow Figure 2. For the Yinshania clade phylogenetic 
relationships are shown to demonstrate the position of Yinshania qianningensis [У. acutangula sensu Al-Shehbaz] (acc. 


no. 1) (refer to Fig. 2). 


of fruit compression, fruit shape, development of 
papillae on the fruit valve, and trichome type, all 
are unreliable in dividing the complex into the gen- 
era Yinshania, Hilliella, and Cochleariella. In fact, 
no single character or set of characters can be re- 
lied upon to subdivide the complex into generic or 
infrageneric taxa. The recognition of a single genus 
is, therefore, taxonomically expedient, as there is 
no single character setting one clade apart from 
another. Even pustules on the testa surface of the 
valves, which were assumed to be a good character 
to separate Yinshania (pustules present) from Hil- 
liella (Zhang & Xu, 1990), appear not to be con- 
gruent, failing to provide a good argument to split 
Yinshania into several genera (Al-Shehbaz et al., 
1998). 


MORPHOLOGICAL CLUSTER ANALYSIS 


A cluster analysis of morphological and ecolog- 
ical characters has been performed by Zhang and 


Xu (1990). This analysis considered distribution, 
altitude, and habit, but also morphological char- 
acters describing hairs, inflorescences, flower de- 
tails, silicles, seeds, and cotyledons. They conclud- 
ed that minute pustules on the testa surface of the 
valves were a good character to separate Yinshania 
from Hilliella. Comparison of our molecular phy- 
logenies with this cluster analysis based on 32 mor- 
phological and 3 ecological characters demonstrat- 
ed the lack of congruency between morphological 
and molecular evolution. This cluster analysis sep- 
arated Yinshania sensu Ma and Zhao (1979) from 
Hilliella sensu Zhang and Li (Zhang, 1986) (a sim- 
ilar resolution to the ITS data). Taxa such as 

paradoxa, H. lichuanensis, and H. changhuanensis 
are closely related to each other (as confirmed by 
molecular data by acc. nos. 11, 13, and 15 herein). 
However, this morphological cluster analysis also 
grouped H. guangdongensis (acc. no. 12 herein) 


270 


Annals of the 
Missouri Botanical Garden 


into this group (У lichuanensis clade, Fig. 2). For 
both ITS and trnL intron sequence, H. guangdon- 
gensis is very divergent from others grouping with 
the Y. lichuanensis clade (Fig. 2). Hilliella alatipes 
var. micrantha and H. sinuata are separated with 
both molecular markers (Fig. 2), but are combined 
as a well-supported group in morphological cluster 
analysis (Zhang & Xu, 1990). In summary, within 
Hilliella there is little morphological agreement 
with our molecular data: (1) H. paradoxa, H. chan- 
ghuaensis, and H. lichuanensis are closely related 
to each other (as Y. lichuanensis clade, Fig. 2); (2) 
H. hunanensis and H. rupicola grouped together on 
the ITS tree (but not based on trnL data, Fig. 2). 
No further correlation could be observed. None of 
the three data sets (ITS, trnL intron, morphology) 
is powerful enough to elucidate phylogenetic signal 
for the whole species complex. However, significant 
correlations could be observed when cpDNA- and 
nrDNA-derived phylogenies were compared, divid- 
ing the Hilliella/Cochleariella clade into several 
subgroups (Fig. 2) clearly separating Yinshania rel- 
atives from Hilliella/Cochleariella with the sole ex- 
ception of Y. qianningensis. 


HYBRIDIZATION, INTROGRESSION, CHLOROPLAST 
CAPTURE, AND CONCERTED EVOLUTION 


The phylogeny based on plastid trnL intron se- 
quence data reflects the maternal lineages because 
plastids are inherited maternally in most angio- 
sperms, including the Brassicaceae (Harris & In- 
gram, 1991; Reboud & Zeyl, 1994). Introgression 
of a chloroplast type characteristic for the Hilliella/ 
Cochleariella clade into Yinshania qianningensis 
demonstrates possible gene flow between both 
groups. Because of the highly polyploid genomes of 
H. yixianensis, H. paradoxa, H. changhuaensis, and 
H. shuangpaiensis (represented by acc. nos. 2, 15, 
11, and 4, respectively, in Fig. 2) with multiple 
rDNA loci, and the assumed hybridization within 
the Hilliella/Cochleariella clade and even with the 
Yinshania clade, there is a high probability of con- 
certed ITS sequence evolution. In principle, there 
are three different ways that two different ITS cop- 
ies evolve within a single individual: (1) unidirec- 
tional concerted evolution leads to the loss of one 
copy and fixation of the second (detected in H. rup- 
icola acc. no. 14, herein; and in Gossypium, Wendel 
et al., 1995a); (2) both ITS copies are still present, 
which might be mostly the case in young hybrido- 
genous taxa (detected in H. fumarioides acc. no. 17, 
and С. zhejiangensis acc. no. 21, Fig. 2; in Krigia, 
Kim & Jansen, 1994; in Arabidopsis, O’Kane et al., 
1996); and (3) concerted evolution leads to a new 


ITS type that represents a mixture of the two orig- 
inal ITS sequences (in Gossypium, Wendel et al., 
1995b; in Microseris, van Houten et al., 1993; in 
Microthlaspi, Mummenhoff et al., 1997). The third 
type of concerted evolution might have happened 
in H. sinuata (acc. no. 3). This accession showed a 
plastome type more similar to H. shuangpaiensis 
acc. no. 4). However, ITS sequence types from pu- 
tative parental ITS sequence types from H. sinuata 
(acc. no. 9) and H. shuangpaiensis (acc. по. 4) ex- 
hibit some additive features found in H. sinuata 
(acc. no. 3). Comparing these three sequences, 
there are 39 variable nucleotide positions (21 in 
ITS1 region, and 18 in ITS2 region). Within the 
1151 region, 17 out of 21 variable nucleotide po- 
sitions (81%) are identical among the two H. sin- 
uata accessions (nos. 3, 9); H. shuangpaiensis 
shared only three mutations with H. sinuata (acc. 
no. 9) and one mutation with H. sinuata (acc. no. 
3). Within the ITS2 region both H. sinuata acces- 
sions have only one nucleotide position out of 18 
variable nucleotide sites invariant, but H. shuang- 
paiensis (acc. no. 4) shared 15 positions (8396) with 
H. sinuata (acc. no. 3) and two positions with H. 
sinuata (acc. no. 9). These findings indicate that 
ITS type of H. sinuata (acc. no. 3) consists of a 
mixture of ITS types from relatives of H. shuang- 
paiensis (mostly ITS2 region) and H. sinuata (1151 
region). The overall sequence divergence between 
H. shuangpaiensis (acc. no. 4) and H. sinuata (acc. 
no. 9) 18 7.896. 

Concerted evolution of ITS DNA loci has been 
shown several times to occur in the Brassicaceae 
O'Kane et al., 1996; Mummenhoff et al., 1997; 
Koch et al., 1998b; Franzke et al., 1998) and other 
families (Wendel et al., 1995a, b; Buckler et al., 
1997). Sequence divergence values of ITS types 
from putative parents, giving rise to hybrids in 
which concerted evolution has been observed, 


~ 


— 


ranged from 3.196 (Microthlaspi natolicum vs. M. 
perfoliatum, Mummenhoff et al., 1997), 5.096 (Car- 
damine amara vs. C. rivularis auct., Franzke et al., 
1998), to 6% (Arabidopsis thaliana vs. A. arenosa, 
O'Kane et al., 1996) comparable to a value of 7.896 
found among H. shuangpaiensis versus H. sinuata. 

We conducted a split decomposition analysis to 
visualize conflicting phylogenetic signal indicating 
concerted evolution in groups among the Hilliella/ 
Cochleariella clade (Fig. 6). This analysis clearly 
indicates hybridization with subsequent concerted 
evolution of ITS regions in H. sinuata (acc. no. 3). 
Concerted evolution of ITS sequences greatly influ- 
ences any interpretation of the ITS phylogeny. 
Since diploid members of the Brassicaceae such as 
Arabidopsis thaliana typically show 2 NOR loci, 


Volume 87, Number 2 
2000 


Koch & Al-Shehbaz 
Chinese Yinshania 


one could assume that in hexaploid Hilliella/Coch- 
leariella taxa at least 6 major NOR loci are present. 
Therefore, sequencing of two individual ITS clones 
is not a sufficient survey to find all putative ITS 
types from a single individual. This undersampling 
leads to an underestimation of ITS type variation 
as well as the degree of hybridization and concerted 
evolution. 

We found evidence suggesting concerted evolu- 
tion and described three examples (H. fumarioides 
acc. no. 17, H. rupicola acc. no. 14, H. sinuata acc. 
no. 3) of possible hybridization and subsequent 
concerted evolution of ITS sequence. The overall 
amount of sequence divergence between taxa in 
this study demonstrates a relatively high age for the 
different lineages. In fact, sequence divergence val- 
ues are much higher when compared to those ob- 
tained in infrageneric studies of closely related spe- 
cies of other Brassicaceae (e.g., Cochlearia s. str. 
< 1.75%, Koch et al. (19992); Noccaea < 4.5%, 
Mummenhoff et al. (1997); Cardamine < 4.5%, 
Franzke et al. s Within the Yinshania clade, 
seq e dist values range up to 696, but with- 
in the ‘HilliellalCochleariella clade they range even 
higher, up to 


Literature Cited 
Al-Shehbaz, I. A. & G. Yang. pu = on Chinese 
Cardamine. Harvard Гар. Bot. 3: 
m u & T. Y. ы 1998. Delimi- 
SR of the dor genera Yinshania, res and 
Cochleariella (Brassicaceae). Harvard Pap. Bot. 3: 79— 
94. 


Tos V. 1983. The е | the family Brassica- 
e. Bot. Zhurn. 68: 1297 

Baldwin, B. G., M. ae » M. Porter, M. F. е 
jeiechowski, С. S. Campbell & M. J. Donoghue. 199 
The ITS region of nuclear ribosomal DNA: A E 
source of evidence on Aris gi phylogeny. Ann. Mis- 
souri Bot. Gard. 82: 2 

Bandelt, H.-J. & A. W. s de 1992. Split decompo- 
sition: А new and useful approach to ји "e 
ysis of distance data. Molec. Phylogen. Evol. 
252. 


Baum, D. A., K. J. Sytsma & P. С. Hoch. 1994. А phy- 
mo ии of Epilobium beg ines hased on 
uclear ribosomal DNA sequences. Syst. Bot 


Böhle, U., H. Hilger, R. Cerff & W. F. Martin. 1995. Non- 
coding n DNA for plant systematics at the in- 
frageneric level. Pp. 391—403 in B. Schierwater, B. 
Streit, С. P. Wagner & R. DeSalle (editors), Molecular 
Ecology and Evolution: Approaches and Applications. 
Experientia үш Basel. 

Bremer, K. 1988. The limits of amino acid sequence data 
in morus phylogenetic reconstruction. Evolution 
42: 79-803. 

Buckler, Е. S., A. Ippolito & T. P. Holtsford. 1997. The 
evolution of ribosomal DNA: Divergent paralogues and 
phylogenetic implications. Genetics 145: 821—832. 

Campbell, C. S., M. J. Donoghue, B. С. Baldwin & M. Е. 


Wojciechowski. 1995. Phylogenetic relationships in 
Maloideae (Rosaceae): Evidence from sequences of the 
internal transcribed spacers of nuclear ribosomal D 
and its congruence with morphology. Amer. J. Bot. 82: 
903-918. 

Downie, S. R. & D. S. Katz-Downie. 1996. A molecular 
phylogeny of Apiaceae subfamily Apioideae: Evidence 
from nuclear ribosomal DNA internal transcribed spac- 

ме: 51. 


1987. A rapid DNA isolation 

procedure for за Еа оѓ fresh leaf tissue. Phy- 
tochem. Bull. 5. 

Dvorák, F. tA s the evolutionary relationship in the 
family Brassicaceae. Feddes Repert. 82: 357-372. 

Endress, P. K. 1 Evolution and floral diversity: The 
phylogenetic surroundings of EL and Antirrhi- 

nt. J. Pl. Sci. 153: 5106–512 
sess 1. 1985. Confidence Es on phylogenies: 
proach using the bootstrap. Evolution 39: 783— 


— 
е 


Fianzke. A., K. Pollmann, W. Bleeker, R. Kohrt 
Hurka. 1998. Mole cular systematics of Cardamine dud 
allied genera (Brassicaeae). ITS and non- ds chlo- 
roplast DNA. Folia E D Ue 33: 22 
Th 


Слеђу, L. & P. Taberlet. use of itus M 
DNA to resolve plant M рай Noncoding versus 
rbcL sequences. Molec. Biol. Evol. 11: 769-777 


Ham, R. C. H. J. van, H. Hart, T. Ё 
Sandbrink. 1994. Molecular evolution of noncoding ге- 
gions of the chloroplast genome in the Crassulaceae and 
related ареста: Curr. Genet. 25: 558- 

Harris, S. Н. & R. dvd 1991; Chloroplast DNA and 
biosy stematics: The effects o 
plaid transmission. Taxon 40: 303-412. 

yek 1911. Enturf eines Cruciferen Systems 
auf рогати ther Grundlage. Вей. Bot. Centralbl. 
27: 127-3 

Houten, ce . N. Scarlett & K. Bachmann. 1993. 

Nuclear DNA nale. of the пуча à tetraploid Mi- 
croseris scapigera and its Nort an diploid rela- 

. Theor. Appl. Genet. 87: 498505. 

Huson, D. H. 1998, SplitsTree: Analyzing and visualizing 
evolutionary data. Bioinformatics 14: 68—73. 

42. Das System der Cruciferen. Oesterr. 

Ж 


ty and 


m bg E. ч 
91: 

Кип, Г + & 3 K. Jansen. 1994. Comparisons of phy- 
logenetic hypothe sis among different data sets in dwarf 
dandelions (Krigia): Additional information from inter- 
nal transcribed spacer se es of nuclear ribosomal 
DNA. РІ. Syst. Evol. 190: 157-185 

Kimura, M. 1980. A simple method for estimating evo- 
lutionary rates of base substitutions through compara- 
tive studies of nucleotide sequences. J. Molec. Evol. 16: 

—120. 

Koch, M., Н. Hurka & К. Mummenhoff. 1996. Chloro- 
plast DNA restriction site variation and RAPD-analyses 
in Cochlearia (Brassicaceae): Biosystematics and spe- 
ciation. iut s Bot. 16: 585—603. 

——— luthmann & H. Hurka. 1998a. Isozymes, 
spec ‘ation = evolution in the polyploid complex Сосћ- 
learia L. (Brassicaceae). Bot. Acta 111: 411-425. 

‚ К. Mummenhoff & Н. Hurka. 1998b. Molecular 

р ити and evolution of Microthlaspi perfoliatum 
s.l. polyploid complex (Brassicaceae): Chloroplast DNA 

‘ind nuclear ribosomal ET A restriction site variation. 


Canad. J. Bot. 76: 382—396 
———, ——— 4; 


3 19994. Molecular phyloge- 


272 


Annals of the 
Missouri Botanical Garden 


netics of Cochlearia L. and allied genera based on nu- 


ar ribosomal IT sequence analysis contradict 
ational concepts of He evolutionary relationships. 
Pl. Syst. Evol. 216: 207 


———, Т Bishop & T. eet Olds. 1999b. Molecular 
systematics and evolution of Arabis and Arabidopsis. Pl. 

Zhao. 1979. Yi пзћата, а new genus 

. Conspectus der "Thlaspi"-Arten Eu- 

ropas, Afrikas und Vorderasiens. Feddes Repert. 84: 

449—470. 

—————. 1979. Kritische Revision der "Thlaspi"-Arten 
Europas, Afrikas und Vorderasiens. I. Geschichte, Mor- 
phologie ves Chorologie. Feddes Repert. 90: 129-154. 

——. 199 eed-coat anatomy as a character for a 

ew а of Thlaspi. Fl. Veg. Mundi 9: 9-15. 

Mie о А och. 1994. Chlo ор restric- 
tion site variation and phylogenetic relationships in the 
genus Thlaspi sensu lato (Brassicaceae). Syst. Bot. 19: 
73-88. 


nzke & M. Koch. 1997. Molecular phy- 

logenetics of Thlaspi s.l. (Brassicaceae) based on chlo- 

roplast DNA restriction site variation and sequences of 

the internal transcribed spacers of nuclear Ше. 
NA. Canad. J. Bot. 75: 469-482. 

O’Kane, S. L., B. A. Schaal & I. A. Al-Shehbaz. 1996. 
The origin of Arabidopsis suecica (Brassicaceae) as in- 
dicated by nuclear rDNA sequences. Syst. Bot. 21: 

9-566. 

Peer, Y. van de & R. de Wachter. 1994. Treecon for win- 
dows: A software package for the construction and 
drawing of evolutionary trees for the Microsoft Windows 


cruciferarum. Novit. Sist. Vyssh. Rast. 5: 136-139. 
. 1970. Revisio generis Cochlearia L., 2. Novit. 

Sist. Vyssh. Rast. 7: 167-195. 

Price, R. A., J. D. Palmer & I. A. Al-Shehbaz. 1994. 
Systematic relationships of Arabidopsis: A molecular 
and morphological к Рр. 7-19 in Е. М. Меу- 

i merville (editors), Arabidopsis. Cold 

Spring Harbor Don Press, New York 

Nucleotide sequence 


of the 185—255 spacer region from mustard DNA. Nucl. 
: 2 


923. Eine neue сее der Gattung Сосћ- 
earia L. Notizbl. Bot. Gart. Berlin-Dahlem 8: 544—557. 

. 1936. Cruciferae. /n: A. Engler & K. Prantl (ed- 
SES Nat. Pflanzenfam., inm 2, uo 227—658. Verlag 


993. PAUP: Phylogenetic Analysis Us- 
ing Parsimony, version 3.1. A computer program dis- 


tributed by the Illinois Natural History Survey, Cham- 
paign, Illinois 
Sytsma, к. J. 1990. DNA and morphology: a of 
lant phylogeny. Trends Ecol. Evol. 5: 104-11 
Taberlet, P., L. Gielly, С. Pautou & J. Bouvet. on. Uni- 
versal primers for amplification of three non-coding re- 
gions of chloroplast DNA. Pl. Molec. Biol. 17: 1105- 
109 


1 ; 

Warwick, S. I., L. D. Black & I. Aguinagalde. 1992. Mo- 
lecular калын o of Brassica and allied genera (sub- 
tribe Brassicinae, Brassiceae)—Chloroplast DNA vari- 
ation Aa the genus Dpi Theor. Appl. Genet. 
83: 839. 

Wendel, J. T A. Schnabel & T. Seelanan. 1995a. 
rectional interlocus concerted evolution и allo. 
ploid "nd iation in cotton (Gossypium). Proc. Natl. 
Acad. Sci. U.S.A. 92: 280-284. 

Я & 1995b. An unusual ribosom- 

al DNA sequence from Cossypium gossypioides reveals 

ancient, cryptic, intergenomic introgression. Molec 
l 


, T. Burns, S. Lee & J. Taylor. 1990. Ampli- 
fication and direct сае чы of fungal ribosomal RNA 
genes for а Saat Pp. 315-322 т M. A. Innis, 
D. H. Gelfand, J. J. Sninsky & T. J. White (editors), 
PCR p. a Academic Press, New York. 

Wojciechowski, M. F., M. J. Sanderson, B. G. Baldwin & 

1993. Monophyly of aneuploid As- 
tragalus (Fabaceae): Evidence from nuclear ribosomal 
DNA internal transcribed spacer sequences. Amer. 
Bot. 80: 711-722. 

ens Y. H. 1985. Cochleariopsis—A new bed of Chi- 

ese Cruciferae. Acta Bot. Yunnan. 7: 143-145 

. 1986. Hilliella, a new genus of Cruciferae. Acta 

Bot. Yunnan. 8: 397—406. 

9 A revision of ү Yinshania (Crucifer- 

ae). Acta | Роа, Sin. 25: 204-219. 
new species of Yinshania with a dis- 

cussion ut the е and origin of the genus. Acta 

Bot. Yunnan. 15: 364—368. 

1 


comparison of chromosome numbers 

and ойна e zymograms of Yinshania and Hilliella. 

J. Pl. Resources Environm. 4(2): 27-31. 
& 1989. Observation on the genera 

Kakani Hilliella, Cochleariella, and Cochlearia T 

ciferae) by SEM. Acta Bot. Boreal.-Occid. Sin. 9: 2 

231. 


K. Xu. 1990. A numerical taxonomic study on 

two genera Yinshania and Hilliella (Cruciferae). J. Wu- 

han Bot. Res. 8: 317-324. 

. 1992. А taxological revision on Cochlearia, 
gir Hilliella, and Cochleariella in China. Acta 

Univ. Intramongol. 23: 561—573. 

Zunk. K., K. Mummenhoff, M. Koch & H. Hurka. 
Phylogenetic relationships of Thlaspi s.l. 
Thlaspidinae, Lepidieae) and allied genera based on 
chloroplast DNA restriction site variation. Theor. Appl. 
Genet. 92: 375-381 


bias 


MOLECULAR CONFIRMATION 
OF UNIDIRECTIONAL 
HYBRIDIZATION IN BEGONIA 
X TAIPEIENSIS PENG 
(BEGONIACEAE) FROM 
TAIWAN! 


Ching-I Peng? and Tzen-Yuh Chiang? 


ABSTRACT 


An unusual Begonia that sheds staminate flowers prematurely at bud stage was collected from several localities in 


northern Taiwan. Observations on morpho 


logy, pollen stainability, and seed set of 
hybrid origin. Morphological comparisons, distribution patterns, chromoso 


this species initially suggested a 
me cytology, and experimental hybridization 


showed that such ite аге F, hybrids (2n = 41) between Begonia formosana (Hayata) Masamune (п = 30) and В 


ia X taipeiensis Pen 


between the putative parents in the wild resulted in a formation of B. Х pius: is No natural hybri 
ed. Abortion caused by a 
ternal parent. Low pollen а of Е, hybrids indicates gun he natural hybrid is uisu ак 


with a maternal origin from B. aptera have been dete 
aptera was used as a mate 
Ы recurrent hybridization between the parental speci 
Key words: atpB-rbcL п 
directional hybridization, аа hybrid, Taiwan. 


11), both of which are a in Taiwan and sympatric in most of their 


ranges. These hybrids 


F, а вя only when В. formosana was used as the femal lecular 
8 а! BÉ hybridization 
1 УМ i 


dem tion barrier 


oncoding spacer, Begonia x жы ын В. formosana, В. aptera, chloroplast DNA, cytology, 


Natural hybridization, one of the most influential 
processes that increases species diversity and sta- 
bilizes genetic complexity via genetic recombina- 
tion or introgression (Arnold, 1992, 1993), occurs 
frequently in plants (P. S. Soltis & D. E. Soltis, 
1991; P. S. Soltis et al., 1992; Arnold, 1997). Stud- 
ies on the origins of natural hybrids and their ge- 
netic composition frequently raise fundamental 
questions concerning reproductive barriers, survi- 
vorship and fitness of the hybrids, and mechanisms 
of reticulate evolution. In many cases (e.g., Loui- 
siana Irises; Arnold & Bennett, 1993) natural se- 
lection may favor hybrid plants of specific maternal 
origin. It is essential to study the genetic makeup 
of natural hybrids in order to elucidate evolutionary 
processes and the ecological adaptation of these 
plants. 

It has been abundantly demonstrated that natural 
hybridization plays a major role in the evolution of 
species groups or complexes (Ludwigia: Peng, 

988, ; Iris: Arnold et al, 1990; Glycine: 
Doyle et al., 1990; Helianthus: Rieseberg et al., 


1990, Rieseberg, 1991; Gossypium: Wendel et al., 
1991; Senecio: Harris & Ingram, 1992; Allium: 
Ohsumi et al., 1993; Leucaena: Hughes & Harris, 
1994; Arabidopsis: Mummenhoff & Hurka, 1995; 
Argyroxiphium: Baldwin, 1997; and Argyranthe- 
mum: Francisco-Ortega et al., 1997). Natural hy- 
bridization occurs frequently in Taiwanese Begonia 
(Y. K. Chen, 1988; Peng & Chen, 1991; Peng & 

ue, 2000), which may have resulted in the high 
level of endemism of this genus on the island 
(66.7%; cf. C. H. Chen, 1993), while causing dif- 
ficulties in clarifying phylogenetic relationships 
(see Rieseberg & Morefield, 1995) 

Begonia X taipeiensis Peng was recently de- 
scribed as a new intersectional hybrid (Peng & Sue, 
2000). Geographical distribution, low level of pol- 
len stainability, and intermediate morphological 
characters between B. formosana (Hayata) Masa- 
mune and B. aptera Blume initially suggested such 
an origin. Begonia aptera [sect. Sphenanthera 
(Hassk.) Warb.] is distributed in northern Taiwan, 


' This study was supported by a National Science Council grant 87-2311-B-001-086, Taiwan, to Ching-I Peng and 
Tzen-Yuh Chiang. The authors thank Chian-Yi Sue for assistance in figure preparation and cytological work and Chen- 


Fang Chiang for assistance in DNA sequencing. 
2 Institute of Botany, Academia Sinica, Nankang, Taipei 1 


15, Taiw 


* Department of Biology, National Cheng-Kung University, Tainan ТОО, Taiwan. 


ANN. Missouri Bor. GARD. 87: 273-285. 2000. 


274 


Annals of the 
Missouri Botanical Garden 


while B. formosana [sect. Platycentrum (Klotzsch) 
A. DC.] is widespread in northern and eastern Tai- 
wan. Both species occur in wide altitudinal ranges 
(50-2000 m). They frequently co-occur throughout 
their range of overlap (Fig. 1). The flowering peri- 
ods are March to December for B. formosana, June 
to August for B. aptera, and April to December for 
B. X taipeiensis. Two discrete populations of Be- 
gonia X taipeiensis were found mixed with its pu- 
tative parents in the Taipei Basin of northern Tai- 
wan. Begonia formosana is characterized by having 
creeping rhizomes with ascending to erect flowering 
stems, broadly ovate, lobed leaves, 2-locular ova- 
ries, and unequally winged capsules, the abaxial 
one being very pronounced. In contrast, B. aptera 
is a tall, cane species lacking horizontal rhizomes, 
has lanceolate leaves, 3-locular ovaries, and round- 
ed, wingless capsules. Begonia X taipeiensis has 
short rhizomes, intermediate plant height and leaf 
shape, and 2-locular ovaries (these occasionally 
highly reduced and without locules) with diminu- 
tive wings. Like all other species of Begonia, B. X 
taipeiensis is monoecious and interflorally protan- 
drous. At anthesis, it produces abundant flowers of 
both sexes. All staminate flowers observed develop 
normally up to their late bud stages and then drop 
off before they open. All Begonia of Taiwan are 
frequently pollinated by blowflies (Calliphoridae), 
hover flies (Syrphidae), and honeybees (Apidae) (Y. 
K. Chen, 1988). Plants of Begonia often grow along 
moist stream banks. Their light seeds float and are 
carried easily by water currents. Seeds of Begonia 
are primarily dispersed with the aid of wind (van 
der Pijl, 1972). 

Traditionally, morphological examination and 
chromosome cytology have provided useful infor- 
mation in revealing putative parents. Nevertheless, 
such data are not sufficient to determine the direc- 
tion of hybridization and to detect genetic variabil- 
ity. Recently, many convincing studies using mo- 
lecular techniques such as comparison of RFLP 
patterns of ribosomal DNA (Arnold et al., 1990; 
Stein & Barrington, 1990; Rieseberg, 1991; P. S. 
Soltis & D. E. Soltis, 1991; Hughes & Harris, 1994; 
Mummenhoff & Hurka, 1995) and those of organ- 
elle DNA (D. E. Soltis & P. S. Soltis, 1989; De- 
Marais et al., 1992; Garcia & Davis, 1994; Wang 
& Szmidt, 1994; Brochmann et al., 1998), coupled 
with experimental hybridizations (Hodges et al., 
1996), have been conducted to address reticulate 
or polyploid evolution (D. E. Soltis & P. S. Soltis, 
1993). Chloroplast and mitochondrial genomes, be- 
ing maternally inherited in most organisms (Chiu 
& Sears, 1985; D. E. Soltis et al., 1990), have been 


useful in elucidating the direction of gene flow be- 


tween populations (Whittemore & Schaal, 1991) 
and in detecting the maternal origin of natural hy- 
brids (Wendel et al., 1991). Thus, in this study, a 
noncoding spacer region between the genes rbcL 
and atpB of the chloroplast genome, which has 
been used as a molecular marker at the species 
level (Golenberg et al., 1993; Chiang, 1994; Manen 
& Natali, 1995; Chiang et al, 1998), was se- 
quenced. 

In this study the following questions will be ad- 
dressed: (1) Did B. X taipeiensis originate from in- 
terspecific hybridization between B. aptera and B. 
formosana? (2) Do hybrids with alternate maternal 
origins have the same survivorship? (3) Do hybrids 
backcross with putative parents and produce viable 
offsprings? (4) Is the frequency of occurrence of 
natural hybrids a reflection of the ease of experi- 
mental crosses? 


MATERIALS AND METHODS 
1. MATERIALS SAMPLED 


Major herbaria in Taiwan (HAST, NTUF, TAI, 
TAIF, TNM) were consulted to determine the dis- 
tribution and phenology of Begonia X taipeiensis 
and its putative parents, B. formosana and B. ap- 
tera. Living plants of the hybrid and its associated 
putative parents were collected from two localities 
of Taipei County, Hsichih and Wulai (Fig. 1). Ad- 
ditionally, a plant each of B. formosana and B. ap- 
tera from Chiufen and Nankang, respectively, where 
B. X taipeiensis was not found, were collected for 
comparison (Fig. 1). All plants were grown in the 
experimental greenhouse of the Institute of Botany, 
Academia Sinica, Taipei, for cytological and mo- 
lecular examination and for experimental hybrid- 
ization. Begonia palmata D. Don, a congener col- 
lected from Hsitou, Nantou County, was used as an 
outgroup. Herbarium vouchers were deposited at 


HAST (Table 1). 


2. EXPERIMENTAL POLLINATION 


Experimental self-pollinations of B. aptera and 
B. formosana were done. Reciprocal crosses were 
carried out between B. formosana and B. aptera 
from the same locality and from different localities 
in the greenhouse. Begonia X taipeiensis collected 
from the wild was used as a pistillate parent for 
backcrossing with both putative parents. The fact 
that Begonia X taipeiensis shed staminate flowers 
prematurely precludes the possibility of using it as 
a pollen donor. Seeds (F, progeny) were grown to 
maturity to examine the fertility of pollen grains 
and for cytological study. 


275 


Peng & Chiang 


Volume 87, Number 2 


2000 


Begonia x taipeiensis Peng 


‘ugjniyy ‘p :Suexuew ‘E ‘Чон ‘g My ‘Г (5115) sisuaiadin) x; ^g pue *(so[jdueun) 2127840 'g *(so[o1to) рирхошлој "Я :seipnis 1e[noo[our pue јеотдојој А о 10} 
(t-1) senieoo| шолу ројозјј0оо suauiroads әҷі JO поцеоој pue :ueAreg ut (se[dueu1) piajdp ‘g pue (so[o1ro) рирзошалој 711099 јо (jesut) uonnquisip әш JUIMOYS dew С] ainsi 4 


— 


276 Annals of the 
Missouri Botanical Garden 
Table 1. 


barium vouchers are deposited at HAST. 


Materials of Begonia X taipeiensis and its putative parents for cytological and molecular analysis. Her- 


EMBL 
Taxa Localities Vouchers accession no. 

В. X taipeiensis 1. Wulai, Taipei Co. Peng 13899 AJ009601 
Peng 15106 AJ009602 
2. Hsichih, Taipei Co. Peng 16320 AJ009600 
B. aptera l. Wulai, Taipei Co. Peng 16276 AJ223092 
Peng 16153 AJ009606 
2. Hsichih, Taipei Co. Peng 16321 AJ009605 
3. Nankang, Taipei Co. Peng 16292 AJ009604 
B. formosana 1. Wulai, Taipei Co. Peng 13915 AJ009599 
2. Hsichih, Taipei Co. Peng 16319 AJ009598 
3. Chiufen, Taipei Co. Leu 867 AJ009597 
B. palmata l. Chitou, Nantou Co. Peng 16081 AJ007745 
2. Tengchi, Kaohsiung Co. Peng 16831 AJ242856 

Experimental hybrid: 
Bf Xa-1: B. formosana X B. aptera = Leu 867 X Peng 16153 AJ242857 
Bf Xa-2: B. formosana X B. aptera = Peng 13915 X Peng 16153 AJ242858 


3. CHROMOSOME CYTOLOGY 


Flower buds to be examined for meiotic behavior 
were fixed in a 3 : 1 mixture of 95% ethanol and 
glacial acetic acid and stored in the refrigerator. 
Prior to staining, the buds were hydrolyzed for 5- 
8 min. at 60°C using a 1 : 1 mixture of concentrated 
HCI and 95% ethanol. They were then squashed in 
FLP orcein (Jackson, 1973). Somatic chromosome 
counts were obtained from actively growing root 
tips pretreated for 3 to 4 hr. in 8-hydroxyquinoline, 
then fixed as above for at least 10 min. The root 
tips were then hydrolyzed in 1 N HCI for 8-10 min. 
at 60°C and squashed in FLP orcein. All analyzable 
chromosome configurations (mostly diakinesis or 
metaphase I) were documented with camera lucida 
drawings or photomicrographs using Kodak Pana- 
tomic-X films. Negatives and drawings are depos- 
ited at the Institute of Botany, Academia Sinica, 
Taipei. 


4. MOLECULAR METHODS 


DNA isolation, PCR, and nucleotide sequencing. 
Fresh tissue of young shoots was ground in liquid 
nitrogen and stored at —70°C for DNA extraction. 
Genomic DNA was extracted following a CTAB 
methodology (Doyle & Doyle, 1989). PCR was per- 
formed in a volume of 100 wl reaction using 10 ng 
of template DNA, 10 ш of 10X buffer, 10 у MgCl, 
(25 mM), 10 pl dNTP mix (8 mM), 10 pmole of 
each primer, 10 pl of 10% NP-40, and 2 U of Тад 
polymerase (Promega, Madison, U.S.A.; cf. Chien 
et al., 1976). The reaction was programmed on an 


MJ Thermal Cycler (PTC 100) as one cycle of de- 
naturation at 95?C for 4 min., 30 cycles of dena- 
turation at 92°C for 45 sec., annealing at 52°C for 
min. 15 sec., and extension at 72°C for 1 min. 
30 sec., followed by a 10 min. extension. Template 
DNA was denatured with reaction buffer, MgCl,, 
NP-40, and ddH2O for 4 min. (first cycle), and 
cooled on ice immediately. Primers [rbcL-1, 5'-AA- 
CACC A(G)JAATCCAA-3'; atpB-1, ACA- 
TCT(G)AA(G)TACT(G)GGACCA ATA A-3'; (Chiang 
et al., s, and Taq polymerase were 
added to the bove ice-cold mix. Reaction was re- 
started at the first annealing at 52°C. PCR frag- 
ments were eluted using High Pure PCR Product 
Purification Kit (Boehringer Mannheim, Germany 
cf. Vogelstein & Gillespie, 1979). PCR grues 
were ligated to a pT7blue T-vector (Novagen, Mad- 
ison, U.S.A.; cf. Marchuk et al., 1991) and cloned 
in competent E. coli DH5a. Plasmid DNA was ех- 
tracted from transformed cells and sequenced with 
%P labeling extension/termination reaction (Sanger 
et al., 1977) using fmol DNA Sequencing System 
(Promega, Madison, U.S.A.; cf. Murray, 1989). For 
completing sequencing, with about 75 base pair 
overlaps from both ends, two additional primers, 
ie. rbcL-2 (5’-ССТТССТТСТТТСААСАТСС-3') 
and atpB-2 (5’-СССТССААСССААТССААТТС- 


3'), were synthesized. 


DNA alignment and phylogenetic analy- 
sis. Alignment of nucleotide sequences was per- 
formed by Clustal V program (Higgins et al., 1992) 
and improved by eye. Parsimony phylogenetic anal- 


Volume 87, Number 2 
2000 


Peng & Chiang 
Begonia X taipeiensis Peng 


277 


yses were performed by using the Phylogenetic 
Analysis Using Parsimony Program (PAUP, version 
3.1.1., Swofford, 1993) with the branch and bound 
algorithm. Neighbor-Joining (NJ) analysis by cal- 
culating Kimura’s (1980) two-parameter distance 
was also performed using Molecular Evolutionary 
Genetics Analysis Program (MEGA, version 1.01, 
Kumar et al., 1993). Confidence of clades recon- 
structed was tested by bootstrapping (Felsenstein, 
1985) with 1000 replicates (Hedges, ) 
nodes with bootstrap values greater than 0.70 are 
significantly supported with > 95% probability 
(Hillis & Bull, 1993). In NJ analysis, complete- 
and-partial (CP) bootstrap technique was used to 
correct the bias (conservativeness) of the standard 
bootstrap approach (Li & Zharkikh, 1995). A gl 
test (Huelsenbeck, 1991) of skewed tree-length dis- 
tribution was calculated from 10,000 random trees 
generated by PAUP in order to measure the infor- 
mation content of the data. Critical values of the g1 
test were obtained from Hillis and Huelsenbeck 

992). The fit of character data on phylogenetic 
hypotheses (Swofford, 1991) was evaluated by the 
consistency index, CI (Kluge & Farris, 1969), and 
the retention index, RI (Farris, 1989). The statis- 
tical significance of the CI was determined accord- 
ing to the method of Klassen et al. (1991). The 
number of nucleotide substitution, which is the 
number of transitional and transversional substitu- 
tions per site, was calculated following the meth- 
odology of Wu and Li (1985) 


Ф 


RESULTS 
EXPERIMENTAL SELFING AND HYBRIDIZATION 


In all experimental geitonogamous selfing at- 
tempts made on Begonia formosana and B. aptera, 
fruits with 95-100% viable seeds were consistently 
obtained. Plants of B. formosana required 30—45 
days for fruit maturation, whereas those of B. aptera 
required 90-150 days. Begonia formosana and B. 
aptera were crossed reciprocally in the experimen- 
tal greenhouse. Nearly all crosses using B. formo- 
sana as pistillate parent were successful, producing 
mature fruits with 80-90% plump seeds 35—45 
days after pollination. Such seeds were viable and 
flowering artificial F, hybrids were readily obtained 
from them. The level of stainable pollen in these 
F, plants was extremely low (ranging from 0 to 5%), 
which agrees with that of the naturally occurring B. 
X taipeiensis. When B. aptera was used as the pis- 
tillate parent in the crosses, however, precocious 
fruit drop occurred ca. 60 days after artificial pol- 
lination. 

Experimental hybrids between B. formosana and 


B. aptera were grown to maturity. They flowered 
annually and have persisted in the mist greenhouse 
since the summer of 1995. The plant height in 
these artificial hybrids was within the variation 
range of naturally occurring B. X taipeiensis. The 
artificial hybrids closely resembled B. Х taipeiensis 
in habit and details of vegetative as well as floral 
characters. When plants of Begonia X taipeiensis 
collected from the wild were backcrossed (as pis- 
tillate parent) to B. formosana and B. aptera, re- 
spectively, fruit set was successfully obtained. 
However, five backcrossing attempts made with B. 
formosana and 11 such attempts with B. aptera pro- 
duced fruits with zero or negligible plump seeds 
that failed to germinate. 


CHROMOSOME CYTOLOGY 


Previous studies revealed a meiotic chromosome 
number of n = 11 in B. aptera, n = 30 in B. 
formosana, and 2n = 41 in B. X taipeiensis (Peng 
& Sue, 2000). Like B. Х taipeiensis, experimental 
hybrids of B. formosana X B. aptera преса. gn. 
have a somatic chromosome number о 
Also, abnormalities in chromosome el shat hes 
were observed. Meiotic chromosome configurations 
of both B. X taipeiensis and B. formosana X B. 
aptera typically consisted of some sticky, often dis- 
oriented bivalents, univalents, and multivalent as- 
sociations (Fig. 2). 


MOLECULAR DATA 


Sequences with a consensus length of 854 base 
pairs of the atpB-rbcL spacer were obtained from 
B. X taipeiensis and its putative parents. Fifteen 
variable sites were found between sequences (Table 
2). That this chloroplast spacer has, on average, 
34.0% A and 36.1% T agrees with one of the com- 
mon properties, i.e., AT-rich, of most noncoding 
spacers (Li, 1997). Differences in the rate of nu- 
cleotide substitution (Table 3) among species of Be- 
gonia we sampled ranged from 0.0017 (between B. 
aptera and B. formosana) to 0.0056 (between B. 
aptera and B. palmata) (mean = 0.0033). 

Of the three individuals of B. formosana we ex- 
amined, two had identical nucleotide sequences 
(Tables 1, 2). The third collection from Chiufen 
(Leu 867) differed at two different positions (bp 191 
and 762). Interpopulational variation was present 
in B. aptera at bp 275 and 794. In our study, se- 
quences of this chloroplast spacer for all three spe- 
cies were highly conserved, with 15 variable sites 
(2.1% of 854 bp) (Table 2). Begonia formosana 
shared with B. aptera 7 derived characters (at sites 
43, 44, 251, 252, 280, 568, 743). Two autapomor- 


278 Annals of the 
Missouri Botanical Garden 


ай 2. Meiotic chromosome spreads of Begonia.—A. Metaphase I, polar мем. —B. Metaphase I, equatorial 
w. —C. Anaphase I. —D. Telophase II. Bar equals 10 jum (A, Begonia X taipeiensis, from Peng 13899; B-D, B. 
Fou X B. aptera, from Leu 867 X Peng 16153). 


Table 2. Variable sites of the nucleotide of the atpB-rbcL spacer region of the chloroplast DNA of Begonia 
species. Dots indicate that the nucleotide е es are identical to those for Begonia aptera 16321. 

Taxa/Sites 43 44 191 251 252 270 275 286 298 371 376 568 743 762 794 
В. aptera 16321 C G T T A A A A G T T C T T G 
B. aptera 16276 . 5 i А Я ; : | : А А А : 3 : 
B. ap 16153 ; 5 : ; | 3 G : А : А : А С 
В. аріега 16292 : А А К 
В. taipeiensis 15106 A G C 
B. taipeiensis 13899 A G C 
B. taipeiensis 16320 A G C 
Bf Ха-: : ; З А G C : 
Bf Xa- о б À G С С 
В. formosana 13915 : A G C 
B. formosana 16319 A G C : 
B. formosana 867 C A G С ; C 
B. palmata 16081 T A C A C T C A T A 
B. palmata 16831 T A C C 1 C A T A 


Volume 87, Number 2 
2000 


Peng & Chiang 279 


Begonia x taipeiensis Peng 


Table 3. 
diagonal). 
B. taipeiensis 13899; 7 


1. B. aptera 16321; 2. B. aptera 16276; 3 
7. B. taipeiensis 16320; 8. Bf Xa- 


Pairwise comparisons of the numbers of nucleotide subsitutions per site (K) in Begonia species (above 
"ie gs 16153; 4. 
9. Bf Xa-1; 10. B. гелии 13915; 11. В. јогтоѕапа 16319; 


аріега 16292; 5. В. taipeiensis 15106; 6. 


12. В. formosana 867; 13. В. palmata 16081; 14. В. Ро 16831. 


|| 2 3 4 5 6 7 8 9 10 11 12 13 14 

1 0.0000 0.0011 0.0000 0.0017 0.0017 0.0017 0.0017 0.0028 0.0017 0.0017 0.0028 0.0056 0.0050 
2 0.0011 0.0000 0.0017 0.0017 0.0017 0.0017 0.0028 0.0017 0.0017 0.0028 0.0056 0.0050 
3 0.0011 0.0028 0.0028 0.0028 0.0028 0.0039 0.0028 0.0028 0.0039 0.0067 0.0061 
4 0.0017 0.0017 0.0017 0.0028 0.0017 0.0017 0.0028 0.0056 0.0050 
5 0.0000 0.0000 0.0000 0.0011 0.0000 0.0000 0.0011 0.0061 0.005 
6 0.0000 0.0000 0.0011 0.0000 0.0000 0.0011 0.0061 0.0056 
7 0.0000 0.0011 0.0000 0.0000 0.0011 0.0061 0.0056 
8 0.0011 0.0000 0.0000 0.0011 0.0061 0.0056 
9 0.0011 0.0011 0.0000 0.0061 0.0056 
10 0000 0.0011 0.0061 0.0056 
11 0.0011 0.0061 0.0056 
12 0.0061 0.0056 
13 0 
phies, at positions 371 and 376, in B. formosana DISCUSSION 


distinguished it from B. aptera. 

The sequences of Leu 867 and Peng 13915, both 
B. formosana, differed at two sites. The geneti 
uniqueness of the two collections was кашы 
to the Е, offspring, i.e., В/Ха-1 and BfXa-2, ге- 
есер. when they меге used as maternal parent 
in experimental crosses. We sampled two collec- 
tions of Begonia X taipeiensis, Peng 13899 and 
15106, both associated with B. formosana, Peng 
13915, from the same locality. Both Begonia X 
taipeiensis specimens were found to have identical 
sequences (Table 2) with the experimental hybrid 

a- 

Parsimony analysis identified two equally parsi- 
monious trees of 16 steps (Fig. 3), a CI of 0.938 (P 
< 0.01), and an RI of 0.958. A gl statistic of 
— 1.468 indicated а significant signal (P < 0.01). 
The Neighbor-Joining tree (Fig. 4), recovered by 
MEGA, based on the Kimura's two-parameter dis- 
tance (Table 4), is consistent with the parsimony 
trees, but with higher resolution. Three major 
clades in both parsimony and NJ trees (Figs. 3, 
obtained in these analyses were supported signifi- 
IC p E um the clade of B. formosana, 

B. eiensis, an experimental hybrid 
РИЙ 2 po clade ш В. formosana (Leu 867) and 
an experimental hybrid (В/Ха-1), and the clade of 
B. aptera. Within the first clade the monophyly of 
the subgroup composing the maternal parent (B. 
formosana, Peng 16319), the hybird offspring 
(BfXa-1), and another B. formosana (Peng 13915) 
was also significantly supported. 


HYBRID ORIGIN IN B. X TAIPEIENSIS: EVIDENCE FROM 
MORPHOLOGY, CYTOLOGY, AND DISTRIBUTION 


Based on morphological criteria, B. formosana 
and B. aptera were initially suggested as the pu- 
и parents of B. X taipeiensis (Peng & Sue, 

000). Cytological data showed that both the ex- 
| hybrids between B. formosana (n = 30) 
and B. aptera (n = 11) and the naturally occurring 
B. X taipeiensis have the same chromosome num- 
ber of 2n = 41. Begonia formosana is the only 
species with n = 30 in Taiwan, which is the highest 
chromosome number among these Begonia. Chro- 
mosome numbers of n = 11, 15, 18, and 19 are 
known for other members of Begonia on the island 
Y. K. Chen, 1988). A chromosome number of n = 
11 is documented in only two species, B. aptera 
and B. palmata (Y. K. Chen, 1988; Peng & Chen, 
1991). Begonia palmata was excluded as a candi- 
date for a putative parent of B. X taipeiensis based 
on the geographic distribution of the species. Ex- 
tensive fieldwork and examination of herbarium 
material revealed that the ranges of B. aptera and 
B. formosana largely overlap and that plants of B. 
X taipeiensis usually co-occur with these two spe- 
m in elevation. On the other 


— 


с1ез а! са. 60-200 
hand, В. palmata, МА а por species of 
higher eda (ca. 6 00 m), is geographi- 
cally and altitudinally эй Based on data from 
experimental crosses, cytological observation, and 
geographical distribution, we conclude that B. х 
taipeiensis represents F1 progeny from natural hy- 
bridization between B. formosana and B. aptera. 


280 


Annals of the 


Missouri Botanical Garden 


Figure 3. 


B aptera 16321 


B aptera 16276 


B aptera 16153 


B aptera 16292 


B taipeiensis 15106 


B taipeiensis 13899 


B taipeiensis 16320 


Bfxa-2 


B formosana 13915 


B formosana 16319 


Bfxa-1 


B formosana 867 


B palmata 16081 


B palmata 16831 


One of the parsimonious trees recovered by PAUP from nucleotide sequences of the atpB-rbcL spacer of 
chloroplast DNA rooted at Begonia palmata. Numbers at nodes are bootstrap values. 


Volume 87, Number 2 
2000 


Peng & Chiang 
Begonia x taipeiensis Peng 


99 B aptera 16276 


99 B aptera 16321 


B aptera 16292 


B aptera 16153 
B formosana 13915 


B formosana 16319 


Bfxa-2 


B taipeiensis 16320 


B taipeiensis 13899 


B taipeiensis 15106 


| sedes] Bfxa-1 


100 L———— B formosana 867 
B palmata 16081 
г р 


ова В palmata 16831 


ure 4. Neighbor-Joining tree recovered by MEGA from nucleotide sequences of atpB-rbcL spacer of chloroplast 
DNA rooted at уе palmata. Numbers at nodes аге complete-and-partial (CP) bootstrap values. 


UNIDIRECTIONAL HYBRIDIZATION GIVING RISE TO B. 
X TAIPEIENSIS: EVIDENCE FROM MOLECULAR DATA 
AND EXPERIMENTAL HYBRIDIZATION 


Identical sequences of the chloroplast atpB-rbcL 
spacer between B. X taipeiensis and B. formosana 
suggested that the former had a maternal origin 
from the latter. Unidirectional hybridization giving 
rise to the natural hybrid B. X taipeiensis is con- 
gruent with the results from reciprocal crosses in 
which viable Е, were obtained only when B. for- 
mosana was used as the pistillate parent. Experi- 
mental crosses using В. aptera as the maternal раг- 
ent resulted in precocious fruit drop, possibly as a 
result of genetic disharmonies. A similar observa- 
tion was made on Lousiana irises (Arnold & Ben- 
nett, 1993), in which unidirectional hybridization 
was documented using cpDNA haplotypes. Natu- 
rally occurring unidirectional hybridization sug- 
gests that hybrids of reversed parentages may have 
different survivorship, because organelle genomes 
may contribute genetic information that critically 
affects the survivorship of their progeny. 
Compared to other organisms, such as in Hylo- 
comiaceae (K = 0.012; Chiang, 1994) and Rubi- 
aceae (K = 0.027; Manen & Natali, 1995), the 
chloroplast genetic variation between species of Be- 
gonia appears to be low. The distinctness of B. for- 


mosana supported by two derived sites (371 and 
376) and that of B. aptera supported by a shared 
derived site (298) indicated, however, that this 
spacer is an appropriate marker at species level. 
The low level of interspecific variation in chloro- 
plast DNA in Taiwanese Begonia may be ascribed 
to the abundant and recent natural hybridization 
and introgression events (Liu, 1999), which facili- 
tated the morphological evolution and evolution of 
nuclear DNA via genetic recombination (based on 
our preliminary assessment of RAPD and nrDNA 
ITS sequence data). The possibility that the evo- 
lutionary rate of morphological changes may have 
surpassed that of the chloroplast DNA was similarly 
demonstrated in oaks with frequent natural hybrid- 
ization (Whittemore & Schaal, 1991). 


FIT OR UNFIT, AND RECURRENT HYBRIDIZATION OF 
В. X TAIPEIENSIS 


Natural hybrids may be considered to be impov- 
erished genetically (cf. D. E. Soltis & P. S. Soltis, 
1993; Arft & Ranker, 1998). Whether natural hy- 
brids are more or less fit relative to their parents is 
a controversial issue (Arnold & Hodges, 1995). Ex- 
perimental hybridization in this study revealed that 
there are very few if any pre- and post-zygotic re- 
productive barriers between these Begonia. We 


Annals of the 


282 


Missouri Botanical Garden 


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Volume 87, Number 2 
2000 


Peng 4 Chian 283 


g 
Begonia X taipeiensis Peng 


would expect natural hybrids when species of Be- 
gonia, such as B. aptera and B. formosana, co-oc- 
cur and have overlapping flowering periods. Bego- 
nia taxa are also well known for their capability for 
vegetative propagation: they easily proliferate from 
fragments of stems, rhizomes, or leaves. Hybrid 
populations of B. X taipeiensis, however, are of spo- 
radic occurrence (Fig. 1). Furthermore, such hy- 
brids drop staminate flowers precociously and are 
completely seed-sterile. Their sterility, small pop- 
ulation size, and rare occurrence suggest that they 
are less fit than the parental species, B. aptera and 
B. formosana. Many recent studies also indicate 
that natural hybrids are usually less competitive 
than their parental species, unless a novel niche 
can be explored (Arnold, 1993; Arnold & Hodges, 
1995). 


Begonia X taipeiensis was not known until very 
recently (Peng & Sue, 2000). It is seed-sterile, of 
limited distribution, and co-occurs with its parental 
species. Because it sheds staminate flowers preco- 
ciously, we used it as an ovule donor to make ex- 
perimental backcrosses with both of its parental 
species to test the possiblilty of introgression. Such 

ttempts consistently failed to produce viable 
seeds. This, plus the sterility in B. X taipeiensis, 
led us to suggest that it can only persist through 
recurrent hybridization. Many recent molecular 
studies also suggest that recurrent hybridization 
events may occur over short spans of time (Ashton 
& Abbott, 1992; D. E. Soltis & P. S. Soltis, 1993; 
Arft & Ranker, 1998). 

Although B. X taipeiensis is highly sterile, sig- 
nificant genetic variability could have been incor- 
porated and accumulated into this hybrid from ge- 
netically distinct parental species (cf. Arft & 
Ranker, 1998). In nature, B. X taipeiensis with low 
frequency of fertility, which may not have been de- 
tected due to limited sampling, may occur. Simi- 
larly, the low level of viable seeds in backcrossings 
did not rule out the possibility of introgression in 
wild populations. Further study with wider and 
more intense samplings will be able to provide in- 
sight into the impact or potential of rarely occurring 
events on the evolution of Taiwanese Begonia. 

In conclusion, when analyzed in concert, the 
data suggest that the formation of the natural hybrid 
B. X taipeiensis occurs via pollen transfer from B. 
aptera to the maternal species, B. formosana. Uni- 
directional hybridization suggests that differential 
survivorship exists between hybrids with reversed 
maternal origins. Even considering the ease with 
which experimental crosses are obtained, natural 
hybrids appear less fit than the parental species in 
this study, based on their sterility. 


Literature Cited 


Arft, A. M. & T. A. Ranker. 1998. Allopolyploid origin 
and population genetics of y ip orchid Spiranthes 
diluvialis. Amer. J. Bot. 85: 

Arnold, M. L. 1992. Natural Melun as an evolu- 
tionary process. Ann. Rev. Ecol. Syst. 23: 237-261. 
—— ———. 1993. Iris nelsonii penat Origin and ge- 
netic composition of a homoploid hybrid species. Amer. 

J. Bot. > 577-583 
997. Natural Hybridization and Evolution. Ox- 
ford Js Press, New Yor 
& Bennett. 1993. Natural hybridization in 

Louisiana DE Genetic variation and ecological de- 

terminants. Pp. 115-139 in R. G. Harrison (editor), Hy- 

brid Zones and the Evolutionary Process. Oxford Univ. 

Press, New Yor 

& 5 


А Hodg es. 1995. Are natural hybrids fit or 
unfit relative to ud parents? Trends Ecol. Evol. 10: 
67-71. 


B. D. Bennett & E. A. Zimmer. 1990. 
hybridization between Iris fulva and I. he 
tern of riboso 

21 


Natural 
exagona: Pat- 
mal DNA variation. Evolution yn 1512- 


Ashton, P. & R. J. Abbott. 1992. Multiple origins and 
genetic diversity in the newly arisen allopolyploid spe- 
cies, Senecio cambrensis Rosser (Compositae). Heredity 


Baldwin, B. C. 7. Adaptive radiation of the Hawaiian 


Sytsma (editis) Мі 0 Evolution and Adaptive Ra- 
diation. Cambridge Univ. Press, Cambridge 
Brochmann, C., Q. Y. Xiang, S. J. Brunsfeld, р. Е. Soltis 
& P. А. Soltis. 1998. Molecular evidence = polyploid 
origins in Saxifraga eee) The narrow Arctic 
. svalbardensis a as allies. 
Amer. J. Вог. 85: 135-143. 
Chen, C. H. 1 Begoniaceae. Pp. 845—854 in Edito- 
rial Committee of the Flora of Taiwan, Flora of Taiwan, 
Vol. 3. Editorial Committee of the Flora of 


Chen, Y. K. 1988. A pu d of Begonia L. 
(Begoniaceae) of Taiwan s Thesis, Graduate 
School of International De span of China, Section 


sertation, b dps of Biology, Washington Universi- 
ty. SL L 


С. 1 Ping & B. A. Schaal. 1998. Universal prim- 
ers for amplification and sequencing of atpB-rbcL non- 
coding spacer of chlroplast DNA. Bot. Bull. Acad. Sin. 
39: 245-250. 

Chien, A., D. B. Edgar & J. M. Trela. 1976. Deoxyribon- 


ucleic acid polymerase from the extreme thermophile 


W. & В . 1985. Жобо богдай 
chloroplast DNAs does not occur in sexual crosses of 
Oenot hera. Molec. m Genet. 198: 525. 
owling, M. E. Douglas, W. L. 
rsh. 1992. Origin of Gila semi- 


at ions for evolution an consentiat. 
S.A. 89: 2747—2751. 
Doyle, 1. J. & J. L. Doyle. 1989. A rapid DNA isolation 


Annals of the 
Missouri Botanical Garden 


procedure for small ae of fresh tissue. Phyto- 

chem. Bull. 19: 

,J.L.D ОГ ИН 8 А. H. D. Brown. 1990. 

Multiple n of polyploids in the Glycine tabacina 
complex inferred from chloroplast DNA polymorphism. 
Proc. Natl. od Sci. U.S.A. 87: 714—717. 

Farris, J. S. 1989. The retention index and the rescaled 
cons Dinor index. Cladistics 5: 417—419 

— J. 1985. Confidence limits on ТЕТЕ" 
An approach using the bootstrap. Evolution 39: 783- 


Francisco-Ortega, 1. D. J. Crawford, A. Santos-Guerra & 
R. K. Jansen. 1997. Origin and evolution of Argyran- 
themum (Asteraceae Anthemideae) i in Macaronesia. Pp. 
407—431 in T. J. Givnish & K. J. Sytsma (editors), Mo- 
lecular Evolution and Adaptive Radiation. Cambridge 

ambridge. 

‚К. Davis. 1994. Evidence арт з 

ybrid zone in the grass shrimp Palaemonetes kadi- 
akensis (Palaemonidae) as ен by пина a 
markers. Evolution 48: 376-391. 

Golenberg, E. M., M. T. Clegg, M. L. Durbin, J. Doebley 
& . Ма. 1993. Evolution of a noncoding region of 
the chloroplast genome. Molec. Phylogen. Evol. 2: 52- 


Harris, S. A. & R. Ingram. 1992. Molecular systematics 
of the genus Senecio L.: 1. hybridization in a British 
polyploid complex. Heredity 69: 1-10 

Hedges, S. B The number of replications needed 
for accurate estimation of the bootstrap P value in phy- 
logenetic "rur Molec. Biol. Evol. 9: 366—369. 

Higgins A. J. Bleasby & R. Fuchs. 1992. CLUS- 
TAL V: Dae software for multiple sequence align- 
ment. CABIOS 8: 189-191. 

Hillis, D. M. & J. J. Bull. 1993. An empirical test of 
bootstrapping as a method for assessing confidence in 
phylogenetic analysis. Syst. Biol. 42: 182-192. 

— а J. Р. Huelsenbeck. Signal, noise, and 
reliability in „ЧЕ. родго analyses. J. Hered. 

are 


Hodges, S. A., J. M. Burke & M. L. = Id. 1996. Nat- 
ural Сабой of Iris hybrid: Experimental evidence on 
"e establishment of hybrid zone. Evolution 50: 1504— 

1509. 


Huelsenbeck, J. P. 1991. Tree-length distribution skew- 
ness: Ап indicator of phylogenetic information. Syst. 
Zool. 40: 257-270. 

Hughes, C. E. & S. A. Harris. 1994. The characterisation 
and identification of a naturally occurring hybrid in the 
genus Leucaena (Leguminosae: Mimosoideae). Pl. Syst. 
Evol. 192: 177-197. 

Jackson, R. C. 1973. Chromosomal evolution in Haplo- 
pappus gracilis: А centric transposition race. Evolution 
27: 243-256 

1980. A simple method for estimating evo- 

utionary rates of base substitutions through compara- 

tive studies of nucleotide sequences. J. Molec. Evol. 16: 
111-120 

Klassen, G. J., R. D. Mooi & A. Locke. 1991. Consis- 
tency indices and nds data. Syst. Zool. 40: 446— 
451. 


Kluge, A. G. & J. S. Farris. 1969. мис me 
and the dira or anurans. Syst. Z 1 

Китаг, Р. а & M. T ei. 1993. nar Mo- 
lecular Evolutio енн Ce netics Analysis, version 1.01. 
The Pennsylvania State University, Pennsylvania. 


Li, W. H. 1997. Molecular Evolution. Sinauer Associates, 
мечуи Massachusetts. 
Zharkikh. p^ о dE tests of DNA 
о Syst. Biol. 44 
Liu, S. L. 1999. Multiple dies oí Begonia formosana 


A ITS. Master's е. Ма- 
попа] du qw о Tainan, Taiw 

Manen, J. F. & A. Natali. . Comparison of the evo 
lution of ribulose-1, 5- не carboxylase (rbcL) 
and atpB-rbcL noncoding spacer sequences in a recent 
plant dece ES tribe Rubieae (Rubiaceae). J. Molec. 
Evol. 41: А 

Marchuk, | eae A. Saulino & F. S. Collins. 
1991. ан of T-vectors: A rapid and general 
system for direct cloning unmodified PCR products. 
Nucl. Acids Res. 19 

Mummenhoff, K. & H. Шыл 1995. еи origin 
of Arabidopsis suecica (Fries) Norrlin: Eviden 
chloroplast and nuclear genome markers. Bot. Acta 
108: 449-4. 

Murray, V. 1989. Improved double-stranded DNA se- 
quencing using the linear polymerase chain reaction. 
Nucl. Acids Res. 17: 9. 

Ohsumi, C., A. Kojima, K. Hinata, T. Etoh & T. Hayashi. 
1993. а ево Ше hybrid between Allium A and Al- 
lium sativum. Theor. Appl. Genet. 85: 969-975. 

biosystematics of d sect. 

Microcarpium (Onagraceae). Ann. Missouri Bot. Gard. 

75: 970-1009. 

990. Ludwigia X taiwanensis (Onagraceae), a 

new species from Taiwan, and its origin. Bot. Bull. 

343-350. 


Acad. Sin. 31: 


. Chen. 1991. Hybridity “ = of 

dice тата Vana ген ae) from Tai- 

wan. Ann. Missouri Bot. Gard. 7 1001. 

С. У. Sue. 2000. e X taipeiensis + 

goniaceae), a cet natural hybrid in Taiwan. Bot. Bul 
Acad. Sin. 41: —158. 

Pijl, L. van der. Des Principles of Dispersal in Higher 
Plants. Springer Verlag, Berlin and Heidelberg. 

Rieseberg, L. H. . Homoploid -o evolution in 
Helianthus аи от ribosomal 


Evidence 
218-1237. 
995. Character expression, 
phylogenetic reconstruction, and the detection of retic- 
ulate pee Pp. 333-353 in P. C. Hoch & A. G 
Stephenson (editors), К реу and Molecular Ap- 
proaches 10 Plant Biosystematics. Monogr. Syst. Bot. 

Missouri pon ard. 2. 

r& na. 1990. Molecular tests of 
the mue a eer origin of two diploid Helianthus 
spp. (Asteraceae). Evolution 44: 1498-1511. 

Sanger, F., S. Nicklen & А. R. Coulson. 1977. DNA se- 
quencing with rer па не inhibitors. Proc. Natl. 
Acad. Sci. U.S.A. 5463-5467. 

Soltis, D. E. & P. S. is 1989. Allopolyploid пана 
іп Е ae from chloroplast DNA. Am 
J. Bot. 76: 11 

& 


3 . Molecular data and dynamic 

nature of agis d E Rev. Pl. Sci. 12: 243-273. 

, Ness. 1990. Maternal inheri- 
tance of the db genome in Heuchera and Tol- 
miea (Saxifragaceae). J. Hered. 81: 168—170. 

Soltis, P. S. & D. E. Soltis. 1991. Multiple origins of the 
allotetraploid Tragopogon mirus ЗНО rDNA 
evidence. Syst. Bot. 16: 407-413 


Volume 87, Number 2 
2000 


Peng & Chiang 285 


Begonia x taipeiensis Peng 


—— oyle & D. E. Soltis. 1992. Molecular data 
and d evolution in plants. Pp. 177-201 in P. S. 
Soltis, D. E. Soltis & J. J. Doyle (editors), Molecular 
Systematics of Plants. Chapman and Hall, New York. 

Stein, D. B. & D. S. Barrington. 1990. Recurring hybrid 
formation in a population of Polystichum potteri: Evi- 
dence from chloroplast DNA comparisons. Ann. Mis- 
souri Bot. Gard. 77: 334—339. 

Swofford, D. L. 1991. When are phylogeny stimates from 
molecular and morphological data incongruent? 

i Cracraft (editors), 

Phylogenetic Analysis of DNA Sequences. Oxford Univ. 

Press, New York, Oxford. 

993. PAUP: Phylogenetic Analysis Using Par- 
dig, Version 3.1.1. Computer program distributed by 
the Illinois Natural History Survey. Champaign, Illinois. 


Vogelstein, B. & D. Gillespie. 1979. Preparation and an- 

alytical purification of DNA from agarose. Proc. Natl. 
d. Sci. U.S 

„Х.Е. & A. E. Szmidt. 1994. Hybridization and 


les compiex 


wart & J. H. Rettig. 1991. Mo- 
a homoploid bi m evolution 
among Austell species of Gossypium. Evolution 45: 


69 

Whittemore, A. T. & B. A. Schaal. 1991. Interspecific 
gene flow in Tür НИ oaks. Proc. Natl. Acad. Sci. 
U.S.A. 88: 2540- 

Wu, C. I. & W. H. 2 1985. Реке (ог higher rates of 
renti substitution in rodents than in man. Proc 


atl. Acad. Sci. U.S.A. 82: 1741- "1745. 


VEGETATION OF LIMESTONE 
AND DOLOMITE GLADES IN 
THE OZARKS AND MIDWEST 
REGIONS OF THE 

UNITED STATES! 


Jerry M. Baskin? and Carol C. Baskin? 3 


ABSTRACT 


Literature on the vegetation of limestone and dolomite (cedar) glades i in the Ozarks of Missouri and Arkansas and 


in the midw estern United States (Illinois, Indiana, Ohio, 
С, к prairie grasses, of which li 
Wi 


thout removal of invading woody plants by fire or other m 
orest differ from 
(primarily d vaginiflorus 
maintain them. We suggest that the a 


onsin) is reviewed. Dominant plants in in these glades are 


ttle bluestem (Schizachyrium scoparium ne hx.) Nash) is the most important. 


means, succession in these rocky, calcare 


nthro ogenic, prairie-grass-dominated openings in the Ozarks and Midwest be 


called xeric кйш (ог dolomite) prairies and that the term cedar sac be s for an edaphic climax dominated 


by C, summer annual g 
En: words: 


ium, woody plant invasion, xeric limestone prairie 


rasses in natural openings on limestone or dolom 
annual bora olus species, a (U.S.A.) scio Kn Ozark glades, Schizachyrium scopar- 


Although the climatic climax vegetation in the 
eastern United States is forest (Shreve, 1917; Braun, 
1950; Küchler, 1964), long-persisting plant com- 
munities dominated by herbaceous angiosperms and/ 
or cryptogams occur in areas where bedrock is ex- 
posed and/or soil depth in most places is too shallow 
to support trees or shrubs. Examples of these edaph- 
ic climax rock ошсгор communities include the mid- 
Appalachian shale barrens (Platt, 1951; Keener, 
1983; Braunschweig et al., 1999), granite outcrops 
of the southeastern piedmont (Oosting & Anderson, 
1939; McVaugh, 1943; Keever et al., 1951; Bur- 
banck & Platt, 1964; Palmer, 1970; Shure, 1999), 
and the cedar (limestone) glades of the Central 
(Nashville) Basin in Tennessee (Quarterman, 1950; 
Baskin & Baskin, 1985; Somers et al., 1986; Drew, 
1991) and of the d United States in gen- 
eral (Baskin & Baskin, 1 

The terms “седаг wires iud "limestone (or do- 
lomite) glades" are also used to describe grass/forb- 
dominated openings over shallow limestone or do- 
lomite soil on ridgetops and side slopes in the 
Ozarks (Steyermark, 1940; Erickson et al., 1942; 
Kucera & Martin, 1957; Ladd & Nelson, 1982; 
Nelson & Ladd, 1983; Nelson, 1985; Fig. 1) and 
Midwest (e.g., Curtis, 1959; Kurz, 1981; Aldrich et 
al., 1982; Heikens & Robertson, 1995). Thus, the 


use of these terms in the Ozarks (Arkansas, Mis- 
souri), Midwest (Illinois, Indiana, Ohio, Wisconsin), 
and Southeast (see fig. 12.2 (map), p. 208 in Baskin 
& Baskin, 1999) has been interpreted to mean that 
cedar glade vegetation is the same in the three re- 
gions (cf. Curtis, 1959; Küchler, 1964). However, 
cedar glades of the southeastern United States are 
dominated by C, summer annual grasses, primarily 
Sporobolus vaginiflorus, and those of the Ozarks/ 
Midwest are dominated by C, perennial grasses, 
primarily Schizachyrium scoparium (Baskin et al., 
1994, 1995). In a recent review of the literature on 
cedar glades of the southeastern United States, Bas- 
kin and Baskin (1999) concluded that: (1) Sporo- 
bolus vaginiflorus is the most important species in 
this limestone/dolomite rock outcrop vegetation 
type; and (2) neither Schizachyrium scoparium nor 
any other perennial grass species is an important 
component of the vegetation. No such review of 
quantitative information has been published on the 
vegetation of calcareous glades in the Ozarks and/ 
or Midwest. 

Thus, the purpose of this paper is to review the 
literature on the vegetation of limestone and dolo- 
mite glades in the Ozarks and Midwest. In partic- 
ular, quantitative and/or qualitative evidence is 
presented that, indeed, Schizachyrium scoparium is 


' We thank Kayri Havens and several anonymous reviewers for their very helpful comments on various drafts of the 


manuscript. 


? School of Biological Sciences, University of Kentucky, Lexington, Kentucky 40506-0225, U.S.A. 
* Department of Agronomy, University of Kentucky, Lexington, Kentucky 40546-0091, U.S.A. 


ANN. Missouni Bor. GARD. 87: 286—294. 2000. 


Volume 87, Number 2 Baskin & Baskin 
2000 Limestone Glade Vegetation 


" 
Amy жЕ 
Wise Reni АЕ 


а ДИ ceras 


Figure 1. Тор, Valley View Glade, Jefferson County, Missouri. Bottom, а glade in the Hercules Glades Wilderness 
Area, Taney County, Missouri. Photographs taken by Carol С. Baskin on 24 July 1996 (top) and 26 July 1993 (bottom). 


288 


Annals of the 
Missouri Botanical Garden 


the dominant plant species of these calcareous 
glades and that annual grasses are relatively un- 
important. In addition, this review contrasts the cal- 
careous glades of the Ozarks and Midwest with 
those of the southeastern United States. 


THE OZARKS 


Steyermark (1940; see also Steyermark, 1959) 
recognized five climax vegetation associations (sen- 
su the polyclimax theory) of the Missouri Ozarks 
and qualitatively described stages in primary suc- 
cession leading to each of them. He also recognized 
several subclimaxes in these seres. However, these 
presumed successional sequences probably repre- 
sent vegetation sequences along environmental gra- 
dients that are unrelated to succession per se 

The open limestone and dolomite glades of the 
Missouri Ozarks are the first two stages of what 
Steyermark (1940) presented as a six-stage sere be- 
ginning on bare, rocky slopes and ending with a 
sugar maple—white oak climatic climax: (1) Boute- 
loua curtipendula (Michx.) Torr.-Rudbeckia mis- 
souriensis Engelm.; (2) Rhus aromatica Ait.—Dios- 
pyros virginiana L.—Juniperus virginiana L. (with a 
redcedar subclimax persisting on eroded limestone 
slopes and knobs); (3) Bumelia lanuginosa (Michx.) 
Pers.—Viburnum rufidulum Raf.; (4) Ulmus alata 
Michx.-Rhamnus caroliniana Walt.; (5) Quercus 
muhlenbergii Engelm.—Fraxinus americana L. (with 
Q. muhlenbergii forming a subclimax on southern 
and western exposures); and (6) Acer saccharum 
Marsh.-Quercus alba L. Climax vegetation in the 
White River region of southwestern Missouri may 
be Q. muhlenbergii—Cotinus obovatus Raf. Thus, the 
pioneer stage in this sere is a limestone (or dolo- 
mite) glade (sensu Steyermark, 1940: 392) “ 
with largely a component of prairie species... . 
The dominants are Bouteloua curtipendula and 
Rudbeckia missouriensis (B. curtipendula, Psorali- 
dium tenuiflorum (Pursh) Rydb., and Silphium la- 
ciniatum L., or Andropogon gerardii Vit. and B. cur- 
tipendula in some parts of the Ozarks). In addition 
to A. gerardii and B. curtipendula, other important 
C, grasses on these glades are: Panicum virgatum 
L., Sorghastrum nutans (L.) Nash, Schizachyrium 
scoparium, and Sporobolus neglectus Nash. Impor- 
tant forbs include Agave virginica (L.) Rose, Allium 
stellatum Roth., Aster oblongifolius Nutt., Calam- 
intha arkansana (Nutt.) Shinn., Dalea purpurea 
Vent., Echinacea pallida (Nutt.) Nutt., E. paradoxa 
(Norton) Britt. var. paradoxa (an Ozark cedar glade 
endemic), Hedyotis nigricans (Lam.) Fosb., Heli- 
otropium tenellum (Nutt.) Torr., Oenothera macro- 
carpa Nutt. subsp. macrocarpa, and Polytaenia nut- 


” 


tallii DC. Berchemia scandens (Hill) K. Koch is a 
common woody vine in this pioneer stage in the 
southwestern Ozarks. 

A second seral stage begins when Rhus aroma- 
tica, Diospyros virginiana, Juniperus virginiana, 
and other woody plants invade these limestone (or 
dolomite) glades. Juniperus virginiana typically be- 
comes the most conspicuous plant, especially on 
eroded slopes and “bald knobs,” forming what 
Steyermark referred to as “cedar glades,” “red ce- 
dar glades,” or “red cedar balds.” These terms in- 
dicate scattered redcedar trees in a matrix of her- 
baceous vegetation dominated by prairie grasses, in 
particular A. gerardii, P. virgatum, and S. scopar- 
ium. In the White River region of southwestern 
Missouri, J. virginiana and/or D. virginiana may 
invade limestone glades and become the dominant 
species. Sassafras albidum (Nutt.) Nees may co-oc- 
cur with D. virginiana as a pioneer woody species. 

Other investigators also have found that prairie 
grasses are the dominants of cedar glades in the 
Ozarks of Missouri. Kucera and Martin (1957) re- 
ported Schizachyrium scoparium as the dominant 
species (85% frequency, 51% of herbaceous cover) 
in cedar glades in the Ozarks of southwestern Mis- 
souri. Locally dominant grasses included A. gerar- 
ди, B. curtipendula, P. virgatum, Sorghastrum пи- 
tans, Sporobolus heterolepis (A. Gray) A. Gray, 
Tridens flavus (L.) A. Нисћс., and the С, summer 
annual Sporobolus neglectus. Hedyotis nigricans was 
the most important herbaceous dicot, Rhus aro- 
matica the most common shrub, and Juniperus vir- 
giniana the most common tree. Other typical woody 
species included Bumelia lanuginosa, Cotinus 
obovatus, Diospyros virginiana, Ilex decidua Walt., 
Walt. Hicks’s (1981) re- 
sults of a vegetational analysis of seven stands of 
open cedar glades with < 30% woody plant canopy 
cover (stages 1 and 2 in Steyermark’s scheme of 
xerarch succession to a sugar maple—white oak cli- 
max) in the Hercules Glades Wilderness Area in 
southwestern Missouri were quite similar to those 
of Kucera and Martin (1957). Schizachyrium sco- 
parium was the dominant species, and H. nigricans, 
Rhus aromatica, and J. virginiana were the most 
important herbaceous dicot, shrub, and tree, re- 
spectively (Table 1). Schizachyrium scoparium had 
the highest percent importance value (%I.V.) in all 
seven stands sampled by Hicks. 

Hall (1955) determined frequency of herbs + 
seedlings and of sprouts of woody plants 0.27 m or 
less in height and the number of Juniperus virgi- 
niana seedlings/saplings in an open dolomite glade 
in the Missouri Botanical Garden's Shaw Arbore- 
tum, near the northeastern border of the Ozark Pla- 


and Rhamnus caroliniana 


Volume 87, Number 2 Baskin & Baskin 289 
2000 Limestone Glade Vegetation 
Table 1. Average percent importance values (%1.У.) Table 1. Continued. 
for herbs, shrubs/vines, and trees in seven stands of open 
cedar glades in the Ozarks of southwestern Missouri. Only 6 
plants with a %I.V. of = 1.0% are listed. Percent соп- Species constancy %I.V 
stancy and %1.У. were calculated from data in J. L. Hicks Rhamnus Г Walt. 86 4.77 
(1981). Celtis tenuifolia Nutt. 100 4.43 
Fraxinus americana L. 71 3.67 
Cornus drummondii C. Meyer 57 1.96 
Species constancy %I.V Cercis canadensis L. 43 1.54 
HERBS Quercus prinoides Willd. 29 1.30 
Schizachyrium scoparium (Michx.) 6 other species 3.09 
Nas 100 29.21 Total 100.09 
Hedyotis nigricans (Lam.) Fosb. 100 10.47 
Sporobolus neglectus Nash 100 9.47 
Carex sp. 100 7.37 
Rudbeckia missouriensis Engelm. 100 7.30 teau. Understory species with the highest frequen- 
Panicum virgatum 100 640 cies were Sporobolus neglectus (100%), Carex 
Croton capitatus Michx/C. monan- crawei Dewey (90%), Schizachyrium scoparium 
thogynus Michx. 100 5.10 (80%), Hedyotis nigricans (75%), Euphorbia corol- 
Sorghastrum nutans i Nash 100 38! lata Engelm. (60%), and Rudbeckia missouriensis 
a 219 (55%). The total number of juniper seedlings in the 
Nagia betonicifolia Nut HS rama: glade was 6694, and 99.7% of these were in the 
sa Torr. 100 1.73 E | 
С ИРИНА с, 71 151 0.46-m or less height class. Average number of ju- 
Sporobolus heterolepis (A. Gray) A. niper seedlings per m? was 0. all (1955: 177) 
ray 86 1.44. stated, “The Glade is a ‘prairie’ association with 
Palafoxia callosa (Nutt.) Torr. & Andropogon scoparius [= S. scoparium] and Rud- 
A. Gray 71 1.41 beckia missouriensis contributing most to its aspect 
Evolvulus nuttallianus Roemer & and Andropogon scoparius and Sporobolus neglectus 
Schultes | 57 1.31 contributing most to cover.” Hall concluded that 
Heliotropium tenellum (Nutt.) Torr. 86 1,31 this dolomite glade was an edaphic subclimax, as 
Oenothera macrocarpa Nutt. subap. did Erickson et al. (1942) for the dolomite glades 
ee = и of the northeastern Ozarks in general. 
Dalea purpurea Vent. 86 1.11 s . 
104 other Бра 9.18 | The flora and qualitative community ecology of 
Total 101.87 limestone and dolomite glades (as well as that of 
1 | " sandstone, chert, and igneous glades) of the Mis- 
PERDE ENS. | souri Ozarks have been studied extensively by Nel- 
Кын aromatica Ait 100 5877 — son and Ladd (1982, 1983) and Nelson (1985). Nel- 
3 bes о жеен % 971 son and Ladd (1982: 5) stated, "Characteristic 
Berc niani scandens (ИШ) K, Koch "m 794 dominant vascular plants of these [dolomite] glades 
Rosa setigera Michx. 57 540 include Andropogon scoparius, Bouteloua curtipen- 
Andrachne phyllanthoides (Nutt.) J. dula, and Sorghastrum nutans”; and for limestone 
Coulte l4 534 glades, “Dominant vascular plants on these glades 
Mimosa ud L. 29 499 e Andropogon scoparius and Bouteloua curtipen- 
Vitis aestivalis Michx. 57 4.84 dula” (Nelson & Ladd, 1982: 5). Nelson (1985) list- 
Smilax bona-nox L. 57 2.10 ed А. scoparius and B. curtipendula as the dominant 
Parthenocissus quinquefolia (L.) plants on limestone glades and A. scoparius, B. cur- 
— = ын tipendula, and Sporobolus heterolepis as dominants 
Toxicodendron radicans (L.) Kun- on délonits BIN 
- zd a Ver Hoef et al. (1993) quantitatively sampled the 
vegetation of 32 glades on Eminence and Gasconade 
иша dolomites in southeastern Missouri. Schizachyrium 
Juniperus virginiana L. 100 4117 scoparium clearly was the most important species in 
Cotinus obovatus Rat. KOO: ЖАН these rocky forest openings. This C, perennial grass 
hie Ae PM Pers. m pen had a constancy of 10096 in glades on each of the 


two geologic formations, and it was in the highest 
cover class (4, average 96 cover — 31.6) in all 32 


290 


Annals of the 
Missouri Botanical Garden 


i.e., 10–100% cover class, geometric mean 
— 31.6. Other important species on the dolomite 
Pis were Andropogon gerardii, Calamintha ar- 
nsana, Carex meadii, Coreopsis lanceolata L., Ech- 
inacea pallida, Fimbristylis puberula (Michx.) M 
l var. puberula, Hedyotis nigricans, Liatris aspera 
Michx., Liatris cylindracea Michx., Panicum virga- 
tum, Rudbeckia missouriensis, Silphium terebinthin- 
aceum Jacq., Smilax bona-nox L., Sorghastrum nu- 
tans, Sporobolus clandestinus (Bichler) A. Hitchc., 
and S. vaginiflorus. The most important woody spe- 
cies on these glades were Juniperus virginiana, Bu- 
melia lanuginosa, Diospyros virginiana, Pinus echin- 
ata Miller, Quercus prinoides Willd., Q. stellata 
Wangenh., and Ulmus alata. 

Keeland (1978) Lair dd ie ni the veg- 
etation of 13 calcareous gla n the Ozarks of 
northwestern Arkansas. He pierde four general 
community types, which in an apparent succession- 
al sequence are: (1) grass and cedar; (2) cedar; (3) 
cedar-hardwood; and (4) hardwood. Keeland (1978: 
12) described the grassland-cedar type as “... a 
grassy slope with a few scattered woody species." 
Further, it *. . . is the typical Ozark cedar glade as 
described by Hall (1955) and Kucera and Martin 
(1957) in Missouri and Hite (1959) in Arkansas" 
(Keeland 1978: 9). For the representative stand of 
this type described by Keeland, Juniperus virgini- 
ana and Quercus stellata were the most important 
overstory trees, Rhus glabra L. the most important 
shrub, and Schizachyrium scoparium the most im- 
portant herb. Overstory tree basal area on this ce- 
dar glade was low, ca. 4.8m?/ha. Schizachyrium sco- 

parium also was the most important species in 
jin cedar glades quantitatively sampled by 
Logan (1992) in the Buffalo National River area in 
the Ozarks of northwestern Arkansas (Table 2). 

Skinner (1979) placed 0.01-m? and 0.1-m? quad- 
rats around individual plants of the Ozark cedar 
glade endemic Penstemon cobaea Nutt. var. pur- 
pureus Pennell, Centaurium texense (Griseb.) Fern., 
and Stenosiphon linifolius (Nutt.) Heynh. to deter- 
mine species associates (as well as other site char- 
acteristics) of these three rare plant species in lime- 
stone glades of the southwestern Missouri Ozarks. 
Sporobolus neglectus had the highest percent oc- 
currence in quadrats of both sizes placed around 
Penstemon and Centaurium, and Schizachyrium 
scoparium had the highest percent occurrence in 
those of both sizes placed around Stenosiphon, 
which grows in deeper soil (15.7 = 6.1 cm, mean 
= SD) than Penstemon (10.9 + 5.6 cm) or Сеп- 
taurium (5.3 + 2.8 cm). Percent occurrence of S. 
scoparium was considerably higher in the 0.1-m? 
than in the 0.01-m? quadrats: 22 vs. 5, 37 vs. < 5, 


Table 2. The most important species in 20 limestone 
glades studied by Logan (1992) in northwest Arkansas. 
Only species with a presence of at least 50% and/or an 
1.00 are included in the 
list. Percentage presence and average abundance values 
were calculated from data in Logan (1992) 


average abundance of at least 


Species 


Schizachyrium scoparium 


(Michx.) Nash 4.60 95 


Juniperus virginiana L. 2.60 70 
Helianthus hirsutus Raf. 2.05 70 
Andropogon gerardii Vit. 2.00 55 
Quercus stellata Wangenh. 1.95 60 
Ulmus alata Michx. 1.65 70 
Quercus prinoides Willd. 1.40 60 
Croton monanthogynus Michx. 1.35 60 
Opuntia humifusa (Raf.) Raf. 1.25 60 
Eupatorium altissimum L 1.25 45 
Coreopsis tinctoria Nutt. 1.15 30 
Dichanthelium acuminatum (Sw.) 

Gould & Clark 1.15 40 
Fraxinus americana L. 1.15 50 
Ruellia humilis Nutt. 1.15 60 
Lespedeza capitata 1.10 40 
Sporobolus ee жу (Torr. ex 

Gray) Wood 1.10 30 
Dalea purpurea Vent. 1.10 55 
Echinacea pallida (Nutt.) Nutt. 1.05 40 
Euphorbia corollata Engelm. 1.00 55 
Rhus aromatica 1.00 55 
Desmanthus ilinoensis (Michx.) 

MacMillan ex Robins. & Fern. 1.00 35 
Bumelia lanuginosa (Michx.) 

Pers. 0.90 50 
Celtis tenuifolia Nutt. 0.85 50 


undance scale from 1 to 5: 1 = rare (only one or 
two pueri image 2 = occasional; 3 = frequent; 4 
= common; and 5 = abundant (widespread with high cov- 
er, a dominant or ete dara ant). 


and 59 vs. 16 for Penstemon, Centaurium, and 
Stenosiphon, respectively. The increase in percent 
occurrence of S. neglectus with increase in size of 
quadrats was less dramatic: 46 vs. 32, 58 vs. 33, 
and 11 vs. 10 for Penstemon, Centaurium, and 
Stenosiphon, respectively (Skinner, 1979). 

In a dolomite glade in the Ozarks of southeastern 
Missouri, two contiguous 7 Х 7-m sample plots 
within a whole-glade sample unit of 100 contiguous 
7 X 7-m plots dominated by Schizachyrium sco- 
parium and classified as “glades” (i.e., open glades) 
were divided into 200 70 X 70-cm plots (Ver Hoef 
et al., 1993). At this small scale of sampling, the 
glade was divided into rocky glade, shallow soil 
glade, glade, and deep soil glade zones. Sporobolus 


Volume 87, Number 2 
2000 


Baskin & Baskin 
Limestone Glade Vegetation 


vaginiflorus was the dominant species in the rocky 
glade and shallow soil glade zones, and S. scopar- 
ium in the glade and deep soil glade zones (Ver 
Hoef et al., 1993). However, in two other dolomite 
glades in southeastern Missouri sampled by Ver 
Hoef et al. (1993), as described above, S. vagini- 
florus was not an important specie 
Results of studies by Skinner 1979) and Ver 
Hoef et al. (1993) support a statement made by 
Kucera and Martin (1957: 290) in their study of 
glades in the Ozarks of southwestern Missouri: 
“Andropogon scoparius [= Schizachyrium scopar- 
ium] was the principal dominant; Sporobolus neg- 
lectus was generally frequent and locally abun- 
dant.” Hall (1955), Hicks (1981), and Logan (1992) 
also found that either Sporobolus neglectus or S. va- 
giniflorus was a relatively important species in the 
Ozark glades they studied. Nelson (1985) listed S. 
neglectus as a characteristic plant of dolomite 
glades, but not of limestone glades, in the Ozarks 
of Missouri. Thus, both quantitative and qualitative 
studies show that annual Sporobolus species can be 
locally dominant within the larger prairie-like com- 
munity on limestone and dolomite glades in the 
Ozarks. As such, cedar glades of the type described 
for the southeastern United States (Baskin & Bas- 
kin, 1999) occur in microhabitats within xeric lime- 
stone prairies of the Ozarks. 
chizachyrium scoparium also is the dominant 
plant in Ozark glades in terms of dry mass produc- 
tion. In the Ozarks of southwestern Missouri, total 
annual herbage production on glades ungrazed for 
4 and 21 years was 312.2 and 241.5 g/m’, respec- 
tively, according to a study by Buttery (1960). Pro- 
duction by 5. scoparium on these two exclosures 
(the term used by Buttery to mean areas fenced off 
to prevent cattle grazing) was 175.2 and 112.3 g/ 
m’, respectively, and by Sporobolus neglectus 24.7 
and 7.9 g/m?, respectively. In the 4-year exclosure, 
production by Andropogon gerardii and Sorghas- 
trum nutans was 52.2 and 34.8 g/m?, respectively, 
and in the 21-year exclosure 29.2 and 74.7 g/m’, 
respectively. Thus the C, perennial grasses S. sco- 
parium, A. gerardii, and S. nutans produced 84% 
and 89% of the total herbage on the 4- and 21- 
year exclosures, respectively (Buttery, 1960). Re- 
garding herbage production on grazed glades, But- 
tery (1960: 235) stated, "In 1956, as now, the 
glades outside the exclosures were producing about 
400 pounds of oven dry herbage per acre [44.9g/ 
m?], mostly the less desirable baldgrass [S. neglec- 
tus] and black-eyed Susan (Rudbeckia hirta L.) with 
a scattering of the more desirable little bluestem 
(Andropogon scoparius Michx.) and Indiangrass [5. 


nutans]." Thus, annual Sporobolus can be the dom- 
inant grass on overgrazed Ozark glades. 


THE MIDWEST 


In the Midwest, cedar or limestone glades have 
been reported in Wisconsin, Illinois, and Indiana. 
Schizachyrium scoparium was the most prominent 
ground-layer species in what Curtis (1959) called 
cedar glades in Wisconsin, with a presence of 
100% and an average frequency of 38%. Other 
species in the ground layer with relatively high val- 
ues for presence and average frequency were the 
C, herbaceous perennial forbs Aquilegia canadensis 
L., Anemone cylindrica A. Gray, Dalea purpurea, 
Tradescantia ohioensis Raf., Euphorbia corollata, 
Arenaria stricta Michx., Solidago nemoralis Dryan- 
der, Antennaria neglecta E. Greene, Viola pedata 
L., and Scutellaria leonardii Epling; the C, peren- 
nial grasses Andropogon gerardii, Bouteloua hirsuta 
Lagasca, and Bouteloua curtipendula; and the C, 
perennial grass Koeleria pyramidata (Lam.) P. 
Beauv. (Curtis, 1959: table XVI-10, appendix, p. 
573). The most important tree species was Junipe- 
rus virginiana (Curtis, 1959: table XVI-11, appen- 
dix, p. 574). 

Schizachyrium scoparium had a frequency of 80— 
100% in 30 of 32 limestone glades sampled by Kurz 
(1981) in Illinois. Other important grasses were the 
C, perennials Bouteloua curtipendula, Sorghastrum 
nutans, and Sporobolus aspera (Michx.) Kunth. The 
important forbs were the CAM leaf succulent Agave 
virginica; the C, perennials Aster oblongifolius, 
Brickellia eupatorioides (L.) Shinn., Echinacea pal- 
lida, Euphorbia corollata, Hedyotis nigricans, and 
Physostegia virginiana (L.) Benth.; and the C, sum- 
mer annual Croton monanthogynus Michx. Schiza- 
chyrium scoparium also was the most important spe- 
cies in the limestone glades of southern Illinois 
studied by Heikens and Robertson (1995). 

Limestone glades have been reported from five 
counties in southern Indiana (Aldrich et al., 1982; 
Bacone et al., 1983; Homoya, 1987; Maxwell, 
1987), and they obviously are dominated by peren- 
nial prairie grasses. For example, referring to a 
limestone glade in Perry County, Bacone et al. 
(1983: 368) stated, “Indian grass (Sorghastrum nu- 
tans), big bluestem (Andropogon gerardii) and little 
bluestem (A. scoparius) are common grasses.” Re- 
ferring to two cedar glades in Clark County, Max- 
well (1987: 413) stated, “Little bluestem dominates 
both glades as a xeric, bunch-grass surrounded by 
patches of rocky pavement.” 

The Ozarks and Midwest limestone and dolomite 
glades described above are similar vegetationally to 


Annals of the 
Missouri Botanical Garden 


Table 3. e percent frequency of native plant 
taxa in 147 1 X l-m quadrats in five stands of xeric prai- 
rie vegetation on P Peebles dolomite in Adams County, 
Ohio. Only plants with a frequency of =20% are listed 
(calculated from data in Braun, 1928). 


Species % frequency 

Schizachyrium scoparium (Michx.) Nash 79.4 
Euphorbia corollata Engelm 53.6 
Blephilia ciliata (L.) Benth. 50.8 
Lithospermum canescens (Michx.) Lehm. 45.2 
Agave virginica (L.) R 43.2 
Ruellia humilis (Nees) Lindau 41.8 
Silphium trifoliatum L. 38.2 
Bouteloua curtipendula (Michx.) Torr. 32.4 
Thaspium sp. 32.0 
Pycnanthemum ае че (Walt.) Britton, 

Sterns & Poggenb. 29.8 
Senecio pies Nutt. 29.4. 
Sorghastrum nutans (L.) Nash 29.0 
Comandra umbellata (L.) Nutt. 21.6 
Helianthus hirsutus Raf. 27.6 
Andropogon gerardii Vit. 26.2 
Zizia aptera (A. ic Fern. 25.4 
Fragaria vesca 25.2 
Prunella vulgaris iE 23.4 
Delphinium exaltatum Ait. 23.2 
Lobelia spicata Lam. 22.8 
Scutellaria parvula Michx. 21.4 
Solidago nemoralis Ait. 20.0 


what Braun (1928) called xeric prairies on Silurian 
Dolomite in Adams County in southern Ohio. Here, 
prairie grasses, primarily Schizachyrium scoparium, 
also are the dominant plants (Table 3). Interesting- 
ly, Braun pointed out that although there is some 
overlap in species composition between the xeric 
prairies of Adams County and the cedar glades of 
the Nashville Basin, the vegetation is different. 
Neither annual species of Sporobolus nor of other 
grasses seem to be an important component of ce- 
dar or limestone glades that have been sampled 
quantitatively in Wisconsin (Curtis, 1959) or Illi- 
nois (Kurz, 1981; Heikens & Robertson, 1995). 
However, in one of the xeric prairies, Agave Ridge, 
on Silurian dolomite in Adams County, Ohio, sam- 
pled by Braun (1928), S. vaginiflorus had a fre- 
quency of 6896 in 25 1-m? quadrats. The dominants 
of this prairie were Schizachyrium scoparium 
(100% frequency) and Bouteloua curtipendula 
(84% frequency). Braun referred to Agave Ridge 
and one of the four prairies she sampled on Peebles 
Dolomite as being more xerophytic than the other 
three. However, the average frequency of S. vagin- 
iflorus on the five Peebles dolomite prairies was 
only 14.896. Neither is S. vaginiflorus an important 


component of the vegetation of limestone glades in 
Indiana (Aldrich et al., 1982; Bacone et al., 1983; 
Homoya, 1987; Maxwell, 1987). 


Woopy PLANT INVASION INTO OZARK AND 
MIDWEST GLADES 


Martin (1955: 106) stated that “... even the 
thin-soiled areas [in the Ozarks of southwestern 
Missouri] are being invaded by eastern redcedar 
(Juniperus virginiana), winged elm (Ulmus alata), 
and associated drought-tolerant trees.” Using 1938 
and 1975 USDA aerial photographs showing lime- 
stone and dolomite glades in southwestern Missou- 
ri, Kimmel and Probasco (1980) found a dramatic 
decrease in glade area with 0—1596 woody cover 
and a corresponding increase in glade area with 
50–100% woody cover. Lowell and Astroth (1989) 
used USDA aerial photographs to study natural 
succession of glades to forest in the Hercules 
Glades Wilderness Area (southwestern Missouri) 
from 1938 to 1986. They found that “Even those 
areas most favorable for glades [Gasconade soil, 
305-365 m (a.m.s.l), south or southwest slopes] 
continue to convert to forest” (Lowell & Astroth, 
1989: 78). The area most favorable for glades and 
occupied by glades decreased from about 285 ha 
to about 200 ha (30%) via conversions of glade land 
to forest (see fig. 3 in Lowell & Astroth, 1989). 
Using 1955, 1966, and 1984 aerial photographs, 
Ver Hoef et al. (1993) also showed that the sizes of 
dolomite glades in the Ozarks of southeastern Mis- 
souri had decreased due to woody plant invasion. 

Kimmel and Probasco (1980) concluded that the 
most important reason for the increase in woody 
plant cover on open glades between 1938 and 1975 
was that the U.S. Forest Service had not used fire 
to manage glade (cattle) range. Lowell ph is 
(1989: 78) thought that their study “. ars to 
support the theory held by some ЖК ы (USES, 
pers. comm.) that the glades are not a naturally 
occurring ecotype[?] which can maintain them- 
selves independent of fire and/or human interven- 
tion.” Likewise, Ver Hoef et al. (1993) attributed 
woody plant invasion into calcareous glades in 
southeastern Missouri to fire suppression. Fire fre- 
quency was high in the Missouri Ozarks in preset- 
tlement times (Guyette & McGinnis, 1982), and fire 
probably played an important role in the origin and 
maintenance of limestone and dolomite glades in 
that area. 

According to Curtis (1959), cedar glades in Wis- 
consin: (1) may have originated by invasion of red- 
cedar into a dry prairie on sites protected from fire; 
and (2) would in time succeed to oak forest. 


Volume 87, Number 2 
2000 


Baskin & Baskin 
Limestone Glade Vegetation 


293 


Heikens (1991) speculated that, in the absence of 
fire, the perennial grass-dominated limestone 
hac in southern Illinois would succeed to bar- 
rens (“savannas”) and then to forest. 

By studying aerial photographs of limestone 
glade areas in southern Indiana, Aldrich et al. 
(1982) and Bacone et al. (1983) determined that 
these openings in the forest are decreasing in size. 
Aldrich et al. (1982: 484) stated, “Aerial photo- 
graphs document the continuing shrinkage of these 
glades [in Harrison County], as they were nearly 
double their present size in the Os.” Bacone et 
al. (1983: 372) stated, “The aerial photographs 
show a remarkable decrease in size [of the forest 
openings in Perry County] in the last forty years 
due to encroachment of woody species.” 

Braun (1928: 425) thought the xeric prairies in 
Adams County, Ohio, “... antedate settlement by 
white man and are undoubtedly primary." However, 

A aerial photographs taken in 1938, 
1950, 1965, and 1971, Annala et al. (1983) and 
Annala and Kapustka (1983) found that prairie has 
succeeded (is succeeding) to forest. For example, 
prairie openings in the Lynx Prairie Preserve, 
where the soils are “shallow and poorly developed" 
(Annala et al., 1983: 22), covered 4796 of the Pre- 
serve in 1938 but only 1696 in 1971 (Annala et al., 
1983; also see Foré & Guttman, 1996). Based on 
lack of difference in opal (phytolith) mass between 
soils presently under prairies and those presently 
under cedar-hardwood forests on dolomite in Ad- 
ams County, Boettcher and Kalisz (1991: 127) con- 
cluded that *... the long-term vegetative history 
has been generally uniform over all areas on do- 
lomite regardless of present occupancy by prairie 
or forest." Further, *. . . the distinction between pri- 
mary and secondary prairies has little meaning 
since prairies only occur on areas of dolomite, and 
prairies and forest have alternated over time on 
these areas" (Boettcher & Kalisz, 1991: 127) 


CONCLUDING REMARKS 


The vegetation of limestone and dolomite glades 
in the Ozarks and Midwest is dominated by C, pe- 
rennial prairie grasses (primarily Schizachyrium 
scoparium), and burning or some other method for 
removal of woody plant invaders is required to pre- 
vent succession to forest in these rocky, calcareous 
openings. Thus, they differ from cedar glades in the 
southeastern United States, in which the dominants 
are C, summer annual grasses (primarily Sporobolus 
vaginiflorus), and succession to forest does not oc- 
cur in the absence of burning or other forms of 


management (Baskin & Baskin, 1999). The struc- 


ture of the vegetation of the limestone and dolomite 
glades in the Ozarks and Midwest is much more 
similar to the xeric prairies on Silurian dolomite 
described by Braun (1928) in Adams County, Ohio, 
and to the xeric prairies on Mississippian limestone 
described by Baskin et al. (1994) on and adjacent 
to the Kentucky Karst Plain, than it is to what tra- 
ditionally has been called cedar glades in the 
southeastern United States. Küchler's (1964) vege- 
tation map of the United States shows the same 
vegetation type for cedar glades in the Ozarks and 
in middle Tennessee-northern Alabama. However, 
ample evidence is available in the literature to 
show that the glade vegetation of these two regions 
is quite different. Thus, we suggest that the rocky, 
calcareous, anthropogenic, prairie-grass-dominated 
openings in the Ozarks and Midwest be called xeric 
limestone prairies, in contrast to cedar glades, 
which are an edaphic climax dominated by C, sum- 
mer annual grasses. 


Literature Cited 


Aldrich, J. R., J. A. Bacone & M. D. Hutchison. 1982. 
Limestone glades of Harrison County, Indiana. Proc. In- 
diana F^ Sci. d 0—485. 

Annala, у A. Kapustka. 1983. Photographic 
history p foit MB НЕ in several relict yen 
of the edge of ваар preserve system, Adam 
County, | очи Sci. 83: 109-114. 

s & L. A. У itn 1983. Е 
lost М aida А За ћи ooh x m sites in Adam 
County, Ohio. Ohio J. Sci. 83: 2 

Bacone, J. A., L. A. Casebere & M T TCR 1983. 
Glades and barrens of Crawford and Perry counties, In- 
diana. Proc. Indiana Acad. Sci. 92: 367-373. 

Baskin, J. M. & C. C. Baskin. 1985. Life cycle ecology 
of annual plant species of southeastern United States. 
Pp. 371—398 in J. White (editor), The Population Struc- 
ture of Vegetation. Dr. W. Junk, Dordrecht, The Neth- 


erlands 


—. 1999. Cedar glades f southeastern 

United States. Pp. 206—219 in R. C. Anderson, J. 5. 

Fralish & J. M. Baskin (editors), Savannas, Barrens, and 

Rock Outcrop Plant Communities of North America. 

Cambridge Univ. Press, Cambridge 

E. W. Chester. 1994. "The Big Barrens 

Region of ка and Tennessee: Further observa- 

tions and и ri Castanea 59: 226- 

H. Webb & C. C. Baskin. 1995. A floristic 
plant fasc e study of the limestone glades of northern 
Alabama. Bull. ee Bot. Club 122: 226-242. 

Boettcher, S. E. & P. J. Kalisz. 1991. The prairies of the 

. Lucy Braun Preserve, I County, Ohio: A soil 
study. ra Sci. 91: 122-12 

Braun, E. L. 1928. The bd or of the Mineral Springs 
Region of Adams peg ad Ohio Biol. Surv. Bull. 
15 (Vol. "E No. 5): 3 

1950. d gn y а of Eastern North 
вагіна. пау Philadelphia 

Braunschweig, S. H., T. Nilsen & T. Е Wieboldt. 1999. 
The mid- pete a oe om barrens. Pp. 83-96 in R. 
C. Anderson, J. S. Fralish & J. M. Baskin (editors), 


294 


Annals of the 
Missouri Botanical Garden 


Savannas, Barrens, and Rock Outcrop Plant Commu- 
nities Ay North America. Cambridge Univ. Press, Cam- 
ri 

байар, М. P & R. B. Platt. 1964. Granite "n 


communities of "d Piedmont. Plateau i in Georgia. Ec 


. Grazed җе can grow good grass. 

Range Managem. 13: 234-235 
e Vegetation of Wisconsin. Univ. 
Wisconsin Press, Madison. 

Drew, M The Role of Tennessee Coneflower, 
Echinacea tennesseensis, in its Native Habitat: The Veg- 
etation an aphic Analysis. M.S. Thesis, Uni- 
versity =“ вани "Knox xvi ES 

Erickson, R. O., L. G. Brenner & J. Wraight. 1942. Do- 
lomite glades of east-central Missouri. Ann. Missouri 
Bot. Gard. 29: 89-101. 

Foré, S S. I. Guttman, 1996, Spatial and temporal 
genetic structure of Asclepias: verticillata (whorled milk- 

in a forested landscape. 


A. McGinnis. 1982. Fire history of an 
Ozark glade in Missouri. Trans. Missouri Acad. Sci. 16: 
—93. 
Hall, M. T. 1955. Comparison of Juniper populations on 
an Ozark glade and old fields. Ann. Missouri Bot. Gard. 


. Classification of the Natural Forest 

Openings i in авы Illinois. Ph.D. 

Southern Illinois University, Carbondale. 

. A. Robertson. 1995. Classification of bar- 
rens and other forest openings in southern Illinois. Bull. 
Torrey Bot. Club 122: 203-214. 

Hicks, J. L. 1981. A Vegetative Analysis of Hercules 
Glades Wilderness. M.S. Thesis, Southwest Missouri 


Dissertation, 


gfie 
The Мао of Lowe Hollow, Wash- 
ington ee Жр . M.S. Thesis, University of Ar- 


987. A floristic survey of a limestone 

glade i in Versailles State Park, Ripley County, Indiana. 
Proc. Indiana Acad. Sci. 96: 407-412. 

Keeland, B. D. 1978. oo and Soils in Calcareous 
Glades in Northwest Arkansas. M.S. Thesis, University 


983. Distribution and biohistory of the 
mid- Appalac m siii barrens. Bot. Rev. 49: 65-115. 

Keever, C., H. J. О g & L. E. Anderson. 1951. Plant 
succession on inae granite of Rocky Face Mountain, 
о Ang North Carolina. Bull. Torrey Bot. 
Club 78: 

Kimmel, V. À a G. E. Probasco. 1980. Change in woody 
cover on limestone glades between 1938 and 1975. 
Trans. о m Sci. 14: 69-74. 

Kucera, C. . Martin. 1957. Vegetation and soil 
relationships in dd glade region of the southwestern 
Missouri Ozarks. Ecology 38: 285-291. 

Küchler, A. W. 1964. Potential natural vegetation of the 
conterminous United States. i hd accompanying 
manual. Amer. Geogr. Soc. Publ. 

Kurz, D. R. 1981. Flora of imesione glades in Illinois. 
Pp. 183-185 in R. L. Stuckey . Reese (editors), 
The Prairie Peninsula—In e 'shadow" of Transeau. 
Proceedings of Er sixth North American Prairie Con- 
ference, The O dau Univ., Columbus. Ohio Biol. 
Surv. Biol. Notes No. 1 


Ladd, D. & Р. ien s Ecological synopsis of Mis- 
souri glades. Pp. 1-20 in E. A. McGinnes, Jr. (editor), 
ag ud o the Cedar Glade Symposium. Occas. 

Pap. pae в Sci. 

Logan, J. M. . The G lados of the Buffalo National 

M Adams M.S. Thesis, lowa State University, 


Am 
tail n J. H. Astroth, 18 1989. Vegetative suc- 


cession b? controlled fire in 


graphical Information Systems 3: 69 
Martin, S. С. 1955. The place of range livestock j in the 
Missouri Ozarks. J. Range Managem. 8: 105-1 
Maxwell, R. H. 1987. мышы! о area remnants within the 
Indiana Army Ammunition Plant, Charlestown, opis 
The little bluestem glades. сөң Indiana Acad. 5 
1 


Mc Vaugh, R. 1943. The vegetation of the granite flat 
rocks of the southeastern United States. Ecol. Monogr. 
13: 121-166. 

Nelson, P. 1985. The Terrestrial Natural Communities of 
Missouri. Missouri Department of Natural Resources, 
Jefferson City. 

& D. Ladd. 

souriensis 3(4): 6-1 

en Preliminary report on the iden- 

distribution and classification of Missouri 

C. L. Kucera (editor), Proceedings 

of the Seventh North American Prairie Conference. 
Southwest Missouri State adip Springfield. 

Oosting, H. J. & L. E. Anderson. . Plant succession 
on pos rook in eastern North бы Bot. Gaz. 
100: 750—768. 

Palmer, T G. 1970. The vegetation of Overton Rock out- 

crop, Franklin County, North Carolina. J. Elisha Mitch- 

ell Sci. Soc. 66: 80-86. 

51. An ecological study of the mid-Ap- 

palachian shale barrens and of the plants endemic to 

them. Ecol. Monogr. 21: 269-300 

n, E 50. Major plant communities of Ten- 

nessee cedar glades. Ecology 31: 234—254 

Shreve, F. 1917. A map of the vegetation of the United 
end Geogr. Rev. 3: 119-125 + plate III disci 


Vial Missouri glades: Part II. Mis- 


tification, 


Shure. D. Ј. 1999. Granite пе of the southeastern 
United States. Pp. 99-118 in R. C. Anderson, J. S. 
Fralish & J. M. Baskin alice): Savannas, Barrens, and 
Rock Outcrop Plant Communities of North America. 


ds е Requirements of 
Three ge ig A ipiis Missouri Glade Plant 


Species. M.S О of Missouri, Columbia. 
Somers, Sn P. B. Hammel & E. L. Bridges. 
1986. ша. lee of plant communities and 


о changes i п кү: ge in middle Tennessee. 
A. Bull. 33: 92. 
ин i A. us ae on the vegetation of Mis- 
1—1. Natural plant associations and succession in 
the Ozarks of Missouri. Field Mus. Nat. Hist. Bot. Ser. 
9: 348-475. 
59. е же, of the Ozark forest. 
Univ. irai: E Vol. 31: 1-138 
Ver Hoef, Ј. M., t Reiter & D. C. Cann: Lewin. 1993. 
Mer of the ua Scenic Riverways, Missouri. A re- 
o the National Park Service, U.S. Department of 
e NAR Midwest Region, Omaha, Nebraska. N 
Cooperative Agreement CA6640-8-8008. 


Volume 87, Number 2 
2000 


Statistical Summary 


STATISTICAL SUMMARY OF SOME OF THE ACTIVITIES IN THE MISSOURI BOTANICAL GARDEN HERBARIUM, 1999 


Vascular Bryophyte Total 
Acquisition of specimens 
Staff collections 31,644 2,532 34,176 
Purchase 21,173 0 21.173 
Exchange 28,038 5,026 33.064 
Gifts 5,975 1,219 6,794 
Total acquisitions 86,430 8,777 95,207 
Mountings 
Newly mounted 49,199 20,424 69,623 
Mounted when received 21.178 0 21,173 
Total specimens filed 70,372 20,424 90,796 
Repairs 
Specimens repaired 32,134 n/a 32.134 
Specimens stamped 1.585 n/a 1,585 
Total repairs 33.719 0 33,719 
Specimens sent 
On exchange 8,242 5,254 13,496 
As gifts 11,212 637 11,849 
Total 19.454 5.891 25,345 
Loans sent 
Total transactions 375 421 
Total specimens 32,876 2,959 35,835 
To U.S. institutions 
Transactions 198 22 220 
Specimens 16.810 1.408 18,218 
То foreign institutions 
Transactions 177 24 201 
Specimens 16,066 1,551 17.617 
To student investigators 
"Transactions pp 8 80 
Specimens 11,244 967 12.211 
To professional investigators 
Transactions 303 38 341 
ecimens 21,632 1,992 23,624 
Loans received 
"Transactions 278 53 331 
ecimens 32,303 4,062 36,305 


The “Mounted when received” vascular plants are specimens of Chinese plants purchased directly from China. 


From U.S.A. 


From abroad 


Total 


581 


507 


1,088 


The total number of mounted, accessioned specimens in the herbarium on 1 January 2000 was 5,002,128 (4,672,010 
) 


vascular plants and 330,118 bryophytes). 


296 Annals of the 
Missouri Botanical Garden 


The Garden’s herbarium is closely associated with its database management system, TROPICOS. The charts below 
summarize some of the statistics from TROPICOS both for the calendar year 1999 and as year-end totals. Note that 
the specimen records in TROPICOS are primarily based on MO specimens, meaning that about twenty-four percent of 
the bryophytes (an increase of about four percent over 1998) and twenty-nine percent of the vascular plants (an increase 
of about two percent) in the herbarium are now computerized, with an overall total of about twenty-eight percent (an 
increase of about one percent). 


TROPICOS records—1999 additions 


Bryophytes Vascular Plants Total 
Specimens 18,638 95.617 114,255 
Names Е 25,016 26,306 
Synonyms 2,135 18,828 20,963 
Distributions 550 28,816 29,366 
Types 63 18,660 18,723 
Bibliography 1,343 2,333 3,676 


TROPICOS records— Year-End 1999 Totals 


Bryophytes Vascular Plants Total 

Specimens 79,489 1,339,708 1,419,197 
Names 97,184 741,614 844,798 
Synonyms 61,084 370,118 431,202 
Distributions 36,991 740,385 777,376 

ypes 6,826 245,157 251,983 
Bibliography ` 20,606 58,025 78,631 
Specimens in herbarium 330,118 4,672,010 5,002,128 
Percent computerized 24 29 28 


In TROPICOS, literature-based Synonymy is always linked to a reference in Bibliography and directly with at least 
two records in Names, the synonym, usually a basionym, and the correct name. Additional synonymy may be derived 
from these direct links, e.g., all other combinations of a basionym treated as a synonym of a given name are also 
synonyms of it. With the completion of A Checklist of the Mosses (www.mobot.org/mobot/tropicos/most), which required 
recording synonymy and acceptance of the 58,000 valid names of moss species (about 13,000 species are recognized), 
the filing of the mosses in the Bryophyte Herbarium will be adjusted to this new standard. 


—Marshall R. Crosby 


Volume 87, Number 2, pp. 127-296 of the ANNALS OF THE MISSOURI BOTANICAL GARDEN 
was published on June 30, 2000. 


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Reproductive Biology of Erythrina crista-galli (Fabaceae) 
-———--- Leonardo Galetto, Gabriel Bernardello, Irene C. Isele, José Vesprini, 
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Volume 87 
Number 3 
2000 


Annals 
of the 
Missouri 
Botanical 


Garden 


NZ 


THE REDISCOVERY OF A 
MALAGASY ENDEMIC: 
TAKHTAJANIA PERRIERI 
(WINTERACEAE)! 


George E. Schatz? 


ABSTRACT 


The history and rediscovery of Takhtajania perrieri (Capuron) Baranova & J.-F. Leroy, the only extant representative 


of the posite family Winteraceae in the Africa/Ma 


Key words: 


dagascar E 
discovery, Madagascar, Takhtajania, Winterace 


IS recounte 


For most of the 20th century, Takhtajania perrieri 
(Capuron) Baranova & J.-F. Leroy stood as the Holy 
Grail of Malagasy botany. Numerous expeditions 
searched in vain for the only possible living mem- 
ber of the ancient family Winteraceae in the Afri- 
ca—Madagascar region at the Manongarivo Special 
Reserve in northwestern Madagascar, where, in 
1909, Henri Perrier de la Báthie collected the only 
known specimens. With hundreds of square kilo- 
meters of intact forest protected within the Man- 
ongarivo reserve, the species must surely still exist 
there! Nevertheless, the dozens of botanists who as- 


cended the slopes of Mt. Antsatrotro to the 1700- 
meter elevation cited on Perrier de la Báthie's 
handwritten labels, passing through what is infa- 
mously acknowledged to be the most dense zone of 
terrestrial leeches in all of Madagascar, have all 
failed to relocate Takhtajania. 

y the spring of 1997, I commenced work on the 
miis Tree Flora of Madagascar (Schatz, in 
press). The Tree Flora will serve as a revision and 
expansion of René Capuron’s 1957 Essai 
d'Introduction à l'Etude de la Flore Forestière de 
Madagascar, a work existing only in mimeographed 


At the Missouri Botanical Garden, I thank P. H. Raven and P. P. Lowry II for the ПИ to study the Malagasy 
i: 


E and V. Hollowell and P. P. Lowry II for helpful comments on the manuscript. In 


1. Morat and his staff 


have always extended the most cordial hospitality during my visits to the Laboratoire de бш niani In Madagascar, 
fieldwork was conducted under collaborative agreements between the Missouri Botanical ( pe n and the Parc Botanique 


et Zoologique de 


thanks go to c Garreau and Dési 


n Mar 
extended by ы асай of shone ar i не Générale 
otégées. This r 


fecun Nationale pour la Gestion des Aire 


Tsimbazaza and the Direction de la Recherche Forestiere et Piscicole, F 
agascar. pipe would have been Sag without the assistance of the World Wide Fund for Nature; 
é Ravelonarivo at their Andapa office. I gratefully acknowledge е 

е 


OFIFA, Antananarivo, Mad- 


de la Gestion des Ressources Forestieres) and 
‘h was conducted with support from U.S. Nat ronal 


nce Foundation grants DEB-9024749 ax DE B-9627072, iud santa from the National Tropical Botanical Garden, 


Kauai, and the National Geographic Society. 


? Missouri Botanical Garden, P.O. Box 299, St. Louis, Missouri 63166-0299, U.S.A. 
ANN. Missouni Вот. GARD. 87: 297—302. 2000. 


298 


Annals of the 
Missouri Botanical Garden 


form. Although it would be another six years before 
he described Bubbia perrieri [= Takhtajania perri- 
ert] (Capuron, 1963), by 1957 Capuron had already 
determined that Perrier de la Báthie 10158 consti- 
tuted the first and only record of the family Win- 
teraceae in Madagascar, including a description of 
the family in his Essai, as represented in Mada- 
gascar by “Un seul genre et une seule espéce: Bub- 
bia Perrieri R. Cap." (p. 96), with the further note 
that "La famille est nouvelle pour Madagascar; 
l'espéce provient du massif du Manongarivo" (p. 
104). In Paris during March of 1997, I decided to 
write the description of Takhtajania for the Tree 
Flora, and, that to do so, it was high time that I 
examine the type specimen. After several inquiries, 
two sheets of Perrier de la Báthie 10158 were lo- 
cated in the office of Thierry Deroin. He, with the 
late Jean-François Leroy (1915—1999), had recent- 
ly studied the vasculature of the ovary utilizing the 
type material (Deroin & Leroy, 1993). Over the 
course of several days as I wrote my generic de- 
scription, I stared periodically at those enigmatic 
specimens collected nearly 90 years earlier. While 
wondering whether the species might still exist, I 
had to conclude my treatment of the genus with the 
unsatisfactory statement that despite numerous at- 
tempts, Takhtajania had not been seen again. Two 
months later, back in St. Louis, with the images of 
those type specimens still very fresh in my mind's 
eye, I opened the enfolding newspapers of a col- 
lection tentatively determined as “Rhamnaceae,” 
made in 1994 from the Anjanaharibe-Sud Special 
Reserve by the Malagasy collector Fanja Rasoa- 
vimbahoaka, and, eureka, there it was! In an in- 
stant, I realized that Takhtajania had indeed been 
seen again, but not where we had been looking for 
it. Set into motion that momentous day were a series 
of events culminating in the following papers. 


FROM “INDET.” To POLITICAL CARTOON: ONE 
SPECIES’ HISTORY 


The first documented human contact with Takh- 
tajania occurred in May 1909 on the Manongarivo 
Massif in northwestern Madagascar, a chance event 
preserved for all time as two herbarium specimens 
(Perrier de la Báthie 10158) deposited at the Mu- 
séum Nationale d'Histoire Naturelle in Paris. Both 
Capuron (1963) and Leroy (1978, 1993) tran- 
scribed the locality data on Perrier de la Báthie's 
handwritten label as “Massif du Manongarivo, au 
bord des ruisseaux, sur schistes liasiques, vers 
1,700 m d'altitude." It seems likely, however, that 
Perrier de la Báthie's altimeter was inaccurate, in- 
sofar as he attributed some of his collections from 


the same trip to 2000 m, despite a maximum alti- 
tude for the massif of 1876 m at the summit of Mt. 
Antsatrotro. Given this, and that Takhtajania is now 
known to range from 1100 to 1550 m altitude at its 
second known locality within the Anjanaharibe-Sud 
Special Reserve, this may partially explain why it 
has never been relocated at Manongarivo: we may 
well have been looking for it at too high an eleva- 
tion! Accompanying the two type sheets enclosed 
within a red Type folder are Perrier de la Báthie's 
field notes. These were reproduced in full by Leroy 
(1978), and reveal that Takhtajania first entered our 
collective botanical consciousness as a "family in- 
det."; based upon its leaf, stamen, and ovary char- 
acteristics, Perrier suggested both *Annonaceae?" 
and "Dilleniaceae? (Tetracera)," believing the sin- 
gle carpel to correspond to a carpel of a flower with 
multiple, free carpels. A later annotation (with a 
preprinted partial date of *193 ," the specific year 
left blank) affixed to one of the sheets by J. Ghes- 
quiere, a specialist in Annonaceae, rejected it from 
that family by noting simply *Magnol.," perhaps as 
an ordinal placement. Leroy (1978) cited undated 
notes referring to the winteraceous genera Bubbia 
and Drimys by H. Humbert; although he may have 
been the first to recognize the correct family place- 
ment, these notes no longer accompany the type 
specimens. Nevertheless, Takhtajania remained 
nameless for 54 years until Capuron (1963) for- 
mally described it as Bubbia perrieri. Such a lag 
time between field collection and its actual descrip- 
tion serves as a useful illustration of the occasion- 
ally slow pace of botanical studies. 

Having finally received a name, Bubbia perrieri 
soon became the focus of additional studies on pol- 
len (Straka, 1963; Lobreau-Callen, 1977) and leaf 
anatomy (Baranova, 1972; Bongers, 1973). These 
studies reported colpate or trichotomocolpate (tri- 
chotomosulcate) pollen apertures and an anomo- 
cytic stomatal cell arrangement, both features 
anomalous within the Winteraceae (neither condi- 
tion is corroborated herein by Keating or Sampson, 
respectively). Stimulated by these novel findings, 
Leroy (1977) expanded upon Capuron's observation 
of a bilobed stigma for Bubbia perrieri, and daringly 
hypothesized a compound unilocular ovary com- 
prised of two united carpels; he also presciently 
suggested a close alliance to Canellaceae. For Ler- 
oy (1978), such radical differences clearly merited 
distinct higher-taxon status, and thus a new genus 
was described. Bubbia perrieri Capuron became 
Takhtajania perrieri (Capuron) Baranova & J.-F. 
Leroy, and the new genus became the type of a 
wholly new subfamily Takhtajanioideae. When 


Volume 87, Number 3 
2000 


Schatz 
Rediscovery of Takhtajania perrieri 


299 


Vink’s (1978) studies of the ovary concurred with 
his bicarpellate hypothesis, Leroy felt sufficiently 
vindicated against Tucker and Sampson’s (1979) 
cautionary skepticism in the journal Science that 
his enthusiasm knew no bounds. In 1980, he raised 
his subfamily Takhtajanioideae to the rank of fam- 
ily: Takhtajaniaceae, indeed, from a “family indet.” 
to a family apart! 

Meanwhile, numerous botanists failed to relocate 
Takhtajania at the Manongarivo Special Reserve. 
Capuron collected there extensively in 1954, in- 
cluding the upper slopes of Mt. Antsatrotro, but 
then he apparently never returned. In , he 
stayed closer to the main road from Малина и to 
Ambanja that passes ca. 10 km to the west of the 
Reserve boundary, and fully 25 km west of the up- 
per slopes of Antsatrotro. Vink (1978) recounted 
that Capuron told him that the type locality was 
completely deforested, a curious remark given the 
vast area of intact forest present still today (Gautier 
et al., 1999). Several years later, Josef Bogner (M) 
searched for Araceae on the southern slopes of Ant- 
satrotro north of Bejofo, and the late Al Gentry 
walked in from the road to as high as 300 m alti- 
tude in 1974. Then, at the behest of Leroy, Ray- 
mond Rabevohitra (TEF) once again searched the 
upper slopes of Antsatrotro in 1979, but found no 
trace of Takhtajania. Beginning in May of 1989 
(Schatz, 1989), the Missouri Botanical Garden has 
conducted numerous expeditions to Manongarivo, 
and since 1994, Laurent Gautier (1997, 1999) has 
headed Geneva’s ongoing inventory efforts there. 
For all who made the arduous pilgrimage to Man- 
ongarivo, Takhtajania would remain elusive. 

Some 150 km to the southeast of Manongarivo, 
also in 1994, Fanja Rasoavimbahoaka began work 
as a local collector at the Marojejy Strict Nature 
Reserve (now a national park) and Anjanaharibe- 
Sud Special Reserve in conjunction with an AN 
GAP (the National Association for the Management 
of Protected Areas)/World Wide Fund for Nature 
protected areas project. That May, he ascended a 
trail used by local gem and crystal hunters leading 
west from the abandoned village of Andranotsarabe, 
which had served as a camp, gravel depository, and 
water source during the building of the road con- 
necting Andapa and Bealanana that bisects the An- 
janaharibe-Sud Reserve. Unbeknownst to him, he 
collected Takhtajania in flower (Fig. 1; additional 
images of Takhtajania may be found on the web at 


http://ww g gasc/winterac 
html), exactly 85 years to the month after Perrier 
de la Báthie's original collection from Manongarivo. 
Later that same year, a multi-disciplinary study of 
altitudinal variation, sponsored by WWF and head- 


ed by Steve Goodman (1998), used this same trail 
to access the summit of Anjanaharibe-Sud. In fact, 
they established their 1200-m “Camp 2” right 
amidst the Takhtajania population. Expedition bot- 
anist Désiré Ravelonarivo collected fruiting Takh- 
tajania both in September and November, and all 
of the expedition members must have walked by 
Takhtajania numerous times along the trail. 

This brings us forward to that fateful day in May 
1997, when I opened the newspapers containing Е 
Rasoavimbahoaka 303. The first person to whom 1 
showed the collection was Peter H. Raven, who had 
launched MBG's activities in Madagascar several 
decades earlier, partly in hopes of finding Takhta- 
jania again. He suggested that I immediately pre- 
pare an announcement of the rediscovery for the 
journal Nature (Schatz et al., 1998). I then dis- 
patched e-mails to Paris and Madagascar, sounding 
the call to action. Several weeks later, members of 
our Malagasy staff including Sylvie Andriambolo- 
lonera, Jeannie Raharimampionona, and Pierre Jules 
(*Coca") Rakotomalaza accompanied Désiré back 
to the Takhtajania locality at Anjanaharibe-Sud, 
photographing its flowers and collecting the first lot 
of specialized research materials that would be uti- 
lized in the following new studies. The following 
month, a specimen for the Komorov Institute in St. 
Petersburg was presented to Armen Takhtajan at a 
special birthday celebration for him in St. Louis; 
born just one year after Perrier de la Báthie first 
collected Takhtajania, he had waited 87 years for 
its rediscove 

Since then, ‘Тана has been revisited nu- 

merous times. The journey from Andapa to An- 
dranotsarabe by 4-wheel-drive vehicle takes three 
hours, if it has been relatively dry the previous 
days. However, subjected to over 3600 mm of rain- 
fall per year, portions of the road are deteriorating 
rapidly, and landslides threaten continued access 
to the population. Having arrived at the trailhead, 
one must gently climb little more than 200 m in 
elevation through beautiful forest. Even when it is 
raining—which it often is—the leech population 
does not even begin to compare with that of the 
slopes of Mt. Antsatrotro at Manongarivo. In less 
than an hour, one enters Takhtajania's realm, the 
narrow, flattened ridges followed by the trail, and 
the uppermost portions of the steep slopes falling 
away on either side of the ridges. With its large, 
glossy, dark green leaves clustered toward the apex 
of vertical stems, Takhtajania stands out in the un- 
derstory, its slender trunk often leaning severely 
after surviving the impact of fallen debris. Extend- 
ing over eleven square kilometers, this second re- 
corded Takhtajania population appears to be thriv- 


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Volume 87, Number 3 
2000 


Schatz 301 
Rediscovery of Takhtajania perrieri 


ing; Birkinshaw and colleagues (1999a) have 
estimated a minimum of 42,020 individuals repre- 
senting all size classes, including abundant seed- 
lings. 
In his 1998 New Year’s address to representa- 
tives of the diplomatic community, the President of 
Madagascar, Didier Ratsiraka, heralded Takhtaja- 
nia as a national treasure that dated back to the 
time of the dinosaurs. His remarks immediately 
drew the attention of other national figures. Minis- 
ters, at that time, of the Environment, and Waters 
and Forests, C. Vaohita and R. Rajonhson, respec- 
tively, accompanied by the Directors of Biodiversity 
alorization, and Ecotourism at A А ага- 
malala and J. Rakotoarisoa, respectively, all made 
the long trek to see Takhtajania. Shortly thereafter, 
a political cartoon appeared in one of the daily An- 
tananarivo newspapers depicting a Malagasy villag- 
er exclaiming deliriously his good fortune in the 
first panel: T suis riche! Riche! Riche! ... Ја: 
ai...” (“I am rich! Rich! Rich! ... 
.”), and ion! in the second panel, ae a Р 
" seedling identified as Takhtajania perrieri, “J'ai 
faim ...” (“I am hungry”). Ironically, Takhtajania 
holds absolutely no value whatsoever for the villag- 
ers living in the immediate area. When shown the 
plant, they had neither a use nor a vernacular name 
or it. Nevertheless, Takhtajania continues to shine 
prominently on the political landscape, having been 
chosen as the emblem for Madagascar's exhibit at 
O-2000 in Hanover, Germany (during the sum- 
mer of 2000), highlighting the government's con- 
servation and sustainable development programs. 
Rediscovery of Takhtajania has provided won- 
derful new research opportunities, the first results 
of which are reported in the following collection of 
papers. James Doyle sets the stage by reviewing the 
fossil history of Winteraceae. Sherwin Carlquist 
confirms the vesselless nature of Takhtajania wood, 
whereas Taylor Feild and his colleagues examine 
the condition of vesselless wood from an ecophys- 
iological perspective. Reexamining foliar anatomy, 
Richard Keating reports mostly brachyparacytic 
stomata, and thus discounts the original reports of 
an anomocytic arrangement that contributed to the 
initial justification of generic status for Takhtajania; 


— 


furthermore, he summarizes a unique nodal anato- 
my in Zakhtajania. Peter Endress and his сој- 
leagues detail its floral morphology, by virtue of 
which Takhtajania conforms most closely to Pseu- 
dowintera and Zygogynum, in part, due to its small, 
early-rupturing involucre and apical anther sacs. It 
was these two features that induced Capuron (1963) 
to describe the species originally in the genus Bub- 
bia, which is now included by some authors in Zyg- 


ogynum s.l. Further exploring the flowers, Andrew 
Doust examines the ontogeny of perianth parts in 
Winteraceae. His comparative sequential data sug- 
gest that the similarities of an early-rupturing in- 
volucre and perianth arrangement exhibited by 
Takhtajania and Zygogynum reflect the plesiomor- 
phic state, and that petal fusion in the two genera 
is non-homologous. His results further imply that 
the enveloping late-rupturing involucre of Tasman- 
nia and Drimys has evolved independently in each 
genus. Delving deeper into the flower, Bruce Samp- 
son reports round or slightly oval apertures in pol- 
len of Takhtajania, thus dispelling earlier reports 
of colpate and trichotomocolpate (trichotomosulca- 
te) apertures; he also finds possible evidence for 
asynchronous pollen mitosis within a tetrad similar 
to that in Drimys and Pseudowintera. Hiroshi Tobe 
and Bruce Sampson report embryological features 
indistinct from other Winteraceae, but well distin- 
guished from probable sister Canellaceae. Thierry 
Deroin's examination of fruit vascular anatomy re- 
veals that the nutritive role is transferred from the 
median carpellary bundle to the lateral bundles in 
maturing ovaries during fruit development. At the 
cytological level, Friedrich Ehrendorfer and M. 
Lambrou document a “diploidized,” possibly paleo- 
tetraploid, chromosome number of 2n — 36, which 
can only be understood in relation to the paleopo- 
lyploidy exhibited by other Winteraceae (n — 

n — 43) by presuming numerous extinction gaps. 
An analysis of molecular sequence data by Ken 
Karol and colleagues provides phylogenetic con- 
text, firmly placing Takhtajania basal and sister to 
remaining Winteraceae, a result consistent with 
Leroy's subfamilial designation. Reported else- 
where are studies of the habitat of Takhtajania (Bir- 
kinshaw et al., 1999b), and an assessment of its 
risk of extinction (Birkinshaw et al., 19992). Later 
this year, Rivolala Andriamparany concludes his 
studies of the reproductive biology of Takhtajania 
perrieri at the University of Antananarivo under the 
direction of Elizabeth Rabakonandriana and Len 
Thien of Tulane University. 


Literature Cited 


Baranova, M. 1972. Systematic anatomy of the leaf epi- 
dermis in the Magnoliaceae and some related families. 
Taxon 21: 447-469. 

Birkinshaw, C., D. Ravelonarivo, R. Andriamparany, S. 
Rapanarivo, E. Rabakonandriana, G. E. Schatz & L. B. 
Thien. 1999a. Risque d'extinction du en per- 
rieri. Кароол Final 1., September, Antananarivo 

| 1999Ь. $ habitat du Takhtajania p perrieri. Rap- 
port Final II., October, Antanan 

Bongers, J. M. 1973. Epidermal “leaf characters of the 
Winteraceae. Blumea 21: 381—411 


302 


Annals of the 
Missouri Botanical Garden 


Capuron, R. 1957. Essai d'Introduction à l'Étude de la 
Flore Forestière de Madagascar. Inspection Générale 
des е А Fórets, Antananarivo. 

. Contributions à l'étude de la flore de Mad- 
agascar. aL Présence а Madagascar d’un nouveau ге- 
présentant (Bubbia perrieri R. Capuron) de la famille 
des Wintéracées. Adansonia, n.s., 17: 373-378. 

Deroin, T. & J.-F. Le ‘егоу. 1993. Sur l'interprétation de la 
оа ЕНЕ de Takhtajania О 

cad. Sci. Paris, Sér. 3, Sci. Vie 316 


Gautier, L. 1997. Inventaire floristique de la Réserve 
Spéciale de Manongarivo dpa Ouest de + аг): 
| Documents EFB n° 5, С 

. Inventaire foristique de la Песне Spé- 

ciale де NA. (Nord-Ouest de “~ ar): Di- 

cotyledonae. Documents EFB n° 6, Genev 

‚ C. Chatelain & R. Spichiger. 1999. Déforesta- 
tion, altitude, pente et aires protégées: Une analyse 
diachronique des на ents sur Је pourtour де la 
le Manongarivo (NW de Madagascar). 
Pp. 255-279 i in H. Hurni & J. Ramamonjisoa (editors), 
African Mountain Development in a Changing World. 
African ти Association & Сеортарћса Bernen- 
sia, Antananariv 

Goodman, 5. yarn 1998. A floral and faunal inventory 
of the Réserve Spéciale d'Anjanaharibe-Sud, Madagas- 
ni With reference to elevational variation. Fieldiana, 
Zool., n.s., 90: 1-246. 

Leroy, J.-F. 1977. A compound ovary with open carpels 
in Winteraceae (Magnoliales): Evolutionary implica- 
tions. Science 196: 977-978 

1978. Une sous-famille monotypique de Win- 
teraceae endémique a Madagascar: Les Takhtajanioi- 
deae. Adansonia, n.s., 5. 

980. Nouvelles A remarques sur le genre Takh- 

tajania nur sadi. Adansonia, 

n.s., 20. 


— 


d Е et Evolution des Plantes а Fleurs. 


Mas 

ee e. Dp. 1977. Le pollen du Bubbia perrieri 
R. Cap. Rapports palynologiques avec les autres genres 
de Wintéracées. Bie pan n.s., 16: 445—460. 

Schatz, G. E. 1989. The search for Takhtajania (Winter- 
aceae). Bull. ad. Trop. Bot. Gard. 19(4): 117-118 

—— ———. In press. Generic Tree Flora of Mudapaxc ar. 


Royal Botanic Soi Kew and Missouri Botanical 
Garden, St. Lou 


MT & A. Ramisamihantanirina. 1998. 
Takhtajania етн rediscovered. Nature 391: 133— 


Straka, H. 1963. Uber die mégliche phylogenetische Be- 

deutung = Polen Morphologie der madagassischen 
Bubbia perrieri R. Cap. (Winteraceae). Grana Palynol. 

| 355 5-360. 

Tucker, S. C "^ B. Sampson. 1979. The gynoecium of 
оства Е planis Science 203: 920-921. 

Vink, W. 1978. The Winteraceae of the Old World. Ш. 
Notes on the ovary of Takhtajania. Blumea 24: 521- 
525. 


Note added in proof. It happened again! On the very day 
all of the manuscripts were sent to Allen Press, Madagas- 
car Program coordinator Kendra Sikes and I were looking 
through backlogged local collector specimens still in their 
i ein A collected on the Masoala Peninsula 
, 1996, by Јао Aridy. There was Takhtajania in 
fruit! As "before, I immediately dispatched E-mail alert to 


of Takhtajania, aiek included жзг b е Ar- 


иш ош d aya of агс duous, wet searc paio they found 


e adult, and o 


ca. 110 km satel of the Anjanaharibe-Sud Special Re- 

serve population i 

of Mt. Ambohitsitondroina, Rivolala Andriamparany, the 
he Anj : 


Зид на jeporis that the flowe 


tial impression from the original Jao Aridy collection that 
the leaves are less thick and coriaceous, and moreover, 
lack a revolute margin Along ma the significantly lower 
ease tion (790 m vs. 1 100-1: 500 m at Anjanaharibe-Sud), 
the ntial dpt logical differences tantalizingly sug- 

gest the КЫШ of a second, distinct taxon within Takh- 

tajani 


PALEOBOTANY, 
RELATIONSHIPS, AND 
GEOGRAPHIC HISTORY OF 
WINTERACEAE! 


James A. Doyle? 


ABSTRACT 


In combination with ро eee analyses of morphological and molecular data from modern plants, fossils allow 


improved reconstruction of the his 
among Magnoliales, Laurales, and Pi суас laco 
обе и ecords, from the Barremian mad А 


ry of Winteraceae. Phylogenetic analyses link Winteraceae with Can 
g that thei 


tp ie (Early essai of Northern Gondwana (Gabon, Israel), are 


ellaceae, nested 
r lack of vessels is derived rather than primitive. The 


к жалы tetrads vee with an Е underlain by thickened endexine but finer sculpture than moder 


bution of Walkeripollis implies that the winteraceous line originated i 

of modern ellaceae, rather than equable temperate and upland tropical habitats a. thas wher 
a n-gro interaceae, first represented in the 

reticulate tetrads in Australia and vesselless wood in Алганга апа 1 

арре 

vessels were lost as an adaptation to cooler es 

could have dispersed to 


represented by the basal genus Takhtajania) and India. 


ikely represent the stem-lineage leading to crown-group Winteraceae. Earl ies m 
y pr 5 5 group Y 
ve also been compared with 


Winteraceae, are probably not related. The distri- 
opical, possibly dry, envimnments, like those 


a and Africa, 


ou 
Australasia via either South America and Antarctica or Madagascar (where the family is 


y words: angiosperms, biogeography, Cretaceous, paleobotany, palynology, phylogeny, Tertiary, Winteraceae. 


Winteraceae have long attracted students of an- 
giosperm evolution and biogeography because of 
their putatively primitive morphology and disjunct 
Southern Hemisphere distribution (Australasia, 
South America, Madagascar). Emphasizing their 
vesselless wood and plicate (conduplicate) carpels, 
Thorne (1974) considered Winteraceae the most 
primitive living angiosperm family. Others (Walker 
& Walker, 1984; Cronquist, 1988; Takhtajan, 1997) 
argued that Magnoliales such as Degeneria are 
more primitive: although these have vessels, they 
also have plicate carpels, plus gymnosperm-like 
monosulcate pollen with a continuous tectum and 
granular infratectal structure, whereas Winteraceae 
have putatively more advanced pollen shed in per- 
manent tetrads, with a round distal pore (ulcus), 
coarsely reticulate sculpture, and columellar infra- 
tectal structure. After concluding that angiosperms 
arose in the area of Southeast Asia and Australasia 
(cf. Takhtajan, 1969), Smith (1973) postulated that 
Winteraceae originated in Malesia and migrated 
south through Australasia and Antarctica to South 
America and west to Madagascar. This scenario was 
challenged by the theory of plate tectonics and rec- 


ognition that Southeast Asia and Australasia are 
juxtaposed portions of two supercontinents, Laura- 
sia and Gondwana, which were widely separated 
during the rise of angiosperms in the Early Creta- 
ceous. Since only two species of Winteraceae occur 
outside the former limits of Gondwana (Drimys 
granadensis L. f. in Central America, Tasmannia 
9 (Hos . f.) Miers in the Philippines), Te 
could be “spillovers” after South America and A 
ыш: ‘collided with Laurasia in the n Win- 
teraceae were soon offered as one of the best ex- 
ples of a taxon with a Gondwanan distribution 
(Schuster, 1972, 1976; Raven & Axelrod, 1974). 
aleobotanical discoveries and phylogenetic 
analyses of the past two decades have shed new 
light on the history of Winteraceae and other an- 
giosperm taxa. The purpose of this paper is to re- 
view the fossil record of Winteraceae in the context 
of the phylogenetic results, and thus to present an 
updated synthesis of the evolutionary and geo- 
graphic history of the family, as background for dis- 
cussion of Takhtajania. Of special interest is fossil 
pollen evidence extending the winteraceous line 
into the Early Cretaceous, which indicates that 


am grateful to Rosemary Askin and Mary Dettmann for help in locating literature, Sharma Gaponoff, Jean-Michel 


Groult, Bob Hill, Sarah Mathews, Imogen Poole, H 


Hervé Sauquet, Vincent Savolainen, 


George Schatz, Alex Troitsky, and 


Liz Zimmer for use of арав data, Neelima Sinha for help with illustrations, and Else Marie Friis and an 


anonymous reviewer for useful comments on the manu 


script. 


? Section of Evolution per Ecology, University of California, Davis, California 95616, U.S.A. 
ANN. Missouri Вот. GARD. 87: 303-316. 2000. 


Annals of the 
Missouri Botanical Garden 


Winteraceae originated in more tropical environ- 
ments than they inhabit today. 


PHYLOGENETIC CONTEXT 


Phylogenetic analyses have confirmed the posi- 
tion of Winteraceae among magnoliids, now recog- 
nized as a basal paraphyletic grade of angiosperms 
below monocots and eudicots (the 95% of dicots 
with tricolpate and derived pollen). However, they 
call into question the primitive status of vesselless 
wood, at least in Winteraceae 

Based on a morphological айий analysis of 
primitive angiosperms, Young (1981) concluded 
that Winteraceae and other vesselless taxa are nest- 
ed well within the angiosperms, so it is more par- 
simonious to assume that vessels arose in the com- 
mon ancestor of angiosperms and were lost in the 
vesselless taxa. This was confirmed by the analysis 
of Donoghue and Doyle (1989), which placed Mag- 
noliales in a restricted sense, with granular exine 
structure, at the base of the angiosperms, with the 
remaining groups (including Winteraceae) united 
by columellar structure. Winteraceae were linked 
with Illiciales (Illicium, Schisandraceae) and in 
some trees Canellaceae, based on palisade exotesta, 
a similarity of Winteraceae, Illiciales, and Canel- 
laceae noted by Corner (1976). Winteraceae and 
Illiciales had also been associated by Walker 
(1976), based in part on their coarsely reticulate 
pollen sculpture, and were subsequently linked in 
the morphological cladistic analysis of Loconte and 
Stevenson (1991). Another similarity of Wintera- 
ceae and Canellaceae is their irregular, “first rank” 
leaf venation (Hickey, 1977), with secondary veins 
that are poorly differentiated from higher vein or- 
ders and loop repeatedly inside the margin (Hickey 
& Wolfe, 1975). However, this may be a symple- 
siomorphy rather than evidence for direct relation- 
ship; similar venation occurs in Early Cretaceous 
angiosperm leaves and has been considered prim- 
itive (Wolfe et al., 1975; Doyle & Hickey, 1976; 
Hickey, 1977). 

Since vessels are often viewed as unambiguously 
advantageous in allowing more efficient conduction, 
these cladistic results stimulated debate on the 
functional plausibility of loss vs. multiple origins of 
vessels. Donoghue and Doyle (1989) argued that 
loss might be less deleterious than assumed, since 
vesselless wood would be derived from wood with 
very primitive vessels, not advanced ones. Winter- 
aceae and other vesselless angiosperms usually oc- 
cur in cool, wet habitats; Carlquist (1975) argued 
that this is the only environment where they can 
persist, but Donoghue and Doyle (1989) noted that 


it might also be the environment where loss of ves- 
sels would be least disadvantageous. Donoghue and 
Doyle and Feild et al. (2000) suggested that vessel 
loss might even be advantageous in cool climates, 
since embolisms caused by freezing would be re- 
stricted to one tracheid rather than expanding to fill 
a whole vessel. This is especially plausible for Tro- 
chodendrales (nested among lower eudicots with 
vessels: Chase et al., 1993; Hoot et al., 1999; Sa- 
volainen et al., 2000), which were common in the 
Late Cretaceous and Tertiary of northern Laurasia 
(Crane et al., 1991). Carlquist (1983, 1987, 1988, 
1996) presented several arguments against the loss 
of vessels and for multiple origins: the parallel na- 
ture of advancement trends within vessels; the im- 
plausibility of scenarios for loss of vessels as a re- 
sult of movement from dry to mesic (Young, 1981) 
or from aquatic to terrestrial habitats; the fact that 
vessels are almost but not completely lost in Ephe- 
dra, even in extreme habitats; and the lack of tra- 
cheid dimorphism in vesselless angiosperms. How- 
ever, one argument of Bailey and Nast (1944) and 
Carlquist (1975), that the characteristic stomatal 
plugs of Winteraceae are a compensation for lack 
of vessels, appears to be incorrect. Feild et al. 
(1998, 2000) showed experimentally that plugs do 
not reduce transpiration and are more likely a de- 
vice to promote runoff of water in cloud forest hab- 
itats. 

Many aspects of earlier cladistic schemes now 
need revision as a result of molecular phylogenetic 
analyses. Studies of partial rRNA sequences (Ham- 
by & Zimmer, 1992; Doyle et al., 1994) rooted an- 
giosperms not in woody magnoliids but in “paleoh- 
erbs," with Nymphaeales basal, as did more recent 
morphological analyses (Doyle et al., 1994; Doyle, 
1996). Much more extensive studies of rbcL (Chase 
et al., 1993) placed Ceratophyllum at the base of 
the angiosperms. However, this rooting was quickly 
suspected to be a long-branch effect (Qiu et al., 
1993; Donoghue, 1994). This view has been con- 
firmed by analyses of 188 rDNA (Soltis et al., 
1997), cpITS (Goremykin et al., 1996; A. V. Troit- 
sky, pers. comm. 1998), phytochrome genes (Ma- 
thews & Donoghue, 1999), and atpB (Savolainen et 
al., 2000), all of which root angiosperms among a 
series of taxa including not only Nymphaeales but 
also several woody magnoliid taxa: Amborella, Aus- 
trobaileya, Illiciales, and (based on studies of rbcL 
by Renner, 1999) Trimeniaceae. Significantly, these 
taxa occur together as a clade in rbcL trees (Chase 
et al., 1993). Endress (1987) and Donoghue and 
Doyle (1989) associated Amborella and Trimeni- 
aceae with Chloranthaceae, but Chloranthaceae 


Volume 87, Number 3 
2000 


Doyle 305 
Paleobotany of Winteraceae 


now appear to be an isolated line, located above 
the basal grade. 

All molecular analyses that have included the 
relevant taxa have separated Winteraceae from Il- 
liciales and linked them with Canellaceae (Chase 
et al., 1993; Soltis et al., 1997; Mathews & Dono- 
ghue, 1999; Savolainen et al., 2000), making up a 
group that I will call Winterales. All genes except 
185 (Soltis et al., 1997), for which the sampling of 
magnoliids was less complete, place Winterales in 
a clade with three other orders as defined by APG 
(1998): Laurales (Calycanthaceae, Monimiaceae 
s.l., Gomortega, Hernandiaceae, Lauraceae), Mag- 
noliales (Eupomatia, Himantandraceae, Degeneria, 
Myristicaceae, Magnoliaceae, Annonaceae), and Pi- 
perales (Piperaceae, Saururaceae, Aristolochiaceae, 
Lactoris). Based on rbcL (Chase et al., 1993) and 
rbcL plus atpB (Savolainen et al., 2000), the sister 
group of Winterales is Magnoliales; based on phy- 
tochrome genes (Mathews & Donoghue, 1999) and 
atpB (Savolainen et al., 2000), their sister group is 
Piperales. 

Like the morphological trees, the molecular trees 
imply that Winteraceae are secondarily vesselless, 
although interestingly they suggest that the absence 
of vessels in Amborella is primitive. They also call 
into question the view that the carpels of Winter- 
aceae are primitive: the basal lines (Amborella, 
Austrobaileya, etc.) have ascidiate, not plicate car- 
pels (Endress & Igersheim, 1997), implying that 
the plicate condition is derived. Igersheim and En- 
dress (1997) also refuted statements that the car- 
pels of Winteraceae and Degeneria are not com- 
pletely closed (Bailey & Swamy, 1951; 
1961); actually, their margins are postgenitally 
fused. 

These molecular challenges to morphological 
cladistic results appear to be supported by prelim- 
inary analyses of an expanded morphological data 
set of magnoliids and basal monocots and eudicots 
(Doyle & Endress, in prep.), incorporating gynoe- 
cial data of Endress and Igersheim (1997, 1999) 
and Igersheim and Endress (1997, 1998). These 


analyses also move Illiciales near Austrobaileya and 


Eames, 


associate Winteraceae with Canellaceae, based on 
palisade exotesta and truncate stamen connective. 

Phylogenetic analyses have also led to explicit 
hypotheses on relationships within Winteraceae. 
Vink (1988) presented two morphological trees of 
the family, which differed in rooting: one with Tas- 
mannia basal, on the standard assumption that the 
ancestral chromosome number is n — 13 (as in Tas- 
mannia) and n — 43 (as in other members) is de- 
rived; the other with Takhtajania basal, based on 
its elongate inflorescences, and Tasmannia and 


Drimys linked as a clade, which he preferred. Be- 
cause Vink assumed that Takhtajania has п = 43, 
based on its large pollen size (Praglowski, 1979), 
he suggested that n — 13 in Tasmannia is not an- 
cestral but derived from n — 43. This poses a prob- 
lem if the closest outgroup is Canellaceae, which 
have n = 11, 13, and 14 (Kubitzki, 1993). In both 
trees, Bubbia, Belliolum, Zygogynum, and Exosper- 
mum formed a clade, linked with Pseudowintera. 
Two species of Zygogynum (Z. balansae Tiegh., Z. 
pomiferum Baill.) differ from the rest of the family 
in having monad pollen grains (Sampson, 1974); 
the position of Zygogynum within the family im- 
plies that these are secondarily derived from tet- 
rads. 

The first limited rbcL data (Chase et al., 1993) 
were more consistent with the first of Vink's (1988) 
trees in placing Tasmannia below Drimys and Bel- 
liolum, but they did not address the position of 
Takhtajania. Analyses of rDNA ITS sequences by 
Suh et al. (1993) showed unusually low divergence 
within Winteraceae, indicating either an implausi- 
bly recent age for the family or a slowdown in ITS 
evolution, and the presence of two ITS sequences 
in Bubbia, Belliolum, Zygogynum, and Exosper- 
mum, apparently reflecting an unusual persistence 
of polymorphism in ITS. Although unrooted, the 
tree obtained was consistent with the basal position 
of Tasmannia and the existence of a Bubbia-Bel- 
liolum-Zygogynum-Exospermum clade 

The rediscovery of Takhtajania bas allowed 
placement of this genus and resolution of the ap- 
parent conflicts concerning chromosome number. 
Karol et al. (2000) included Takhtajania in analy- 
ses of ITS and trnL-F spacer sequences from Win- 
teraceae and Canellaceae. Analyses of trnL-F in- 
dicated that either Takhtajania or Tasmannia could 
be basal in Winteraceae, but Takhtajania was basal 
in trees based on ITS and the two data sets com- 
bined. Other relationships within the family were 
the same as those found by Suh et al. (1993). These 
results are consistent with Vink's (1988) view that 
the inflorescences of Takhtajania are primitive. 
However, the finding that the chromosome number 
of Takhtajania may be n = 18 ыы & Гат- 
brou, 2000) refutes his aeration that n = 13 in 
Tasmannia is derived from n = 43, which does 
appear to be a synapomorphy of Drimys and the 


remaining groups. 


FossıL RECORD 


There are several old reports of fossil wintera- 
ceous leaves, but these were not based on a critical 
analysis of leaf architecture, and they have not 


306 


Annals of the 
Missouri Botanical Garden 


been reexamined recently. As noted above, Win- 
teraceae have irregular “first rank” venation (Hick- 
ey, 1977), which has been considered primitive for 
angiosperms (Hickey & Wolfe, 1975), but otherwise 
their leaves are not very distinctive. Cuticle anal- 
ysis might allow better evaluation of these deter- 
minations, for example by showing the stomatal 
plugs characteristic of most genera (Bailey & Nast, 
1944; Feild et al., 1998). 

Some leaf reports are from areas where Winter- 
aceae occur today and winteroid pollen is known 
in the fossil record (cf. below). Berry (1938) de- 
scribed Drimys patagonica from the early Miocene 
of Argentina, said to resemble Winteraceae in hav- 
ing a “papillose” lower surface; the venation is con- 
sistent with Winteraceae, though probably not di- 
agnostic. Dusén (1908) described Drimys 
antarctica from the Paleocene (Askin, 1992) of Sey- 
mour Island on the Antarctic Peninsula; the irreg- 
ular spacing and angles of the secondary veins are 
consistent with Winteraceae. Deane (1902), miscit- 
ed by Praglowski (1979) as Card (1902) and pos- 
sibly the source of an unreferenced remark by Ber- 
ry (1938), reported Drimys leaves from the Tertiary 
of New South Wales, but the fragment illustrated 
has no distinctive features. Leaves of Winteraceae 
have not been recognized in more recent studies of 
rich Australian Tertiary floras (Carpenter et al., 
1994; Christophel, 1994; McLoughlin & Hill, 
1996; R. S. Hill, pers. comm. 1999), which have 
used more critical methods of leaf identification. 

More convincing is a report by Poole and Francis 
(2000) of vesselless wood described as Winteroxy- 
lon jamesrossi 1. Poole & J. E. Francis from the 
mid-late Santonian—early Campanian of James Ross 
Island on the Antarctic Peninsula. Although the ex- 
act combination of pitting, ray, and parenchyma 
features does not occur any modern genus of Win- 
teraceae, all are found within the family, and other 
vesselless angiosperm taxa (Amborella, Trochoden- 
drales) are more different. 

A few other megafossil reports are from outside 
the geographic range of modern Winteraceae and 
fossil records of winteroid pollen. Chaney and San- 
born (1933) described Oligocene leaves from 
Oregon as Drimys americana R. W. Chaney & San- 
born, but this is questionable, since the venation 
differs from that of Winteraceae in having thicker, 
more distinct secondary veins. Page (1979) com- 
pared vesselless wood from the Late Cretaceous 
(Maastrichtian) of California with Old World Win- 
teraceae; she separated it from New World Drimys 
and Trochodendrales based on its abundant paren- 
chyma. Gottwald (1992) described vesselless wood 


from the Eocene of Germany as Winteroxylon mun- 


dlosi, which was similar enough to the Antarctic 
wood of Poole and Francis (2000) that they as- 
signed their material to the same genus. If these 
fossils are winteraceous, they would imply that the 
family extended into Laurasia, as in Mexico and 
Malesia today. However, the possibility that they 
represent extinct vesselless lines not directly relat- 
ed to Winteraceae should also be considered, given 
the absence of more diagnostic winteroid pollen, 
the abundance of Trochodendrales in the Early Ter- 
tiary of Laurasia (Crane et al., 1991), and the pres- 
ence of leaf cuticles with similarities to Amborella 
in the lower Potomac Group (Upchurch, 1984). Ex- 
tinct vesselless lines should be more common if the 
lack of vessels in Winteraceae is primitive, since 
this would imply that the ancestors of many other 
taxa lower in angiosperm phylogeny were also ves- 
selless. 

The record of Winteraceae has been solidified 
and greatly extended by palynology, based on ul- 
cerate tetrads closely comparable to the family 
(Figs. la, 2). Unlike the wood, the distinctive fea- 
tures of the pollen are clearly derived and thus 
more indicative of this particular clade. Winteroid 
tetrads are a persistent but minor element in latest 
Cretaceous and Tertiary rocks of Australasia (Dett- 
mann & Jarzen, 1990), consistent with the low pol- 
len production and subordinate ecological status of 
Winteraceae today. Such pollen was tentatively re- 
ported (without illustration) by Cranwell (1959) 
from the Paleocene of Seymour Island, and figured 
by Couper (1960) from the Oligocene of New Zea- 
land as Pseudowintera sp. Couper’s material was 
named Pseudowinterapollis by Krutzsch (1970); 
similar tetrads from the latest Cretaceous through 
Miocene of southeastern Australia were named Ge- 
phyrapollenites (with three species) by Stover and 
Partridge (1973), who were apparently unaware of 
Krutzsch’s article. Martin (1978) indicated that the 
closest match for the Australian fossils is Tasman- 
nia. Mildenhall and Crosbie (1979) extended the 
range of Pseudowinterapollis, which they consid- 
ered most similar to Pseudowintera, from the latest 
Cretaceous through the Pleistocene of New Zea- 
land. As noted by Suh et al. (1993), since phylo- 
genetic analyses indicate that Pseudowintera is 
nested within the family, these data imply that 
crown-group Winteraceae (i.e., the clade consisting 
of all derivatives of the most recent common an- 
cestor of the living members: Doyle & Donoghue, 
1993) originated before the end of the Cretaceous. 
These grains were accepted as Winteraceae by 
Muller (1981) in his critical review of pollen evi- 
dence for extant angiosperm families. In Australa- 
sia, the oldest record is from the mid-Campanian 


Volume 87, Number 3 
2000 


Doyle 
Paleobotany of Winteraceae 


307 


nt and Early Cretaceous pollen tetrads, SEM. — 


ce 
b. “уы еч gabonensis (Doyle et al., 


of the Otway Basin of southeastern Australia (Dett- 
mann & Jarzen, 1990). Winteraceae are also known 
from the Maastrichtian or Paleocene through the 
Miocene of central Australia (Twidale & Harris, 
1977; Harris & Twidale, 1991; Macphail et al., 
1994) and the Oligocene of Tasmania (Macphail & 
Hill, 1994). However, since the early report of 
Cranwell (1959), Winteraceae do not seem to have 
been observed in Tertiary pollen floras from Ant- 
arctica (Truswell & Drewry, 1984; Truswell, 1991; 
Askin, 

Mildenhall and Crosbie (1979) also reported 
loose ulcerate tetrads and monads, named Harrisi- 
pollenites, from the Oligocene through Pleistocene 
of New Zealand, which they compared with the mo- 
nads of Zygogynum species (Sampson, 1974). 
Since phylogenetic analyses indicate that monad- 
producing Zygogynum is one of the most apical 
branches in the family (Vink, 1988; Suh et al., 
1993; Karol et al., 2000), this record is evidence 
that crown-group Winteraceae had diversified to a 
high level by the Oligocene. These considerations 
would further support the view of Suh et a 
that the low divergence of ITS sequences within 
Winteraceae is due to a slowdown in molecular evo- 
lution, rather than a recent origin of the crown- 


Two types of winteroid tetrads are also known 


a. Drimys winteri (cultivated, Davis, California). — 


1990a), Zone С-УП (late Barremian? ), Gabon. Scale bar = 5 um. 


from Tertiary beds (considered early Miocene) of 
the Cape region in South Africa (Coetzee, 1981; 
Coetzee & Muller, 1984; Coetzee & Praglowski, 
1988), where Winteraceae are now extinct. This im- 
plies that Takhtajania is the only survivor of a for- 
merly more widely distributed assemblage of Win- 
teraceae in the African-Madagascan region. The 
same floras contain other taxa that no longer occur 
in mainland Africa but persist in Madagascar: As- 
carina (Chloranthaceae), Casuarina, е 
Sapindaceae), and Sarcolaenaceae. However, ас- 
cording to Coetzee and Muller (1984) nd Coetzee 
and Praglowski (1988), 
ilar not to pollen of Takhtajania, but rather to that 
of the Australasian genera Tasmannia (“Drimys” pi- 
perita) and Bubbia (“Zygogynum” queenslandian- 
um). 

Although pollen of Drimys occurs at low fre- 


these fossils are most sim- 


quencies in the Quaternary of Chile (e.g., Heusser, 
1981), there have been few reports of winteroid tet- 
rads from older sediments in South America, al- 
though such r the been extensively studied 
(cf. Askin & Baldoni, 8). Apparently the oldest 
are grains identified м Baldoni (1987) as Gephyr- 
apollenites calathus Partridge from the Paleocene- 
Eocene of Argentina. Barreda (1997) reported 
Pseudowinterapollis couperi Krutzsch from the Oli- 
go-Miocene of Argentina; she provided SEM figures 


308 


Annals of the 


Missouri Botanical Garden 


Northern 
Gondwana 
Southern 

Gondwana 


Early 
Cretaceous 


Late 
Cretaceous- 
Tertiary 


Pre- т occurrences of "iid tetrad pollen related to Winteraceae and vesselless wood of Poole 
Bot 


. Top, Early VP ee аи 
iary коа тар 50 Му, Кос 
Oligocene, Miocene, Pliocene. See text e E Ru 


showing a tetrad closely comparable to Drimys in 
sculpture and presence of a well-defined annulus. 

The stratigraphic and geographic range of win- 
teroid pollen was extended dramatically by Walker 
et al. (1983), based on tetrads in two cores from 
the late Aptian-early Albian of Israel. At this time 
(Fig. 2), Israel was part of the Northern Gondwana 


у, Barremian-Aptian: Scotese, 1997). Bottom, Late Creta- 


MEA 1997); 1: аи. Maastrichtian; 2: Paleocene, Eocene; 3: 


province of Brenner (1976), which straddled the 
Early Cretaceous equator and included all but the 
southern portions of Africa and South America. In 
contrast, Late Cretaceous, Tertiary, and Recent oc- 
currences of Winteraceae (except those in Central 
America and Malesia) are in regions that belonged 
to Brenner’s Southern Gondwana province in the 


Volume 87, Number 3 
2000 


Doyle 
Paleobotany of Winteraceae 


309 


Early Cretaceous. The Israeli tetrads, designated 
Walkeripollis sp. A by Doyle et al. (1990a), resem- 
ble pollen of modern Winteraceae in having a 
round ulcus consisting of a central pore surrounded 
by a thicker annulus, underlain by a safranin-stain- 
ing ring that Walker et al. (1983) assumed was 
thickened endexine, as in many modern Wintera- 
ceae (Praglowski, 1979). However, Walker et al. in- 
terpreted them as more primitive than living Win- 
teraceae because of their finer, foveolate-reticulate 
sculpture. 

Still older winteroid pollen was described by 
Doyle et al. (1990a, b) from the Cretaceous of Ga- 
bon, as Walkeripollis gabonensis J. A. Doyle, Hotton 
& J. V. Ward (Fig. 1b). These tetrads appear to be 
even more primitive in having a slightly elliptical 
aperture (presumably transitional from a sulcus) 
and foveolate sculpture, with small tectal perfora- 
tions only. Transmission electron microscopy (TEM 
confirmed that the endexine is thickened under the 
annulus. These grains were found in only one sam- 
ple, from an interval (Zone C-VII) near the end of 
filling of the nascent South Atlantic rift with con- 
tinental sediments, but they are fairly common in 
this sample. Their significance was overlooked by 
Doyle et al. (1977, 1982), who assumed that they 
were tetrad-producing variants of Tucanopollis cri- 
sopolensis (Regali, Uesugui & A. S. Santos) Regali, 
a common angiosperm in these beds, which also 


м 


has a round, sculptured aperture. The age of Zone 
С-УП and its equivalents in Brazil was originally 
considered early Aptian (Doyle et al., 1977, 1982), 
but Regali and Viana (1989) and Doyle (1992) ar- 
gued that it is late Barremian, based in part on 
reports of two associated new groups, Afropollis and 
tricolpate pollen, in independently dated late Bar- 
remian rocks in Morocco (Giibeli et al., 1984) and 
England (Penny, 1989) (and more recently Israel: 

. J. De Haan, unpublished Ph.D. thesis, Botany, 
Univ. California, Davis, 1997). 

Compared with modern Winteraceae, both Walk- 
eripollis gabonensis and W. sp. A are anomalous in 
being calymmate (Van Campo & Guinet, 1961), 
with the tectum partially continuous between ad- 
jacent monads of the tetrad. In this respect, they 
appear more advanced than the acalymmate tetrads 
of the modern taxa, where the tectum stops at the 
junction between monads. Since the circular aper- 
ture and coarser reticulum of Walkeripollis sp. A 
suggest that this species is phylogenetically closer 
to modern Winteraceae than W. gabonensis, Doyle 
et al. (1990b) inferred that the Ронни condition 
arose in the common ancestor of the two species 
and reversed to acalymmate in modern Wintera- 
ceae, along with coarsening of sculpture. The fact 


that Walkeripollis is calymmate may be grounds for 
caution in relating it to Winteraceae, but W. sp. А 
is almost perfectly intermediate between W. gabo- 
nensis and modern Winteraceae, and both species 
are only weakly calymmate, implying that the re- 
versal required would be minor. The fact that both 
Walkeripollis species have a conspicuous annulus, 
like Takhtajania, Drimys, Belliolum, and Zygogyn- 
um species with monads (Praglowski, 1979), sug- 
gests that the lack of a well-differentiated annulus 
in Tasmannia, Pseudowintera, Bubbia, Exosper- 
mum, and Zygogynum species with tetrads is de- 
rived. Absence of an annulus seems loosely corre- 
lated with smaller aperture size. Exospermum and 
Zygogynum are most like Walkeripollis in having 
fine sculpture, but their position in morphological 
and molecular phylogenies (Vink, 1988; Suh et al., 
1993; Karol et al., 2000) implies that this condition 
is secondarily derived. 

Winteroid tetrads may be more widespread in the 
Early Cretaceous than reported so far. In the middle 
Albian Khafji member of the Wasia Formation of 
offshore Saudi Arabia, S. L. Gaponoff (pers. comm. 
1990 & 1999) found a single tetrad, which is sim- 
ilar to Walkeripollis sp. A in sculpture but looser 
and apparently acalymmate, suggesting that it is 
still more closely related to crown-group Wintera- 
ceae. But so far there is a gap in the record of such 
pollen through the first half of the Late Cretaceous. 

A possibly related pollen group consists of fo- 
veolate-reticulate, ulcerate monads from the early 
Aptian to Albian of Israel, described by Brenner 
and Bickoff (1992) as Retimonoporites operculatus 
G. J. Brenner & Bickoff. Their sculpture is roughly 
similar to that of Walkeripollis sp. A; the ulcus is 
covered by an operculum and underlain by a dark- 
staining area interpreted as endexine. Because 
these monads predate W. sp. A in Israel, Brenner 
(1996) interpreted them as representing a more 
primitive, pre-tetrad state; however, they are prob- 
ably younger than W. gabonensis. It is possible that 
these grains are related to Winteraceae, but there 
are enough differences to make this view somewhat 
speculative. They are much smaller than the mo- 
nads of Walkeripollis, the ulcus is proportionally 
smaller, and the endexine forms a solid patch rather 
than a ring around the border of the ulcus. 

Doyle et al. (1990a, b) also suggested that two 
other Cretaceous pollen groups may be related to 
Winteraceae: Afropollis, which is abundant from the 
late Barremian through the early Cenomanian of 
Northern Gondwana, and Schrankipollis, from the 
Aptian of Egypt and Maryland. Afropollis varies 
from operculate or zonasulculate in the oldest spe- 
cies to inaperturate in younger ones, with a loose 


Annals of the 
Missouri Botanical Garden 


reticulum surrounding a central body, and the op- 
erculates have t 
below the reticulum. Schrankipollis is zonasulculate 
but elliptical and more finely sculptured. The com- 
parison with Winteraceae was based primarily on 
the fact that Afropollis, Schrankipollis, and Walker- 
ipollis gabonensis all have finely segmented muri 
and tend toward circular shape and a round aper- 
ture, with the zonasulculus presumably derived by 
broadening of an operculum. Based on a cladistic 
analysis of these fossils and pollen of living Win- 
teraceae and Illiciales, Doyle et al. (1990b) inferred 
that Afropollis and Schrankipollis belong to an ex- 
tinct sister group of Walkeripollis, Winteraceae, and 
Illiciales. The two Walkeripollis species were suc- 
cessive branches of the stem-lineage leading to 


ical angiospermous columellae 


both Winteraceae and Illiciales, based on the ellip- 
tical aperture of W. gabonensis and the finer sculp- 
ture of both species. The trichotomosulcate (“зуп- 
tricolpate") monads of Illiciales were linked with 
modern winteraceous tetrads by their coarse sculp- 
ture, implying that they are secondarily derived 
from tetrads. Because this analysis placed Walker- 
ipollis on the stem-lineage to Winteraceae and Il- 
liciales, and Illiciales have vessels, Doyle et al. 
(1990b) speculated that the plants producing Walk- 
eripollis still had vessels. 

his scheme can no longer be defended, since 
only the tetrads seem securely related to Wintera- 
ceae, and molecular evidence against a relationship 
of Illiciales and Winteraceae has become over- 
whelming (cf. above). Doyle et al. (1990a, b) ac- 
knowledged that the winteraceous affinity of Afro- 
pollis and Schrankipollis was more speculative. 
They noted that Afropollis is anomalous in having 
a thick endexine all around the grain, as in gym- 
nospermous seed plants, rather than under the ap- 
erture only, as in Winteraceae and most other mag- 
nolid angiosperms. Because other characters of 
Afropollis seemed so angiospermous, and because a 
few magnoliids do have a thick endexine, they sug- 
gested that the endexine character may not rule out 
angiosperm affinities. However, Friis et al. (1999) 
have found Afropollis in separate pollen sacs with 
no evident angiosperm features. A similar combi- 
nation of gymnospermous endexine and angiosper- 
mous columellae and sculpture also occurs in the 
Late Triassic Crinopolles pollen group, which Cor- 
net (1989) interpreted as angiospermous, but which 
Doyle and Hotton (1991) and Doyle and Donoghue 
(1993) suggested is related to the angiosperm 
crown-group but more primitive—i.e., on the an- 
giosperm stem-lineage. By the same reasoning, Af- 
ropollis might also be a side-line of the angiosperm 
stem-lineage that persisted into the Cretaceous. 


These doubts do not apply to Schrankipollis, since 
it has only a little endexine under the aperture, but 
without Afropollis to link it with Winteraceae, there 
is little reason to associate it with the family. 

Scenarios for origin of the tetrad pollen of Win- 
teraceae also need revision in light of molecular 
and morphological evidence that Canellaceae are 
the sister group of the family. Pollen of Canellaceae 
is small, round, and monosulcate, with occasional 
trichotomosulcate variants (Wilson, 1964; Walker, 
1976; J.-M. Groult, unpublished D.E.A. thesis, Mu- 
séum National d'Histoire Naturelle, Paris, 1998). 
Its exine structure varies from granular with a con- 
tinuous scabrate tectum (Capsicodendron) to colu- 
mellar and either foveolate (most genera) or retic- 
ulate (Cinnamosma). Walker (1976) assumed that 
the granular extreme is primitive, as part of a gen- 
eral trend in angiosperms as a whole. This would 
imply that columellae arose independently in Ca- 
nellaceae and Winteraceae. The idea that granular 
structure is primitive in angiosperms was supported 
by the analysis of Donoghue and Doyle (1989), 
which placed Magnoliales at the base of the angio- 
sperms, but in more recent analyses, where Mag- 
noliales are nested within woody magnoliids, their 
granular structure is a reversal. Canellaceae are 
rooted differently in trees based on ITS and trnL- 
F (Karol et al., 2000), but the unrooted tree is the 
same, and Capsicodendron is not basal in either 
tree. This suggests that the common ancestor of 
Winteraceae and Canellaceae had small, round, 
monosulcate pollen with columellae and foveolate 
sculpture, a type common in the Early Cretaceous. 
Such pollen may go back to the first angiosperms; 
for example, it occurs in the near-basal genus Aus- 
trobaileya (Endress & Honegger, 1980). These data 
also remove the basis for the conjecture that the 
parent plants of Walkeripollis had vessels (Doyle et 
al., 1990b). Based on pollen morphology, a phylo- 
genetic analysis would place Canellaceae below 
Walkeripollis, so vessels could have been lost at any 
point on the line leading to crown-group Wintera- 
ceae, either before or after Walkeripollis. 


GEOGRAPHIC HISTORY AND ECOLOGICAL 
EVOLUTION 


Fossil data suggest that the stem-lineage leading 
to Winteraceae, represented by Walkeripollis, orig- 
inated in the tropical zone of Northern Gondwana. 
This is consistent with the view of Raven and Ax- 
elrod (1974) that angiosperms as a whole originated 
in this region. The Early Cretaceous climate in the 
classic areas of Gabon and Brazil has been inter- 
preted as hot and dry (Brenner, 1976; Doyle et al., 


Volume 87, Number 3 
2000 


Doyle 
Paleobotany of Winteraceae 


1977, 1982), based on the low frequency of spores, 
abundance of Classopollis (the conifer family Chei- 
rolepidiaceae, noted for its xeromorphic vegetative 
morphology) and ephedroid pollen (related to mod- 
ern Gnetales, of which Ephedra and Welwitschia are 
desert plants), and the presence of thick Aptian salt 
deposits, which mark the first influx of marine water 
into the rift. However, there is evidence for wetter 
conditions in the Middle East and northern South 
America, which were near the equator rather than 
about 15 degrees to the south: less common Clas- 
sopollis and ephedroids, abundant fern spores and 
Araucariaceae, and occasional coals (Doyle et al., 
1982; McCabe & Parrish, 1992; Brenner, 1996) 
Interestingly, Walkeripollis occurs in both areas, 
suggesting that its parent plants could tolerate some 
range of rainfall. The fact that W. gabonensis is from 
a rift sequence raises the possibility that its parent 
plants were growing in cooler upland areas flanking 
the rift. However, models for rift evolution predict 
that relief would decrease with time, and although 
the presence of bisaccate podocarpaceous pollen 
suggests that there were high elevations near the 
rift in the earliest Cretaceous, such pollen had dis- 
appeared by the time Walkeripollis appeared in the 
Barremian (Doyle et al., 1982). 

This scenario also fits the present distribution of 
probable outgroups of Winteraceae. Canellaceae 
are entirely African, Madagascan, and A 
and Walker (1971) and Raven and Шы. (1974) 
interpreted them as of Northern Gondwanan origin. 
Ecologically, they are more tropical than Wintera- 
ceae, and they extend into drier environments (Ku- 
bitzki, 1993). The second outgroup to Winteraceae, 
whether Magnoliales or Piperales, is less well es- 
tablished. However, if Magnoliales are the second 
outgroup, it may be significant that rbcL, atpB, and 
the two genes combined indicate that the basal 
branch in this group is Myristicaceae (Chase et al., 
1993; Savolainen et al., 2000), which have a geo- 
graphic distribution that also suggests a Northern 
Gondwanan origin (Walker, 1971; Raven & Axel- 
rod, 1974). Myristicaceae are also a lowland trop- 
ical group, and although their species diversity 18 
highest in Asia, Walker and Walker (1981) argued 
that their most primitive members are the Mada- 
gascan genera Mauloutchia and Brochoneura, 
which have helical rather than whorled stamens (a 
view confirmed by a morphological cladistic anal- 
ysis by H. Sauquet, unpublished D.E.A. thesis, 
Univ. Pierre et Marie Curie, Paris, 1999). 

Thus both fossil distributions and phylogenetic 
results suggest that the temperate, Southern Gond- 
wana distribution of crown-group Winteraceae 
came about by southward migration and radiation 


of plants derived from a stem-lineage in the lowland 
tropics. In the Early Cretaceous, Southern Gond- 
wana was characterized by Podocarpaceae, Arau- 
cariaceae, and abundant spore-bearing plants 
(Brenner, 1976; Herngreen et al., 1982; Dettmann, 
1994), suggesting cool, wet conditions like those 
where Winteraceae grow today. These data may fit 
the hypothesis discussed above that vessels were 
lost in cool, wet environments because of their sus- 
ceptibility to embolisms caused by freezing (Don- 
oghue & Doyle, 1989; Feild et al., 2000). 
major unresolved problem is when exactly 
Winteraceae moved into the temperate zone, and 
by what route. After the records of Walkeripollis in 
the Early Cretaceous, the line is not known until 
the Santonian-Campanian of Antarctica, represent- 
ed by Winteroxylon wood (Poole & Francis, 2000), 
and the Campanian-Maastrichtian of Australia and 
New Zealand (Stover & Partridge, 1973; Mildenhall 
& Crosbie, 1979; Dettmann & Jarzen, 1990), rep- 
resented by coarsely reticulate tetrads resembling 
crown-group Winteraceae. These tetrads seem well 
established in Australasia, and there are no records 
of them in the tropics (although given the rarity of 
earlier reports the significance of such negative ev- 
idence can be questioned). Migration to Southern 
Gondwana would pose little problem in the Earl 
Cretaceous, when the continents were still largely 
connected (Fig. 2). However, by the time the crown- 
group is first recognized, ocean barriers had be- 
come much wider, especially between Northern 
Gondwana and Australasia. Possible scenarios may 
be considered in terms of continental positions ca. 
6 My before the Campanian (Fig. 3; Scotese, 1997). 
One route to Australasia might be through South 
America and Antarctica, which was attached to 
Australia until late in the Cretaceous. On paleo- 
geographic grounds, it is more likely than not that 
the stem-lineage occurred in South America. In the 
Early Cretaceous, Brazil and Gabon were two sides 
of the same rift valley, and given the rarity of Walk- 
eripollis, the lack of reports from Brazil is probably 
not significant. However, if the crown-group spread 
from South America to Australasia in the Late Cre- 
taceous, it would also have to disperse across the 
South Atlantic in one direction or the other to ex- 
plain its Tertiary and Recent occurrences in Africa 
(Coetzee & Muller, 1984) and Madagascar (Takh- 
tajania). This scenario implies that near-basal lines 
of Winteraceae occurred in South America in the 
Late Cretaceous, which may conflict with the rela- 
tively nested position of the American genus Dri- 
mys in the family and the rarity of reports of pre- 
Quaternary winteroid pollen in South America 


312 


Annals of the 
Missouri Botanical Garden 


Early 
Late Cretaceous 


Figure 3. 
to Australasia (90 My, 
Madagascar-India. 


(despite numerous palynological studies: cf. Askin 
& Baldoni, 1998), as compared with Australasia. 
Anche: route to Australasia might be through 
Madagascar. Gondwana began to split into East 
Gondwana (Madagascar, India, Antarctica, Austral- 
asia) and West Gondwana (Africa, South America) 
in the Middle Jurassic (Rabinowitz et al., 1983). 


ince in the Early Cretaceous (Brenner, 1976; Hern- 
green et al., 1982; Dettmann, 1994), formed a block 
that might have acted as a stepping stone between 
Africa and Australasia and/or Antarctica (Fig. 3). 
This scenario might fit molecular evidence that 
Takhtajania is basal in Winteraceae (Karol et al., 
2000): Takhtajania would represent a branch that 
stayed near the original Northern Gondwana area, 
while the rest of the family was derived from a 
branch that dispersed to Australasia. Drimys could 
be a line that dispersed later from Australasia to 
South America via Antarctica (Raven & Axelrod, 
1974), consistent with its position in the family. 
If, however, Winteraceae in the Tertiary of South 
Africa were related to modern Australasian genera 
(Coetzee & Muller, 1984; Coetzee & Praglowski, 
1988), more complex scenarios must be envisioned, 
with a wider Cretaceous distribution of the crown- 
group and many local extinctions. The discovery of 
winteraceous wood in the Late Cretaceous of Ant- 
arctica (Poole & Francis, 2000) also suggests it 


Paleogeography near the time of inferred dispersal of crown-group Winteraceae from Northern Gondwana 
Turonian: Scotese, 1997), showing alternative routes through South America-Antarctica and 


would be premature to favor one scenario over an- 
other: it implies that Winteraceae reached Antarc- 
tica very early, from which they could easily spread 
to either Australasia or South America. Better ev- 
idence on the phylogenetic position of known Cre- 
taceous and Tertiary fossils, or discoveries of new 
fossils, could greatly clarify the geographic history 
of the family. 


CONCLUSIONS 


The stem-lineage leading to Winteraceae is one 
of the oldest recognizable angiosperm lines, ex- 
tending back to the Barremian stage of the Early 
Cretaceous in Northern Gondwana. Modern Win- 
teraceae appear to be derived from a line that 
spread from the tropics into the southern temperate 
zone, where Winteraceae were widespread in the 
latest Cretaceous and Tertiary, but the details are 
uncertain. Major priorities are to fill in the Late 
Cretaceous record and to clarify the time of ap- 
pearance of crown-group Winteraceae in Africa- 
Madagascar, Antarctica, and South America. Hope- 
fully this paper will alert palynologists and 
paleobotanists working in the Gondwana continents 
to winteraceous pollen and megafossils, and thus 
contribute to a more detailed reconstruction of the 
history of this important family of primitive angio- 
sperms. 


Volume 87, Number 3 
2000 


Doyle 313 
Paleobotany of Winteraceae 


Literature Cited 


APG (Angiosperm Phylogeny Group). 1998. An ordinal 
classification for the families ie а plants. Ann. 
Missouri je hor 85: 531 
Askin, R. te es cid: Tertiary Ant- 
acti outc гор ner но | m к and climates. 
Pp. 61-73 in J. P. Kenn D. rnke (editors), 
The жеры, S киса ма ent: rspective on 
Global Change. American Geophysical Union, Vash- 
ington, D.C 

——— & A. Baldoni. 1998. The Santonian through Pa- 
leogene record of Proteaceae in the ав Жуз 
Атепса- а Peninsula region. Austral. Sys 
11: 373- 

Bailey, 1. т & С. С. Nast. 1944. The comparative тог- 
phology of the Winteraceae. У. Foliar аи and 
sclerenc Te J. Arnold Arbor. 25: 342-34 

& B. G. L. Swamy. 1951. The ае ИЕ саг- 
pel of E and its initial trends of specializa- 

tion. Amer. J. Bot. 38: 373-379. 

Baldoni, A. M. 1987. Nuevas descripciones palinológicas 
en el área de Collón ie (Terciario inferior) Provincia 
del Neuquén, Argentina. IV Congreso Latinoamericano 
de еони. Bolivia (1987)1: 399-414 

Barreda, V. . Palynomorph assemblage ui the Chen- 
que Formation, ide Oligocene?-Miocene from Golfo San 
Jorge basin, Patagonia, Argentina. Part 4. Polycolporate 
and porate pollen. Ameghiniana 34: 145-154. 

Berry, E. W. 1938. Tertiary flora from the Rio Pichile 
Argentina. Special Pap. Geol. Soc. Amer. 12: 1-14 

Brenner, G. J. 1976. Middle Cretaceous floral provinces 
and early migrations of angiosperms. Pp. 23-47 in C. 
B. Beck че Origin and Early Evolution of Angio- 
sperms. Columbia Univ. Press, New York. 

— iai for the earliest stage of angio- 

b pollens evolution: A d section from 
be Pp. 91- n D. W. Taylor & L. J. Hickey (ed- 
itors), Flowering Plant PLE Evolution & Phylogeny. 

Chapman & Hall, New Yor 

& I. S. Bickoff. 1992. Fiho and age of the 

Lower Cretaceous basal Kurnub Group from the coastal 

plain to the northern Negev of Israel. Palynology 16: 

137-185. 


Card, G. W. An vig nid pbi breccia from the 
Bingera аади field. Кес. Geol. Surv. New South 
Wales 7: 29-101. 


Carlquist, S. 1975. Ecological Strategies of Xylem Evo- 
lution. Univ. California Press, Berkeley. 
983. Wood anatomy of Bubbia (Winteraceae), 
with comments on aie of vessels in dicotyledons. 
к Bot. 70: 578— 
Presence en ен in wood of Sarcandra 
(Chloranthaceae) Comments on vessel origins in angio 
$ Amer. J. Bot. T4 3205-1071, 
Near sness in Ephedra and its 
a Зари pa J. Bot. 15: 598-601. 
1996. Wood anatomy of primitive angiosperms: 
New Desi and syntheses. Pp. 68-90 in D. W. 
aylor & L. J. ds (editors), Flowering Plant Origin, 
y. Chapman & Hall, New York. 
ill & G. J. Jordan. 1994. Ce- 
smania: Macrofossil evidence. 
Pp. 276-298 in R. S. Hill (editor), History of the Aus- 
tralian Vegetation: нн, to Recent. Cambridge 
Univ. Press, Cambridg 
Chaney, R. W. & Е. 1. en 1933. The Goshen flora 


of west central Oregon. Publ. Carnegie Inst. Wash. 439: 
103. 


eu M. W., D. E. Soltis, R. G. grain D. Morgan, 
all, 


, W. D. Clark, M. 
„ге , R. K. Jansen, К.-Ј. Kim, С. Е 
Wines: J. F. "Ба, С. К. Furnier, S. Н. Strauss, Q.- 

Xiang, С. М. Plunkett, Р. 5. Soltis, 5. 


genetics of seed plants: An Апар d nuc Jeotidé se- 

quences from the plastid gene rbcL. Ann. Missouri Bot. 

Gard. 80: 526- 

i . The early Tertiary mac 'rofloras 
il 


of the Australian Vegetation: Cretaceous 

to Recent. Cambridge Univ. Press, Cambridg 

Coetzee, J. A. 1981. A palynological record of very prim- 
шуе р in Tertiary deposits of the south-west- 


~ 


ern Cape Province, South Africa. 5. African J. Sei. 77: 
343. 


& J. Muller. 1984. The phytogeographic signifi- 

cance of some extinct Gondwana pollen typ 

Tertiary of the southwestern Cape (South Africa). Ann. 

Missouri Bot. Gard. 1 

г J. Praglowski. Winte eraceae pollen from 
the Miocene of the NE Cape (South Africa): 
Relationship to modern taxa and phytogeographical sig- 
nificance. Gran “ 24 27-37. 

Corner, E. 976. The Seeds of the Dicotyledons. 
Сана E Press, Cambridge 

Cornet, В. 1989. Late Triassic angiosperm-like pollen 
from the Richmond rift basin of Virginia, U.S.A. Pa- 
bsp deas Abt. B 213: 37-8 


es from the 


Couper, R. 1960. New Zealand Mesozoic and Caino- 
7010 кн "y digi. cs New Zealand Geol. Surv. Pa- 
leontol. sm 


Crane, Р. i Manchester & D. L. Dilcher. 1991. 
nis e and t f Nordenskioldia 
(Troc hodendracese} 4 a sim dicotyledon from the 
Early Tertiary of the Northern Hemisphere. Amer. J. 

4. 


Cranwell, L. M. 1959. Fossil epe н Seymour Island, 
Antarctica. Nature 184: 1782-17 

C ronquist, А. 19 ће ^E e and Classification of 
Flowering Plants, 2nd ed. New York Botanical Garden, 
t ronx. 

Deane, H. 1902. Notes on fossil leaves from the Tertiary 
deposits of Wingello and Jum. Кес. Geol. Surv. 
New South we 7: 59-6 

Dettmann, M. E. 1994. Ру ae vegetation: The mi- 
crofossil record. Pp. 143-170 in R. S. Hill (editor), His- 
tory of the Australian Vegetation: Cretaceous to Recent 
Cambridge Univ. Press, Cambri 

en. 1990. The Amare Austrian 
rift valley: Late pi dp cradle of northeastern Aus- 
tralasian relicts? Rev. Palaeobot. Palynol. 65: 131-144. 

Donoghue, M 1994. Progress and prospects in recon- 
structing plant pirdifgens. Ann. Missouri Bot. Gard. 81 

5-418. 


& J. A. Doyle. 1989. Phylogenetic analysis of 
ie e p and the relationships of Hamamelidae. Pp. 


17-45 in P. R. Crane & S. Blackmore (editors), Evo- 


314 


Annals of the 
Missouri Botanical Garden 


lution, Systematics, and Fossil History of the Hama- 
melidae, Vol. 1. Clarendon Press, Oxford. 

Doyle, J. A. 1992. Revised palynological correlations a 
the lower Potomac Group (USA) and the Cocobeach s 
quence of Gabon (Barremian-Aptian). Cretaceous Res. 
13: 337-349, 

96. Seed plant phylogeny and the relation- 

ships of Gnetales. Int. J. Pl. Sci. 157 (6, Suppl.): 53- 
39. 


Donoghue. 1993. Phylogenies and ап- 

giosperm Пи ‘ation. о 19: 141-167 
& L. J. Hickey. 1976. Pollen and leaves inr the 
mid-Cretaceous Potomac eie and iud bearing on 
early a еса evolution. Рр. 139-206 in С. В. Beck 
(editor), Origin and Early Evolution of ш ос 
Columbia Univ. Press, New York. 

С. Hotton, 1991. 

angiosperm pollini in a cladistic context. Pp. 
in S. Blaka & S. H 


Diversification of early 
169-195 
. Barnes (editors), Pollen and 
: Patterns of Diversification. Clarendon Press, 


A. Doerenkamp & 5. Jardiné. 1977 
fer nee iG from the pre-Albian Cretaceous of 
Equatorial Africa ww Centr. Rech. Explor.-Prod. Elf- 
Aquitaine 1: 451 

jene ки & E. A. Zimmer. 1994. Inte- 
gration a morphologic ral and ribosomal RNA data on 
the origin of angiosperms. Ann. Missouri Bot. Gard. 81: 

9-450. 


tton & J. V. Ward. 1990a. Early Cre- 
taceous ane zonasulculate pollen, and Winteraceae. 
I. p pu p and ultrastructure. Amer. J. 
Bot. 44—15 


& 90b. Early Cretaceous tet- 
rads, нз ulate polen: and Winteraceae. П. Cladis- 
tic analysis and implications. Amer. J. Bot. 77: 1558- 

568. 


5. Jardiné & A. Doerenkamp. 1982. Afropollis, 
a new quan of early angiosperm pollen, with notes on 
(ће Cretaceous palynostratigraphy and po 
ments of Northern Gondwana. Bull. Centr. Rech. 
plor.-Prod. Elf-Aquitaine 6: 39-117 
Dusén, Р. 1908. Uber de tertiüre Flora der Seymour-Insel. 
iss. Ergebn. Schwed. Südpolar-Exped. 1901—1903, 
3(3): 1-27. 
Eames, A. J. 1961. 
Graw-Hill, New Y 
Ehrendorfer, F. & M. Lambrou. 2000. Chromosomes of 
Takhtajania, other Winteraceae and Canellaceae: з 
logenetic implications. Ann. Missouri Вог. Gard. 
13 


Morphology of the Angiosperms. Mc- 
‘ork. 


Endress, P. K. 1987. The Chloranthaceae: ов 
structures phylogenetic position. Bot. Jahrb. Syst 
109: 153- 


А МИҢ 1980. The pollen of Austrobail- 
eyaceae and its phylogenetic significance. Grana 19: 
177-182. 


Igersheim. 
— е of Laurales 
16 


1997. Gynoecium Diversity and 

. Bot. J. Linn. Soc. 125: 93-168. 
. Gynoecium diversity and s 

tematics of the basal eudicots. Bot. J. Linn. Soc. 


1 30: 


30 : 

Feild, T. S., M. A. Zwieniecki, M. J. Donoghue & N. M 
Holbrook. 1998. Stomatal plugs of Drimys winteri 
гы ee protect leaves ЈЕ mist but not drought. 
Proc. Natl. Acad. Sci. U.S : 14256-14259. 

——_, — —— & N. M. Peru 2000. Winteraceae 


evolution: An ecophysiological perspective. Ann. Mis- 
souri Bot. Gard. 87: 323-334. 

Friis, E. M., K. R. Pedersen & P. R. Crane. 1999. Early 
angiosperm diversification: The diversity of pollen as- 
sociated with angiosperm re productive structures in 
Early uu eous floras from Portugal. Ann. Missouri 
Bot. Gard. 86: 259-296. 

Goremykin, V., а“ Bobrova, J. Pahnke, А. Troitsky, A. An- 
tonov & W. Martin. 1996. Noncoding sequences from 
the slowly ‘évalving chloroplast inverted repeat in ad- 
dition to rbcL data do not support етен affinities 
of angiosperms. Molec. Biol. Evol. —396. 

Gottwald, H. 1992. Hólzer aus marinen ы, Чез ођ- 
eren Eozün von Helmstedt (Niedersac hsen). Palaeonto- 


oo i B 25: 27-10. 


Gübeli, A. Hochuli & W. Wildi. 1984. Lower 
с КШ sediments from the Rif chain 
(Northern Marocco)—Palynology, stratigraphy Er pe 


laeogeographic setting. Geol. Rundschau 73: 
1114. 

pes В. К. & E. A. Zimmer. 1992. Ribosomal ја 

a phylogenetic tool in plant systematics. Pp. 50— 

in LP S. Soltis, D. E . J. Doyle (editors), in 
lecular Systematics of Plants. Chapman and Hall, New 
York. 

Harris, W. K R. Twidale. 


. & С 1991. Revised age for 
yers Rock jd the Olgas. 
5: 


Trans. Roy. Soc. South Aus- 


. Randrianasolo & J. W. Verbeek 

982. Mic paleo ‚ of Albian to A strata in 
и, paleontology 28: 9 

Heusser, С. 1. А үе: of the ie: interglacial- 
glacial cycle in ы of when Chile. Quatern 
Res. 16: 293-321. 

Hickey, L. J. 1977. Stratigraphy and berger d of the 
Golden Valley Formation (Early Tertiary) of western 

Amer. 150: 1– "m 

5. The bases of angiosperm 
Ловепу: + morphology. Ann. Missouri Bot. 

Gard. 62: е 

Hoot, 5. В., S. Magallén & P. R. Crane. 1999. ео 
of basal pum p De. on three molecular data sets 
jon rbcL, and 185 nuc d a DNA sequences. 

. Missouri Bx Gard. 86: 1–32. 

ioe A. & РК. 

ч апа ое 
Bot. J. Linn. 


kota. Mem. Geol. Soc. 


1997. Gynoecium diver- 
s of the oo and winteroids. 
24: 21: 


a 


1998. йел ium diversity and sys- 
289— 


tematics of the йерей. Bot. J. Linn. Soc. 127: 
370. 


Karol, K. G., Y. Suh, G. E. Schatz & E. A. Zimmer. 2000. 
Molecular evidence Lg the de ios position of 
Takhtajania in the Winteraceae: Evidence from nuclear 
ribosomal and corp gene spacer sequences. Ann. 
Missouri Bot. Ga 4432. 

Krutzsch, W. Kenntnis fossiler disperser Te- 
РА Paliiontol. Abh. Abt. B 3: 399—433. 

Kubitzki, K. „1993. Canellaceae. Pp. 200–203 in K. Ku- 
bitzki, J. G. Rohwer & V. Bittrich ке The Fami- 
lies and аи of Vascular Plants, Vol. 2. Flowering 
Plants—Dicotyledons. Magnoliid, Hamamelid and Car- 
yophyllid Families. Springer-Verlag, 

D. n. i, ‘Cladistics of the 


; . 5. Hill. Du K-Ar dated paly- 
Tasmania 1: Early Oligocene, фени 
UD Zone ie НЦ Wilmot Dam, northwes 


Моште 87, Митбег 3 
2000 


Ооује 315 
Paleobotany of Winteraceae 


ern Tasmania. Pap. & Proc. Roy. Soc. Tasmania 128: 
© Alley, E. M. Truswell & I. В. K. Sluiter. 
4. Early пина b ar on: Evidence from spores 
and pollen. Pp. 1 61 in R. S. Hill editor). к 
of the Australian pis Cretaceous to Rec 
Cambridge Univ. Press, Cambridge. 
Martin, H. A. 1978. Evolution of the Australian flora and 
vegetation through the Tertiary: Evidence from pollen. 
202. 


& M. J. Donoghue. 1 root of angio- 
sperm ор. inferred from alee phytochrome 
enes. Science 286: 947—950. 

McCabe, P. J. & J. T. Parrish. 1992. Tectonic е 
controls on the distribution and quality of С eous 
coals. In: P. J. McCabe & J. T. Parrish (editors), ‘cil 
on the Distribution and Quality of Cretaceous Coals. 
Special Pap. Geo Amer. 267: 1-15. 

McLoughlin, S. & R. Hill. 1996. The succession of 
Western о cur terrestrial floras. Pp. 
61—80 in S. D. Hopper et al. (editors), колы Нег- 
itage: Past, Present and Future of the Western Austra- 
lian Biota. Surrey Beatty & Sons, Chipping Norton, 


N.S 
Mildenhall, D. С. & Y. M. Crosbie. 1979. Some ош 
. New 


Zealand J. Geol. Geophys. 22: 499-508. 

1981. Fossil pollen records of extant angio- 
sperms. Bot. Rev. 47: 1-142. 

Page, V. M. 1979. Dicotyledonous wood from the Upper 
Cretaceous of central California. J. Arnold Arbor. 60: 
323-349 

Penny, J. H. J. 1989. New Early Cretaceous forms of the 
angiosperm p pollen genus Afropollis from England and 

Rev. Palaeobot. Palynol. 58: 289-299. 

. E. Francis. 2000. The first record of fossil 
wood of Winteraceae from the Upper Cretaceous of Ant- 
arctica. Ann. Bot. 85: 307-315. 

Praglowski, J. Winteraceae Lindl. World Pollen 
and Spore Flora 8: 1-36. Almqvist & Wiksell, Stock- 
holm 


Qiu, Y.-L., M. W. Chase, D. H. Les & C. R. Parks. 
Molecular phylogenetics of the Magnoliidae: е, 
nalyses of nucleotide sequences of the pases gene 
nd Ann. Missouri Bot. Pu 80: 587—606. 
Rabinowitz, P. D. n & D. ы. 1983. Тће 
separation of hee d Africa. Science 220: 67— 
69. 


Raven, P. H. & D. I. Axelrod. 1974. Angiosperm bioge- 
ography and past continental movements. Ann. Missouri 
Bot. Gard. 61: 539-673. 

Regali, M. S. P. & C. К. Мапа. 1989. Late Jurassic-Early 
Cretaceous in Brazilian Sedimentary Basins: Correlation 
with the International Standard Scale. Petrobrás, Rio de 
Janeiro. 

Renner, S. S. 1999, Circumscription and phylogeny of 
the Laurales: Evidence from molecular and morpholog- 
ical data. Amer. J. Bot. 86: 1301-1315. 

1974. A new pollen type in the Winter- 
5. 


Savolainen, V., M. W. Chase, S. B. Hoot, C. M. n, 
D. E. Soltis, C. Bayer, M. F. Fay, A. Y. De Bru UE 
Sullivan & Y.- L. Qiu. 2000. Phylogenetics of flowering 
plants based on combined analysis of plastid atpB and 
rbcL gene sequences. Syst. Biol. 49: 306-362 

Schuster, В. M. 1972. Continental movements, “Wal- 
lace's Line" and Indomalayan-Australasian арена! 


of land plants: Some eclectic concepts. Bot. Rev. 38: 


3-86 


1976. Plate tectonics and its bearing on the geo- 
graphical origin and dispersal of angiosperms. Pp. 4 
13 . B. Beck (editor), Origin and Early Evolution 
of Angiosperms, Columbia Univ. Press, New York. 

Scotese, C. R. 1997. Paleogeographic Atlas, PALEOMAP 
Progress Report 90-0497, Department of Geology, Uni- 
versity of Texas at Adlington: Arlington, Texas. 

Smith, A. C. 1973. Angiosperm evolution and the rela- 
tionship of the floras D Africa гөн а epe Pp. 
49–61 in B. J. Meggers, E. S. u & W. D. Duck- 
worth (editors), Tropical DL ВЫ in Africa 
and South America: A Comparative Review. Smithson- 

ian rwr E m С. 

; Bs; 


Soltis, Satis. DL. Nickrent, L. A. Johnson, 
re 


| Ур) А. 
S.-M. Chaw, L. J. Gillespie, W. J. Kress & K. J. Sytsma. 
1997. Angiosperm phylogeny inferred from 185 ribo- 
somal DNA sequences. Ann. Missouri Bot. Gard. 84: 
149. 


Stover, L. E. & A. D. Partridge. 1973. Tertiary and Late 
Cretaceous spores and pollen from the Gippsland Basin, 
southeastern Australia. Proc. Roy. Soc. Victoria 85: 

- T 

Suh, H. E. Reeve & E. Zimmer. 
1993. e da atn and phylogenetic implic a- 

n Winteraceae. Amer. J. Bot. 80: 

Takhtajan, A. L. 969. Flowering Plants: а and Dis- 
persal. evelli Institution, Washingt 

1997. Diversity and Classification й Flowering 
Plants. Columbia Univ. Press, New Yor 

Thorne, R. F. 1974. A phylogenetic classification of the 
doge) Aliso 8: 147—209. 

Truswell, E. 1991. Antarctica: A history of terrestrial 
vegetation. y^ 499—537 in R. J. Tingey (editor), The 
Geology of Antarctica. Clarendon Press, Oxford. 

D. J. Drewry. 1984. Distribution and prove- 
nance Ы recycled palynomorphs in surficial sediments 
s Sea, Ad tarctica. Mar. Geol. 59: 187-214 

с W. К. Harris. 1977. The age of Alois 
Rock and the Olgas, central Australia. Trans. Roy. Soc. 
South Australia 101: 45—50 

Upchurch, G. R. 1984. Cuteular anatomy of angiosperm 
leaves from the Lower Cretaceous Potomac Group. I. 
Zone I leaves. Amer. J. Bot. 71: 192-202 

Van Campo, M. & P. Guinet. 1961. Les sino composés. 
ze des Mimosacées. Pollen & Spores 3: 201- 


Vink, b 1988. Taxonomy in Winteraceae. Taxon 37: 


кыт. y. 1971. Pollen morphology, phytogeography, 
and њу of the Annonaceae. Contr. Gray Herb. 
202: 31. 


А Ta .E volutionary significante of the exine in 


Muller (editore 
ny Significance of the Exine. Academic Press, 
London. 
& A. G. Walker. 1981. Comparative pollen mor- 
phology of the Madagascan genera of Myristicaceae 
(Mauloutchia, Brochoneura, and Haematodendron). 
Grana 20: 1-17. 

—— ———. 1984. Ultrastructure of Lower Creta- 


316 


Annals of the 
Missouri Botanical Garden 


ceous angiosperm pollen and the origin and early evo- 
lution of flowering plants. Ann. Missouri Bot. Gard. 71: 
521. 


E: J. Brenner & A. G. Walker. 1983. Wintera- 
ceous pollen in the Lower Cretaceous of Israel: Early 
evidence of a magnolialean angiosperm family. Science 


220: 1273-1275 


Wilson, T. K. 1964. ч morphology of the Са- 
nellaceae. III. Pollen. Bot. Gaz. 125: 192-197. 
Wolfe, J. A., J. A. Doyle & V. M. Page. 1975. The bases 
of angiosperm Б оар Paleobotany. Ann. Missouri 
Bot. Gard. pes 01—824. 

Young, D. A . Are the angiosperms primitively ves- 
selless? Syst. Те 6: 313–330. 


WOOD AND BARK ANATOMY 
OF TAKHTAJANIA 
(WINTERACEAE); 
PHYLOGENETIC AND 
ECOLOGICAL IMPLICATIONS! 


Sherwin Carlquist? 


ABSTRACT 


Wood and bark anatomy of Takhtajanie are newly reported on the basis of woody stems to 4.5 cm in diameter, a 


se. Multiseriate 


upright cells except for a small number of procumbent cells 
'thereal oil pun 


are absent in rays. Bark contains vain nests and e 
] imys, in agreement with с 
of Winteraceae other than 


nclusions iua d o 
Ta. khtaj Ba are lacking. Wood features of Takhtajania agree with those of subtropical 


multiseriate rays. E 
Wood features closely match those of Tas- 
on the basis of molecular data. Comparative data on bark 


species by t features found i in temperate Winteraceae (growth rings, warted tracheid surfaces, helical thick- 


enings in trac 
ey words: 


uud wood anatomy, Magnoliidae, Takhtajania, tracheids, vessellessness, Winteraceae. 


Wood of Winteraceae has been surveyed at the 
species level for all genera except Takhtajania. The 
herbarium specimen of Takhtajania hitherto avail- 
able did not have sufficient secondary xylem for 
satisfactory study; twigs are often of limited value 
in determining the nature of wood anatomy of a 
species. Rediscovery of Takhtajania has been de- 
scribed in an accompanying paper of this series 
(Schatz, 2000 this issue). 

Winteraceae have traditionally been regarded as 
retaining numerous character states primitive for 
dicotyledons (e.g., Dahlgren, 1975; Takhtajan, 
1987; Thorne, 1992). Recent molecular studies 
(e.g., Qiu et al., 1993) place Magnoliales, which 
contain Winteraceae, at a low level of advancement, 
but place other orders of flowering plants (notably 
paleoherbs) as basal to Magnoliales. А primitive 
status among dicotyledons for paleoherbs has been 
advanced by Taylor and Hickey (1992). Wood of 
Takhtajania is of potential interest in assessing the 
position of Winteraceae. The position of Takhtaja- 
nia within Winteraceae can be assessed on the ba- 
sis of wood anatomy, because wood character states 
within the family are sufficiently diverse that a key 
could be constructed on the basis of wood data 


alone (Carlquist, 1989). 


Another phylogenetic question regarding Winter- 
aceae has been raised by the assertion (e.g., Young, 
1981) that woody groups of vesselless dicotyledons 
(Amborellaceae, Tetracentraceae, Trochodendra- 
ceae, Winteraceae) may be secondarily vesselless 
rather than primarily so. The latter view (e.g., Bai- 
ley, 1944) has been widely propagated. 

Ecological questions also are of significance. The 
presence of growth rings, warted inner wall surfaces 
of tracheids, and helical thickenings in secondary 
xylem tracheids characterize more temperate spe- 
cies of Drimys (Carlquist, 1988a), Pseudowintera 
(Patel, 1974; Meylan & Butterfield, 1978), and Tas- 
mannia (Carlquist, 1989). These features are lack- 
ing in species that occupy subtropical or frost-free 
sites, as shown by Belliolum (Carlquist, 19832), 
Bubbia (Carlquist, 1983b), Exospermum (Carlquist, 
1982a), and Zygogynum (Carlquist, 1981). The 
ecology of Takhtajania, which grows along a moist 
ridge in montane tropical Madagascar, has been de- 
scribed in an accompanying paper (Feild et al., 
2000 this issue). 

Information is offered on bark anatomy of Takh- 
This information may serve for com- 
parison when studies of bark anatomy, currently 


о 
lacking (Metcalfe, 1987), are offered. 


tajania here. 


! The writer is grateful to George E. Schatz of the Missouri Botanical Garden and to the collectors cited for providing 


the material of Takhtajania. 


? Santa Barbara Botanic Garden, 1212 Mission Canyon 


Road, Santa Barbara, "pw 93105, U.S.A. Address 


correspondence to author at 4539 Via Huerto, Santa Barbara, EN не 93110-2323, U.S 


ANN. Missouni Bor. 


GARD. 87: 317—322. 2000. 


318 


Annals of the 
Missouri Botanical Garden 


MATERIALS AND METHODS 


The first collection of Takhtajania perrieri (Ca- 
puron) Baranova & J.-F. Leroy made available to 
me is documented by the collection Pierre Jules 
Rakotomalaza et al. 1342 (MO). The collection lo- 
cality is in the Anjanaharibe-Sud Special Reserve 
southwest of Andapa in northeastern Madagascar 
(Schatz, 2000), along a ridge in which many of the 
Takhtajania trees are semi-prostrate and covered 
with lichens. This collection provided liquid-pre- 
served material of a woody stem 3.5 cm in diameter 
and a portion of a stem with only a little secondary 
growth. A second collection from this locality, Chris 
Birkinshaw 483 (MO), provided a liquid-preserved 
stem 4.5 cm in diameter and a root 4 cm in di- 
ameter. All of these materials were originally pre- 
served in formalin-acetic alcohol and were trans- 
ferred to 50% aqueous ethanol. 

Sections of the smaller stem and the larger stems 
and the root (all with both bark and wood) were 
prepared according to the schedule of Carlquist 
(1982b). In addition, sections of the stem 3.5 cm 
in diameter, the stem 4.5 cm in diameter, and the 
root 4 cm in diameter were prepared on a sliding 
microtome. Some of these sections were dried be- 
tween glass slides, sputter-coated, and examined 
with scanning electron microscopy (SEM). Other 
sliding microtome sections were stained with a saf- 
ranin-fast green combination and mounted in Can- 
ada balsam. Macerations of the wood of the larger 
stems and of the root were prepared with Jeffrey’s 
Fluid and stained with safranin. Mean tracheid di- 
ameter is based upon tangential diameter of tra- 
cheids. Thickness of radial walls is used for mea- 
surements of wall thickness. Means are based on 
25 measurements except for tracheid wall thick- 
ness, in which wall portions judged to be typical 
were used. 


ANATOMICAL RESULTS 
WOOD (FIGS. 1-9) 


Data are based on mature wood of the stem 3.5 
cm in diameter unless otherwise indicated. Quan- 
titative data for the two large stems were not con- 
sidered significantly different from those of the 
large root (except for tracheid length), so only data 
on the stem 3.5 cm in diameter form the basis for 
the description below. 

Growth rings absent (Fig. 1). Tracheary elements 
all tracheids (Figs. 1, 2, 5—9). Mean tracheid length 
(largest stem), 3002 jum. Mean tracheid length 
(root), 3821 jum. Mean tracheid lumen diameter, 38 
шт. Mean tracheid wall thickness, 4.3 jum. Lateral 


walls of tracheids with scattered circular bordered 
pits (Figs. 7, 8). End walls of tracheids with bis- 
eriate circular bordered pits or scalariform pits 
(Figs. 5, 6), the latter sometimes more crowded than 
shown in Figure 5. Pit membranes present in end 
wall pits (Fig. 6), both in stems and in root. Tra- 
cheids with scalariform pits less common than 
those with circular pits on end walls formed in 
small groups with no apparent pattern of distribu- 
tion within the wood. Tracheids with scalariform 
end-wall pitting somewhat more common in the root 
studied than in the stems. Circular pits with nar- 
rowly elliptical apertures (Figs. 8, 9), pit cavities 
about 9 jum in 
small number of tracheids with characteristic elon- 
gate elliptical thin areas oriented transversely to 
helically on inner wall surfaces; these are not like 
helical thickenings reported in Pseudowintera and 
one species of Tasmannia (Carlquist, 1989). Warts 
absent on tracheid wall inner surfaces (Figs. 5, 8). 


iameter as seen in face view. А 


Pitting generally more abundant on radially orient- 

walls, but end-wall pitting may be seen on ra- 
dially oriented and diagonally oriented tracheid 
walls. Axial parenchyma very scarce (Fig. 7, left), 
distributed in a diffuse fashion and composed of 
strands of five or six cells. Rays multiseriate and 
uniseriate (Fig. 2), about equally frequent (if bis- 
eriate rays are included among multiseriate rays). 
Multiseriate rays are more common than the pho- 
tograph of Figure 2 suggests because there are long 
uniseriate wings on most multiseriate rays. Multis- 
eriate rays up to five cells in width at widest point 
(Fig. 2), mean multiseriate ray width 2.5 cells. 
Mean multiseriate ray height, 4176 jum; mean un- 
iseriate ray height 1473 рт. Uniseriate rays com- 
posed of erect cells; multiseriate rays composed 
mostly of square to erect cells, a few procumbent 
cells present (Fig. 3), and the ray type of the spe- 
cies thus intermediate between Heterogeneous I 
and Paedomorphic I (Carlquist, 1988b: 179). Ray 
cells with thick lignified walls, most pits bordered 
as seen either in sectional (Fig. 4) or face view. 
Borders well developed on pits of tangentially ori- 
ented ray cell walls. Some ray cells with dense, 
granular, dark-staining contents interpreted here as 
tannins (Figs. 3, 4). Ethereal oil cells and sclereids 
absent in rays. Wood nonstoried. 


BARK 


At the periphery of the phloem, a sheath of fibers 
develops (Fig. 10). These can be termed protophlo- 
em fibers. Fibers were not observed within the sec- 
ondary phloem. In bark of the larger specimens, 
nests of thick-walled brachysclereids (not illustrat- 


Volume 87, Number 3 Carlquist 319 
2000 Wood and Bark Anatomy 


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Figures 14. Sections of wood of Takhtajania эне t — 1. Transection, showing vesselless nature and lack he 
ићи rings. —2. Tangential section; both multiseriate and "ise rays are present. —3. Radial section; most r 
cells are upright; a few square and proc 'umbent cells are near t prp —4. Portion of ray cells from radial 
section, кш bordered pits in sectional view and јерм и contents. Figs. 1, 2, magnification scale above Fig. 
1 (divisions = 10 рт); Fig. З, scale above Fig. З (divisions = 10 рт); Fig. 4 stale Ta Fig. 4 (divisions = 10 
pm). 


320 Annals of the 
Missouri Botanical Garden 


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Figures 2-9. SEM photographs (5, 6, 8) and light microscope photographs (7, 9) of wood sections of Takhtajania 


perrieri. —5. End wall of tracheid from radial section, showing both scalariform and circular pits. —6. Scalariform and 
oval pits from ‘nd wall of tracheid of radial section, “to show presence of pit membranes, which have ја iio tured 
because of и or preservation method. —7. Radial section, with ray cells at right and a strand of axial parenchyma 


Inside surface of two tracheids from radial section; walls are smooth. —9. Tracheids from radial section; 
sparse circular pits in tracheids at left; tracheid to right of center has laterally гч elliptical thin areas. Figs. 5, 
6, 8, magnification scales at upper left (bars = 10 jum). Fig. 7, scale above Fig. 3; Fig. 9, scale above Fig. 4. 


Моште 87, Митбег 3 
2000 


Carlquist 
Wood and Bark Anatomy 


Figures 10, 11. 


Transections of bark of Takhtajania p 
ortex at right, to show sheath of protophloem fibers (dark) 


с 
cortical cells (below) and р 


hellem (above); lamellate wall 
cation scale above Figure 3. 


ed here) are present in bark tissue that represents 
tangentially stretched cortical cells. Scattered ethe- 
real oil cells are also present in this bark tissue. 
hellem is notably thick on the older stems stud- 
ied (Fig. 11). As seen at higher magnifications, the 
walls of the phellem cells are clearly lamellate. 


ECOLOGICAL CONCLUSIONS 


Presence of growth rings, presence of warts on 
inner walls of tracheids, presence of helical thick- 
enings in tracheids, and to a certain extent, nar- 
rowness of tracheids have been interpreted as re- 
lated to coldness in habitats of particular species 
of Winteraceae (Carlquist, 1989, and papers cited 
therein). Takhtajania lacks all of these features. 
From this evidence alone, one could conclude that 
Takhtajania grows in a frost-free zone, in contrast 
to the habitats of, for example, Pseudowintera. 

Length of vessel elements is related to ecology 
in vessel-bearing dicotyledons (Carlquist, 1975). In 
vesselless angiosperms, as in vesselless gymno- 
sperms, tracheid length is primarily related to di- 
ameter of axis from which a wood sample was taken 


е 


5 


rrieri. —10. Section from younger stem, with phloem at left, 
— 11. Section from older stem, to show tangentially stretched 


may be seen in some phellem cells. Figs. 10, 11, magnifi- 


(Carlquist, 1975: 141). The tracheid length record- 
ed here for Takhtajania is consonant with that con- 
cept, because the sample was taken from a rela- 


the root as compared 
accordance with data by Patel (1965) that root ves- 
sel elements are longer than stem vessel elements 
in dicotyledons at large. Sections and macerations 
suggested that scalariform end-wall pitting on roots 
was somewhat more abundant in tracheids of roots 
as compared to those of stems. 


PHYLOGENETIC CONCLUSIONS 


In the summary of woods of Winteraceae (Ca- 
паша, 1989), a key based on wood of the genera 
other than Takhtajania was presented. Now that 
Takhtajania wood has become available, one can 
place this genus in the key. Takhtajania falls clos- 
est to Тазтапта. In both genera, scalariform pit- 
ting appears in some tracheids (at least in the ear- 
years of xylem formation), 
multiseriate rays average between 2.1 and 4.1 cells 


lier secondary 


at widest point, procumbent cells are scarce in mul- 


322 


Annals of the 
Missouri Botanical Garden 


tiseriate rays, and ethereal oil cells are absent in 
rays. The wood of Drimys is similar, differing in 
having more abundant procumbent cells in rays. 
These results are interesting in that the molecular 
data now at hand (Karol et al., 2000 this issue) 
show greater pui between Takhtajania and 
Tasmannia or ys than between Takhtajania 
and either Belliolum. Bubbia, Exospermum, Pseu- 
dowintera, or Zygogynum. In view of the fact that 
molecular results (Karol et al., 2000) show Takh- 
tajania to be basal within ашын wood fea- 
tures of Takhtajania, as well as those of Drimys and 
Tasmannia, can be expected to exhibit more nu- 
merous primitive character states than those of the 
other genera. 

Bark anatomy of Takhtajania cannot be com- 
pared to bark of other Winteraceae because few 
data have been accumulated on bark anatomy in 
the family (Metcalfe, 1987). Esau and Cheadle 
(1984) have described secondary phloem of Win- 
teraceae in detail, but were not concerned with 
broader aspects of bark in the family. 


Literature Cited 


Bailey, 1. У. 1944. The development of vessels in angio- 
sperms and its significance in morphological research. 
Amer. J. Bot. 311: 421—428. 

Carlquist, 5. 1975. Ecological Strategies of Xylem Evo- 
lution. Univ. California Press, Berkeley and Los Ange- 
es. 

Wood anatomy of Zygogynum Sis cin 
севе); Field observations. Bull. Mus. Natl. His . Nat. 

Paris, B, Adansonia, sér. 4, 8: 281-292. 

. 1982a. Exospermum stipitatum (Winteraceae): 
Observations on wood, leaves, flowers, pollen, and fruit. 
Aliso 10: S. 

——— . The use of ethylene diamine in softening 
hard plant pu tures for paraffin sectioning. Stain Tech- 
nol. 57: 311— pod 

od anatomy of Belliolum (Wintera- 
ceae) and a note on чен J. Arnold Arbor. 64: 161— 


169 


. 1983b. Wood anatomy of Bubbia (Winteraceae). 


Takhtajan, A. 


Taylor, D. W. & L. J. H 


Thorne, 


Young, D. 


with comments on origin of vessels in dicotyledons. 
Amer. J. Bot. 70: 578—590. 

— ———. 1988a. Wooa ши of Drimys 8. 8. (Winter- 
mm Aliso 12: 81-95. 

1988b. oe Wood Anatomy. Springer- 
Verlag, pos and Heidelberg. 

———. 19 ood е of Tasmannia; Summary of 
vnd: a of Winteraceae. Aliso 12: 257-27! 

Dahlgren, R. M. T. 1975. A Не он af он oe of 
the angiosperms to be used to demonstrate the 

bution of characte ers. Bot. Not. 128: 119-147. 

Esau, K. & У. 1. Cheadle. 1984. Anatomy of the second- 
ary phloem in Winteraceae. 1. A. W. A. Bull., N.S. 5: 
13-43. 


distri- 


Feild, T. S., M. A. Zwieniecki & М. M. Holbrook. 2000. 
Winteraceae evolution: Ап ec ophysiologica perspec- 
tive. Ann. жар Вог. Gard. 87: 323— 

Karol, K. G., Y. Suh, С. E. Schatz & E. А. v mmer. 2000. 

lar о. for the phylogenetic position of 
Takhtajania in the Winteraceae: Inference from nuclear 
ribosomal and chloroplast gene spacer sequences. Ann. 
Missouri An Gard. 87: 414—432. 
s calfe, 87. Anatomy of Dicotyledons, Ed. 2, 
. 3. Magnoliales, Illiciales, and Laurales. Clarendon 
Paes 1978. The structure 
В. Bull. 222: 1-250. 


965. A comparison of the anatomy of the 
secondary xylem in roots and stems. Holzforschung 19: 
72-79. 


. 1974. Wood anatomy E the dicotyledons indig- 

enous to o Zealand. 4. Winteraceae. New Zealand J. 
Bot. 11: 587—598. 

Qiu, Y.-L., M. W. Chase, D. H. Les & C. R. Parks. 1993. 
Molecular phylogeneties of the Макош, Cladistic 
тыге of nucleotide sequences of the plastid gene 
rbcL. Ann. Missouri Bot. Gard. 80: 587 “606, 

Schatz, G. E. 2000. The rediscovery of a Malagasy en- 
demic: ПИ Lea Winteraceae. Ann. Missouri 
Bot. Gard. 87: 297 

1987. Sans И Oficina 

*'NAUKA," St. Petersbur 

ic Sn 1992. Phylogenetic evi- 
dence for the herbaceous origin of angiosperms. Pl 

Syst. Evol. 180: 137-156. 

В. К. 1992. Classification T кр of the 

flowering eer Bot. Rev. 58: 225- 

A. 


Editoria ‘ 


Are the anie ees ves- 


selless? Syst. "Bor 6: 313- 


WINTERACEAE EVOLUTION: 
AN ECOPHYSIOLOGICAL 
PERSPECTIVE! 


Taylor S. Feild,? Maciej А. Zwieniecki,? 
and N. М. Holbrook? 


ABSTRACT 


Winteraceae have long been regarded as the js p living family of angiosperms. The justification for 


The 
this view partly arises from their lack of xylem vessels. 


ecological history than can be deduced from their current distribution. These obs 


been considered a key innovation 


0 their current “| dominance, Winteraceae have been portrayed as declining 


of the functional significance and selective pressures underlying the pattern and direction of character change of the 
distinctive features of this family. In particular, physiological studies on stomatal plugs and xylem hydraulic parameters 


are consistent with their ecological success in 


wel, temperate 


environments. 


words: stem water transport, tracheids, Takhtajania, Winteraceae, xylem evolution. 


The first angiosperms have been traditionally 
portrayed as slow-growing trees characterized by 
"primitive" morphological features and confined to 
wet tropical environments (Bailey, 1944; Takhtajan, 
1969; Carlquist, 1975; Cronquist, 1981). According 
to this woody-tree hypothesis, Winteraceae are the 
least modified descendents of this hypothetical an- 
giosperm ancestor (van Tieghem, 1900; Thompson 
& Bailey, 1916; Bailey & Thompson, 1918; Bailey, 
1944; Smith, 1945; Takhtajan, 1969; Thorne, 
1974). A major reason for this association is the 
absence of xylem vessels in Winteraceae (van 
Tieghem, 1900; Bailey & Thompson, 1918; Bailey, 
1944; Cronquist, 1981; Carlquist, 1983, 1987, 
1996; Gifford & Foster, 1989), although the pres- 
ence of other putatively primitive morphological 
characters has been used to reinforce this view 
(Bailey & Swamy, 1951; Thorne, 1974; Hickey & 
Wolfe, 1975). A logical extension of this perspec- 
tive is to view Winteraceae as an evolutionary dead 
end, confined to wet forest refugia where primitive 
aspects of their morphology and ecology would not 
be disadvantageous (Bailey, 1944, 1953; Bailey & 
Nast, 1945; Smith, 1945; Takhtajan, 1969; Carlqu- 
ist, 1975, 1983). In particular, Winteraceae have 
been described as limited to understory habits well 


suited to their presumably inefficient xylem, with 
their survival further enabled by the evolution of a 
distinctive leaf-level feature, 1.е., waxy plugged sto- 
mata (Bailey, 1944, 1953; Bailey & Nast, 1944; 
Baranova, 1972; Carlquist, 1975, 1996; Cronquist, 


— 
2 
со 


Recent findings оп the phylogeny of angiosperms 
and the paleoecology and physiological ecology of 
Winteraceae challenge the idea that Winteraceae 
represent an unchanged lineage (Young, 1981; 
Donoghue & Doyle, 1989; Doyle et al., 1990; Chase 
et al., 1993; Crane et al., 1995; Soltis et al., 1997; 
Nandi et al., 1998; Feild et al., 1998; Doyle, 1998, 
2000; Mathews & Donoghue, 1999). Our goal in 
this paper is to review Winteraceae’s ecological 
characteristics in relationship to historical changes 
in their distribution and climatic associations. In 
particular, we focus on how physiological studies, 
combined with paleoecological and biogeographic 
information, alter our perspective on two of the 
most distinctive features of Winteraceae: stomatal 
plugs and vesselless wood. The impact of the re- 
discovery of Takhtajania perrieri (Capuron) Вага- 
nova & J.-F. Leroy (Schatz et al., 1998) on our un- 
derstanding of the ecological evolution of the 
Winteraceae will also be considered. 


Support was provided by a Deland Grant from the Arnold Arboretum, a Graduate Research Training Grant in Plant 


Systematics (NSF D 


DGE-9554522), and a Dissertation Improvement Grant (NSF IBN-9902239). We gratefully thank 


Fakhri Bazzaz, Chuck Bell, Tim Brodribb, Alex Cobb, Michael Donoghue, Jim Doyle, H. S. Feild, Tanguy Jaffré, Greg 


Jordan, Eithne O’Brien, Porter P. Low 


ry П, George Schatz, Leonard Thien, and Matt Thompson 


? Department of Organismic and чы им. Biology, Cambridge, Massachusetts 02138, U. 5. A. Author for corre- 
d 


spondence, TSF: tfeild@oeb.harvard.edu. 


ANN. Missouni Вот. GARD. 87: 323—334. 2000. 


324 Annals of the 
Missouri Botanical Garden 
Table 1. Classification and geographic distribution of modern Winteraceae. Nomenclature and distributions follow 


Smith (1942, 1943a, b). Ecological distributions were compiled from Feild (unpublished obs. in Australia, Costa Rica, 
New Caledonia, and New Zealand), G. Schatz (pers. comm. 1998), Smith (1942, 1943a, b), and Vink (1970, 1977, 
1985) 


Genus Distribution 


Ecology 


Belliolum (6—7 species) 
Bubbia (?33 species) 
Islands | 


Drimys (6 species) 


Exospermum (2 species) New Caledonia 
Pseudowintera (2 species) 


Takhtajania (1 species) 


New Zealand 
Madagascar 


Tasmannia (5-30+ species) 


s, New Сите 


Zygogynum (6 species) New Caledonia 


New Caledonia, Solomon Islands 
Australia, Lord Howe Island, New 
1 


Jaledonia, New Guinea, Solomon tropical prem 


Central America, Mexico, South 
merica 


gone Borneo, Indonesia, Philip- 


Understory and subcanopy treelets to trees 
in subtropical lowland rainfor 

Shrubs, treelets, to large trees (15 m+) in 

ntane, montane cloud for- 
est; in New Csledonja some species on 
serpentine soils 

Subcanopy and canopy shrubs to trees in 
tropical montane cloud forest, maritime 
temperate rainforest, páramos 

Large trees in subtropical lowland rainforest 

Subcanopy trees, alpine shrubs 

Treelets to subcanopy trees in montane 
cloud forest (~1000 m) 

Diverse growth forms in alpine and lowland 
temperate rainforests, tropical lowland 
forest, tropical montane cloud forest 

Subcanopy trees in subtropical lowland 
rainforest and montane cloud forest 


ECOLOGICAL CHARACTERISTICS OF LIVING 
WINTERACEAE 


Winteraceae, with 65 species in four to six gen- 
era (Smith, 1942, 1943a, b; Vink, 1988; Suh et al., 
1993), are a diverse evergreen assemblage -— 
growth forms that include minute-leaved epiphytes, 
terrestrial alpine shrubs and scramblers, large- 
leaved understory treelets, and medium-sized (— 
20 m tall) trees (Smith, 1942). Although generally 
limited to wet habitats (rainfall > 1200 mm/year), 
they can be found in a variety of habitats ranging 
from subtropical montane mossy rainforests to cool 
coastal and montane temperate rainforests, and 
subalpine and alpine shrubberies. Tasmannia lan- 
ceolata (Poir.) A. C. Smith is an exception that oc- 
curs as a small tree in moist sites such as gullies 

and stream margins in dry sclerophyll-dominated 
woodlands (rainfall about 800 mm/year) in Tasman- 
ia. The current distribution of Winteraceae follows 
a southern Gondwanan geographic pattern with 
most species and genera in Australasia (Table 1; 
Raven & Axelrod, 1974; Schuster, 1976; Raven, 
1980), excepting Drimys granadensis L. and Tas- 
mannia piperita Hook., which currently extend into 
regions outside of Gondwana. Drimys is the sole 
genus represented in the New World, where it oc- 
curs from cold, wet Magellanic rainforests in south- 
ern Chile and Argentina to the tropical and tem- 
perate highlands of Mexico (Veblen et al., 1995). 
Finally, the monospecific genus Takhtajania is 


found only in cool montane forests of northeastern 
Madagascar (Schatz et al., 

interaceae are most diverse in subtropical re- 
gions (Vink, 1970, 1977, 1985, 1993), but ecolog- 
ically more successful in terms of their abundance 
in temperate environments. Within the subtropics, 
Winteraceae are found in areas with abundant pre- 
cipitation and/or low evaporative demand such as 
along rivers, in understory habitats, and in everwet 
mossy montane forests (Smith, 1942, 1943b; Vink, 
1970, 1977, 1985, 1993; Jaffré, 1995). Populations 
of many Winteraceae such as most Bubbia and 
Zygogynum species in the subtropics consist of 
only a few individuals (Smith, 1942; Vink, 1977; 
1990). A potential contributing fac- 
tor to this low density may be the tendency for lim- 


Pellmyr et al., 


ited seed production due, in part, to infrequent pol- 
linator visits (Sabatinca moths, Palontus weevils; 
Pellmyr et al., 1990). Most species of subtropical 
Winteraceae are tolerant to shade and occur as un- 
derstory treelets (1—4 m tall) and infrequently as 
subcanopy trees (to 10 m tall) that become estab- 
lished late during succession (Vink, 1993; T. S. 
Feild, unpublished obs.). Many of these treelets 
have exceptionally large leaves (15-30 cm in 
length is not uncommon especially in Belliolum) 
compared to other Winteraceae from cooler cli- 
mates, which cause plants to appear “top heavy" 
and susceptible to breakage (Vink, 1970, 1993; T. 
S. Feild, unpublished obs.). Another characteristic 


Volume 87, Number 3 
2000 


Feild e 


al. 325 
Раа Ecological Evolution 


of some Winteraceae is their ability to reproduce 
vegetatively by stem sprouting. Тазтапта piperita 
in New Guinea is reported to reproduce primarily 
by stem sprouts, while other subspecies are known 
to reproduce by subterranean stolons (Smith, 
1943b; Vink, 1970). Profuse stem sprouting, re- 
sulting in multiple-branched plants, occurs in Bub- 
bia, Drimys, Pseudowintera, Takhtajania, and most 
species of Tasmannia (Vink, 1970; Raleigh et al., 
1994; T. Feild, unpublished obs.; G. Schatz, pers. 
comm. 1998). 

The view of Winteraceae as an ecologically re- 
stricted group is difficult to reconcile with its high 
abundance in wet, temperate rainforest habitats (ca. 
3000 mm a year, with frequent frost and some 
snow). In temperate areas, Winteraceae species can 
dominate the understory and subcanopy as well as 
grow in exposed habitats. For example, Drimys win- 
teri forms dense subcanopy thickets in coastal Chi- 
lean temperate rainforests and occurs frequently as 
a large canopy tree, 19 m tall and up to 65 cm 
trunk diameter (Lusk, 1993). Winteraceae are also 
abundant as individuals in subalpine and alpine 
communities of Australia, Chile, New Guinea, and 
New Zealand, where freezing can occur at any time 
of the year (Vink, 1970; Barry, 1980; Kirkpatrick, 
1983, 1997; Veblen et al., 1995). All four major 
clades within Winteraceae (Bubbia, Drimys, Pseu- 
dowintera, and Tasmannia, as inferred from molec- 
ular phylogenetic analyses using ITS rDNA; Suh et 
al., 1993) contain species that grow as small-leaved 
(< 1-3 cm long) shrubs and small trees in high- 
altitude wet montane communities (Smith, 1943a; 
Hope, 1980; Nunez et al., 1996). In Tasmania, Tas- 
mannia lanceolata occurs as an abundant prostrate 
shrub (e.g., approx. 45% cover) on rocky scree 
slopes well above the eucalypt and Nothofagus tree 
lines, where it is found alongside conifers such as 
Diselma (Cupressaceae) and Microstrobus (Podocar- 
paceae) (Gibson et al., 1995; Kirkpatrick & Bridle, 
1999). Drimys granadensis 
branched shrub in Espeletia-(Asteraceae) dominat- 
ed wet páramos of Colombia (Smith, 1943a). In 
New Zealand, Pseudowintera colorata and P. trav- 
ersii grow in the company of conifers on alpine pla- 


grows as a multiple- 


teaus and mountain tops, where they are gregarious 
colonizers of landslides (Stewart & Harrison, 1987). 

Within these temperate habitats, Winteraceae 
appear to be good competitors. Rebertus and Veb- 
len (1993) reported that Drimys winteri produced a 
substantial rain of viable seeds, and seedlings grew 
ri in response to forest gap formation, imped- 

ing the recruitment and growth of Nothofagus 
(Nothofagaceae) and other vessel-bearing angio- 
sperms. Drimys apparently requires large-scale dis- 


turbances, such as large gaps produced by wind 
damage and multiple treefalls, to become estab- 
lished (Rebertus & Veblen, 1993). However, with 
distance from the coast, Drimys abundance de- 
creases rapidly, perhaps reflecting a reduction in 
water availability (Rebertus & Veblen, 1993). Early 
succession recruitment of Winteraceae occurs in 
other temperate taxa. Tasmannia lanceolata is an 
abundant colonizer of open fields on the central 
basalt plateau of Tasmania, which has frequent frost 
in the winter (Read & Hill, 1983). This species 
forms dense thickets in grasslands, understories of 
dry sclerophyll woodlands (— 800 mm rainfall a 
year), and on slopes of wet eucalypt gullies (Vink, 
1970; Raleigh et al., 1994). Tasmannia ргрегиа 
dominates forest edges in subalpine rainforests and 
penetrates into frost-stricken grasslands of high al- 
titudes (> m) in New Guinea (Vink, 1970; 
Hope, 1980). Tasmannia piperita is also common 
on landslide-exposed soils in montane habitats 


(Hope, 1980). 


PALEOECOLOGICAL AND PALEOGEOGRAPHIC 
HISTORY OF WINTERACEAE 


To understand Winteraceae's ecological success 
(in terms of abundance of individual plants) in wet 
temperate versus subtropical environments, histor- 
ical patterns of Winteraceae distribution, continen- 
tal movements, and climatic changes must be con- 
sidered. The extensive recovery of Winteraceae 
fossil pollen (tetrads) indicates that Tertiary and 
Late Cretaceous Winteraceae shared a similar 
Southern Hemisphere temperate distribution with 
modern Winteraceae (Coetzee & Muller, 1984; 
Coetzee & Praglowski, 1988; Dettmann & Jarzen, 
1990; Doyle et al., 1990; Dettmann, 1994; Mac- 
phail et al., 1994; Doyle, 2000). The oldest Aus- 
tralasian pollen (collected in southeastern Austra- 
lia) is from the Campanian (Dettmann & Jarzen, 
1990), indicating that, at 80 Ma BP, the geographic 
range of Winteraceae corresponded to the Southern 
Gondwana flora province of Brenner (1976). This 
region was a moist, cool temperate region dominat- 
ed by conifers (Podocarpaceae, Araucariaceae) and 
later included the austral angiosperms Nothofagus 
and Proteaceae (Axelrod, 1984; Dettmann & Jar- 
zen, 1991; Spicer et al., 1993; Srivastava, 1994; 
Hill & Scriven, 1995). Specht et al. (1992) sug- 
gested that during the Late Cretaceous, Wintera- 
ceae, along with Trimeniaceae, Пех (Aquifoliaceae), 
and Proteaceae, were understory shrubs growing in 
temperate lowland forests (mean annual tempera- 
ture — 12?C) dominated by podocarps and protea- 
ceous trees (e.g., Knightia, Macadamia). These for- 


Annals of the 
Missouri Botanical Garden 


Winteraceae 


Canellaceae 
Takhtajania 
Tasmannia 
Pseudowintera 
Bubbia-Zygogynum 
complex 


Drimys 


? Walkeripollis 


Figure 1. о hypothesis for the relation- 
ships among Winteraceae, based on the molecular phy- 
logeny of Suh et al. (1993). the fossil pollen genus Walk- 
eripollis (Doyle et al., 1990), and Canellaceae (Chase et 
al., 1993). Node A is the split from Canellaceae, while 
node B represents the ind that pave rise to all extant 
Winteraceae as well as Late Cretaceous and Tertiary fossil 
pollen taxa distributed in мүн млн (Doyle et = ., 1990). 
Walkeripollis may represent an extinct side-branch, found 
ч Northern conan, that pre-dates the origin d node 

B (Doyle et al., 1990). 


est associations occurred in what is now southern 
Australia and Antarctica; however, no modern an- 
alog of this forest community exists today (Specht 
et al., 1992; Dettmann, 1994; Hill & Scriven, 


Understanding of the climatic and geographic 
distribution of Winteraceae was greatly enhanced 
by the discovery of considerably older winteraceous 
pollen (Late Barremian-Early Albian, 125-105 Ma 
BP) in several localities well to the north of South- 
ern Gondwana (e.g., Israel and Gabon (Walker et 
al., 1983; Doyle et al., 1990; Brenner, 1996)). Cla- 


distic studies of these fossils indicate that the 


plants that produced them were more primitive than 

e common ancestor of Late Cretaceous, Tertiary, 
and modern genera of Winteraceae (Fig. 1; Doyle 
et al., 1990). Surprisingly, these putatively related 
tetrads occurred within the Cretaceous subarid 
tropical belt of northem Gondwana (Doyle et al., 
1990; Srivastava, 1994; Brenner, 1996; Doyle, 
2000). A major implication of these fossils is that 
the Late Cretaceous Winteraceae and their extant 
descendents are a temperate branch from an ini- 
tially tropical lineage that spread southward from 
Northern Gondwana (Fig. 1; Doyle et al., 1990). 
Thus, the Australasian regions that include the cur- 
rent center of Winteraceae diversity, such as New 
Caledonia, (Vink, 1993), are not the cradle of Win- 
teraceae evolution (Рештапп & Jarzen, 1990; 
Doyle et al., 1990; Dettmann, 1994 

A Northern Gondwanan origin ЕА that early 
Winteraceae were likely to have experienced sub- 
stantial changes in climate during their migration 
into the Southern Hemisphere. Two migratory 
routes into Australasia appear plausible, based on 


Cretaceous (Coetzee & Praglowski, 1988) before 
dispersing first to Madagascar, and then via over- 
water dispersal into temperate Australia with Ant- 
arctica providing an overland route (Fig. 2B). Pres- 
ently, it is not clear which of these scenarios is 
more probable because of the large temporal gap 
~ 25 Ma) in fossil pollen data between the first 
appearance of winteraceous po in northern 

Gondwana and the occurrence of fossils resembling 
modern Winteraceae in southern Australia (Doyle 
et al, 1990; Dettmann, 1994; Macphail et al., 
1994; Doyle, 2000). Local extinction of Wintera- 
ceae, implied by fossil pollen in Africa and central 
Australia, makes interpretation of their biogeogra- 
phy especially problematic (Coetzee & Muller, 
1984; Macphail et al., 1994). Regardless of the 
route taken to reach Australasia, the fossil pollen 
record indicates that extant Winteraceae were de- 
rived from an originally tropical lineage that mi- 
grated into relatively cold areas. This suggests that 
distinctive features of this family might be associ- 
ated with success in wet temperate environments. 


T 


STOMATAL PLUGS: KEEPING WATER IN ок Ойт? 
Despite the occurrence of most Winteraceae in 
wet environments, their leaves possess several 


Volume 87, Number 3 
2000 


Feild et al. 
Winteraceae Ecological Evolution 


327 


A. South American Origin 


B. African Origin 


Land 


X Center of origin for modern Winteraceae | 


Figure 2. Two мани hypotheses for the center of origin and routes of migration of Winteraceae into 


gur 
annos The arran 


In the first. route, 


nt of continental areas is from the Late Cretaceous (Ear 
fossil pollen closely alin modern “crown-group” Winteraceae first appeared (Dettman 
Winteraceae originated in temperate regions of southern South Am 


y Campanian, 80 Ma BP) when 
nn & Jarzen, 1990). —A. 
erica following dispersal from 


Northern Gondwana (1). To account for Winteraceae pollen in Africa and Takhtajania in Madagascar, ed to Africa 


and then 


and 4). —B. An alternative route may have involved t 


southern areas) with Takhtajania as a и descendant from this radiation. Wir 


Madagascar must be invo g (2). Winteraceae would then have moved a 


cross Antarctica into Australasia (: 


—. 
> 


overwater ог possibly overland into Antarctica (1) and finally into other ea regions (2 2 and 3). Stars indicate 


the centers of origin for node B Winteraceae (see Fig. 


seemingly xeromorphic features. To varying de- 
grees, Winteraceae leaves have thick cuticles and/ 
or the stomata are sunken into the abaxial leaf sur- 
face (Bailey & Nast, 1944; Baranova, 1972; Bon- 
gers, 1973). Perhaps the most striking manifesta- 
tion of Winteraceae’s apparent xeromorphy is the 
obstruction of each stomatal pore with a granular 
“plug” composed of cutin and wax (Fig. 3; Bailey 
& Nast, 1944; Baranova, 1972; Bongers, 1973; 
Feild et al., 1998). These structures are responsible 
for the characteristic white reflective appearance of 
the undersurfaces of Winteraceae leaves (Bailey & 
Nast, 1944). 

The diversity of stomatal ornamentation found in 
Winteraceae is enormous (Bongers, 1973). In Takh- 
tajania and some species of Tasmannia, stomatal 
plugs are absent (Fig. 3A; Baranova, 1972; 


gers, 1973). In others (most Tasmannia species), 
the stomata are only partially occluded by the pres- 
ence of small crystalline wax rodlets confined to the 
970; Bongers, 1973). The 
abaxial epidermal surfaces of Drimys brasiliensis 
Miers and some entities of Тазтапта piperita are 
covered with dense overarching papillae formed 
from extensions of the cuticular layer си 
1943a; Bailey & Nast, 1944; Bongers, 1973). 
nally, in some Bubbia and Zygogynum species, the 
stomatal apparatus is completely buried under a 
mat of encrusting wax rodlets and an additional lay- 
er of cuticle such that the stomata cannot be seen 
(Bongers, 1973; Metcalfe, 1987; Vink, 1993). 

A long-held belief has been that stomatal plugs 
and the other apparently xeromorphic leaf features 
of Winteraceae function to restrict water loss (Bai- 


guard cell rims (Vink, 


328 


Annals of the 
Missouri Botanical Garden 


е 3. es of stomatal ornamentation in Winteraceae. — 


Fig 
ашаа, free of waxy obstruction. —В. Epi 
cuticle and ушай махе s. The stom "ied "шеп 


аге лівае ii the dark clefts, r 


nt ep idermis, "ic h has a den ^nse mat о 
oncentration of e (possibly composed of wax or cutin associated specifically with the 


stoma of Takhtajania perrieri, end is 
dermal surface of Drimys granadensis, which is covered with p 


cutic Шар papillae (visible between the 


stomatal dicendi Se ne bars аге: 5 wm (A & С) and 20 pm (B & D 


ley, 1944, 1953; Bailey & Nast, 1944; Baranova, 
1972; Carlquist, 1975; Cronquist, 1981). As anti- 
transpirants, these structures were proposed to have 
evolved to compensate for the presumed inferiority 
of a tracheid-based transport system (see below; 
Bailey, 1944, 1953; Bailey & Nast, 1944; Barano- 
va, 1972; Carlquist, 1975; Cronquist, 1981; Sperry, 
1995). Stomatal plugs also occur in most Southern 


Hill, 1989; Stockey et 
al., 1992; Carlquist, 1996; Stockey & Atkinson, 
1993; Stockey & Frevel, 1997; Brodribb & Hill 
1998; Stockey et al., 1998). 

The hydraulic compensation argument has been 
applied to Winteraceae for many years, despite the 
absence of any information on how stomatal plugs 
affect rates of leaf gas exchange. In contrast to what 


their name suggests, stomatal plugs are not solid, 
but riddled with air-filled pores formed between 
tubes of cutin and granular wax crystals (Bongers, 
1973). Although a porous construction is necessary 
for CO, uptake, this fact seems to have been over- 
looked in arguments of presumed transpiration-re- 
tarding effects. Recently, Feild et al. (1998) ex- 
amined how these structures influence water loss 
rates from Drimys winteri var. chilensis (DC.) A 

Gray leaves. Comparisons of plugged and “un- 
plugged” leaves (leaves from which stomatal plugs 
and associated epicuticular waxes were experimen- 
tally removed) demonstrated that under high rela- 
tive humidity (85%) plugs posed a eei small 
resistance to leaf water loss (Feild e 98). 
Maximum stomatal conductances of Drima leaves 
with stomatal plugs were about 10% lower than 
leaves from which the stomatal plugs had been ex- 
perimentally removed. Maximum conductances of 
plugged leaves (approx. 100 mmol H,O m^? s~!), 
although lower than many lowland rainforest trees 


Моште 87, Митбег 3 
2000 


Feild e 


al. 329 
ies Ecological Evolution 


and crop plants, were similar to those reported from 
other montane cloud forest angiosperms (Feild et 
al., . With exposure to progressively greater 
evaporative demand, the water loss rates of Drimys 
leaves with plugs compared to ones without plugs 
markedly diverged. Unplugged leaves exhibited a 
70% decline in stomatal conductance with decreas- 
ing ambient relative humidity (from 105 to 35 mmol 
H,O m^? s^! over 85% to 40% relative humidity 
drop). This degree of stomatal closure in response 
to dry air is similar to that reported for a wide va- 
riety of plants lacking stomatal occlusion. In con- 
trast, stomatal conductances of plugged leaves de- 
creased 10% over the same drop in relative 
humidity, from 100 to 90 mmol H,O m ? ѕ ! (Feild 
et al., 1998). The mechanism by which stomatal 
plugs prevent stomatal closure is unclear. It is pos- 
sible that plugs may interfere with stomatal move- 
ments by maintaining a water vapor concentration 
around the guard cells that uncouples the guard 
cells from ambient conditions, or plugs may retard 
stomatal closure by physically interfering with 
guard cell movements (Feild et al., 1998). Regard- 
less of the exact mechanism, these results provide 
physiological evidence that stomatal plugs in Dri- 
mys cannot be considered adaptations for drought. 

In sharp contrast to their traditionally assigned 
role as anti-transpirants, the functional significance 
of stomatal plugs appears to be related to the oc- 
currence of Winteraceae in areas that are generally 
wet (rainforests and cloud forests). One of the in- 
evitable consequences of frequent rainfall and 
cloud cover is prolonged wetting of leaf surfaces 
(Brewer & Smith, 1997). Because CO, diffuses 
through water 10,000 times more slowly than in air, 
water films on leaf surfaces can create a high re- 
sistance to CO, diffusion into leaves for photosyn- 
thetic carbon gain. Photosynthetic measurements of 
Drimys leaves exposed to mist demonstrated that 
stomatal plugs allow relatively unperturbed aei 
synthetic activity when leaves are exposed to mist 
(Feild et al., 1998). Specifically, ренина 
electron transport rates decreased approximately 
40% following misting of Drimys leaves that lacked 
stomatal plugs. In contrast, electron transport rates 
of plugged leaves were unaffected by misting. Sto- 
matal-associated waxes and cutin allow for contin- 
ued gas exchange when leaves are wet because wa- 
ter droplets are repelled from the stomatal 
apparatus (Feild et al., 1998). These water-epider- 
mis interactions keep the stomatal pore and inter- 
cellular spaces from being filled with water, which 
can occur when leaves are exposed to fog (Brewer 
& Smith, 1997). Further support for the efficacy of 


stomatal waxes in shedding excess water has been 


observed in Tasmannia lanceolata. Small trees of 
T. lanceolata that occur in eucalypt gallery forest 
understory and subcanopy, where they encounter 
frequent leaf wetting from canopy drip, have wax 
rodlets associated with their guard cells. In con- 
trast, shrubby forms of this species growing on ex- 
posed ridge tops in alpine heathlands, which could 
presumably benefit from transpiration restriction, 
lack any epicuticular waxes associated with the sto- 
matal apparatus (Feild, unpublished obs.). 

These functional considerations provide a new 
perspective of the selective forces that may have 
driven the evolution of stomatal occlusion in Win- 
teraceae. The observation that stomatal plugs, at 
least in Drimys, do not apparently protect leaves 
from drought is not consistent with the idea that 
these structures are adaptations that have played 
an important role in the persistence of a “primitive” 
vesselless phenotype. Further support for the new 
interpretation of stomatal plugs is that similar 
structures have evolved in other plants with vessels 
such as some Myristicaceae and Epacridaceae; 
Koster & Baas, 1981), and without vesse!s (as in 
many rainforest conifers). All these plants occur in 
wet forest habitats. In addition, other vesselless an- 


— 


giosperms such as Amborella, Tetracentron, Takh- 
tajania, and many conifers from drier habitats, lack 
stomatal plugs or other waxy structures obstructing 
the stomatal apparatus (Fig. 3A; Metcalfe, 1987; 
Brodribb & Hill, 1998). 


VESSELLESS Woop: HYDRAULIC LIMITATION OR 
ENHANCED FREEZING TOLERANCE? 


Perhaps the best-known feature of Winteraceae 
is the lack of water-conducting xylem vessels in 
their wood (Bailey & Thompson, 1918; Cronquist, 
1981; Gifford & Foster, 1989). Instead, the water- 
conducting system of Winteraceae is composed en- 
tirely of tracheids. Vessels are also absent in a 
number of other angiosperm groups such as Am- 
borella (1 species), Tetracentron (1 species), and 
Trochodendron (1 species). Tracheids are xylem 
cells that lack cytoplasm such that their lumen pro- 
vides an unimpeded path for water flow. Water 
movement between tracheids, however, requires 
that water must traverse cell walls (Zimmermann, 
1983). The primary path for water movement be- 
tween tracheids are pit depressions in the second- 
ary wall, which although porous in construction, 
provide some resistance to flow (Zimmermann, 
1983). In contrast, xylem vessels are water-con- 
ducting tubes generally larger than tracheids both 
in length and diameter (Carlquist, 1975; Zimmer- 
mann, 1983). Vessels consist of files of cells, with 


330 


Annals of the 
Missouri Botanical Garden 


each termed a vessel element, that unlike tracheids 
have substantially modified axial walls or perfora- 
tion plates that allow for a more open pathway of 
water movement (Bailey, 1944; Carlquist, 1975; 
Zimmermann, 1983; Gifford & Foster, 1989). Ves- 
sel elements were apparently derived by a modifi- 
cation of the developmental program giving rise to 
tracheids, such that the pit membranes were hy- 
drolyzed between the end walls of adjacent cells 
(Gifford & Foster, 1989). Vesselless wood has tra- 
ditionally been considered a retained ancestral fea- 
ture in angiosperms (van Tieghem, 1900; Bailey & 
Thompson, 1918; Bailey, 1944, 1953; Carlquist, 
1975, 1983; Cronquist, 1981). What has propelled 
this idea is the assumption that vessels represent 
such an adaptive advantage that their subsequent 
loss is highly improbable (Carlquist, 1975). With 
the evolution of xylem vessels, botanists have ar- 
gued that the early angiosperms were able to exploit 
habitats with uncertain water supplies and develop 
growth-enhancing traits such as large, undissected 
leaves and greater gas exchange rates (Carlquist, 
1975; Doyle & Donoghue, 1986; Bond, 1989). By 
this perspective, hydraulically compromised plants 
such as Winteraceae have escaped extinction only 
by being restricted to wet cloud forest habitats 
(Carlquist, 1975, 1983, 1987). 

The view that Winteraceae are primitively ves- 
selless has been challenged by phylogenetic anal- 
yses (Young, 1981; Donoghue & Doyle, 1989; 
Chase et al., 1993; Mathews & Donoghue, 1999). 
Cladistic analyses indicate that the first angio- 
sperms had vessels (albeit primitive ones), which 
were subsequently lost in several early angiosperm 
lines (Young, 1981; Donoghue, 1989; Donoghue & 
Doyle, 1989; Doyle, 1998). Although molecular 
phylogenies imply that Amborella and Nymphaeales 
are likely to be basal angiosperm branches and 
thus primitively vesselless (Mathews & Donoghue, 
1999), both Winteraceae and Trochodendrales are 
nested within clades containing plants with vessels. 
Winteraceae are most closely related to Canella- 
ceae, and Trochodendrales are included within eu- 
dicots (Chase et al., 1993; Soltis et al., 1997). To 
assume irreversible vessel evolution requires more 
evolutionary steps by additional independent ori- 
gins of vessels in angiosperms (Young, 1981; Don- 
oghue & Doyle, 1989). While the parsimony debt 
incurred by assuming that vessels cannot be lost 
was not overwhelming, this conclusion implies that 
the functional arguments for such an assumption 
should be examined (Donoghue & Doyle, 1989; 
Mathews & Donoghue, 1999). The cladistic argu- 
ments supporting vessel loss in Winteraceae have 


received much skepticism, primarily on the 


grounds that no viable mechanism could drive the 
evolutionary loss of vessels, other than a shift to 
aquatic environments, as this would entail a shift 
to an inefficient hydraulic system (Carlquist, 1983, 
1996). However, claims about the likelihood of ves- 
sel loss in Winteraceae need to be evaluated in a 
physiological context. 

In support of this view, Donoghue and Doyle 
1989) argued that little support exists for the view 
that vessel loss in Winteraceae is likely to have 
been selected against. First, the forests with low 
evaporative demand where most Winteraceae cur- 


~ 


rently occur were suggested to be the type of hab- 
itats where any decrease in xylem transport capac- 
ity by vessel loss would have a relatively small 
effect (Donoghue, 1989; Donoghue & Doyle, 1989). 
Donoghue (1989) also suggested that the first ves- 
sels, hypothetically present in an ancestor of Win- 
teraceae, were presumably of a primitive morphol- 
ogy containing numerous scalariform partitions, 
limited in number, and in a background of func- 
tional tracheids (Donoghue, 1989; Donoghue & 
Doyle, 1989). At this stage of low evolutionary spe- 
cialization, the loss of vessels may not have been 
difficult, entailing only the retention of pit mem- 
branes. For example, the structural distinction and 
therefore the potential hydraulic consequences of 
having only tracheids versus having both tracheids 
and vessels is less obvious when Winteraceae are 
compared with their closest extant relatives, Ca- 
nellaceae. Most Canellaceae vessel elements are 
similar in size and overall shape to tracheids of 
Winteraceae (Wilson, 1960). Tracheid diameters 
(16—69 рт; Patel, 1974; Meylan & Butterfield, 
1982; Carlquist, 1981, 1982, 1983, 1988, 1989) of 
Winteraceae overlap with the lower range of vessel 
element diameters reported from Canellaceae (20— 
140 pm; Wilson, 1960; Metcalfe, 1987). Xylem 
conduits in Winteraceae and Canellaceae differ in 
that the pit membranes are retained in tracheids of 
Winteraceae, while in vessel elements of Canella- 
ceae scalariform perforation plates are developed. 
Some Winteraceae species show an intermediate 
condition where the tracheid pit membranes are 
partially dissolved, forming relatively large porosi- 
ties (0.05 jum diam.) as in some species of Bubbia 
(Carlquist, 1983). However, the comparative hy- 
draulic properties of Canellaceae and Winteraceae 
xylem are not known. 

Although it is difficult to determine the impor- 
tance of specific selective pressures in the past, 
functional and physiological studies can indicate 
what factors might have been involved. Combined 
with information from historical biogeography, func- 
tional analyses also provide a context toward inter- 


Моште 87, Митбег 3 
2000 


Feild e 


al. 331 
Pa зый. Ecological Evolution 


preting phylogenetic inferences about the past eco- 
logical and climatic associations of a lineage (Doyle 
et al., 19 ne common characteristic of the cur- 
rent distribution, ecological abundance, and paleo- 
migratory patterns of Winteraceae is an association 
with cool, wet temperate environments. Paleocli- 
matological evidence suggests that at the time Win- 
teraceae moved into Australasia, these southern 
high-latitude regions may have been colder than 
previously suspected, perhaps including some re- 
gions with persistent snow cover and frozen ground 
(Rich et al., 1988; Ditchfield et al., 1994; Sellwood 
et al., 1994; Stoll & Shrag, 1996). Where freezing 
temperatures are common, tracheids appear advan- 
tageous over vessels in terms of their resistance to 
freezing-induced cavitation (Hammel, 1967; Sucoff, 
1969; Sperry & Sullivan, 1992; Sperry et al., 1994; 
Tyree et al., 1994; Davis et al., 1999). When stems 
freeze, the insolubility of dissolved gases in ice re- 
sults in air bubbles. Upon thawing, these bubbles 
act as nucleation sites for the formation of air-em- 
bolisms blocking the movement of water through 


Ф 
= 
c 


probability of a freeze-thaw event resulting in xy- 
lem embolism correlates with conduit volume 
(Sperry & Sullivan, 1992; Sperry, 1995; Davis et 
al., 1999). This is because a greater conduit volume 
results in both more numerous and larger air bub- 
bles, which can easily expand to fill the entire xy- 
lem conduit during the thaw of xylem sap. 

Most conifers are resistant to freeze-thaw-in- 
duced cavitation irrespective of the number of 
freezing events experienced because of their tra- 
cheid-based vascular system (Hammel, 1967; Su- 
coff, 1969; Robson et al., 1988; Sperry & Sullivan, 
1992; Sperry et al., 1994). In tracheids, the bubbles 
produced in frozen sap are sufficiently small that 
they are collapsed by surface tension during thaw- 
ing. In contrast, many vesselled angiosperms, and 
especially those with large conduit volumes, em- 

olize extensively e one or more freeze- 
thaw events (Sperry, 1995; Pockman & Sperry 
1997; Davis et al., 1999). The tradeoff between "a 
nerability to freezing-induced cavitation and con- 
duit size may have influenced the direction of xy- 
lem evolution in Winteraceae (Donoghue & Doyle, 
1989; Donoghue, 1989). Indeed, Winteraceae have 
been documented to be relatively more frost toler- 
ant compared to co-occurring plants with vessels. 
Sakai et al. (1981) demonstrated that the freezing 
resistance of buds, leaves, and stems of Tasmannia 
lanceolata was greater than most co-occurring an- 
giosperms that had vessels (e.g., Nothofagus, Eu- 
calyptus, and several Proteaceae) and was similar 


to that of co-occurring conifers such as Diselma, 
Microcachrys, and Phyllocladus. Drimys winteri in 
southern Chile is also reported to be more tolerant 
to frost than Nothofagus species growing in the 
same environment (Alberdi et al., 1985). One part 
of the observed tolerance of Winteraceae to freezing 
may be the ability of a tracheid-based xylem to 
avoid freezing-induced limitations on stem water 
transport. 

Despite the central position of xylem structure in 
discussions on the evolution of Winteraceae, no 
published reports exist for the hydraulic perfor- 
mance in the field. Clearly, comparative studies of 
xylem properties and water flux rates under field 
conditions are needed. However, the selective forc- 
es on wood evolution are not confined to hydraulic 
efficiency. Safety, with respect to avoidance of air 
embolisms, and mechanical strength are two addi- 
tional selective pressures on the direction and pat- 
tern. of xylem evolution in plants (Tyree et al., 
1994). Certainly smaller xylem conduits offer high- 
er resistance to water flow. Nonetheless, in colder 
climates tracheids appear to increase the overall 
hydraulic capacity by minimizing embolisms by 
freeze-thaw events (Sperry et al., 1994; Davis et al., 
1999). This advantage for small-diameter xylem 
conduits for evergreen woody plants subject to 
freezing temperatures may explain the loss of ves- 
sels and a return to a tracheid-based vascular sys- 
tem in Winteraceae as indicated by phylogenetic 
analyses. 


CONCLUSIONS 


Studies of ecology, historical biogeography, and 
physiology challenge the long-held belief that Win- 
teraceae are an unchanged and relictual angio- 
sperm lineage. Further investigations into the struc- 
ture and function of Winteraceae morphological 
features with respect to their natural environments 
will continue to inform discussions on their evolu- 
tionary history. 


Literature Cited 


Alberdi, M., M. Romero, D. Rios & H. Wenzel. 1985. 

Altitudinal gradients of seasonal frost resistance of 

Nothofagus communities of southern Chile. Acta Oec- 
dua 6: 


Axelrod, D. I. 1984. An interpretation of Cretaceous and 
Tertiary biota in polar regions. Palaeogeogr. Palaeocli- 
matol. зүм» о]. 45: 105—147. 

Вапеу, 1. The development of vessels in angio- 
sperms ur its ми Ме а ance on morphological research. 
Amer. J. Bot. 31: 421-428. 

1953. р» of the tracheary tissue of land 

plants, Amer. J. Bot. 40: 4–8. 

— & C. С. Nast. 1944. The comparative morphol- 


Annals of the 
Missouri Botanical Garden 


ogy of the Winteraceae V. Foliar epidermis and scler- 

encyhma. J. Arnold Arbor. 25: 342-348. 

945. The comparative morphology 
of Winteraceae. Үп. Summary and conclusions. J. Ar- 
nold Arbor. 26: 37-47. 

a Swamy. 1951. The conduplicate car- 
pel of diui и ата and its trends of specialization. 
Ame ie 38: 373-379. 

P. Thompson. 1918. Additional notes upon 
the angiosperms Tetracentron, Trochodendron, and Dri- 
mys, in which vessels are absent from the wood. Ann. 
Bot. 32: 503—512 

— M. 1972. Systematic anatomy of the leaf epi- 
dermis а eae and some related families. Tax- 
on 21: 447-469 

Barry, J. B. 1980. Minniin climates of New Guinea. Pp. 
75-109 in P. van Royen (editor), Alpine Flora of New 
Guinea. о Verlag, Germany. 

Bond, W. J. . The tortoise aad the hare: Ecology of 
spem pu uda and gymnosperm persistence. 

Biol. J. Linn. Soc. 36: 227-249, 
Bongers, J. M. 19 73. Epidermal Pos haracteristics of the 
interaceae. Blumea 21: 381—4 

Brenner, С. J. 1976. Middle ыл eous floral na 
and early migrations of angiosperms. Pp. 23-4 Я 
В. Beck (editor), Origin and Early Evolution of rene 
sperms. Columbia Univ. Press, New York. 

———. 1996. Evidence for the earliest stage of angio- 
sperm pollen evolution: A paleoequatorial section from 
Israel. Pp. 91-115 in D. W. Taylor & L. J. Hickey (ed- 
itors), Flowering Plant Origin, Evolution and Phylogeny. 

n & Hall, New Yor 

W. К. Smith. 1997. Patterns of leaf sur- 
face wetness for montane and subalpine plants. РІ. Cell 
Environm. 20: 

Brodribb, T. & R. 5. Hill. 1998. Imbricacy and stomatal 
wax plugs reduce maximum is conductance in South- 
ern Hemisphere conifers. Austral. J. Bot. 45: 657—668 

Carlquist, S. 1975. d Strategies of Xylem Evo- 
ution. Univ. California Press, Berkeley 

1 d anatom of Zygog ynum (Wintera- 
севе): Field observations. М nia 3 (new series): 


à 1982. Exospermum | stipitatum (Winteraceae): 
Observation on wood, leaves, flowers, pollen, and fruit. 
Aliso 10: 277-289 

983. Wood anatomy of Bubbia (Winteraceae) 
with comments “ the origin of vessels in dicotyledons. 


Amer. J. Bot. 7 
19 


ча of velle in wood of Sarcandra 
(Chloranthaceae): Comments on vessel origins in angio- 
spe Amer. J. Bot. 74: 1765- 
aa. Wood anatomy of Drimys 5.8 
седо). о 12: 81-95. 
. Wood anatomy of il Summary of 
wood. anatomy а of Winteraceae. Aliso 251-215 
.1 Wood anatomy of ends angiosperms: 
New емма and syntheses. Рр. 68–90 in D. W. 
Taylor & L. J. Hickey (editors), Flowering Plant Origin, 
Evolution and Phylogeny. Chapman & Hall, New York. 
Chase, M. W., D. E. Soltis, R. G. Olmstead, D. Morgan 
L 


5. (Wintera- 


Wimpee, J. F. Smith, G. R. Furnier, S S. H. Strauss, Q.- 


Y. Xiang, G. M. Plunkett, P. S. Soltis, S. W. Swensen, 
S. E. Williams, P. A. Gadek, C. 


Barrett, S. Dayanandan & V. А. Albert. 1993. Phylo- 
genetics of seed plants: An analysis of nucleotide se- 
quences from the = gene rbcL. Ann. Missouri Bot. 
Gard. 80: oss 
& J. Mills: 1984. The гаи 
ге of some extinct Gondwana pollen 
' ry of the southwestern Саре (South Af- 
rica). Ann. Missouri Bot. Gard. 71: 1088-1099 
& J. Praglowski. Winteraceae pollen from 
the Miocene of the southwestern Cape (South Africa). 
Grana 27: 27-37 
Crane, P. R., E. M. Friis & K. R. Pedersen. 1995. The 


origin and early diversification of angiosperms. Nature 
374: 27-33 


Cronquist, A. 1981. An integrated System of Flowering 
Plants. Columbia Univ. Press, New Yor 

Davis, S. D., J. S. Sperry & U. С. Ноќе, 1999. Тһе 
relationship bobo xylem conduit diameter and d 
itation caused by freeze-thaw events. Amer. J. Bot. 
1367-1372 

Dettmann, M 1994. Cretaceous vegetat ion: The m 
crofossil record. Pp. 143-170 in R. S. Hill (editor), His- 
tory of the Australian Vegetation: Стась ^ous to Recent. 
Cambridge s Press, Cambridge 

& D. M. Jarzen. 1990. The Айшен Australian 

rift valley: Late Cretaceous ctadle e of northeastern Aus- 

tralasian relicts? Rev. desine Palynol. 65: 131-144. 

. Pollen evidence for Late Cre- 
taceous Зана ов А о 'eae in southern polar 
forests. Canad. J. Bot. 69: 901—906 

Ditchfield, P. W., J. D. us D. Pirrie. 1994. High- 
latitude paleotemperature variation—New data from the 
Tithonian to Eocene of James Ross Island, Antarctica. 
Palaeogeogr. Palaeoclimatol. Palaeoecol. 107: 79-101. 

Donoghue, M. J. 1989. Phylogenies and the analysis of 

exa 


evolutionary sequences, with mples from seed 
дыст vig us 43: 1137-1155. 
Doyle. 1989. Phylogenetic analysis of 


ни | the relationships of Hamamelidae. Pp. 
17-45 > В. Crane & S. Blackmore (editors), Evo. 
lution, E dts and Fossil bai of the Hama- 
melidae. Clarendon Press, Oxfo 

Doyle, J. A. 1998. Phylogeny of а plants. Ann. 
Rev. eo Syst. 29: 567—599. 

000. о оом and geographic 

history of Winterac . Missouri Bot. Gard. 87: 
303-316 


J. Donoghue. 1986. Seed plant phylogeny 

and the origin of angiosperms: An experimental cladis- 

tic в Вог. Rev. 52: 321-431. 
С. L. Hotton & J. V. W. 1990. Early Creta- 

ceous Пака zonasulcate allen. nd Winteraceae. 2. 

Cladistic analysis and implications. Amer. J. Bot. 
568. 


. A. Zwieniecki, N. M. Holbrook & M. J. 
1998. Paged a stomata of Drimys winteri 
pisi protect leave 1 mist not drought 
Proc. Natl. Acad. Sci. U.S. А. 95: 14256— 
к N., P. С. J. Barker, P. J. Cullen & A. Shápéoit. 
995. Conifers of eue Australia. Pp. 223-251 in 
3 J. Enright & R. Hill (editors), Ecology of the 
Southern Conifers. ie ue Institution Press, Wash- 
ington, D.C 


Volume 87, Number 3 
2000 


Feild e 333 


al. 
ieee Ecological Evolution 


Gifford, E. M. & A. S. Foster. ie жы жы and 
Evolution of Vascular Plants. Freeman Press, New York. 
. & J. J. Sauter. 1996. Xylem inue =a 

winter and recovery ofh in diffus 
porous and g porous ia, Oecologia 105: 435-439. 


ammel, 967. Freezing of xylem sap without cav- 
itation. Pl. ы. 42: 5 
Hickey, L. J. & J. А. Wolfe. 1975. The bases of angio- 


19 
sperm dca Vegetative morphology. Ann. Missouri 
Bot. Gard. 62: 538—589 

Hill, R. S. & L. J. Scriven. 1995. The a Rio dad -dom- 
inated woody —€— * Antarctica—A review. Rev. 
Palaeobot. Palynol. 86: 198. 

Hope, G. S. 1980. New о montane vegetation com- 
munities. Pp. 153—222 in P. van iiie (editor), Alpine 
Flora of New Guinea, Gantner Verlag, German 

Jaffré, T. 1995. Distribution and ecology a де conifers 
of New Caledonia. Pp. 171-196 in N. J. Enright & 

S. Hill (editors), Ecology of the oe fm ane 
Smithsonian Institution Press, a а D.C 
Jeffree, С. Е., Johnson & Р. G. Jarvis, 1972. 
Epicuticular wax in the aed incuba of Stika 
its effects on the diffusion of water vapour 
carbon dioxide. Planta 98: 1—10. 
983. Treeless plant communities of 
e Tasmanian high country. Proc. Ecol. Soc. Australia 
61-71. 


—— ———. 1997. Alpine Tasmania: An Illustrated Guide to 
the Vegetation. Oxfon Univ. Press, Oxfo 

& K. L. Bridle. 1999. Environment and ia 's 
of ten penu alpine Сай ор. Austral. J. Bot 


1— 
Koster, J. & P. Baas. 1981. Alveolar cuticular material 
in Myristicaceae. Рр. 131—137 т D. Culter (editor), The 
Plant Cuticle. Academic Press, San Diego. 

Lusk, C. H. 1993. Stand dynamics of the shade-tolerant 
ney ae nubigena and Saxegothea conspi- 
cua in Chilean temperate rain forest. J. Vegetation Sci. 
1: 549-558.. 

Macphail, M. K., N. F. Alley, E. M. Truswell & I. R. K. 
Sluiter; 1994. Early Teniary M rap Evidence from 
spores and pollen. Pp. 189 in R. S. Hill (editor), 
History of the Australian бо. Cretaceous to Ке- 

ri 


The root of angio- 
I1 phylogeny inferred from duplicate genes. Science 
286: 947-950. 

о С. Е. 1987. Anatomy of the Dicotyledons, Vol. 
Ш. — о Oxfor 

Meylan, B. A. . Butterfield. 1982. Pit membrane 
structure in oa aster less woods of Pseudowintera 

— Pv ur €: Bull. 3: 167-175. 

Nandi, M. e & P. K. Endress. 1998. A 
e uds ladite мы of angiosperms using rbcL 

and non-molecular data sets. Ann. Missouri. Bot. Gard. 
85: 137-212. 

Nunez, M., J. B. Kirkpatrick & C. Nilsson. 1996. Rainfall 
estimation in south-west Tasmania using satellite im- 
ages and phytosociological calibration. Int. J. Remote 
Sensing 17: 1583-1 

4. Wood anatomy of the dicotyledons 
indigenous to xs ir 4. Winteraceae. New 
land J. d 12 | 

Ре туг, O., L. B. ы. z Bergstrom & I. Groth. 1990. 
Pollination d New Caledonian Winteraceae: Opportu- 
nistic shifts or parallel edel with their pollinators. 

РІ. Syst. Evol. 173: 143-157. 


Pockman, W. T. & J. S. Sperry. 1997. Freezing-induced 
xylem cavitation and the northern limit of Larrea tri- 
ya tata. Oecologia 109: 19— 
., P. Y. Ladige, T. Ј. Entwisle & A. N. Drin- 
Morphometric studies s Tas- 
тапта (Жесе in Victoria, маб, “Mulleria 
38: 235-256. 

Raven, P. H. 1980. Plate tectonics and Southern Hemi- 
не Mira aphy. Pp. 3-24 in K. Larsen & L. B. 
Holm-Nielsen Маг ни: Tropical Botany. Academic 
Press. Mes 

—— —— & D. I. Axe 1974. Angiosperm biogeography 
and past рее. movements. Ann. Missouri Bot. 
Gard. 61: 539 

Read, J. & R. ^i Hill 1983. Rainforest ne onto 
елмен old-fields. Austral. J. Ecol. 8: 149—161. 

Rebertus, A. J. & T. T. Veblen. 1993. аид and tree- 
fall gap pies of old-growth Nothofagus forests in 
Tierra del Fuego, Argentina. J. Veg. Sci. 4: 641-654. 

Rich, P. V., T. H. Rich & B. E. Wagstaff 1988. Evidence 
for low temperatures and biologic diversity in Creta- 
ceous high latitudes of Australia. Science 242: 1403— 

1406. 


Robson, D. J., W. J. McHardy & J. A. Petty. 1988. Freez- 
ing in conifer xylem. II. Pit m and bubble for- 
жаса " Exp. Bot. 39: 1617-16 
Sakai, A., D. M. Paton & Р. E 1981. Freezing re- 
кайа : trees of the south temperate zone, especially 
i alpine species of Australasia. Ecology 62: 563- 
57 


. P. Lowry II & A. Ramisamihantanirina. 
e 391: 


Se us G. E., 
1998. Takhtajania perrieri: Rediscovered. Nature 
133-134. 


Schuster, R. M. 1976. Plate tectonics and its bearing on 


Evolution of Апріовретиів. Columbia Univ. Press, New 
ork. 


мыл B. W., С. D. Price & P. J. Valdes. 1994. Cooler 


estimates of Cretaceous temperatures. Nature 370: 453— 
455. 


Smith, A. C. 1942. wv? of Paupasian plants, V. J. 
Arnold Ese 23: 417 

The Vien an species of Drimys. J. Ar- 

nold T 24: 1-33. 

— |943b. Taxonomic notes on the Old World spe- 
cies of о J. Arnold Arbor. 24: 119-1 

— 1945. Geographic distribution of the Wintera- 

eae. J. ke Arbor. 26: 48— 
tis, D. L. 


Nickraak L. A. Johnson, 


S.-M. Chaw, L. J. Gillespie, W. J. Kres 
1997. Angiosperm phylogeny inferred from T ibo. 
somal DNA sequences. Ann. Missouri Bot. Gar 

1-49 


Specht, R. L., M. E. Dettmann & D. M. Jarzen. 1992. 
Community associations and structure in the Late Cre- 
taceous vegetation of southeast Australasia and Antarc- 
tica. Palaeogeogr., Palaeoclimatol., Palaeoecol. 94: 
283-309. 

Sperry, J. S. 1995. Limitations on stem water transport 
and their consequences. Pp. 105-124 in B. L. Gartner 
(editor), Plant Stems: Physiology and Functional Mor- 
phology. Academic Press, San Diego. 

. M. Sullivan. 1992. Xylem embolism in 

response to бш thaw cycles and water stress in ring- 


334 


Annals of the 
Missouri Botanical Garden 


porous, diffuse-porous, and conifer species. Pl. Physiol. 
100: eis 3. 

——. К. `. M. Sullivan & S. Е. Eastlack. 
1994. s embolism 1 in E porous, diffuse-porous, 
and coniferous tree n Utah and interior Alas- 
ka. Ecology i 1736-1752. 

Spicer, R. A., P. M. Ree „ Chapman. 1993. Cre- 
taceous phytogeography a prata signals. Philos. 
Trans. R. Soc. London Ser. B 341: 277-285 

Srivastava, S. K. 1994. Evolution of Cretaceous phyto- 
geoprovinces, continents and climates. Rev. Palaeobot. 
Palynol. 8 

Stewart, С. Н. & J. I 3. J. Harrison. 1987. Plant commu- 
nities, landforms, xd soils of a geomorphically active 
drainage basin, Southern Alps, New Zealanc №. 2. ] 
Вог. 25: oes 

Stockey, R. Atkinson. 1993. Cuticle micro- 
кое of fae ie Salisbury. Int. J. Plant Sei. 154: 
187-2 


= 
~ 


revel. 1997. Cuticle micromorphology 
of Prumnopitys > (Родосаграсеае). Int. J. Plant 
Sci. 158 a 


ү Woltz. 1992. Cuticle micromorphol- 
ogy of Falcatifolium de Laubenfels (Podocarpaceae). 
Int. J. Plant Sci. 153: 589—601. 

Fr revel & P. Woltz. 1998. Cuticle micro- 
morphology of Podocarpus, subgenus Podocarpus, sec- 
tion Scytopodium ma arpaceae) of Maagano ar and 
South Africa. Int. J. Plant Sci. 159: 923—940. 

Stoll, H & D. P. Shrag. 1996. Evidence for glacial 
control of rapid sea | changes in the Early Creta- 
ceous. Science 272: 17 

Sucoff, E. 1969. reins of con iler xylem sap and the 
cohesion-tension theory. Physiol. Pl. (Copenhagen) 22: 
424—431. 

Suh, Y., . E. Reeve & E. A. Zimmer. 1993. 
Molecular ы, and phylogenetic implications of 

il of ribosomal 


Are Tetrace 'entron, 
xin dry gi and Drimys spec vid. or primitive 


? Mem. New York Bot. Gard. 6: 27–: 


Thorne, R. Е. 1974. А oo classification of the 
nnoniflorae. Aliso 8: 209. 

Tieghem, Р. van. 1900. d Гр dicotylédones du groupe 
des ER RT J. a Momi 14: 259-297, 330-361. 

os M. T., S. D. Dav chi 

'al коис maven of ~ m evolutior 

of hydraulic efficiency for ШАКЫ to dysfunction? 
ТАМА Bull. 15: е 

Veblen, T. T., B. Burns, T. Kitzberger, A. Lara & R. 
Villalba. 1995. us lun of the piss : southem 
South America. Pp. 120-155 in N. J. Enright & R. 
Hill (editors), Ecology " the Southern Conifers Aa 


Old World. 1. 
Са апа Drimys—Morphology and taxonomy. 
Blumea 18: 226—354. 

—— The Winteraceae of the Old World. II 
Zygogynum-—Morphology and taxonomy. Blumea 23: 
219-250. 

The Winteraceae of the Old кө S The 

mecs species of Bubbia. Blumea 28: : 

Taxonomy in Winteraceae. e 37: 


691-698. 

1993. Winteraceae. /n: P. Morat & H. S. Mackee 

(editors). СА de Іа Nouvelle-Calédonie 19: ен 
Muséum National d'Histoire Naturelle, Paris. 

Walker, J. W., G. J. ines A. G. Walker. 1983. Win- 
teraceae pollen in the Lower Cretaceous in Israel: Early 
evidence of a И Ш angiosperm family. Science 
220: 1273-1275. 

Wells, P. M. & R. S. Hill. 1989. Leaf d о = 
imbricate-leaved Podocarpaceae. Austral. Syst. 

369-3 

Wilson, Т. K. 1960. The comparative morphology of the 

е | Synopsis of genera and wood anatomy. 
тор. Woods 112: 1-27. 

ras F. & А. Sakai. 1985. Types of Florin rings, dis- 
tribution patterns of epicuticular wax, and their rela- 
das in the genus Pinus. Canad. J. Bot. 63: 2150- 
215 


Yong, D. A. . Are P n primitively vessel- 
less? Syst. Er E r 3- ~ 
о М. aa Structure and the As- 
nt of Sap. nire е New York. 


ANATOMY OF THE YOUNG 
VEGETATIVE SHOOT OF 
TAKHTAJANIA PERRIERI 
(WINTERACEAE)! 


Richard C. Keating? 


ABSTRACT 


Material of the rare monotypic genus Takhtajania has been examined for anatomy of the young vegetative shoot. 
Leaf cuticles are smooth, stomata mostly brachyparacytic and free of alveolar deposits common to other Winteraceae 


Mesophyll structure is weakly differen tiated. Petio 
e nodal level. Splitting of lateral 


appears amon 


traces lea 


Ме bases are vasc — by three independent traces, each of whic ћ 


more or less 5-trace V-pattern at higher levels 


ranalian families, т appear to ђе nearest this level of specialization, mostly on the basis of similarities in 


gs and nodal structu 


words: 


Canellaceae, leaf, nodal anatomy, stem, Takhtajania, vegetative shoot, Winteraceae. 


Phylogenists have long considered Winteraceae 
to be among the least specialized in the angio- 
sperms (cf. Takhtajan, 1997). Among analyses sup- 
porting this conclusion are those based on the pres- 
1983; 
Tieghem, 1900), a mixture of sieve-element plastids 
(Behnke, 1988; Behnke & Kiritis, 1983), plesiom- 

orphic floral structure (Erbar & Leins, 1983; En- 
dress, 1983; , 1988), and paleoantarctic dis- 
ибн (Smith, 1945). Takhtajania, the rarest and 
most recently recognized genus, has been known 
only from dried type material. It is isolated from 
other Winteraceae geographically and morphologi- 
cally. Only recently has liquid-preserved material 
been available in support of a detailed study of its 
morphology and anatomy. The most important ques- 
tions to be initially considered were (1) what is the 
place of the genus among its likely neighbors, and 
(2) what is its level of specialization within the 


ence of vesselless wood (Carlquist, van 


woody Ranales 

The original Takhtajania collections were made 
in Madagascar in 1909 by Perrier de la Báthie. 
They awaited examination at Paris (P) until Capu- 
ron (1963), finding them to represent a new taxon 
of Winteraceae, named the collection Bubbia per- 
rieri. Later study of the leaves by Baranova led to 
reevaluation of B. perrieri and its recognition as the 
monotypic genus Zakhtajania perrieri (Capuron 
Baranova & J.-F. Leroy. At the same time, it was 
placed in a new subfamily Takhtajanioideae by Ler- 


— 


oy (1978). The species remains а a and iso- 
lated from other known genera of the fami 

After repeated attempts over the past 25 years 
to find more material, Malagasy parataxonomist 
Fanja Паола А located a second popula- 
tion of Takhtajania in 1994 about 150 km east of 
the original locality. Then in spring 1997, the new 
locality was revisited by parataxonomist P. J. Rak- 
otomalaza and Chris Birkinshaw who collected liq- 
uid-preserved flowering and vegetative material. 
For more detailed information regarding the inter- 
esting collection history of this taxon, see Schatz 
2000 this issue). 
At the Missouri Botanical Garden, the preserved 
materials were divided among several investigators 


no 
> 


who are contributing observations on wood and re- 
productive organs. This contribution includes ob- 
servations on leaf vasculature from clearings and 
transsections of the leaf lamina and midrib, petiole 
and nodal anatomy, and other anatomy of the young 
vegetative shoot. 


MATERIAL AND METHODS 


Specimens of vegetative and flowering material 
were collected in the Anjanaharibe-Sud Special 
Reserve southwest of Andapa in northeastern Mad- 
agascar as follows: P. J. Rakotomalaza et al. 1342, 
and C. Birkinshaw 483, 14°45'S, 49?28'E, at 1200 
m altitude in a perhumid forest with abundant moss 
and lichens covering the stems. 


! | thank Thomas К. Wilson, Miami University, Oxford, Ohio, for useful discussions and the use of his laboratory 


facilities for preparation of some samples 


? Missouri Botanical Garden, P.O. Box 299, St. Louis, 


ANN. Missouni Bor. 


. L also thank W. Г. 


Missouri 63166, U.S 


Stern and an anonymous rev iewer for thorough reviews. 
y. A. 


GARD. 87: 335-346. 2000. 


Annals of the 
Missouri Botanical Garden 


Young stems and leaves used in this study were 
field preserved in 50% ethanolic FAA and trans- 
ferred to 70% ethanol upon arrival at the Missouri 
Botanical Garden. Leaf clearings were made by an 
improved method (Keating, unpublished) where 
specimens were placed in cold (room temperature) 
5% NaOH and then microwaved. The watt 
magnetron was set on defrost cycle, 1.е., pulsed 
power, 2 seconds on/off for three cycles of 20 sec- 
onds. This promoted uniform clearing and avoided 
boiling, which is quite destructive of soft tissues. 
The NaOH-covered specimens were then placed in 
a 45°C oven for 48 hours during which time clear- 
ing was completed. Specimens were immersed in 

cold (room temperature) 5.2596 sodium hypochlo- 

rite (Clorox®) for 30 minutes until the specimen 
was white. After three gentle tap water rinses, the 
specimen was soaked in LKI (0.2/2.096) (v/v) in 
1596 ethanol for two hours until uniformly brown. 
Without further aqueous rinse, the specimen was 
changed to 20% CaCl, solution and then mounted. 
Within minutes, dark magenta veins contrasted 
against a transparent background in the cleared 
specimens. 

Cross sections of leaf lamina and midrib, petiole, 
node, and young stem were made by hand micro- 
tome as well as free-hand and mounted in CaCl,. 
Some sections were mounted unstained in glycer- 
ine. Most were stained in cresyl violet acetate, to- 
luidine blue, Schiff's reagent, or I,KI. Differentia- 
tion of these dyes is sharp in the strongly ionic 
CaCl, (Herr, 1992; Keating, 1996). Two additional 
specimens of liquid-preserved young stems with at- 
tached leaf bases were paraffin-embedded, serial- 
sectioned transversely on a rotary microtome at 10 
jam, and stained in Safranin-O and Fast Green FCF. 


OBSERVATIONS 
SHOOT APEX 


The young stem expands massively within 5—7 
mm of the shoot tip causing the apical meristems 
to appear somewhat embedded among massive pet- 
iole bases (Fig. 1A, B). From an early stage, petiole 
bases show an obvious cicatrice constriction (Fig. 
1A). Young primordia form a 2/5 phyllotaxy of pet- 
iole bases that show early cell division and expan- 
sion, laterally and dorsiventrally. Packing geometry 
causes petiole bases to remain partially fused at the 
shoot apex level and to form polygonal shapes (Fig. 
1C). The axillary bud is, in two observations, an 
outgrowth of the adaxial side of the petiole of the 
7th primordium (Fig. 1D). No vascular connections 
could be followed, nor was any bud development 
visible in the true axil of more mature nodes. 


LEAF SURFACE AND PARADERMAL ANATOMY 


Leaf outline elliptical (Fig. 2A) with a decurrent 
base (Fig. 2B). Length/width ratio 4—5:1. Cuticle 
smooth. Epidermis: cells rounded-polygonal (1-3: 
1 Мм) (Fig. 2C). In some areas of older leaves cu- 
ticular flanges form thick dividers between cells, 
3—7 pm wide. Stomata abaxial only, guard cell 
pairs with lengths 22-29 jum, widths 18-21 pm, 
most commonly brachyparacytic with four adjacent 
subsidiary cells that differ only in position from 
normal epidermal cells (Fig. 2C). Occasionally ca. 
2096 of stomata have 5 or 6 subsidiary cells, but 
remain brachyparacytic with regard to the presence 
of two epidermal cells always parallel with the 
guard cells. Mesophyll: cells of adaxial palisade 
zone appearing compact and shortly lobed with ca. 
10% air cavity space. Spongy tissue with numerous 
air cavities 2-3X as broad as cell diameters. Air 
space ca. 60%. Venation pinnate, looped brochi- 
dodromous. Intercostal areas irregular. Higher-or- 
der venation showing incomplete and very irregular 
areolation (Fig. 2D), 0—6 vein endings per areole 
(Fig. 2E). Areole sizes: smaller ones ca. 340 jum 
isohedral; larger ones ca. 2610 X 1248 jum. 


LEAF TRANSSECTION 


Structure weakly dorsiventral (Fig. 3A). Midrib: 
outline shallowly concave adaxially, deeply convex- 
rounded abaxially. Cuticle smooth, thickness 5-7 
um (Fig. ЗВ). Сипсшаг flanges absent or narrow 
between epidermal cells, up to 18 jum deep. Epi- 
dermis: cells thin and cuboidal on both surfaces. 
Stomata level with surface. Guard cells small, 
oblique to and oriented toward the surface in re- 
lation to adjacent subsidiary cells, which subtend 
the guard cells internally (Fig. 3D). Outer cuticular 
ledge well developed at stomatal opening; internal 
ledge not present or much smaller. No cuticular 
alveolar deposits occlude the stomatal openings. 
Hypodermis generally absent although adaxial- 
most layer of mesophyll may lack chloroplasts in 
some areas. Mesophyll differentiation weak. Cells 
of the palisade zone mostly small, rounded cells in 
1-3 closely packed layers, occupying up to 25% of 
mesophyll, forming a gradual transition to spon 
tissue. Spongy cells in lower mesophyll horizontally 
elongated with irregular long lobes. Chloroplasts 
most densely packed in adaxial mesophyll cells, 
gradually becoming more dispersed in abaxial me- 
sophyll. Air cavities: most larger cavities abaxial 
and aligned over stomata as deep substomatal cav- 
ities extending above the center of the mesophyll 


(Fig. 3A, D). 


Моште 87, Митбег 3 
2000 


Keating 337 
Vegetative Shoot Anatomy 


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Takhtajania perrieri shoot tip ie ae ‘al meristem re egion. —A. Young shoot tip showing apical mer- 


"s и (A-arrow m cicatrice constric s (С 
ge meristem. Base of 


таи ses of several leaf primordia fused as m 


-arrow). т; 
f leaf primordia | 'rmined by ра 
stem 15 


anssection of young stem tip, ca. 100 рт distal 
deling geometry. —C. Shoot apex at apical meristem 


partially sunken at did summit. —D. Shoot apex са. 
:В.С 


120 wm aia to ilg meristem. Axillary bud attached to leaf uc ce и base. Scale Dn А = 1 ст 


300 um; D = 150 р 


Midrib vasculature usually 5 bundles arranged 
in a V-shape, one abaxial bundle and 2 or 3 lateral 
bundles on each side. Xylem: largest bundles occur 
with 10-12 files of metaxylem facing phloem with 
a convex procambial boundary. Phloem: crescent- 
tion of sieve elements and companion cells. 
show no alignment or regularity of pattern in cross 
section. Bundle sheaths a single layer of thin- 
walled 


Sclerenchyma: lateral-most bundles in midrib have 


cells, little modified from spongy tissue. 


1-2-layered extraxylary fiber cap outside phloem. 


Secondary bundles with two lateral fiber strands at 
phloem and small strand capping xylem. Secretory 
tissue: slightly enlarged oil cells frequent in me- 
sophyll (Fig. 


discontinuou 


3C, E). Tannin cells in midrib area in 
w boundi 


smaller cells within phloem. Starch grains either 


ing phloem; some dark 
common throughout cells of the lamina, or grains 
mostly in spongy cells and not numerous. In mid- 
rib, starch grains packed in ground tissue internally 
adjacent to xylem points. Grains less common but 
often well developed in all ground tissue in vicinity 
of vascular strands. 


338 Annals of the 
Missouri Botanical Garden 


№ EM 


№ 
3 
"X 


Figure 2. Takhtajania ар young leaf structure. —A. Pair of young leaves, . not actually oppos site, at shoot apex. 
—B. Young leaf, adaxial surface showing decurrent lamina. —C. Abaxial surface of ep vidermis showing polygonal 


epidermal cells and stomata. D. Leaf clearing showing small о amd free vein yard) —E. Leaf c earing showing 
large imperfect areole and free vein endings. Scale lines: A, B — ; С = 50 pm: D = 200 um; Е = 300 pum. 


Volume 87, Number 3 Keating 339 
2000 Vegetative Shoot Anatomy 


Wt. 


ПЕР arl A. 
+ "m 
га F 


Figure 3. Takhtajania perrieri е ph petiole structure. —A. Lamina transsection (TS) sl t mesophyll 
in palisade re gion and circular vascular bundle. —B. Lamina TS, hand section showing кА thic k е cuticle. 
— 'oung lamina b one р expansion showing several mature oil cells (arrows). —D. Lamina TS abaxial 
mos 'e. Stomata with guard cells arranged obliquely to epidermal cells. —E. Young lamina showing young oil cells. 

—F. Petiole base s Webs V-pattern of median bundle and divided lateral bundles. Scale bars: A, D — 

E = 50 um; F = 300 рт. 


150 шт; B, С, 


340 Annals of the 
Missouri Botanical Garden 


Figure 4. Takhtajania perrieri stem and nodal structure. —A. Young stem TS outer cortex, epidermis, and cuticle. 
—B. Stem TS pith and starch grains. —C. Node showing median trace and two lateral traces. —D. Node TS hand 
section showing split median trace and one lateral trace. —E. Node TS divided median trace within the stele. Small 


Volume 87, Number 3 
2000 


Keating 341 


Vegetative Shoot Anatomy 


PETIOLE TRANSSECTION 


Outline: flattened adaxially at base, becoming te- 
rete distally. Size: diameter 2.5 mm. Cuticle 
smooth, thickness 3.6-7 шт. Epidermis: 
small and cuboidal, diameter са. 22 jum. Ground 
tissue: cells spheroidal, diameter 26—70 um, loose- 
ly packed but with no organized aerenchyma. In- 
tercellular air spaces at cell corners comprising 
10–20% of ground tissue. Venation: 3 to 7 collat- 
eral vascular bundles arranged in V-shape (Fig. 
3F). Xylem: several protoxylem points lead to or- 
ganized files of metaxylem, up to three cells wide 
per protoxylem point. Xylem production ends at 
straight or broadly convex cambial boundary with 
hemielliptical phloem strand. Phloem: sieve ele- 
ments and companion cells in tiers 1–2 cells wide. 
Tannin cells abundant in ground tissue, these not 
differentiated from ground tissue except by dark- 
staining contents. Tannin cells randomly scattered 
abaxially or in radial rows adaxially. Starch grains 
common in cells adjacent to vascular bundles. 


cells 


YOUNG STEM TRANSSECTION AND LONGISECTION 


Outline terete. Diameter ca. 1 cm. Cuticle 
smooth, thickness, ca 3.5 jum (Fig. 4A). Epidermis: 
cells small and cuboidal. Chlorenchyma: chloro- 
plasts very common in outer cortex. Cortex: ground 
tissue cells mostly spherical, loosely packed with 
numerous air cavities. Cells axially elongate, 2—3 x 
cell diameter in transsection. Air space ca. 20% of 
cortex. Pith cells polygonal and somewhat larger 
than cortical parenchyma cells; air cavities 5-10% 
of pith. Vascular bundles collateral. Xylem: distinct 
protoxylem points at pith boundary but xylem 
quickly develops cambial growth producing a pseu- 
dosiphonostele within 90 jum of pith boundary. 

hloem: sieve element arrangement irregular. Com- 
panion cells not easily distinguished in cross sec- 
tion. Widest pith rays with slight tendency toward 
ray dilation in phloem. In longisection, metaphloem 
sieve element length up to 395 jum. Early second- 
ary sieve element length 215—290 jum. Sieve plates 
horizontal or 5—10? off horizontal and somewhat 
broadened. Companion cells short or mostly as long 
as sieve elements, highly variable in shape and 


Sclerenchyma: no extraxylary fibers present in 
small stems. In one sample, ca. 9 mm diameter, 
occasional single fibers occur around phloem pe- 


rimeter in inner cortex. In longisection, these single 
cells overlap and are contiguous for at least longer 
than 2 cm. In same larger specimen, sclerotic layer 
occurs subepidermally around perimeter. Sclerotic 
cells in 1-3 layers, heavily lignified, with up to 20 
isotropic wall layers especially well developed on 
inner and outer periclinal walls. Tannin cells scat- 
tered and common in all ground tissue. Starch 
grains rounded, single or in clusters (Fig. 4B). 
Grain diameter 9-13 jum. Grains common through- 
out pith and in sheath of 5—10 inner cortical cell 
layers. 


STEM—NODE—LEAF CONTINUUM 


Stem outline in the vicinity of the node is irreg- 
ularly elliptical. In the young stem, the vascular 
procambium appears to be a nearly continuous cir- 
cle with maturation of proto- and metaxylem only 
observed in what will become leaf traces. А char- 
acteristic cut through the node of a stem with a 
young active cambium shows three non-adjacent 
traces vascularizing the leaf base, i.e., the node 
would be called three-trace, tri-lacunar (Fig. 4C) 
using the original siphonostele paradigm begun by 
Jeffrey (1898) and carried into dicot nodal anatomy 
by Sinnott (1914). However, the two specimens ob- 
served here show some intriguing differences. The 
older stem sections show most leaf traces appearing 
double, that is, there are two proto-xylem points 
causing the development of two paired traces. They 
may cause a node to appear 2:2:2, 2:2:1, or 1: 
2:1 (Fig. 4D, E, F). In the younger stem pieces, 
which were paraffin-sectioned at 10 jum, this ob- 
servation was confirmed less frequently. Diagram- 
ming the vasculature through several nodes showed 
mostly late-appearing bifurcations of larger bun- 
dles. 

Nothing was found to indicate that the individual 
leaf traces, especially the median trace, have com- 
ponents that occur in different sympodia of the vas- 
culature. In this material, leaf traces were well de- 
fined through at least four nodes before entering a 
leaf base. New leaf traces appeared to arise from 
undifferentiated interfascicular procambium. More 
material, sectioned at closer intervals, will be need- 
ed to identify vein sympodium branching patterns. 
Following the leaf base distally from the node, the 
three traces are observed to relate in a V appear- 
ance, whether or not they appeared as double at 


— 


incipient traces visible within the procambial cylinder. —F. 
300 


bial cylinder. Scale bars: A, В = 50 jum; C, 


Node TS showing divided lateral traces at level of procam- 
; = 200 p 150 pm. 


Annals of the 
Missouri Botanical Garden 


the nodal level. Distally, in the leaf base, the two 
lateral traces bifurcate to form two pairs of lateral 
traces. Using Sugiyama’s (1979) notation the Takh- 
tajania node can be most usually figured (moving 
distally into the petiole) as L-M'M-L => L-M-L = 
L'L-M-L'L. In the base of the petiole and continu- 
ing up to the lamina, the resulting five traces form 
a marked V of closely associated vascular bundles 
with the median trace forming the base of the V. 

Axillary buds do not appear among the first 4—5 
leaf primordia. They occur removed from the axil, 
high on the adaxial side of young petioles (Fig. 1D 
No vascular connections to axillary buds were de- 
tected in this material. 


— 


DISCUSSION 
THE LEAF 

Leaf venation is typical of other ranalian families 
with elliptic leaves with entire margins. The pin- 
nate, festooned brochidodromous structure, accom- 
panied by imperfect and variable areole structure, 
produces a low first-rank leaf (cf. Hickey, 1977: 


Metcalfe (1987) noted much variability in cuticle 
texture and sculpturing in Winteraceae, and Takh- 
tajania’s smooth, unsculptured cuticle seems di- 
agnostic within the family. Bongers (1973) noted 
that leaf cuticles of Takhtajania and some Drimys 
species lack the alveolar layer present in all other 
Winteraceae. However, Baranova (1972) noted that 
"Group 2" Winteraceae, Bubbia, Belliolum, Pseu- 
dowintera, and Zygogynum, also have ordinary (— 
non-alveolar) or more or less grainy cuticles. Ex- 
isting data are not a useful guide to relationships 
until comparable observations can be made across 
the genera. The known leaf venation trends are not 
sufficiently refined for one to offer an opinon on 
whether Takhtajania is more or less specialized 
than other winteraceous species. 

Leaf surface observations do not confirm Bara- 
nova's (1972) report of the existence of mostly an- 
omocytic stomata in Takhtajania; instead, stomata 
are mostly brachyparacytic as is true of other win- 
teraceous genera. This removes a major argument 
for subfamily-level segregation of the genus. Also, 
stomatal apertures are said to be occluded with al- 
veolar material (Metcalfe, 1987; Bongers, 1973; 
Baranova, 1972), but this is not true in Takhtajania 
or in Drimys (Tasmannia) piperita Hook. (Bongers, 
1973). 

In genera studied thus far, including Takhtaja- 
nia, mesophyll in the family is reported to be not 
clearly differentiated into palisade and spongy me- 
sophyll, e.g., Exospermum (Carlquist, 1982) or Zyg- 


ogynum (Sampson, 1983). This weakly bifacial 
structure is consistent with a relatively low place- 
ment of this leaf structure among the Ranales. Bai- 
ley and Nast (1944b) noted that thicker, more co- 
riaceous leaves in Drimys have large sclereids in 
the mesophyll between vascular bundles, and Rao 
and Das (1979) also reported nests of sclereids in 
the family. By their absence, Takhtajania shows 
simpler structure. In most Old World genera of 
Winteraceae, all foliar vascular bundles are com- 
monly ensheathed by a slender ring of thick-walled 
sclerenchyma cells. This was not observed in Takh- 
tajania. 


THE NODE 


Observations on nodal anatomy are not as sys- 
tematically powerful as they could be, partially due 
to persistent use of the siphonostele paradigm (see 
review by Beck et al., 1982), partially to unsolved 
theoretical concerns regarding the reality of cauline 
vasculature, and partially to the difficulties in re- 
lating nodal traces to stelar vasculature. At least for 
three-trace ranalian genera showing open architec- 
ture, the following discussion predicates that vas- 
cular patterns are best recognized as a cylinder of 
eustelic sympodial bundles of varying number. Fol- 
lowed from a proximal to distal direction, all of 
these bundles become leaf traces at regular inter- 
vals. In terms of their origin, all so-called cauline 
bundles are first identified as leaf traces. They 
probably connect basipetally to existing leaf trace 
sympodia. 

As seen from the contiguous older nodes and two 
short pieces of stem tip available in the present 
study, nodal structure places Takhtajania among 
Benzing's (1967) trilacunar ranalian families. Benz- 
ing noted that nodes in Bubbia sp., Drimys colorata 
Raoul, D. winteri J. R. Forst. & G. Forst., Canella 
alba Murray, and Warburgia ugandensis Sprague 
are so similar that they can be described as one. 
АП five species showed a 2/5 phyllotaxy, which is 
also confirmed in Takhtajania. Benzing's specimens 
showed distinct helicoid sympodia where every 5th 
leaf is supplied by a median trace from the same 
sympodium. This seems possible but could not be 
quite confirmed from currently studied Takhtajania 
material. The relationship of 3 traces vascularizing 
a leaf to similar sets of traces above and below that 
node appears somewhat unpredictable and irregular 
in the available material (Fig. 5). 

About 5 mm below the apical meristem, in a 
region of closely spaced nodes, the vasculature of 
the young stem forms a eustele consisting of pre- 
cociously matured xylem strands, here interpreted 


Volume 87, Number 3 
2000 


Keating 343 


Vegetative Shoot Anatomy 


igure 5. Takhtajania perrieri: diagram of shoot vas- 

cular pattern through five nodes, reconstructed from 10 

um serial dicis Fusteli ic bundles appearing as solid 

lines, each eventually becoming a leaf trace. Incipient 

traces occurring entirely within cambial zone shown as 
edia 


broken lines. n leaf (гасе; L = lateral leaf 


trace. Other inc mh. traces, 2—5 between larger traces, 
are not shown. Leaf trace bifure ations occurring at various 
levels also not shown. 


as leaf traces, embedded in a cylinder of procam- 
bium. Usually, the clearly defined vascular bundles 
occur in pairs or threes, rather than as singles. The 
vascular diagram for Takhtajania (Fig. 5) shows 
that lateral and median leaf traces arise indepen- 
dently. АП of the closely associated trace pairs do 
not arise from different sympodia as far as can be 
followed. All are superficial bifurcations arising 
distal to their identity as single incipient traces in 
the procambial cylinder. 

An urgent need is the diagramming of the vas- 
culature pattern from sections in the 1—3 jum range 
to see if the type of sympodial arrangement of ter- 
minal traces and renewal branches drawn by Benz- 
ing (1967) for a number of ranalian families can be 
confirmed. From the three short stem pieces avail- 
able for this study, such an integrated pattern can- 
not be drawn. 

As noted above, several relevant studies have to 
be translated from the cauline vasculature concept. 
In Bailey and Nast's (1944a: 215) study of the Win- 


teraceae, they suggested that foliar bundles arise 


from “interfascicular parts of the hypothetical, cau- 
line, primary vascular cylinder." Also, Beck et al. 
(1982: 795) stated, “There are 5 protoxylem strands 
in Drimys winteri that apparently represent axial 
bundles ....” Both of these conjectures cannot be 
related to the present observations. In Takhtajania, 
only leaf traces can be discerned when following 
vasculature through 4—5 nodes. There are no sets 
of 5 protoxylem strands at any leve 

Beck et al. (1982: 795) also stated that, while 
there is much “variation in the mode of lateral trace 
production in D. winteri, one lateral trace typically 
diverges from the same axial bundle as the median 
trace, the other lateral may, ultimately arise from 


way. In a similar line of reasoning, Howard (1974 
noted that some workers refer to cauline bundles 
as those that persist in the stele through several 
nodes. This definition appears somewhat arbitrary 
since leaf traces, followed through several nodes, 
are still leaf traces, at least in these ranalian genera 
with open vasculature. 

Nothing resembling the above scenario occurs in 
Takhtajania, nor does it agree with Benzing (1967). 
In that work, it can be seen that the essence of 
vascular sympodia is that all so-called cauline bun- 
dles are leaf traces, and the number of nodes along 
the way between a bundle’s origin and its orienta- 
tion into a petiole base does not change this defi- 
nition. It would help if trace architecture of the 
ranalian shoot were interpreted consistently in 
terms of the primary vascular system in order to 
avoid artificial concepts and terminology (Esau, 


- 


The present observations of a pair of traces re- 
lated to the median gap are a first report for Win- 
teraceae, nearly fulfilling Takhtajan's (1964, 1969, 
1991) conjecture of a hypothetical nodal type. 
which he believed to be ancestral for the angio- 
sperms. But Takhtajan's prediction of a 1:2:1 set 
of traces varies from Takhtajania's potential for 
having pairs at the lateral gaps also (2:2:2, 1:2: 
2). Swamy (1949) demonstrated pairs of bundles 
present in the cotyledonary node of Degeneria (1: 
2:1) and Magnolia grandiflora L. (pairs of median 
bundles and split laterals), but those taxa have mul- 
tilacunar mature nodes. Observations by Marsden 
and Bailey (1955) on Clerodendrum, and by Bailey 
and Swamy (1949) and Dickison and Endress 
(1983) on Austrobaileya described double leaf trac- 
es where the components arose from independent 
portions of the eustele, a condition not found in 
Takhtajania. Other observations by Canright (1955) 
on Magnoliaceae and by Ozenda (1949) on Liriod- 


344 


Annals of the 
Missouri Botanical Garden 


endron, Uvaria, and Anona described three-trace or 
multitrace nodes where the median traces bifurcate 
or branch, tendencies also absent in Takhtajania. 

In Sugiyama’s (1979) broad study of Magnoliales 
nodal anatomy, Drimys (Tasmannia) piperita was di- 
agrammed. Its three-trace node has a median trace 
that produces two small lateral traces which then 
degenerate: L-M-L => L-mMm-L => 'LL-M-L'L. In 
Takhtajania there are no aborted lateral median 
traces, but, otherwise, patterns found in Tasmannia 
are closer than those in most other genera to the 
pattern found here. If one considers multiple in- 
dependent nodal traces to о the least a 
cialized condition for the angiospe s do 
enda (1947), Sugiyama (1979), Neubauer Du 
and I, then t 
angiosperm basal lines in this character. Finally, as 
noted above, because of the different assumptions 
brought to past studies in nodal anatomy, early 
trends of specialization in the Winteraceae will re- 
main tentative until one anatomical study across 
the family generates descriptions based on one cur- 
rent paradigm. 


e Winteraceae are not close to the 


PETIOLE VASCULAR PATTERNS 


Superimposed on the basic three-trace architec- 
ture, several genera of Winteraceae develop quite 
complex petiole vasculatures. Bailey and Nast 
(1945) proposed two independent, plausible trends 
of specialization in the Drimys node: (1) three 
strands leading to numerous derivative bundles, 
and (2) three strands fusing to form a single vas- 
cular arc in the petiole. The three traces found in 
Belliolum, Bubbia, Exospermum, and Zygogynum 
divide to form three abaxial bundles and “numer- 
ous smaller bundles irregularly arranged in an ad- 
axial position" (Metcalfe, 1987: 6). This seems to 
be a development parallel with Bailey and Nast's 
(1945) first Drimys pattern. Likewise, Metcalfe 
(1987: 6) described the bundles as being “amphi- 
cribral, hippocrepiform or appearing as divided 
bundles with their xylem units facing one and 
Carlquist (1982: 281) characterized such a 
in Exospermum stipitatum (Ваш.) Tiegh. ш. 
and midribs as having “peculiar circles of bundles” 
as well as nests of sclereids in the аш both 
characters not found in Takhtaj 

Dehay and Ghestem (1969) frm distal pet- 
iole vasculature for three genera of Winteraceae. 
Exospermum, with its circular or semicircular trac- 
es in an irregular pattern, and Zygogynum, with 
numerous traces in three layers, are both quite dis- 
tinct. Bubbia, with its simple V of 7 bundles in the 
petiole, as well as some species of Drimys, appears 


closest to Takhtajania. Such a pattern fits close to 
the base of Bailey and Nast's (1945) proposed 
trends for the 3-trace node. 

THE STEM 

Stem transsectional histology is quite simple. 
The uniform ground tissue of the cortex and pith is 
unexceptional. The early development of interfas- 
cicular cambium quickly produces an uninterrupt- 
ed cylinder of cambial derivatives. This was termed 
the pseudosiphonostele by Bailey and Nast (1948), 
and the condition is common in woody Ranales. On 
the pith side of this continuous cylinder, the pro- 
toxylem is circumferentially discontinuous. In 
Takhtajania, secondary xylem accumulation initial- 
ly remains limited as the young stem retains a suc- 
culence through the 1 cm diameter stage, the larg- 
est material available in this study. (See Carlquist, 
this volume, for details regarding secondary vas- 
culature development.) 

One unusual histochemical feature involves tis- 
sue lignification. In the presence of metachromatic 
dyes in combination with the highly ionic calcium 
chloride mountant (see Herr, 1992; Keating, 1996), 
tissue sections of most higher dicots and monocots 
show marked differentiation of lignified tissues from 
surrounding ground tissue and phloem. In Takhta- 
jania, the lignification process appears to be poorly 
bounded, spreading beyond the limits of xylem, fi- 
ber walls, and cuticles. Using several detection sys- 
tems (cresyl violet acetate, Toluidine blue, Schiff's 
reaction, iodine-potassium iodide, or phloroglucin- 
ol) few purely cellulosic-hemicellulosic walls were 

ound. There seems to be at least a modest amount 
of lignification on all cells except sieve elements. 


SPECIALIZATION AND RELATIONSHIPS OF 
TAKHTAJANIA 


Takhtajania lacks the complexity of histology 
found in other genera of Winteraceae that have 
been studied in detail. Its lack of cuticular orna- 
mentation and alveolar stomatal plugs, lack of 
sclereid development and sparing formation of ex- 
traxylary fibers, absence of other histological spe- 
cializations, as well as its geographic isolation sug- 
gest that the plant should 
plesiomorphic and not particularly close to other 
winteraceous genera. This is consonant with Vink's 
1988) proposed relationships. In terms of relative 
specialization, it would appear that Drimys (which 
shows quite variable structure) and Тазтапта are 
likely closest neighbors to Takhtajania. 

On the basis of their analysis of rbcL nucleotide 
sequences, Qiu et al. (1993) suggested that the 


e interpreted as 


— 


Volume 87, Number 3 
2000 


Keatin 345 


g 
Vegetative Shoot Anatomy 


Magnoliidae have five major lineages with Magno- 
liales and Nymphaeales being the most derived. 
urther, within the Magnoliales, they found Canel- 
laceae and Winteraceae appearing closely related. 
Conclusions by Wilson (1965) and Benzing (1967) 
upported recognition of similarities between these 
families based on comparative anatomy. However, 
characters in the Canellaceae are variously more 
and less specialized. Advancements include having 
distinctively different floral morphology and anat- 
omy, particularly fusions in the androecium (Wil- 
son, 1966). Vascular structure differences include 
the presence of vessels, considered more special- 


oblique to vertically oriented end walls with mul- 
tiple sieve areas (Wilson, 1965) and are much less 
specialized than those cells found in Takhtajania 
and other Winteraceae (Zahur 1959). 

Some characters are compatible but inconclu- 
sive. Sieve element size ranges are about the same 
in both families (Wilson, 1965). Observations on 
sieve-element plastids (Behnke, 1988) record the 
two families as being variable with overlapping P- 
protein types. Wilson (1965) encountered the same 
difficulties in attempting to trace the obscure ori- 
gins of nodal architecture in the young stem of Ca- 
nella such as occur in Takhtajania. Bailey and Nast 
(1945) emphasized the relative isolation of Winter- 
aceae when they noted that, while variation within 
the family is wide, there is no overlap with other 
ranalian families investigated. In this context, Ca- 
nellaceae could be most closely related. 


Literature Cited 


Bailey, 1. W. & C. С. Nast. 1944a. The comparative mor- 
phology of the Winteraceae. IV. Anatomy of the node 
and vascularization of the leaf. J. Arnold Arbor. 25: 


215-221 


. 1944b. The SS meyin oiy 
of the Winteraceae. V. Foliar Pare and sclere 
chyma. J. Arnold Arbor. 25: 342-3 

& 1945. The лы morphology 
of the Winteraneac id Summary and conclusions. J. 
Arnold Arte 26: 37-4 

. 1948. “Morphol and relationships 

of Шит, сна duds and Kadsura. I. Stem and leaf. 
J. Arnold Arbor. 

& В. 


. The morphology and 
relationships “ak шк ley 3. Arnold Arbor. 30: 
i манија anatomy of the leaf epi- 
e and some related families. 


Baranova, M. 1972. 
d in the сет 
е 447 
Beck, C. ide пик“ & С. W. Rothwell. 1982. Stelar 
and the primary vascular system of seed 
plants. Bot. Rev. 48: 691-815. 
Behnke, H.-D. 1988. Sieve-element plastids, phloem pro- 


tein, and evolution of flowering plants: Ш. Magnoliidae. 
Taxon 37: 699-732. 


tiation of sieve 
Winteraceae. Protoplasma 118: 148-1 

Benzing, D. 1967. Developmental patterns in stem 
primary xylem of woody Ranales. 2. Species with tri- 
ze 'unar and multilacunar nodes. Ame. J. Bot. 54: 813- 


bs J. M. 1973. AF cp n" characters of the 
Winteraceae. Blumea 21: 381—4 

Canright, J. E. 1955. The Eus morphology and 
relationships of the Magnoliacea Wood and nodal 
anatomy. J. paa Arbor. 36: 119-140. 

Capuron, R. 1963. Contributions à l'étude de la T de 
Мадлена Xil Présence à Madagascar d'un nouveau 
аза is (Bubbia perrieri R. Capuron) de la famille 

s Wintéracées. Adansonia n.s. 3: 373—378. 
Gace S. 1982 о stipitatum (Wintera- 
ceae): — on wood, leaves, flowers, and fruit. 
Aliso 10: 277-28 

————. 1983. cte — of Bubbia (Winteraceae) 
vas comments is t in of vessels in dicotyledons. 

Amer. J. Bot. : 578-590 

Dehay, С ` ФА. aoe 1969. Caracteres de l'appareil 

libéro-ligneux foliare we e quen Wintéracées. Bull. 
Soc. ap? France 116: 

Dickison, W. C. & P. K. Kr 1983. Ontogeny о 
stem- АМ leal vascular continuum of pt н 
Amer. J. pe с 

Endress, Р. . The pond floral development of Austro- 
dein in ien 103: 481-497. 

Erbar, C. & P. Leins. 1983. Zur Sequenz von Blütenor- 
ganen T einiger Magnoliiden. Bot. Jahrb. Syst. 103: 


965. Vascular Differentiation in Plants. Holt, 
un & Winston, New Yor 

r, J. M., Jr. 1992. New uses for calcium chloride so- 

vito as a mounting medium. Biotech. Histochem. 67: 

Hic o d J. 1977. Stratigraphy and Paleobotany of the 

Early beoe of Western 

h Dakota. Geol. Soc. Amer. Mem. 

1974. The stem-node-leaf continuum of 

the Dic PETA J. Arnold Arbor. 55: 125-173. 

Jeffrey, E. . The morphology of the central m 
inder in the nid а Trans. Roy. Canad. Inst. 
599-636. 

Keating, R. С. 1996. Anther investigations: A review of 
methods. Pp. 255-271 in W. С. D'Arcy & R. C. Keating 
л The Anther. Form, Function and Phylogeny. 

Cambridge Univ. Press, New York. 

Leroy, J.-F. 1978. Une sous-famille monotypique de Win- 
teraceae endémique а Madagascar: Les Takhtajanioi- 
deae. Adansonia n.s., 17: 383- 395. 

Marsden, M. P. Е. & I. W. Bailey. 1955. A fourth type of 
nodal anatomy in dicotyledons illustrated by Cleroden- 
dron trichotomum. J. Arnold Arbor. 36: 1—51. 

Metcalfe, C. R. Anatomy of the Dicotyledons, ed. 
2, Vol 3. T Illiciales, and Laurales. Claren- 
jn Press, es 

Neubauer, H. Uber Knotenbau und Blattgrund- 
пи be p celos Beitr. Biol. Pflanzen 56: 

–366 


35 ; 
Ozenda, P. 1947. Structure du noeud a et Mag- 
noliacées et des Annonacées. Acad. des . Paris, 


Compt. Rend. 224: 1521-1523. 


346 Annals of the 
Missouri Botanical Garden 
. 1949. Recherches sur les dicotyledones apocar- S B. С. L. 1949. Further contributions to the mor- 
e Publ. Lab. de l'École Norm. Зир., Sér. Biol. hog of the Degeneriaceae. J. Arnold Arbor. 30: 10— 


sc. П. beue Paris. 
Qiu. ҮЛ. . W. Chase, D. H. Les & C. R. Parks. 
Molec dur phylogenetics of the Magnoliidae: и. 


analyses of nucleotide sequences of the plastid gene 
rbcL. Ann. Missouri Bot. Gard. 80: 587—606. 
ТА. as. 1979. Leaf sclereids—Occurrence 


132: 


and distribution in the angiosperms. Bot. Not. 

19-324. 

Sampson, F. B. 1983. A new сз of Zygogynum (Win- 
teraceae). нов 28: 353-360 

5 Тће tees covery of a Malagasy en- 

КОТ perrieri (Winteraceae). Ann. Mis- 

ri Bo t. 7: 297—302 

Teni E. E 19 on. aves он on the phylogeny of 
the angiosperms. I. The anatomy of the node as an 
in the classific sm of angiosperms. Amer. J. Bot. 1 
303-322. 

Smith, A. C. 1945. Geographical "i a of the Win- 
teraceae. J. Arnold Arbor. 26: 

Sugiyama, M. 1979. A ыз - of the nodal 
anatomy in the Magnoliales based on the vascular sy 
tem in the node-leaf continuum. J. Fac. Sci. Univ. Tokyo 
Ш. Botany 12: 199-179, 


=. 
~ 


Ыы. А. L. 1964. Foundations of the Evolutionary 
Morphology of ik pre Akad. Nauka SSSR. Mos- 
cow каш Leningrad [in Russian]. 

1969. Flowering Plants. ien and Dispersal. 

Smithsonian Institution, Washingt ).C. 

1991. Evolutionary Trends in n Flowering Plants. 

Columbia Univ. Press, New York. 

1997. Diversity and Classification of Flowering 

Plants. Columbia Univ. Press, New York. 

Tieghem, P. van. 1900. Sur les Dicotyledones du groupe 
des Homoxylées. J. Bot. 14: 259-297, 330-36 

Vink, Taxonomy in Winteraceae. Taxon 31: 
91–698. 

Wilson, T. К. 1965. 
Canellaceae. П. Anatomy of the yo 
к bn 52: 369—378. 

The comparative morphology of the 

а "W Floral morphology and conclusions. 

Amer. J. Bot. 53: 336—343. 

Даи. M. S. 1959. Comparative study of secondary phlo- 
em of 423 species of woody е reae dn to 
85 oe Cornell Univ. Agric. Exp. Sta. Mem. 

1- 


The comparative morphology of the 
ung stem and node. 


FLORAL STRUCTURE OF 
TAKHTAJANIA AND ITS 
SYSTEMATIC POSITION IN 
WINTERACEAE' 


Peter К. Endress,? Anton Igersheim,? 
B. Sampson,? and George E. Schatz‘ 


ABSTRACT 


Floral structure of Takhtajania perrieri, the sole species of Winteraceae in Afri cal Mar gaccan was studied and 
a 


horizontal, and therefore the gynoec ium i not conform to a usual paraca 


The ovules are larger than those in other Winteraceae. 


arietal placentation. 


rp lu 
ver, the total evidence of floral features clearly shows the 


position of Takhtajania in the Winteraceae. Within the family it fits best in the Pseudowintera/Zygog ynum-clade, which 


is sister to the Tasmannia/Drimys clade 


words: 


androecium, floral anatomy, floral morphology. gynoecium, perianth, Takhtajania, Winteraceae. 


Takhtajania perrieri (Capuron) Baranova & J.-F. 
Leroy is the sole surviving species of Winteraceae 
in the Madagascan/African region. It achieved no- 
toriety because of its strange bicarpellate but uni- 
locular gynoecium, which is unique for Wintera- 
ceae, a feature that was noticed only 70 years after 
the discovery of the plant in Madagascar (Leroy, 
1977, 1978). For almost 90 years the plant was 
known only from the type collection of 1909, which 
contained only scarce floral material, and it was 
thus thought to be possibly extinct. The rediscovery 
(re-collected in 1994 and determined in 1997; 
Schatz et al., 1998) offers the possibility for de- 
tailed studies of the disputed floral structure and a 
comparison with the other genera of Winteraceae. 

Since the unusual gynoecium structure of Takh- 
tajania has puzzled botanists, it seems appropriate 
to give a short introductory survey of the previous 
interpretations. In the original description of T. per- 
rieri, the gynoecium was described as unicarpellate 
(Capuron, 1963). Because of the small floral invo- 
lucre and the apical anthers with subhorizontal the- 


cae, Capuron (1963) associated the plant with Bub- 
bia and placed it into that genus as Bubbia perrieri. 
Baranova (1972) later found that the leaf epidermis 
of the plant differed from other Winteraceae. It was 
her suggestion that it could be a separate genus 
that prompted Leroy (1977, 1978) to restudy the 
flowers (see also Leroy, 1993). To his surprise, he 
found the gynoecium to be bicarpellate, syncarpous 
but unilocular. This was at first questioned by 
Tucker and Sampson (1979), because its external 
shape scarcely differs from single carpels of some 
other Winteraceae. However, Vink (1978 
firmed its bicarpellate nature, but interpreted the 
two longitudinal furrows of the gynoecium as being 
dorsal in each carpel (because they alternate with 
d сарчаи and not lateral, as Leroy (1977) con- 

. This was later also accepted by Leroy 
жен and Deroin and Leroy (1993). 

Study of the scarce floral material of the type 
specimen concentrated on the puzzling gynoecium, 
whereas the other floral organs received less atten- 
tion. The aim of the present study is thus to provide 


con- 


т 


! We are indebted to Р. Н. Raven for coordinating the collection of Takhtajania perrieri in Cpe For material 


of other Winteraceae we thank A. M. Ju 
support during fieldwork with Winteraceae in North 


d T. F. icit P.K.E. one Ear nks B 


uncosa, P. Leins, an 
ern Queensland, and P. 


. P. M. Hyland for his 
Менон. and the late Н. 


Mackee in New Caledonia. We are indebted to G. K. Rickards for kindly кэ теш bile microscope photographs 


of a cleared gynoecium of ni a a We thank U. Jauch for support with the SEM, R. 
e cladistic analysis. F.B.S. t 


sections, and O. Nandi for assisting w 


Siegrist for microtome 
anks the School of Biological Sciences, V.U.W., 


for financial assistance. This study is uh of a project of P.K.E. Миса by the Swiss National Foundation (Nr. 3100- 
94). 


040327. 


2 Institute of Systematic Botany, University of Zurich, Zollikerstrasse 107, 8008 Zurich, Switzerland. 
.O. Box 


* School of Biological Sciences, Victoria University of Wellington 


Eur ME 


ington, New Zealand. 


+ Missouri Botanical Garden, Р.О. Box 299, St. Louis, Mou 63166-0299, U.S. 


ANN. Missouni Bor. 


GARD. 87: 347—365. 2000. 


348 Annals of the 
Missouri Botanical Garden 
Figure l. Takhtajania perrieri. Anthetic flower at female stage. Scale bar = 2 mm. 


a more detailed description of the floral morphology 
and anatomy of Takhtajania and to compare it with 
that of other Winteraceae, especially the putatively 
basal Tasmannia and Drimys 


MATERIALS AND METHODS 


Flowers fixed in FAA were used from the species 
and collections listed in Appendix 1. 

Some of the flowers were embedded in Paraplast, 
sectioned with a rotary microtome at 10 jum, ап 


mbedded in Kulzers Technovit 7100 
-hydroxyethyl methacrylate), sectioned at 6 jum 
or less, and stained with ruthenium red and tolui- 
dine blue (for details of procedure, see Igersheim 
& Endress, 1997). For scanning electron micro- 
scope (SEM) studies, the specimens were dehy- 
drated in ethanol and acetone and subsequently 
critical-point dried. The dried specimens were 
mounted on aluminium stubs and sputter-coated 
with gold. The confocal microscope photographs 
were taken by G. K. Rickards, School of Biological 


Sciences, Victoria University of Wellington, from a 


gynoecium (of a flower bud immediately before an- 
thesis) cleared in 596 KOH at 40? for approximately 
3 days and stained with ethydium bromide (1 mi- 
crogram per m 

MID аи analysis was performed with 
PAUP* vers. 3.1.1, using the heuristic search op- 
tion, Р неча TBR (tree bisection-reconnection 
branch swapping), and MULPARS (retention of all 
equally parsimonious trees) in effect. Consensus 
trees of shortest trees were gained by reweighting 
with a rescaled consistency index (CI) of original 
shortest trees. 


RESULTS 
ANTHETIC FLOWERS 


The flowers of Takhtajania are ca. 1.5-2 cm 
across and have spreading red perianth parts at an- 
thesis (Schatz et al., 1998) (Fig. 1). From the be- 
havior of the stamens (see below) they are probably 
protogynous. 


Моште 87, Митбег 3 
2000 


Endress et al. 349 


Floral Structure 


Figure 2. Takhtajania perrieri. Floral bud, transverse 


section; the numbers in the floral organs indicate whorls 


postgenitally fused in the overlapping area: marked by zig- 
zag line; whorl 7 is one-sided). Scale bar — 0.5 mm. 


NUMBER AND PHYLLOTAXIS OF FLORAL ORGANS 


Floral phyllotaxis in Winteraceae is commonly 
irregular. There have been different attempts to de- 
rive phyllotaxis of particular groups from a spiral 
or whorled pattern (see discussion). In Takhtajania 
only scarce herbarium material has previously been 
studied, and it is not surprising that in each of the 
three publications that dealt with the arrangement 
of floral organs there is a different interpretation of 
floral phyllotaxis and merosity. These include: (1) 
outer six perianth parts decussate, stamens in three 
4-merous whorls, one carpel (Capuron, 1963); (2) 
outer six perianth parts decussate, inner organs spi- 
ral, two carpels (Baranova & Leroy, in Leroy, 1978); 
(3) perianth parts in four 3-merous whorls, stamens 
in three whorls, the two outer ones 6-merous, the 
inner one 4-merous, two carpels (Vink, 1978). 

We studied transverse microtome sections of 
eight floral buds close to anthesis to determine 
phyllotaxis and organ number; an example is illus- 
trated in Figure 2. Organ position is somewhat un- 
stable. However, there is a more or less whorled 
pattern with changing merosity from the outside to 
the center of the flower. The outermost floral organs 
are in dimerous alternating whorls. Then there is a 
switch to 4-merous alternating whorls. The result is 
that after a 2-merous whorl there appear two in- 


stead of one organ in each alternating position, and 
thus there are double positions instead of single 
positions (for discussion of double organ positions 
in flowers, see Endress, 1987, 1996). Then there is 
a further switch to 5-merous whorls by an addition- 
al double position. The innermost whorl of stamens 
may be incomplete on one side because of the 
asymmetric shape of the floral apex. 

These switches in merosity do not always take 
place at the same site. All eight flowers studied 
begin with three 2-merous alternating whorls of te- 
pals, followed by a 4-merous whorl of tepals (by 
double positions on the broader sides). In most 
flowers, the next whorl of tepals is 5-merous. Then 
two 5-merous whorls of stamens follow. The third 
whorl of stamens is either also 5-merous or there 
are only three (Fig. 2) or two stamens on one side 
of the flower (because the floral apex is slightly 
asymmetric). The gynoecium is 2-merous. In two of 
the flowers studied the transition to 5-mery was 
only in the second whorl of the androecium. One 
flower had two and one had three 4-merous whorls 
of tepals, and in both of them the first whorl of 
stamens was also 4-merous. 

Thus the number of floral organs in the eight 
floral buds studied was: tepals (14—)15(-18), sta- 
mens 12-16, carpels 2. The previous counts by Ca- 
puron (1963), Baranova and Leroy (in Leroy, 1978), 
and Vink (1978) are all in this range, except that 
Vink found only 12 perianth parts. 


PERIANTH 


Winteraceae commonly have 2 (rarely 3) outer 
perianth organs that are more or less congenitally 
united and form a tight cover over the floral bud, 
which is often called the involucre or calyptra (e.g., 
Vink, 1988). The other perianth organs are free or 
more rarely some outer ones are (always postgeni- 
tally?) united. Terminology of the perianth organs 
of Winteraceae is not uniform in the previous lit- 
erature. The involucre or calyptra was variously in- 
terpreted to be made up of bracts or sepals. The 
other perianth organs were variously called tepals, 
sepals, or petals (e.g., Nast, 1944; Capuron, 1963; 
Baranova & Leroy in Leroy, 1978; Vink, 1978; 
Gottsberger et al., 1980). There is no clear distinc- 
tion between sepals and petals either in Wintera- 
ceae or in other basal angiosperms. Therefore, we 
prefer to use the term tepals for all perianth organs 
including those of the involucre (Hiepko, 1965; En- 
dress, 1996). 

In Takhtajania, the outer two tepals are congen- 
itally united to form an involucre. Since they are 
much shorter than the other tepals, they do not pro- 


350 


Annals of the 
Missouri Botanical Garden 


tect the inner organs in older buds. Already in bud 
they form a more or less horizontal platform, which 
is somewhat elongated in the plane of its two tepals. 
The next inner four tepals (two pairs) form the to- 
pographical periphery of old floral buds. The outer 
two slightly overlap the inner two. In the overlap- 
ping region they are postgenitally united for about 
half of their length by interdentation of the cuticle 
(Figs. 2, 4A, B). As the flower opens this bond rup- 
tures, and these four tepals are the first to spread. 
On their outer surface these four tepals are smooth 
and have a thick cuticle; on their inner surface they 
are papillate. All the following tepals have papillate 
surfaces. This papillate epidermis is tanniferous, as 
are the one or two cell layers below the epidermis. 
The margins of these tepals are not tanniferous; the 
cells are less vacuolate and have relatively large 
nuclei (Fig. 4C). From their appearance they seem 
to be secretory, and in microtome sections blue- 
staining secreted material is often concentrated 
around the tepal margins. This secretion was also 
observed in the field in June 1998 by G. Schatz. 
In the colored photographs of an open flower in 
Schatz et al. (1998) and in Schatz (2000 this issue), 
the red petals have white margins that correspond 
to this secretory zone. The material investigated 
contained insect (thrips?) larvae in the flower buds. 
It is uncertain whether these insects caused some 
of the secretion. Intercellular spaces in the tepals 
are not extensive. Starch is present especially 
around the vascular bundles. 

The inner tepals commonly have three vascular 
bundles at their base, whereas the outer tepals have 
more. However, there is only one vascular strand 
from each tepal joining the stele in the floral base. 


ANDROECIUM 


At anthesis the stamens have apical extrorse an- 
thers on club-shaped filaments (broadest shortly 
below the anther). Thus the thecae are basally 
spreading and almost horizontal. At the transition 
from the female to the male phase of anthesis, the 
filaments elongate and considerably thicken in the 
upper part, while the anthers shrink as they open 
(Fig. 5). As a result, the proportions of the stamens 
change considerably during anthesis. The filaments 
are more or less circular or slightly broader than 
thick in transverse section. The epidermis is slight- 
ly papillate and more or less tanniferous. Tannif- 
erous tissue also occurs in scattered patches below 
the epidermis but is largely lacking in the center 
of the filament. This tanniferous region extends up 
to the ventral surface of the anther connective. Oil 
cells are present. Cells with oxalate crystals are 


lacking. The stamens contain a single collateral 
vascular bundle. The tissue around the vascular 
bundle contains abundant starch before the fila- 
ment expands. After expansion the starch has dis- 
appeared (Fig. 6). Expansion of the filament goes 
hand in hand with cell enlargement. 


GYNOECIUM 


The gynoecium is disymmetric and club-shaped 
(Fig. 7D, E). It consists of two congenitally united 
carpels (Leroy, 1977; Vink, 1978). In young floral 
buds the tips of the two carpels can clearly be dis- 
tinguished (Fig. 8A, B). At anthesis each carpel has 
a longitudinal furrow on its dorsal side (Fig. 7A, 
D). There are stomata on the outer surface (Fig. 7F). 
The gynoecium has a single locule. In the termi- 
nology of Leinfellner (1950) the entire ovary is sym- 
plicate; there is no synascidiate part at the base. 
The inner surface of the gynoecium is secluded 
from the outside by a completely postgenitally 
fused slit, which at the surface extends as a line 
between the two carpels (Figs. 3A, 9B). In the mid- 
dle this line is crossed by a more or less distinct 
transverse furrow (Fig. 7B). The entire slit is sur- 
rounded by the stigma, which forms a large convex 
cap atop the gynoecium. The stigmatic zone is more 
extended in the commissural region than in the me- 
dian plane of the carpels (Fig. 7D, E). The broadest 
part of the gynoecium, the ovary, is slightly above 
mid-length. Above the ovary there is a massive part 
with the common pollen tube transmitting tract 
(Figs. 3A-D, 9A—E). Below the ovary is a relatively 
long solid base, the common stipe of the two carpels 
(Fig. 3I-K). Although the ovary is unilocular, the 
placentae of the two carpels are obliquely directed; 
they meet in the center of the locule (Fig. 7G). They 
form an arch, which is deepest in the middle (Fig. 


Each carpel has a dorsal vascular bundle, which 
extends up to some distance above the locule, 
where it may branch (Figs. 3A, 7G). The branches 
end about halfway between the level of the placen- 
tae and the stigma. Each carpel also has two or 
more lateral bundles, which are sometimes separate 
from the floral base (Figs. 3D-K, 7G). They flank 
the placentae and serve the ovules. They are con- 
nected with the dorsal bundles outside of the pla- 
centae and may also show connections between 
each other (Figs. 3C, 9C). The dorsal bundles com- 
monly have two xylem portions that are directed 
toward each other and two phloem portions directed 
away from each other, which gives the appearance 
of two bundles (Fig. 3D-K). However, they proba- 
bly originate from a single bundle. The same dou- 


Volume 87, Number 3 Endress et al. 351 
2000 Floral Structure 


Figure 3. Takhtajania perrieri. Gynoeci ium at anthesis, transverse section series (d: dorsal vascular bundle, 1: lateral 


dos bundle). —A. Stigmatic zone, inner surfaces of gynoecium postgenitally fused. —B. Lowermost stigmatic zone, 
inner surfaces not fused. —C. Zone above ovary, lateral carpel vascular bundles "s with dorsal bundles. —D. 
Upper placental zone. —E. Lower placental zone. —F. Ovary with all ovules. —G, H. Ovary base. —I-K. Solid base 
of gynoecium with rearrangement of vascular bundles. Scale bar = 0.5 mm 


Annals of the 
Missouri Botanical Garden 


Figure 4. 


Scale bars in A and 


ble appearance also occurs in strong lateral bun- 
dles. Thus the ovules are served primarily by the 
lateral bundles, and not by the dorsal bundles (in 
contrast to the interpretation by Deroin & Leroy, 
1993; see discussion) (Fig. 7G). 

The stigmatic surface is unicellular-papillate and 
secretory (Fig. 7C). Also the pollen tube transmit- 
ting tissue that lines the inner gynoecium surface 
down to the placentae has large, unicellular papil- 
lae (Fig. 9E). The stigmatic papillae are tanniferous 
and the several cell layers of the tissue below them 
still more so (Fig. 9A, B). Below the region of the 
pollen tube transmitting tissue, the inner surface of 
the ovary is lined with about two layers of tannif- 
erous cells. Throughout the gynoecium there are 
scattered tanniferous cell groups. The gynoecium 
wall also contains ethereal oil cells. Cells with ox- 
alate crystals are present. The inner layers of the 
ovary wall contain abundant starch. Stomata are 
scattered over the surface of the gynoecium, espe- 
cially in its upper part. Stone cells were not found 
in the gynoecium. 

In 31 flowers studied we found (5—)6—7(-8) 
ovules per gynoecium (1 with 5, 9 with 6, 16 with 
7, 5 with 8 ovules). Capuron (1963) and Leroy 
(1978) mentioned 5-11 ovules for the type mate- 


> Takhtajania perrieri.—A. Transverse section of outer tepals in floral bud, showing postgenital fusion of 
overlapping area. —B. The same in higher magnification, showing interdigitation of cuticles of the postgenital fusion 
area. —C. Transverse section of non-tanniferous margin area of inner [ера], surrounded by secreted material (arrow). 
" [4 
5 mm. 


С = 0.1 mm, in В = 0.0 


rial; however, in the figures by Vink (1978) and 
Deroin and Leroy (1993) there are only four ovules 
in a gynoecium. The ovules are pendent and are 
arranged in two lines. They are bitegmic, crassin- 
ucellar, and anatropous (Fig. 10). They are ca. 900 
um long. Ovule width/length ratio is 0.6. The nu- 
cellus is са. 280 рт broad. The micropyle is 
formed by the inner integument (Fig. 10). The outer 
integument is semiannular, and the inner is annular 
(Fig. 10A, B). However, of the 31 gynoecia studied 
we found two in which one of the six ovules was 
orthotropous and both integuments were annular; 
the orthotropous ovule was smaller than the other 
ones and had a long funicle. In transverse section 
the ovules are wedge-shaped because they alternate 
on both sides of the double placenta and are 
wedged together (Figs. 3F, 9F). The outer integu- 
ment is 4—5 cell layers thick, and the inner is 3 
cell layers thick. Tanniferous tissue occurs mainly 
in the periphery of the ovule (outer integument and 
raphe, especially around the vascular bundle). 


DISCUSSION 
NUMBER AND PHYLLOTAXIS OF FLORAL ORGANS 


Lability of floral organ number and phyllotaxis, 
and the tendency toward some irregularity and 


Моште 87, Митбег 3 
2000 


Endress et al. 
Floral Structure 


е 5. Takhtajania perrieri. Inner stamen in female and male phase of anthesis; in the male phase the filament 


is inflated and the anthers are open and shrunken; each pair of pari is at the s 
id D. From above. —( 


—A. Female phase. —B. Male phase. C, 
0.5 mm 


asymmetry, is common in flowers of Winteraceae 
and is also present in Takhtajania. Floral phyllo- 
taxis seems to be predominantly irregularly whorled 
in Winteraceae (Vink, 1970, =a 1978, 1985, 
1993b; E , 1986, 1987 om the record in 


the literature, Dies winteri seems to be ап inter- 


ndress 


esting exception with more or less regular spiral 
floral phyllotaxis (Hiepko, 1966; Erbar & Leins, 
19 wever, Ronse Decraene and Smets (1998) 
mentioned chaotic floral phyllotaxis also for Drimys 
winteri. Doust (1997) shed light on this seeming 
contradiction by his observation that terminal flow- 
ers in Drimys winteri have a more or less spiral 
pattern, while lateral flowers have more chaotic pat- 
terns due to initial asymmetries of the floral apex 


(see also Vink, 1970); Doust (1997) also found that 


ame magnification. A, B. Frc m dorsal 
. Female phase. —D. Male phase. Sc a bars = 


terminal flowers have more floral organs than lateral 


2 


nes. 

Notwithstanding this irregularity, there is a most 
common pattern within the family: The outer floral 
organs are decussate. Then there is a change to 4- 
merous alternating whorls and sometimes, by ad- 
ditional double positions, to whorls with a higher 
number of organs. This was reported to be common 
for Pseudowintera (less common іп Tasmannia) 
Vink, 1970), and for Zygogynum (Vink, 1977, 
1983, 1988). As shown here, it is also present in 
Takhtajania. 


— 


PERIANTH 
In Takhtajania the two outermost, congenitally 
united tepals, which form the involucre, are much 


354 


Annals of the 
Missouri Botanical Garden 


$. 
SS 4 


eet 


EV 
vedere 


Takhtajania perrieri. Transverse sections of filaments of inner stamens immediately below anthers, i 


e 6. 
female and male phase of ant 


was present in the female phase, has disappeared; each pair of figures is at the А 
hase. C, D. Central part of A and B at higher magnification. —C. 


transverse section. —А. Female phase. —B. Male p 


n 
esis; in the male phase the filament is inflated by cell enlargement, and starch, which 


same magnification. A, B. Entire 


Female phase (note starch grains in tissue around vascular bundle: dark dots). —D. Male phase (starch grains no 
l 


longer present). Scale bars in A and B 


shorter than in Drimys and Tasmannia, where they 
are protective organs for the buds. In Drimys and 
Tasmannia the involucre encloses the other floral 
parts until the flower opens, while in Zygogynum 
and Pseudowintera it ruptures very early and the 
next inner tepals attain a protective function (Vink, 
1988). The next inner whorl of tepals is postgenitally 
united in Takhtajania. Such union is also reported 
for some Zygogynum species but not for other Win- 
teraceae (Vink, 1985, 1988; although without indi- 
cation whether it is postgenital or congenital). We 
did not find tepals with secretory margins in Drimys 


= 0.5 mm, in С and D 


= 0.1 mm. 


and Tasmannia. In addition, the epidermis was tan- 
niferous and not papillate in those genera. Tepals 
are white in Drimys and Tasmannia because of large 
intercellular spaces in the mesophyll, which form an 
optical tapetum that reflects incoming light. Takh- 
tajania, in contrast, without an optical tapetum, has 
red tepals, as do some species of Zygogynum, but 
those have much thicker tepals (see Thien, 1980; 
Vink, 19933). In addition to Takhtajania, some other 
Winteraceae contain starch in the tepals, which, at 
least in some, may provide food for pollinators (bee- 


tles, thrips) (Pellmyr et al., 1990; Thien et al., 1990). 


Volume 87, Number 3 Endress et al. 355 
2000 Floral Structure 


ы 
3 


~ 
1 


Figure 7. cin двм perrieri. A-C. Flower in fe pin Line of anthesis. —A. Flower from the side, tepals broken 
off. s From above. —C. Stigmatic ат with sect D-F. Gynoecium, from the side, shortly after anthesis. 
—D. wing ех side of one of the two carpe ls, ih Попа furr 7. Gynoecium rotated at 90°. —F. 
Masi ation of D, show wing stomata on gynoecium surface. —G. Confocal ae photograph showing vasculature (xylem) 

a gynoecium (same view as E), arrows — dorsal vascular aber en arrow-heads — lateral vascular bundles. Scale 
hie in A, В, D, E, and С = 1 mm, in C = 0.05 mm, and in F = 0.1 mm 


Annals of the 
Missouri Botanical Garden 


Figure 8. Takhtajania perrieri. Gynoec 
formation. —B. At beginning of stigma formation. Scale bars = 0.1 r 


ANDROECIUM 


Club-shaped filaments with the thecae on top as 
in Takhtajania also occur in Pseudowintera and 
Zygogynum. In Drimys and even more so in Tas- 
mannia the filaments are thinner and the thecae 
are less terminal and more lateral (Bailey & Nast, 
1943a; Sampson, 1987; Endress & Hufford, 1989; 
Endress, 1994). 

The behavior of the stamens of Takhtajania dur- 
ing anthesis with elongation and especially thick- 
ening and broadening of the filament is also char- 
acteristic for other Winteraceae. It was shown for 
Pseudowintera by Sampson (1980) and Lloyd and 
Wells (1992), and for вв бата mackeei Vink, 2. 
stipitatum, and ncheri by Carlquist (1981, 
1982, 1983). Carlquist (1982) also noted the de- 
crease in starch content in stamens of 2. stipitatum. 
He discussed filament expansion in the context of 
flower opening by pressure of the stamens. How- 
ever, we found the conspicuous filament expansion 
only after flower opening, at the transition from the 
female to the male phase and interpret it as asso- 
ciated with pollen presentation (see also Sampson, 
1980, for Pseudowintera). Loss of starch may be 
correlated with rapid cell growth in this phase, as 
also indicated by Carlquist (1982) for Zygogynum 
stipitatum. It should also be studied whether starch 
loss is here associated with scent production, as 
this often occurs in osmophores (Vogel 1990). Pell- 
myr et al. (1990) discussed the significance of floral 
scents in Winteraceae for pollination but did not 
mention the source of the scents (see also section 
on periant 

Since the thecae in Takhtajania are on top of the 
club-shaped filaments, the position of the thecae is 
highly oblique to almost horizontal (also in Pseu- 
dowintera and Zygogynum). Therefore, in trans- 


verse sections of stamens the thecae are cut 


cia of young floral buds, sid the side. —А. Before beginning of stigma 


obliquely. As a consequence, the endothecium, 
which is one-layered, may appear to be two- or 
more-layered (see Swamy, 1952, for Zygogynum 
baillonii), while in reality it is only one-layered. 


GYNOECIUM 


The gynoecium of Takhtajania is peculiar. Al- 
though it is bicarpellate and unilocular, it is not 
aracarpous in the normal sense with parietal pla- 
centation. The placentae are not vertical but 
oblique to almost horizontal. Therefore, the placen- 
tae of both carpels are separate and are only con- 
tiguous at their morphological bases. Thus, placen- 
tation is not laminar (as opposed to Leroy, 1993) 
but has a normal linear configuration. The stigma 
is topographically apical, but morphologically it 
surrounds the entire (postgenitally fused) entrance 
into the internal space of the two carpels. The stig- 
ma is not commissural either (as opposed to Leroy, 
1980, 1993), because the entire orifice is stigmatic 
and not only the lateral parts, although the stig- 
matic surface is more extended in the commissural 
than in the median region. Thus, it corresponds to 
the stigma extension of most other Winteraceae, 
which have a double-crested stigma with the crests 
confluent at both ends. 

This unique bicarpellate unilocular gynoecium of 
Takhtajania could have evolved from a unicarpel- 
late ancestor. Unicarpellate gynoecia are known 
from species of Tasmannia, Pseudowintera, and 
Zygogynum (Bubbia) (Sampson, 1963; Vink, 1970, 
1983, 1993a; Ueda, 1986). In these species, some- 
times two carpels instead of one carpel develop in 
a flower (Sampson & Kaplan, 1970). In such gy- 
noecia, the available space for two carpels is lim- 
ited so that they may form a unilocular paracarpous 
structure, as Sampson and Kaplan (1970) showed 
for Pseudowintera traversii Dandy. Furthermore, in 


Volume 87, Number 3 Endress et al. 357 
2000 Floral Structure 


Figure 9. Takhtajania perrieri. —A. у at anthesis, in approximately median longitudinal section. B-F. 
Gynoecium at anthesis, transverse section series. igmatic zone, inner “и es of gynoecium postgenitally fused. 
— С. Zone above placenta, inner surface not fus A —р. Upper zone of placenta. —E. Same in higher magnification, 


showing the papillate pollen tube Sui Bi tissue (arrows). —F. Ovary, aia the transversely sectioned wedge- 
shaped ovules. Scale bars — 0.5 


Annals of the 


358 
Missouri Botanical Garden 


Figure 10. 


inner integum ent (arrow), outer in 


Ovule in approximately median longitudinal section. Scale bars in A and 


Zygogynum two (or three?) central stigmas of a gy- 
noecium may be confluent (Z. baillonii, Vink, 

3a). A dorsal furrow in the carpels as in Takh- 
tajania also occurs in Tasmannia lanceolata (Lein- 
fellner, 1965; Vink, 1970; Leroy, 1980). The posi- 
tion of the furrows is dorsal because they alternate 
with the two placentae in the bicarpellate, syncar- 
pous gynoecium of Takhtajania. In the free carpels 
of Tasmannia, the furrow lies opposite the placenta. 
The significance of the furrows is not clear; dehis- 
cence of the mature fruits has not been reported. 
Another, more conventional hypothesis is evolution 
of the gynoecium of Takhtajania from two free car- 


pels. 

In their discussion, Deroin and Leroy (1993) 
mentioned the apical placenta of Takhtajania as 
peculiar. This differs from the gynoecium in para- 
carpous Аппопасеае, with which they made а com- 
parison. However, it is not peculiar within Winter- 
aceae, because the majority of them have "apical" 
placentae (because of their more or less horizontal 
direction). 

How does the gynoecium of Takhtajania compare 
with that in other Winteraceae (apart from its pe- 
culiar syncarpy)? Several authors published studies 


tegument semiannular. —B. 


Ma. ч 
CREE 


PI Tja 

за: 

CM ec 

"2. 
prm 


a =, 


ж эв эы» 


a. ab 
^w" ag) wp omi ыы 
са E ay) = 
i LI 
i NN “= 


лазы ТТ ИИ 
2e. 


Ф –_— 


A 
И à 


Takhtajania perrieri. Ovules at anthesis. —A. Ovule from micropylar side, micropyle formed by the 


Same, micropylar part in нк magnification. —С. 
В = 0.5 


= 0.1 mm, in С 5 mm. 


on more than one genus of the Winteraceae that 
may serve as a comparative basis for this question: 
(1) the studies by Bailey and Nast (1943b, 1945) 
and Bailey and Swamy (1951) especially focused 
on the vasculature; (2) the studies by Tucker (1959, 
1975), Tucker and Gifford (1964, 1966a, b), Samp- 
son (1963), Sampson and Kaplan (1970), and 
Sampson and Tucker (1978) concentrated on the 
morphological and anatomical development of car- 
pels, vascularization, and placentation; (3) the stud- 
ies by Leinfellner (1965, 1966a, b, 1969) primarily 
dealt with the outer and inner morphology and pla- 
centation; (4) the study by Igersheim and Endress 
(1997) focused on morphology and histology of car- 
pels and ovules in comparison with that in other 
ш and winteroids. 

infellner (1965, 1966a, b, 1969) and Tucker 
and Gifford (1966b) found an unusually high vari- 
ability of carpel shapes in Winteraceae from highly 
ascidiate to largely plicate. Non-ascidiate carpels 
as in Takhtajania are only known from Tasmannia 


(see also Frame, 1996). However, these a are not di- 
nan t h 


rectly comparable, because the non 
in Takhtajania may be caused by its рани 
while Тазтапта is apocarpous. 


Моште 87, Митбег 3 
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Endress et al. 359 


Floral Structure 


Gynoecium vasculature of Takhtajania is not dif- 
ferent from that in other Winteraceae. The ovules 
are served primarily by lateral bundles (see also 
Vink, 1978), and not predominantly by the dorsal 
bundles as contended by Deroin and Leroy (1993). 
Carpels in Winteraceae generally have a dorsal vas- 
cular bundle, which has sometimes been charac- 
terized as “double,” or there are two dorsal bun- 
dles, such as in Tasmannia (Tucker & Gifford, 
1964); in addition, there are two ventral (lateral) 
bundles associated with the placentae, which may 
merge into one bundle in the ascidiate basal part 
of the carpel. Dorsal and lateral vascular bundles 
may be connected by secondary bundles later in 
development. In a critical study Tucker (1975) 
showed that ovules are principally served by lateral 


” 


carpellary vascular bundles in species of Drimys 
and Tasmannia. In contrast, Bailey and Nast 
(1943b) had described the ovules as being vascu- 
larized partly by branches of the dorsal strands, and 
partly by anastomoses between dorsal and ventral 
strands. It seems to be a peculiarity that the ovular 
vascular strands differentiate relatively late, when 
the dorsal and ventral vascular bundles are already 
far differentiated. This is probably due to the fact 
that the ovules arise relatively late, when the car- 
pels are already relatively massive and the primary 
vasculature is relatively advanced in development. 
As a consequence, the ovular traces connect with 
secondary vascular bundles between the lateral and 
dorsal main strands that have formed later. How- 
ever, the connection with the lateral vascular bun- 
dles is still there (Tucker, 1975; see also Ueda, 
1978). Likewise, in Takhtajania, the ovules аге 
served by lateral vascular bundles or by connec- 
tions between the dorsal and lateral ones (and not 
by dorsal ones as Deroin & Leroy, 1993, de- 
scribed). 

The ovules of Takhtajania are much larger at 
anthesis than those in all other taxa of Winteraceae 
studied (see list in section “Material and Meth- 
ods”). The ovules of Takhtajania are 900 um long, 
whereas those of the other taxa investigated vary 
between 330 рт in Tasmannia insipida and 625 
um in Zygogynum baillonii. This may be correlated 
with the low number of ovules per ovary in Takh- 
tajania and the different architecture of the bicar- 
pellate unilocular ovary as compared to the ovary 
in free carpels. In morphology and histology (es- 
pecially distribution of tannins), the ovules are sim- 
ilar to those of other Winteraceae (see Strasburger, 
1905; Bhandari, 1963; Sampson, 1963; Bhandari 
& Venkataraman, 1968; de Boer & Bouman, 1974; 
Prakash et al., 1992; Imaichi et al., 1995; Iger- 
sheim & Endress, 1997; Svoma, 1998). 


SYSTEMATICS OF WINTERACEAE AND SYSTEMATIC 
POSITION OF TAKHTAJANIA 


Before recognition of Takhtajania, Tasmannia 
and Drimys were considered to be the basal 
branches in Winteraceae. Tasmannia was favored 
as the basalmost clade because of its low chromo- 
some numbers (Ehrendorfer et al., 1968) and the 
conduplicate carpels (Smith, 1969) long viewed as 
a model for an archaic carpel form (Bailey & Swa- 
my, 1951). Drimys was considered as the closest 
neighbor of Tasmannia because of many morpho- 
logical similarities. In fact, for some time Tasman- 
nia was subsumed under Drimys. However, chro- 
mosome studies by Ehrendorfer et al. 
prompted Smith (1969) to reinstate Тазтапта. The 
ITS studies by Suh et al. (1993) supported the split 
between the two genera. They also supported Tas- 
mannia as sister of the rest of the family, which has 
Drimys in the basal position, followed by Pseudow- 
intera and Zygogynum (the latter including Bubbia, 
Belliolum, and Exospermum, as proposed by Vink 
(1985) on morphological grounds; see also Vink, 
1993b). Kubitzki and Reznik (1967) found a per- 
sistent difference in leaf flavonoids between Drimys 
and Tasmannia. The isolated position of Tasman- 
nia, as well as the unity of the group Bubbia, Bel- 
liolum, Exospermum, and Zygogynum, was empha- 
sized by Williams and Harvey (1982) based on the 
leaf flavonoid patterns. However, they interpreted 
Tasmannia as the most advanced genus in the fam- 
ily. On the basis of leaf epidermis, Baranova (1972) 
considered the basal dichotomy to be between Dri- 
mys/Tasmannia and Bubbia/Belliolum/Pseudowin- 
tera/Zygogynum; further, she emphasized the iso- 
lated position of Bubbia perrieri. 

After Takhtajania was recognized as a separate 
genus, Vink (1988) explicitly proposed a basal po- 
sition in the family for it, followed by a Tasmannia/ 
Drimys clade. A basal position of Takhtajania had 
also been implied by Leroy (1978) by the erection 
of a subfamily Takhtajanioideae and later even a 
separate family Takhtajaniaceae (Leroy, 1980). 
Family status was later not accepted by other au- 
thors and was also rejected by Leroy (1993). Even 
before Takhtajania was erected as a genus and was 
still included in Bubbia as B. perrieri, Bongers 
(1973) found that alveolar material was present on 
the leaf surface of Winteraceae except for Tasman- 
nia and Bubbia perrieri. In contrast, in view of its 
very large pollen tetrads and its particular pollen 
structure (Lobreau-Callen, 1977), which may indi- 
cate polyploidy, Bubbia perrieri was considered to 
be related to Belliolum (Zygogynum) and Drimys 
rather than Tasmannia (for correlation of chromo- 


360 


Annals of the 
Missouri Botanical Garden 


Е 


—- 


Degeneria vitiensis 
Zygogynum pancheri 
Zygogynum baillonii 
Zygogynum stipitatum 
Zygogynum tieghemii 
Takhtajania perrieri 
Pseudowintera axillaris 
Pseudowintera colorata 
Drimys winteri 

Drimys confertifolia 
Drimys granadensis 
Tasmannia piperita 
Tasmannia membranea 
Tasmannia insipida 


Tasmannia lanceolata 


Figure 11. Cladogram of the representatives of Winteraceae studied, with Degeneria as са 4n sed on 13 


representative floral features, showing Takhtajania nested in the Pseudowintera/Zygogynum clade (PA 


1, heuristic 


search, TBR: consensus tree of 3 shortest trees with CI 0.765, RI 0.894, gained by reweighting m res Е CI of 25 


shortes t trees of length 29 with CI 0.621 and RI 0.788). 


Volume 87, Number 3 Endress et al. 361 
2000 Floral Structure 


Canella alba 


Takhtajania perrieri 


Pseudowintera axillaris 


Pseudowintera colorata 


Zygogynum tieghemii 


NE: Zygogynum pancheri 
5 Zygogynum baillonii 
Zygogynum stipitatum 


Drimys confertifolia 


Drimys granadensis 


Drimys winteri 


Tasmannia membranea 


Tasmannia piperita 


EIU Tasmannia insipida 
Tasmannia lanceolata 
Figure 12. Cladogram of the representatives of Winteraceae studied, with Canella as outgroup, bas sed on 13 rep- 
resentative floral features, showing Takhtajania nested in the Pseudowintera/Zygogynum clade (PAUP 3.1.1, heuristic 


search, TBR: consensus tree of 20 shortest trees with CI 0.713, RI 0.868, gained by reweighting wid rescaled СТ of 
70 shortest trees of length 30 with CI 0.600 and RI 0.769). 


Annals of the 
Missouri Botanical Garden 


some number and pollen size in Winteraceae, see 
Hotchkiss, 1955). Praglowski (1979) emphasized 
the special similarity of its pollen tetrads with those 
of Drimys. 

From the results of the present comparative 
study of flowers of Takhtajania and other Winter- 
aceae some new features come into the discussion. 
The particular club-shaped stamens and almost 
horizontal position of the thecae are shared with 
Pseudowintera and Zygogynum. The presence of an 
involucre that is short and protective only in young 
floral buds is shared with Pseudowintera and Zyg- 
ogynum. The presence of whorls of 4 or 5 tepals is 
shared with Pseudowintera and Zygogynum spe- 
cies. The fusion of the tepals following the involu- 
cre is shared with some Zygogynum species. Red 
tepal color is shared with some Zygogynum spe- 
cies. This and the large pollen grains may indicate 
that Takhtajania constitutes a clade with the Pseu- 
dowintera/Zygog ynum group and that a Tasmannia/ 
Drimys group is sister to this clade (see also fig. 
2.2 in Vink, 1988). This is also shown by a clad- 
ogram based on representative floral characters 
(Fig. 11; Appendixes 2, 3) and with Degeneria as 
an outgroup, a genus that tends to come out as 
sister group of Winteraceae in preliminary cladistic 
analyses based on gynoecium structures through all 
families of the basal angiosperms. 

Another scenario, indicated by molecular data 
(Karol et al., 2000 this issue) shows a sister rela- 
tionship of Winteraceae and Canellaceae. But even 
with Canellaceae as an outgroup, Takhtajania ap- 
pears nested in a Pseudowintera/Zygogynum clade 
in the morphological analysis (Fig. 12; Appendixes 
2, 3). If this scenario with Canellaceae sister to 
Winteraceae stands corroborated, the alternative 
view with Takhtajania basal in Winteraceae woul 
be better supported. Takhtajania shares with Ca- 
nellaceae having red flowers with whorled phyllo- 
taxis and only a short involucre of united outer te- 
pals. They also share a bicarpellate, paracarpous 
gynoecium, which, however, is different in detail. 
Canellaceae have vertical parietal placentae and 
campylotropous ovules with zig-zag micropyle, but 
Takhtajania has obliquely horizontal separate pla- 
centae that meet at their morphological base, and 
anatropous ovules with micropyle formed by the in- 
ner integument; see Igersheim & Endress (1997). 
Thus the paracarpous gynoecium is unlikely to be 
a synapomorphy for Takhtajania and Canellaceae. 
In addition to molecular studies, the karyotype of 
Takhtajania will be crucial in a phylogenetic inter- 
pretation. 


Literature Cited 


Bailey, 1. W. & C. С. Nast. 1943a. The comparative mor- 
phology of the Winteraceae. 1. Pollen and stamens. Ј. 
qe Arbor. 24: Pert 346. 

& 1 The comparative morphology 

of the Winterie 'eae. | Carpels. J. Arnold Arbor. 24: 

2-481 


& The comparative morphology 

of the Winteraceae. VII. Summary and conclusions. J 

= bus 26: 37-47. 

Swamy. 1951. The conduplicate саг- 

E "s c ls and its initial trends of specializa- 

. Amer. J. Bot. 38: 373-379. 

Baranova. M. 1972. Systematic anatomy of the leaf epi- 
dermis in the Magnoliaceae and some related families. 
Taxon 21: 447—469. 

Bhandari, N. N. 1963. Embryology of Pseudowintera co- 
lorata—A vesselless dicotyledon. Phytomorphology 13: 


303-316 


Venkataraman. 1968. а of Dri- 
mys winteri. J. Arnold Arbor. 49: 509—: 
„= R. де & Е. Bouman. 1974. oe M studies 
n the Polycarpicae. Ш. Drimys winteri (Winteraceae). 
ree Bot. Neerl. 23: 19-27. 
1973. Epidermal leaf characters of the 

'Taceae. s Blames 21: 381-411. 

Capuron, R. 1963. Présence à Madagascar d'un nouveau 
représentant (Bubbia же К. Capuron) de la famille 
des Wintéracées. Adansonia, n.s., 3: 373-378. 

Carlquist, S. 1981. Wood pee nes of a aa n 
teraceae); Field observation ns. Bull. Nat ist. 
Nat., Paris, Sér. 4 Adansonia: 281 -29 92 

я stipitatum (Winteraceae): 

Observations on a leaves, flowers, pollen, and fruit. 


Aliso 277 


83. «ж чаш а Belliolum (Кинане еае) 
and а note on flowering. J. Arnold Arbor. 64: 161—166 
Deroin, T. & J.-F. Sur l'interprétation de la 
vasc pde ovarienne de Takhtajania (Wintéracées). 
. R. Acad. Sci., Paris, Sci. Vie, 316: 725-729. 

Doubt, A. N. 1997. Variability and pattern in the flowers 
of the Winteraceae (Magnoliidae). Amer. J. Bot. 84(6, 
Abstract): I 

Ehrendorfer, F., Е. Krendl, E. Habeler & W. Sauer. 1968. 
и numbers and evolution in йзгй angio- 
sperms. Taxon 17: 337—353. 

Endress, P. K. 1986. Reproductive structures and phy- 
logenetic d ance of extant primitive angiosperms. 
Р]. Syst. Evol. –28. 

a Floral Pape and floral evolution. 

Bot. Jahrb Syst. 108: 417—438. 

994. 


Shapes, sizes and evolutionary ape in 
ша of Magnoliidae. Bot. Jahrb. Syst. 115: 429— 
4 


. Diversity and Evolutionary Biology of 
Topical Flowers, ed. 2. Cambridge Univ. Press, Cam- 
bridge 
& L. D. Hufford. 1989. The diversity of stamen 
structures and dehiscence patterns among Magnoliidae. 
Bot. J. Linn. Soc. 100: apa 
Erbar, C. & P. Leins. 1983. 
ganen bei einigen es гы n. Bot. Jahrb. Syst. 
433—449. 
Frame, D. 1996. Carpel a cures in princes in- 
sipida Maia eae). Int. J. Pl. Sci. 157: 698- 
Gottsberger, G., Wes cer s ent & Е [MT 


Sequenz von Blütenor- 


103: 


Volume 87, Number 3 Endress et al. 363 
2000 Floral Structure 
dorfer. 1980. ү огон ча biology in the primitive rel- _ Pellmyr, O., L. B. Thien, С. Bergström & I. Groth. 1990. 


$ brasiliensis (Winteraceae). Pl. 


` . J. Witsoe Pp. 125-127 in G. J. 
Harden (editor), Flora of New South Wales 1. Royal 
Botanic Gardens, Sydney 

Hiepko, P. 1965. ud -morphologische und en- 
twicklungsgeschichtliche Untersuchungen über das 
Perianth bei den Polycarpicae. Bot. Jahrb. Syst. 84: 
359-508. 

966. Das Bliitendiagramm von Drimys xod 


2» Е e G. Forst. (Winteraceae). Willdenowia 4: 22 


ij A. T. 1955. Chromosome тн and RS 
tetrad size in the Winteraceae. Proc. Linn. Soc. New 
South Wales 80: 47—53. 

Igersheim, A. & P. K. Endress. 1997. Gynoecium diver- 
sity and systematics of the pum and winteroids. 
Bot. J. Linn. Soc. 124: 213-271 

Kato & H. Okada. 1995. Morphology of 

ee primitive angiosperm fam 

. J. Bot. 73: 1242-1249, 

Karol, K. G., Y. Suh, С. E. Schatz & E. A. Zimmer. 2000 
Molecular evidence for the phylogenetic position of 
Takhtajania in the Winteraceae: Inference from nuclear 
ribosomal and chloroplast gene spacer sequences. Ann. 
ue Bot. Gard. 87: 414—432. 

Kubitzki, K. & H. EE 1967. Flavonoid-Muster der 
Polycarpicae als systematisches Merkmal I. Übersicht 
über die Familien. Beitr. Biol. Pfl. 42: 445—470. 

Leinfellner, W. 1950. Der iu оу des syncarpen Gyn- 

oeciums. е Вог. 7. 97: 4 

965. Wie sind die a ae е tat- 

, Sektion 


mt. gebaut? I. Die Karpelle von Drim 

Tasmannia. Österr. Bot. Z. 112: 554—575. 

966a. Wie sind die Winteraceen- Karpelle tat- 

sitchlich gebaut? II. Über das Vorkommen einer ring- 
fórmigen Plazenta in den А von Drimys, Sektion 

adip m Bot. Z. 113: 95. 

Wie san die Taek Karpelle tat- 

c pore Ш. Die Karpelle von Bubbia, Belliol- 

um, Pseudowintera, Exospermum und Zygogynum. Os- 

terr. € 2 113: 245—264. 

69. Uber die Karpelle verschiedener Magno- 

TRUM Vil Mercer über alle Familien der Ordnung. 

Österr. Bot. Z. 117: 107-127 

977. А Бора ovary with open carpels 

> (Magnoliales): Evolutionary implica- 

tions. Science 196: 977-978 

1978. Une sous- se нац peng ae i de Win- 
teraceae endémique a Madagascar: Les Takhtajanioi- 

deae. Adansonia, n.s., 17: 383-305 

1980. Nouvelles remarques sur le genre Takh- 
солга (Winteraceae—Takhtajanioideae). Adansonia, 

n.s., 20: 9-20. 

1993. Origine et évolution des plantes à fleurs. 
TS Paris. 

Lloyd, D. G. & M. S. Wells. 1992. Reproductive biology 
of a primitive angiosperm, Pseudowintera colorata (Win- 
teraceae), and the evolution of гая systems in 
the n РІ. Syst. Evol. 181: 

Lobreau-Callen, D. 1977. Le pollen | pm perrieri 
R. Cap. Rapports palynologiques avec les autres genres 
de Wintéracées. Adansonia, n.s., 60. 

Nast, C. G. 1944. Comparative manco of Wintera- 
сеае. VI. Vascular anatomy of the flowering shoot. J. 


Arnold Arbor. 25: 454—466 


Pollination of New Caledonian Winteraceae: Opportu- 
nistic shifts or parallel radiation with their pollinators? 
|. Syst. Evol. 173: 143-157. 

Praglowski, J. 1979. Winteraceae. Pp. 1 in S. Nilsson 
(editor), World Pollen and Spore Klo a Almquist & 
Wiksell, Stockholm 

Prakash, N., A. L. Lim & F. B. Sampson. 1992. Anther 
and неми development in Tasmannia (Winteraceae). 
Austral. J. Bot. 40: т 

Е. Е. Smets. 1998. Notes оп the 

evolution of androecial organisation in the Magnolioph- 
ytina Мите Bot. Acta 111: 77-86. 

Sampson, . The floral morphology of Pseudow- 
intera, the NS dus member of the vesselless Win- 
Бы Miei, s 13: 403-423. 

—. 1980. Natural aidien. E oe (Win- 
terac Bed hg: Zealand J. Bot. 

19 ae: venation in Dis Е Blu- 


mea 32: n 

D. R. Я: 1970. Origin and development 
of the terminal carpel in Pseudowintera traversii. Amer. 
J. Bot. 57: 1185-1196 


978. a ae не in Exosper- 

mum обв А, ваи Bot. Сах. (Crawfords- 

ville) p rol 

. Williams & P. S. Woodland. 1988. The 
m dod and taxonomic 2 of Romania par 
cifolia (Winteraceae), a n 
J. Bot. 36: 395—413. 

У 2000. The rediscovery of a Malagasy en- 

She geo fene (Winteraceae). Ann. Mis- 

souri Bot yard. 8 02. 

LE * 


у A. Ramisamihantanirina. 1998. 
аа perrieri rediscovered. Nature 391: ІХ, 133- 
13: 


зтић, А. 1969. А јуни of the genus Tas- 
тапта ж + Tax 86—290. 

Strasburger, Е. 1905 ене anlage von Drimys win- 
teri umd die Е koxp bei Angiospermen. Flo- 
ra 95 и 

Suh, Y., L. B. ate on, Н. E. Reeve & E. A. Zimmer. 1993. 
Macs due and phylogenetic implic ation 
internal transcribed of r 


Syst. Evol. 209: 205-229 
: embryology 
Natl. Inst. Sci. India B 


some Annonaceae. Pl. 
Swamy, B. G. L. 1952. 
of sched bailloni. Proc. 
18: : 
Thien, j | Tue Patterns of pollination in the primitive 
angiosperms. Biotropica 12: 1—13. 
———— ге] Yatsu 


Imyr, L. Y u, G. Bergstróm & G. Mc- 


Some aspects in the em 


Pherson. jn Pulse Бане food-bodies as polli- 
nator rewards in Exospermum stipitatum and other Win- 
teraceae. Adansonia, n.s., 12: 19 


Tucker. S. C. 1959. Ontogeny of the inflorescence and 
the flower in безе winteri var. chilensis. Univ. Calif. 
Publ. Bot. 30: 257-33 

. 1975. С э, vasculature and the ovular vas- 

"mm supply in Drimys. Amer. J. Bot. 62: 191—197. 

& Е. M. Gifford, Jr. 1964. Carpel vascularization 

of rims lanl. оро 14: 197-203. 

ба. Organogenesis in the carpel- 

late flower of n еа Amer. J. Bot. 53: 433- 

442. 


Annals of the 
Missouri Botanical Garden 


—— & . 1966b. Carpel е velopment in Dri- 
mys lanceolata. Amer. J. Bot. 53: 578. 
К. B. Sampson. 1979. The gynoecium of win- 
teraceous plants. Science 203: 920-921. 


Ueda, K. 1978. Vasculature in the carpels of Belliolum 
pancheri (Winteraceae). Acta Phytotax. Geobot. 29: 
9-125. 


1986. Flora and vegetation at the s vis zone 
of Gunung Binaya and С. Owae Puku. Pp. 20-35 in M. 
Kato et al. (editors), Taxonomic Studies of the e of 
Seram Island. Botanical Gardens, University of Tokyo. 

Vink, W. 1970. The Winteraceae of the Old World. I. 
Pseudowintera and Drimys—Morphology and taxonomy. 
Blumea 18: 225-354. 

1977. The Winteraceae of the Old World П. 

23: 


лоне Morphology and taxonomy. Blumea 
219-250. 

1978. The Winteraceae of the Old World Ш. 

Notes on the ovary of Takhtajania. Blumea 24: 521— 


—— The Winteraceae of the Old World E The 
ттар species of Bubbia. Blumea 28: 311— 
1985. The Winteraceae of the Old World ~ Ex- 
ospermum links Bubbia to Zygogynum. Blumea 31: 39— 


. 1988. Taxonomy in Winteraceae. Taxon 37: 691— 
698. 


. 1993a. Winteraceae. Pp. 90-171 in P. Morat & 

H. S. Mac dp (editors), Flore de la Nouvelle-Calédonie 

19. Muséum National d'Histoire Naturelle, гы 8. 

19935. Winteraceae. Pp. 630—638 i Ku- 
еј. J. С. Rohwer & У. Bittrich (editors), The Fami- 
lies and Genera of Vascular Plants II. Springer, Berlin. 

Vogel, 5. 1990. The Role of Scent Glands in Pollination 
(translated by 5. 5. Renner). Smithsonian Institution, 

Washington, D.C. 
Williams, C. A. & W. J. Harvey. 1982. Leaf ioi 


patterns in the Winteraceae. Phytochemistry 21: 
337. 


A 


Appendix L ~ (FAA) specimens examined 
(specimens are housed at 

Drimys confertifolia Phil. Chile. Juan Fernandez Islands, 
Masatierra, Т. F. Stuessy et al. 5474. 

Drimys vip cam L.f. Costa Rica. Volcan Poas, P. K. 
Endress 

Drimys wine . & G. Forst. Switzerland. Cult. Isole 

i о, P. К. usn 6524. 

пендене axillaris (J. R. & G. Forst.) Dandy. New 
Zealai ‚ К. Endress 62 

Preudowiniera colorata (Raoul) Dandy. 0 Cult. 
pie anic Garden, University of Bonn, P К. Endress 


n perrieri (Capuron) Baranova & J.-F. Leroy. 
Madagascar. Anjahanaribe-Sud RS., P. J. Rakotomalaza 
et al. 1342, 13 VI 1997; G. E. Schatz ul VI 1998 
(Fig. 1); D. Ravelonarivo s.n., Ш 1998 (Fi 

Tasmannia insipida R.Br. ex DC. Авне а, a 
Queensland, P. K. Endress 4294. 

Tasmannia lanceolata (Poir. А. C. Sm. New Zealand. 

anical атпен, page! of Canterbury, 

, 23 IX 1992. 

11.) А. С. Sm. 
Northern Queensland, Р К. Endress 4215. 

Tasmannia piperita (Hook. f.) А. С. Sm. Papua Хем Guin- 
ea, P. K. Endress 4137. 


Australia. 


Zygogynum baillonii Tiegh. New Caledonia, P. K. Endress 
6295. 


Zygogynum е (Baill.) Vink. New Caledonia, P. К. 
Endress 6251 

Zygogynum xipitatum Baill. New Caledonia, A. M. Jun- 
COSA S.N., 1. 

Zygogynum tieghemi Vink. New Caledonia, P. К. Endress 
6329. 


Appendix 2. Matrix of 13 representative floral charac- 
ters of Takhtajania pe perrieri and 13 other species out of all 
genera of Winteraceae. 


E 
e 
— 
EY 
E 
N 
> 
о 


Takhtajania perrieri 


Drimys confertifolia 


Drimys granadensis 


Drimys winteri 


P jowint axillaris 


D 4 


| int colorata 


Tasmannia insipida 


Tasmannia lanceolata 


Tasmannia membranea 


о 


e 
o 
— 


Zygogynum stipitatum 


о 
ES 


Zygogynum tieghemii 


Degeneria vitiensis 


ыыы ОО О |- | - м (мм 
о о о-о Оо ооо ооо о/о ~ 


— 
ОО ООо о - |- - оо |- | - що 


Canella alba 


ыыы мо | ооо | — 


о о | | |- |- | ојојојо|- | ооо 
= |-|- |- |- =|= | -_ о ојо | ~ | |- |- |- 
о |“ | | | -" оојо ојојојојојо о 
о | о | |- |- | || о јо |- |- |- о 
| јојојо---ојоојоојоојојо|- 
о -јо|-|- | 5|- ојојо|- |- |о|- о |- 
О |- |- |р | | 5 |о |- |- |- |- |- | - ојо | 


< | - | --5 | - |О | -| | О | | О | О | ~ | = | ~ | ЦД | | 
о = оо go ——|—|—|—|oioooci— 
раҳ 


— 


Volume 87, Number 3 
2000 


Endress 


et al. 


Floral Structure 


— 


~ 


o 


н> 


л 


Appendix 3. Characters used for cladistic analysis. 


Тера! number (involucre not counted) and arrange- 
ment: (0) 0-3, whorled; (1) 4 or more, whorled; (2) 


. Involucre (congenitally united outermost tepals) in 
advanced floral buds (Vink, 1993b): (0) shorter than 
other tepals; (1) as long as or longer than other tepals. 

. Postgenital (2) union of the tepals реше: the in- 

volucre (Vink, 1993a): (0) absent; (1) pres 

Tepal color ees 1990; Vink, 1993a): T white; 

(1) cream or ге 

Theca position on the filament (Sampson, 1987): (0) 

+ vertical; (1) + horizontal. 


Pollen diameter (polar axis) (Praglowski, 1979): (0) 
18 um or less; um or more. 
rpel number per flower Mes а Sampson et 


Т. 
a 71988): (0) 3 or more; (1) 1 


Сезе crystals (including Papi is ovary wall: (0) 
absent; (1) present. 


. Sclereids in ovary wall: (0) absent; (1) present. 
. Ovule number per carpel: (0) 8 or less; (1) 9 or more. 
3 ule oh at anthesis: (0) 600 рт or less; (1 


) more 


А Оше: гене. number of cell layers commonly 


— (0) 3 or less; (1) more than 3 


ба ear number of cell же commonly 
1) 3. 


3. Inner 
онуи (0) 2; (1) 


COMPARATIVE FLORAL 
ONTOGENY IN 
WINTERACEAE! 


Andrew N. Doust? 


ABSTRACT 


The flowers of Winteraceae have often been considered representative of the floral morphology of the earliest angio- 


sperms. The diversity in number a 
examine floral не processes in t 


flowers of eight species from six genera of Winter. 
light m microscopy. Re. 
Leroy, the recently rediscovered specie 


nd arrangement of the (mostly) free floral organs 
s basal angiosperm family. сш ontogeny and morphology of 

e were examined using s 

sults were compared with (he. floral morphology of Takhtajania perrieri (Capuron) Вагапоуа & J-F. 
's of Winteraceae from Madagascar. The 


ives an opportunity to critically 
'anning electron microscopy (SEM) and 


analysis showed that the basic floral 


ао in the family is one of varying numbers of decussate organs followed by whorls with four (ог more) organs. 


Spiral and irregular о also occur, with i 
primordia on an me 
relative to the aie ae bract of the 


ral meristem, producing a anne calyptra bearing the sepal 
туз) ог ruptures while the bud is still young (Zygogyr num, Беира, Bubbia, 

ИЛА aid Takhtajania). "Differences in the position of the sepals relative to the subtending bract and the 
of the petals relative to the sepals create sie ‘es in floral architecture between taxa. Th 


the flo 

until anthesis (Tasmannia anc 

positio 

support + Бете 

separate from Bubbia. = 

found throughout the fan 
Key words: 


кы Drimys being considered sep 


rregularities in floral organ xe RR often the result of initiation of 
rical floral meristem. "The irst organs initiat ed a E pair o 


ese differences 


and Zygogynum and Ех xospermum being considered 


decussate and whorled floral arc оез of Takhtajania perrieri reflects the basic pattern 


floral ин т floral ontogeny, Takhtajania, Winteraceae. 


Winteraceae have long been considered to be 
one of the basal angiosperm families and have 
played an important role in our understanding of 
the evolutionary history of angiosperms (B в, 
1915; Bailey & Nast, 1945; Cronquist, 1988). Thei 
importance can be traced t 
suite of morphological character states that have 
been presumed to be primitive for flowering plants. 
Foremost among these has been a simple flower 


the possession of a 


construction of many free parts and the occurrence, 
at least in some taxa, of conduplicate carpels (Bai- 
ley & Nast, 1943). Conduplicate carpels reflect the 
theory that angiosperms evolved carpels by the 
folding of an ovule-bearing leaf (Bessey, 1915; Bai- 
ley & Nast, 1945; Cronquist, 1988). Tubular (as- 
cidiate) carpels can also be found in Winteraceae 
(Tucker, 1959; Leinfellner, 1965, 1966; Frame-Pur- 
guy, 1996). 

In recent angiosperm phylogenies Winteraceae 
are one of the first branching lineages (Qiu et al., 
1993; Nandi et al., 1998), and the floral morphol- 


ogy of the family possesses many characteristics 


that may elucidate the features of floral evolution 
in basal angiosperms. The recent rediscovery of the 
only Winteraceae from Madagascar, Takhtajania 
perrieri (Schatz et al., 1998), allows examination of 
all extant genera in the family. 

Other papers in this issue of the Annals discuss 
the phylogenetic placement of Takhtajania within 
etail its floral and vegetative 
morphology. Its floral morphology is unusual in the 
family because of the unique syncarpellate gynoe- 
cium (Leroy, 1977), and recent collections have al- 
lowed the structure of the gynoecium to be com- 


Winteraceae and 


prehensively examined for the first time (Endress 
et al., 2000). As a complement to the detailed anal- 
ysis of Takhtajania (other papers, this issue), floral 
morphology and ontogeny in Zygogynum bailloni, 
Bubbia howeana, Pseudowintera axillaris, Pseu- 
dowintera colorata, Tasmannia lanceolata, Tasman- 
nia xerophila, and Drimys winteri are presented. 
Analysis of the mature floral morphology of other 
species of Zygogynum, Bubbia, and Exospermum is 
also presented. 


! | thank Peter Stevens, Simon Malcomber, and Ge rope Schatz for reviewing drafts of this paper; Andrew Drinnan, 


Bruce Sampson, and Wim Vink for valuable discussions 


nd Una Smith, 


Peter Endress, and Victoria Hollowell for 


offering many useful and insightful comments. I thank Соне Schatz (МО) for the gift of buds and flowers of abs a 


perrieri, and Bru 


ruce Sampson (Victoria University of Wellington, New Zealand) for g gifts of buds and flowers of Zygogynu 
bailloni, Exospermum stipitatum, Bubbia howeana, Pseudowintera colorata 


a, and P. axillaris. 


* Department of Biology, University of Missouri-St. Louis, St. Louis, Missouri 63121, U.S.A. adoust@umsl.edu. 


ANN. Missouni Bor. Савр. 87: 366—379. 2000. 


Volume 87, Number 3 
2000 


Doust 367 
Comparative Floral Ontogeny 


Comparative analysis of ontogenetic data can 
help in assessing the possible homologies of mature 
floral structures. Data from floral ontogeny may also 
shed light on two persistent systematic questions 
within the family. One of these is whether the gen- 
era Bubbia, Belliolum, Exospermum, and Zygogyn- 
um are best regarded as separate genera or as one 
large and variable genus. Bubbia and Belliolum 
were combined by Burtt (1936). Vink (1985) com- 
bined Bubbia (including Belliolum), Exospermum, 
and Zygogynum into an expanded Zygogynum s.]., 
with the argument that the characters that separate 
the genera (degree of connation of petals and of 
carpels) were variable and overlapping between 
them. In this paper data have been recorded under 
Bubbia (including Belliolum), Exospermum, and 
Zygogynum s. str. so as to highlight possible dis- 
tinctions between them. The other systematic ques- 
tion is whether Тазтапта should be considered a 
separate genus or a section of Drimys (Smith, 1969; 
Vink, 1970, 1988; Sampson et al., 1988). In this 
paper data on Tasmannia are presented separately 
from that for Drimys. 

These results are part of a more comprehensive 
study of floral development in Winteraceae (Doust, 
1997; Doust & Drinnan, 1999; unpublished data). 
The focus in this paper is on early ontogenetic pat- 
terns, especially of sepal and petal initiation. 


MATERIALS AND METHODS 


Freshly collected buds and flowers were either 
dissected immediately or fixed in FAA (formalin, 
acetic acid, 70% ethanol, 10:5:85 v/v) and stored 
in 70% ethanol. Preparatory to dissection fixed 
buds were dehydrated in an ethanol concentration 
series (70%, 85%, 95%) and stained for greater 
visibility with 1% acid fuchsin in 95% ethanol. Ex- 
cess stain was removed from the buds by daily 
washing in 95% ethanol for three days before dis- 
section. Several changes of 100% ethanol were 
used to rid the specimens of all water before critical 
point drying. After drying, specimens were mount- 
ed on aluminum stubs and sputter-coated with gold 
before being imaged in either a Jeol 840 or Phillips 
XL30 scanning electron microscope. Herbarium 
specimens of a number of species of Zygogynum, 
Exospermum, and Bubbia were examined at MO. 
The number of buds and flowers dissected and a 
list of voucher specimens are given in Table 1. 


RESULTS 


The floral ontogeny and morphology of each spe- 
cies examined are described below. A generalized 


diagram of the pattern of floral morphology for each 
species is given in Figure 1 


ZYGOGYNUM BAILLONI 


The inflorescence of Zygogynum bailloni is one- 
to three-flowered. Both terminal and lateral flowers 
have a tubular calycine calyptra, which encircles 
the young bud but ruptures as the bud expands. 
The remains of this calyptra are thereafter persis- 
tent around the base of the flower. The calyptra 
bears two sepal tips, which are oriented at right 
angles to the subtending floral bract in lateral flow- 
ers but which are variably oriented in terminal 
flowers (Fig. 2A). The next organs to be initiated 
are two opposite decussate sets of petals, the four 
bases of which are valvate and connate while the 
tips are free and imbricate The tips of the first ini- 
tiated pair of these two pairs of petals overlap those 
of the second. The first pair of petals in lateral flow- 
ers is usually oriented parallel (rarely perpendicu- 
lar) to the orientation of the lateral sepals (Fig. 2A). 
It was not possible to discern the orientation of the 
first petal pair relative to the sepals in terminal 
flowers. The two pairs of decussate petals become 
connate basally as they grow. They may also be 
more or less fused post-genitally toward the apices 
of the petals. An adaxial groove is formed at the 
boundary between the connate regions of adjacent 
petals because the adjoining tissue is thinner than 
the petals themselves. When the petal tips are dis- 
sected away from the flower an invagination of the 
adaxial epidermis into the groove can be seen; epi- 
dermal tissue is also clearly evident when the 
boundary region is observed on the longitudinal 
edge of an individual petal (Fig. 2B, C). The four 
outer petals rupture along the boundary grooves 
when the flower expands at anthesis. 

Following the initiation and growth of the outer 
two pairs of petals, two inner tetramerous whorls of 
petals are initiated (Fig. 2B). On rare occasions 
fewer or greater numbers of petals may be found in 
these whorls. The petals in both inner whorls have 
narrow bases and grow throughout as completely 
free structures. The arrangement of the petals is 
mostly regular, although the sizes of the petal pri- 
mordia in the inner two whorls vary, apparently ac- 
cording to the space available for them. The second 
inner whorl of petal primordia is initiated almost at 
the same height on the meristem as the first inner 
whorl, and, with continued expansion of the meri- 
stem, members of the two whorls rapidly become 
indistinguishable and the petal bases appear to be 
in one whorl of eight. However, the middle whorl 
completely overlaps the inner whorl at its apex. 


Annals of the 
Missouri Botanical Garden 


Table 1. Sample size and voucher list. 


Taxon 


Sample size 


(# buds, flowers) 


Selected voucher specimens 


Zygogynum bailloni Tiegh. 


Zygogynum bicolor Tiegh. 


Zygogynum pomiferum 
Baill. 


Zygogynum veillardii Baill. 


a howeana (F. Muell.) 


iegh. 
"ies amplexicaulis 
(Parm.) Dandy 


Bubbia comptonii (E. G. 
Baker) Dandy 


Bubbia crassifolia (Baill.) 
Burtt 

Bubbia pancheri (Baill.) 
Burtt 


Exospermum stipitatum 


(Baill.) Tiegh. ex Morot 


Pseudowintera axillaris (J. 
R. & G. Forst.) Dandy 


Pseudowintera colorata 


(Raoul) Dandy 


Tasmannia lanceolata (Poir.) 
А. С. Smi 


Tasmannia xerophila (Р. 
Parm.) Gray 


Tasmannia stipitata (Vick- 
ry) A. C. Smith 


Tasmannia glaucifolia J. B. 


Williams 


30 


30 


30 


30 


30 


100 


100 


200 


50 


NEW CALEDONIA. Riviere Bleue, Dec. 1996, Doust 927 (MELU), 5 
Dec. 1967, Sampson (WELTU), 7 Dec. 1967, Sampson (WELTU). 

NEW CALEDONIA. Near top of track from Sarraméa to Plateau de 
Dogny, Dec. 1996, Doust 926 (MELU); Old lumber road S of road 
leading from Cascade de Ciu to Koindé and La Foa, 10 Dec. 1993, 
McPherson 6121 (MO). 

NEW CALEDONIA. Forested slopes of watershed of Riviere des Pi- 
rogues, N of Nouméa-Yate road, 15 July 1981, McPherson 3945 
(MO); Thy River valley, ca. 12 air km NE of Nouméa, McPherson 


3082 (MO). 

NEW CALEDONIA. South of Thio on coastal road, between Nimbo 
and Camboui Rivers, 26 Apr. 1984, McPherson 6517 (MO); road N 
of Canala to Prokoméo and Kouaoua, 20 Apr. 1983, McPherson 
5421 | (MO) Prokoméo Region, N of Canala, 28 Dec. 1983, McPher- 
son 6222 (MO). 

AUSTRALIA. Lord Howe Island: 2 Nov. 1963, Green (WELTU), 8 
Dec. 1968, Chinnock (WELTU). 

NEW CALEDONIA. Mt Panié, ca. 20 A km NW of Hienghéne, 31 
Mar. 1981, McPherson 3695 (MO); Mandjélia, above Pouébo, 30 
s 1984, McPherson 6280 (MO); Pu Mont Mandjélia, 6 Mar. 

979, MacKee 3665 ; 

NEW CALEDONIA. Ponerihouen: Mont Aoupinié, 8 Sep. 1976, 
Mackee 31875 (MO); Mt Panié, ca. 20 air km NW of Hienghéne, 
20 July 1980, McPherson 2882 (MO); along old lumber road to top 
of Mt. Me Ori, above Katrikoin, 6 Sep. 1980, McPherson 3045 
(МО). 

NEW CALEDONIA. Massif de Boulinda, са. 8 air km N of Роуа, 22 
May 1980, McPherson 2692 (MO); Mont Taom: Créte Est, MacKee 
38150 (MO). 

NEW CALEDONIA. Haute Amoa, 9 Oct. 1971, MacKee эе a 
along trail from Sarraméa toward Plateau de Dogny, 15 
McPherson 4923 (MO); Duretto 1995 (MEL). 

NEW CALEDONIA. Mt Panié, N of Hienghéne, 3 Nov. 1983, McPher- 
son 5929 (MO); Boguen River Valley, along lumber track above Ka- 
trikoin, ca. 20 air km E of Bourail, 21 Nov. 1979, McPherson 2110 
(MO): Tchamba River valley, 1 Nov. 1983, McPherson 5916 (MO). 

NEW ZEALAND. North Island: Kaitoki Waterworks Reserve, Wel- 
lington, 3 Nov. 1996, Doust 891 (MELU); Akatawara Road, Upper 
Hutt Valley, 4 Nov. 1996, Doust 692, 894, 895 (MELU); Kakanui 
Ridge Road, S. Tararuas, 4 Nov. 1996, Doust 895 (MELU). 

NEW ZEALAND. North Island: Kakanui Ridge Road, S. Tararuas, 4 
Nov. 1996, Doust 896, 897, 899 (MELU); S. Tararuas, 1995, Samp- 
son (WELTU). 

AUSTRALIA. Victoria: Bemm River, East Gippsland, 5 
Doust 832 (MELU); Mt. Macedon, 23 Dec. 1996, Doust 932 
(MELU); Mt. Donna Buang, Yarra Ranges National iet Nov. 1997, 
eni 994—1007 (MELU) 

AUSTRALIA. Victoria: Mt. Donna Buang, Yarra Ranges National 

ark, re 1997, Doust 984—993 (MELU); Lake Mtn., May 1995, 
Doust 782 (MELU). 

AUSTRALIA. New South Wales: Point Lookout, 30 Aug. 1981, 
Sampson & Foreman (NSW); Point Lookout, 15 Sep. 1985, Sampson 
& Williams (NSW). 

AUSTRALIA. New South Wales: Dilgry, 28 Nov. 1983, Williams 
(NSW); Moffat Falls, 8 Nov. 1986, Sampson (NSW). 


E 1995, 


Volume 87, Number 3 
2000 


Doust 369 


Comparative Floral Ontogeny 


Table 1. Continued. 
Sample size 
Taxon (# buds, flowers) Selected voucher specimens 
Tasmannia insipida R.Br. ex 50 AUSTRALIA. New South Wales: Bowens Creek, Blue Mtns., 29 Aug. 
DC 


Foreman (NSW 


1965, Sampson (NSW); Doyles River, 1 Sep. 1981, Sampson & 
). 


Drimys winteri J. R. & € 500 AUSTRALIA. Victoria: grounds of the University of Melbourne, near 
Forst. var. chilensis DC) Zoology building, Oct. 1998, Doust 1127-1130 (MELU) (cult.). 
A. Gray 

Takhtajania perrieri (Capu- 20 MADAGASCAR. Préfecture Antalcha: Sous-préfecture Andapa, 
ron) Baranova & J-F. Ler- Commune Bealampona, Fokontany Befingotra. Ex village Andranot- 
oy sarabe, piste vers Ambatomainty suivant une crête, le pic 


d'Anjanaharibe, Rakotomalaza 1342 (МО). 


Stamen initiation follows directly on from petal 
initiation. There are usually eight stamens in each 
whorl, stamens in the first whorl alternating with 
the whorl of eight petal bases formed from the two 
inner whorls of petals (Fig. 2D). Succeeding whorls 
of stamens alternate with each preceding whorl, 
and there are usually the same number of stamens 
in each whorl. The last whorl of organs initiated 
contains both stamens and carpels, and the larger 
size of the carpels often leads to a reduction in the 
total number of organs in this whorl (Fig. 2D). The 
carpels are connate but there are no connections 
between the stigmatic crests and thus no common 
pollen-transmitting tract (Igersheim & Endress, 
1997 


BUBBIA HOWEANA 


Each flower of Bubbia howeana has a calycine 
calyptra that protects the bud early in development 
ut which ruptures as the bud expands. The re- 
mains of the calyptra persist as remnants of tissue 
around the base of the flower. The calyptra initiates 
as two laterally placed free sepals but attains most 
of its growth by the action of an annular meristem 
underneath the sepal tips. Two outer decussate 
pairs of petals are initiated next, with the first of 
these two pairs of petals oriented perpendicularly 
to the sepal tips. The two outer decussate pairs of 
petals in the flowers of B. howeana are free 
throughout their growth, and they tightly wrap 
around the bud and protect it as it develops (Fig. 
2E). An inner whorl of petals is next initiated with 
four or rarely five petals, followed by a second 
whorl of petals and/or stamens (this whorl is shown 
as the outermost whorl of stamens in Fig. 
Three or four alternating whorls of four or five sta- 
mens are then initiated, followed by three to six 
free carpels arranged either symmetrically or more 
irregularly. 


PERIANTH PATTERN IN ZYGOGYNUM AND BUBBIA 


There is a difference in orientation of sepals and 
petals in Zygogynum bailloni and Bubbia howeana. 
In Zygogynum bailloni the lateral sepals are fol- 
lowed by a pair of lateral petals (Figs. 1A, 2A), 
whereas in Bubbia howeana the lateral sepals are 
followed by a pair of medial petals (Figs. 1B, 2E). 
These contrasting patterns of perianth morphology 


lated genus Exospermum (Vink, 1985), and the re- 
sults are given in Ta ere is a clear 
distinction in perianth pattern between Bubbia on 
the one hand and Zygogynum and Exospermum on 
the other, with lateral flowers of all species of Bub- 
bia examined having lateral sepals and medial first 
petal pair and lateral flowers of all species of Zyg- 
ogynum and Exospermum examined having lateral 
sepals and lateral first petal pair. The two groups 
of species examined also differ in petal connation, 
with species of Bubbia having free outer petals and 
species of Zygogynum and Exospermum (except Z. 
pomiferum and some specimens of E. stipitatum) 
having connate outer petals. 


PSEU AXILLARIS AND P. COLORATA 


No material of the earliest developmental stages 
for either species of Pseudowintera was examined. 
However, at a somewhat later stage the buds are 
enclosed in a calycine calyptra that has two lateral 
sepal tips. F. B. Sampson (pers. comm.) and Vink 
(1970) reported that three sepal tips may be pre- 
sent. The calyptra encloses the bud during early 


persist around the base of the flower. The pattern 
of initiation of the petals is somewhat variable, and 


370 


Annals of the 
Missouri Botanical Garden 


A. Zygogynum 
bailloni 


colorata 


МР 
Е. Тазтапта 
lanceolata (male) 


G. Tasmannia 
xerophila (male) 


I. Drimys winteri 
(terminal) 


s 88 в 


B. Bubbia howeana 


D. Takhtajania 
perrieri 


F. Tasmannia 
lanceolata (female) 


H. Tasmannia 
xerophila (female) 


Й 
, 
s 


J. Drimys winteri 
(lateral) 


нук үзүгү o ee 


“со ~ © 


Кеу: 


Моште 87, Митбег 3 
2000 


Doust 371 


Comparative Floral Ontogeny 


differences in size of the petals may indicate that 
they are initiated sequentially (Fig. 2F). However, 
the petals are often in two decussate pairs with fur- 
ther petals in a whorl inside these first four petals. 
Other patterns, such as spirals and whorls of three, 
are also occasionally seen. 


TASMANNIA LANCEOLATA AND 1. XEROPHILA 


The plants of Tasmannia are dioecious, but the 
pattern of sepal and petal initiation is the same in 
both male and female flowers (although the number 
of organs can differ). In Tasmannia lanceolata two 
sepals are initiated at the lateral poles of the mer- 
istem, followed by the initiation of two more sepals 
situated medially (adaxially and abaxially) (Fig. 3A, 
B). The abaxial sepal is at first only a line of tissue, 
but a flap develops from this region in later devel- 
opmental stages. In Tasmannia xerophila and all 
other species of Tasmannia examined (Table 1) only 
medial sepals are initiated, the adaxial sepal ini- 
tiating before the abaxial sepal (Fig. 3D, E). The 
continued growth of the calyx in both species oc- 
curs via a ring of meristematic tissue that encircles 
the floral meristem, so that the calycine calyptra 
grows as a cylinder, bearing the sepal tips at its 
apex. The calyptra encloses the bud until anthesis, 
when it ruptures and abscises as the flower ex- 

ands. 

In all species of Tasmannia petals are initiated 
soon after the sepals and alternate with them. 
Tasmannia lanceolata four petals alternate with the 
four sepals (Fig. 3B), while in other species of Tas- 
mannia two lateral petals initiate perpendicular to 
the two medially placed sepals (Fig. 3E). In all spe- 
cies further petals may be initiated alternating with 
the previous whorls. (Note that in Drimys piperita 
[Tasmannia] s.l. [as defined by Vink, 1970] stami- 


nate and pistillate flowers of the same entity [spe- 


— 
2 


cies] may have different numbers of petals [P. Е. 
Stevens, pers. comm.]). In staminate flowers in Tas- 
lanceolata and T. xerophila stamen initia- 
m the initiation of the 
petals (Fig 

may be variable, as the shape of the meristem is 
often asymmetric. A single nae carpel is the last 


organ to be initiated. In pistillate flowers of T. lan- 
ceolata a single terminal carpel is initiated after the 
initiation of the petals, while in other species a 
number of lateral carpels are usually initiated (T. 
insipida only initiates one or occasionally two car- 
pels). The position of the first four of these lateral 
carpels alternates with the positions of the medial 
sepals and lateral petals (Fig. 3F). When more than 
two petals are initiated these alternate with the 
*whorl" of the medial sepals and lateral petals in 
the same positions as illustrated for the first four 
carpels (Fig. 1H 


DRIMYS WINTERI 


The inflorescences of Drimys winteri are com- 
posed of a number of racemose uniflorescences with 
a terminal and up to eight lateral flowers. In ter- 
minal flowers the calycine calyptra initiates as a 
ring of tissue around the approximately circular flo- 
ral meristem (Fig. 4A), whereas in lateral flowers 
two sepal tips are initiated laterally with respect to 
the subtending floral bract (Fig. 4B). These sepal 
tips are subsequently borne aloft by the growth of 
a calycine ring of tissue around the floral meristem, 
so that the growth of the calyptra is similar in ter- 
minal and lateral flowers. The calyptra eventually 
encloses the developing flower, and at anthesis the 
calyptra splits into two or three segments and the 
petals unfold. The reflexed calyptra segments even- 
tually wither and abscise. 

In both terminal and lateral flowers there is a 
considerable delay between initiation of the caly- 
cine calyptra and the corolla; petal primordia ap- 
pear only when the calyptra has almost enclosed 
the floral meristem. At this stage the floral meristem 
is large and domed, and petals initiate low down 
on the flank of the dome (Fig. 4C). The initiation 
of stamens follows the petals without delay. А num- 
ber of tiers of stamens are initiated on the almost 
vertical flanks of the floral meristem. The initiation 
of carpels follows the stamens without delay and, 
as the growth of the floral meristem ceases, the car- 
pels eventually develop all over the flattened apex 
of the dome (Fig. 4D). The arrangement of the pet- 


In all 


g ottor sp 
laterally are toward the sides of the diagrams, and organs positioned medially are at the top and bottom of the diagrams. 


und, especially in 
us whorls o 


rimys, Pseudowintera, and in t 


stamens only the first two or three whorls have been shown 


and the others have been indicated by broken circles. Drawings are not drawn to scale relative to each other. 


372 Annals of the 
Missouri Botanical Garden 


gure 2. . Zygogynum bailloni: a terminal and lateral bud, with the lateral bud (left) showing the lateral 
position of both 1 | sepal пр (LS) and the first pair of outer petals (1). The second pair of outer petals, perpendicular 
in position to the first pair, is also visible (2). In the terminal bud the position of the first and second pair of outer 
petals relative to the sepal tips is not clear. Scale = 5 mm. — ailloni: a young bud with calyptra and outer four 
petals removed. The invagination of the adaxial sales rmis тр еп the cars of each of the outer four petals (OP) can 
easily be seen. The two alternating whorls of inner petals (IP1, 1Р2) ‘tes just been initiated. Scale = 200 рт. —C. 
Z. bailloni: a longitudinal view of the basal region of the od e of an outer petal showing the smooth covering of 
epidermis (E) on the adaxial side (ADX) of the petal and the broken cells of the connate portion of the petal toward 
the abaxial side (ABX). Scale = 500 jum. —D. 2. bailloni: late bud stage with calyptra and petals removed, showing 
regular whorls id stamens and the оге р irregular arrangement of the carpels as they begin to за (ош шег etal 
scars = OP, scars of first whorl of 1 Сане petals = 1Р1, scars of second whorl of inner petals = 1Р2). Scale = 50 
—E. Bubbia ae side view о a bud showing the lateral sepal tip (LS) and the first pair of outer Ра (1) опе mee 
perpendicular to the sepal tips. s small section of one of the second pair of outer petals (2) is also visible. Scale — 
1.5 mm. —F. Pseudowintera colorata: a young bud with the calycine ue removed showing the BEN Wiss 
five petals. Petals 1 and 2 (P1, P2) are es each other and petal 3 (P3) is alternate with these two. Petal 4 
is almost opposite petal 3 but s (P5) appears to be either transitional to a spiral pattern or in the first d 
of the next whorl. Scale = 100 jum 


=. 


Z. 


rg 


Volume 87, Number 3 
2000 


Doust 373 
Comparative Floral Ontogeny 


Table 2. 


Perianth pattern and petal connation in some species of Zygogynum, Exospermum, and Bubbia. These 


were observed from the fresh and herbarium specimens detailed in Table 1. Positions noted for the sepals and first 


pair of petals are relative to the subtending floral bract. 


Position of first pair of petals Petal connation 


Taxon Position of sepals 
Zygogynum bailloni lateral 
Zygogynum bicolor lateral 
Zygogynum veillardii lateral 
Zygogynum pomiferum lateral 
Exospermum stipitatum lateral 
Bubbia howeana lateral 
Bubbia amplexicaulis lateral 
Bubbia comptonii lateral 
Bubbia crassifolia lateral 
Bubbia pancheri lateral 


lateral connate 
lateral connate 
lateral connate 
lateral (mostly) free 
lateral (mostly) connate 
medial free 
medial free 
1edia free 
medial (rarely lateral) free 
medial free 


als, stamens, and carpels may either be in more or 
less regular whorls or spirals. 


TAKHTAJANIA PERRIERI 


The flower of Takhtajania perrieri has a two- 
lobed calycine calyptra with the lobes oriented lat- 
erally with respect to the subtending floral bract. 
Although no young buds at early stages were avail- 
able it is likely that the calycine calyptra initially 
encloses the bud before rupturing as the bud in- 
creases in size. Fragments of the calycine calyptra 
are persistent around the floral base in flowering 
and fruiting stages. The initiation of this calyptra 
is followed by initiation of the outer petals in two 
decussate pairs. After the initiation of the outer two 
pairs of petals an inner whorl of four petals is ini- 
tiated, the positions of the inner petals alternating 
with the positions of the four outer petals. This is 
followed by the initiation of a further tetramerous 
whorl of petals. The outer petal pairs are imbricate 
and fused to each other along their lateral overlap- 
ping edges (as shown by Endress et al., 2000), and 
the basal adaxial face of the outer petals is partially 
fused to the abaxial faces of the petals in the first 
inner whorl of petals (Fig. 4E, F). Occasionally 
more than four petals may occur in each of the 
inner two whorls of petals; sometimes this is be- 
cause the fifth petal is in the position of one of the 
stamens of the first androecial whorl. Usually two 
complete whorls of four or five stamens are initi- 
ated, as well as one or two stamens of a third whorl 
before initiation of the gynoecium. The syncarpous 
gynoecium is terminal, and its morphology is fully 
described by Endress et al. (2000). The overall 
structure of the flower comprises three sets of op- 
posite decussate whorls followed by alternating te- 
tramerous or pentamerous whorls, and ending in 
the formation of the terminal gynoecium. 


DISCUSSION 
SEPAL INITIATION 


In all species of Winteraceae the calyx forms a 
tubular calyptra, which completely encloses the 
bud for at least a short time during development. 
The presence of the calyptra appears to be a syn- 
apomorphy for Winteraceae, as neither of the likely 
outgroups of Canellaceae or Degeneriaceae (Suh et 
al., 1993; Nandi et al., 1998; Karol et al., 2000; 
Endress et al., 2000) possess such a tubular struc- 
ture enclosing the bud. 

In Drimys winteri, Tasmannia lanceolata, and T. 
xerophila the calyptra grows with and encloses the 
bud until anthesis, abcising as the flower opens. 
However, in Zygogynum bailloni, Bubbia howeana, 
Pseudowintera axillaris, and P. colorata the calyp- 
tra encloses the bud only during early development 
and is ruptured as the bud enlarges. The remnants 
of the ruptured calyptra persist around the base of 
the flower and can still be seen in flowering and 
fruiting stages. Early stages of the flowers of Takh- 
tajania perrieri were not available for study, but the 
mature flower shows remnants of the calyptra 
around the base of the flower similar to those found 
in Zygogynum, Bubbia, and Pseudowintera. This 
implies that in T. perrieri the calyptra encloses the 
bud only in early development. Other species of 
Zygogynum, Exospermum, Bubbia, and Pseudow- 
intera also have a calyptra that encloses the bud 

only during early development (Sampson, 1963; 
Vink, 1970, 1977, 1983, 1988). 

The first stage in the development of the calyp- 
tra in nearly all flowers in the family is the initi- 
ation of two or more sepals. The exception is in 
the terminal flower of Drimys winteri (and possibly 
other species of Drimys as well), where the calyx 
initiates as an annular primordium surrounding 


374 Annals of the 
Missouri Botanical Garden 


Figure 3. —А. оо А two lateral и (LS) have been initiated, and the adaxial se d (ADS) can 


just be | ial вера! at this stage is is typically merely a line of tissue. Scale = 100 wm. —B. T. lanc ЕЕ 
petals (P) are being ан | in seni alternate with ihe lateral (LS) and medial Gia = ADS, abaxial = 5 
pairs of sepals. Scale = 100 jum. —С. T. lanceolata: semi-mature bud with calyptra removed showing initiation gi 
four teg (P) and a number of ма of stamens (5). A sterile carpel will ђе the last organ to be initiated. Scale = 
200 wm. —D. Тазтапта xerophila: early sepal initiation with only the adaxial sepal e and the а (М) 
visible. The position where the abaxial sepal will initiate is marked in parentheses as (ABS) . Scale 00 рт. —E. 


T. xerophila: both adaxial (ADS) and abaxial (ABS) sepals have been initiated and two petals (P ш пом ђееп 
initiated in lateral positions, alternate with the sepals. Scale = 100 рт. —F. T. xerophila: a young bud with the sepals 
removed, showing four carpels (С) that have = Sua: in positions alternate with the medial sepals and lateral 
petals (P). Note the tilted meristem (M). Scale 


the floral meristem. In the lateral flowers of D. annular meristem. The initiation of sepals lateral 
winteri two sepal tips lateral to the subtending flo- — to the subtending floral bract is also seen in lateral 
ral bract are commonly initiated before the bulk flowers of Zygogynum, Exospermum, Bubbia, and 
of the growth of the calyptra takes place via an Pseudowintera. A calyptra also forms in terminal 


Volume 87, Number 3 Doust 
Comparative Floral Ontogeny 


BSE я м. 


NE winteri: a terminal flower bud at the stage of early calyx initiation showing the annular calyx 
А hue (CX). Scale = 100 —B. D. winteri: a lateral flower bud showing the two lateral sepals (LS). Scale — 
00 um. —C. D. winteri: a ny bud with the calyx removed showing the initiation of көч primordia (Р), with the 
primordia being in no particular relationship to the arrangement of the sepals. Scale = 1 m. —D. D. winteri: a 
late stage bud with the calyptra removed, showing the initiation of stamens and —€— Petals (P), stamens (S), and 
carpels (C) in this flower are arranged in a spiral pattern. Scale 00 шп. —E. Takhtajania poran a transverse 
section of the petal region showing fusion of two outer petals (OP) and an inner petal (IP). Scale = 50 шп. —F. 
perrieri: a close-up of the om of fusion of the petals (from a different specimen from Fig. 4E) showing interdisitation 


of the cuticles. Scale — 


buds and flowers of Takhtajania perrieri indicates 
that two lateral sepals are initiated. Thus the pat- 
tern of initiation of the sepals has three states; two 
lateral sepals, two medial sepals, or two lateral 
sepals followed by two medial sepals. By itself, 


flowers in these genera, but here the orientation 
of the sepals is unclear. In Tasmannia lanceolata 
lateral sepals are initiated as well as a further pair 
of medial sepals. In T. xerophila only two medial 
sepals are initiated. Inspection of semi-mature 


376 


Annals of the 
Missouri Botanical Garden 


the pattern of initiation allies Drimys with Takh- 
tajania, Zygogynum, Bubbia, and Pseudowintera, 
rather than with Тазтапта. The pattern shown by 
Tasmannia lanceolata (two lateral sepals and two 
medial sepals) is intermediate between the flowers 
of other Tasmannia species and those of the rest 
of the genera in the family. 

The pattern of initiation of the sepals in Tas- 
mannia and Drimys may help decide whether these 
two taxa should be considered as one genus or two 
(Smith, 1943a, b; Vink, 1988; Sampson et al., 
1988). Smith (1969) segregated Tasmannia as a 
separate genus from Drimys because of differences 
in chromosome number (Тазтапта, n = 13, Dri- 
mys, n = 43; Ehrendorfer et al., 1968), and sexu- 
ality of the flowers (Drimys is bisexual, Tasmannia 
is unisexual and dioecious). The two genera also 
differ markedly in flavonoid composition (Williams 
& Harvey, . However, Vink 1988, 
1993) disagreed with the segregation of Tasmannia, 
and recombined it with Drimys, citing the calyptra 
that protects the bud until anthesis, the monopodial 
construction of both Tasmannia and Drimys, and 
the observation that sporadic plants of Tasmannia 
have bisexual flowers. In the molecular phylogenies 
presented by Suh et al. (1993) and Karol et al. 
2000) Tasmannia and Drimys do not appear as sis- 
ter taxa, but as sequentially branching lineages on 
the tree. The differences in position of sepal initi- 
ation between the two genera supports the branch- 
ing pattern of the molecular phylogenies, and is 
consistent with Тазтапта being a separate genus 
from Drimys. This implies that the possession of a 
calyptra that encloses the bud until anthesis may 
have evolved independently in these two genera. 


FLORAL ORGAN ARRANGEMENT 


Flowers in Winteraceae generally have organs 
arranged in pairs and whorls. In most cases the 
pattern in the flowers is determined by the position 
of the sepals, and, in general, succeeding pairs or 
whorls of organs alternate with the previous pair or 
whorl. This is the case in Bubbia and Takhtajania, 
where a lateral pair of sepals is followed by medial 
and lateral pairs of petals (Table 2; Figs. 1B, D, 
2E). A number of whorls of petals, stamens, and 
carpels are then initiated. A similar situation oc- 
curs in Zygogynum and Exospermum, except that 
the sepals and the first pair of outer petals are par- 
allel with each other (they are both lateral with re- 
spect to the subtending floral bract) (Table 2; Figs. 
1A, 2A). In the large flowers of Z. bailloni the two 
innermost tetramerous whorls of petals initiate al- 
most simultaneously and act as one pseudo-whorl 


of eight petals (Fig. 2B). Succeeding whorls of eight 
stamens each alternate with the pseudo-whorl of 
eight petals (Figs. 1A, 2D). Irregularities in floral 
organ arrangement are often seen in the larger flow- 
ers of species of Zygogynum and Exospermum 
(pers. obs.; Endress, 1986). These have previously 
been explained as the result of the combination of 
asymmetry of the floral apex and the initiation of 
small primordia relative to the size of the floral 
apex (Endress, 1986). Another cause of irregular 
arrangements is that the changeover from petal to 
stamen and from stamen to carpel production can 
occur within one whorl. The different sizes of the 
primordia in a whorl can then perturb the pattern 
of primordial position 
The difference in sepal and petal initiation in 
Zygogynum and Exospermum (lateral sepals, lateral 
first pair of petals) versus that in Bubbia (lateral 
sepals, medial first pair of petals) was also noted 
by Vink (1977). This difference does not support 
the decision by Vink (1985) to combine Zygogyn- 
um s. str, Exospermum, and Bubbia into an ex- 
panded Zygogynum s.l. Vink made his decision af- 
ter observing that the main characters used to 
separate the genera (degree of connation of the pet- 
als and carpels) were variable within each genus 
and transitional between genera (Vink, 1985). Such 
variability was observed in the present study for 
species of Zygogynum (Table 2) and Exospermum. 
However, the position of the sepals and first pair of 
petals indicates that the three genera fall into two 
groups; (Zygogynum + Ехозреттит) and (Bubbia). 
his suggests that it might be advantageous to use 
floral ontogeny to reexamine the homology of petal 
and carpel connation in the different genera. 


А basic pattern of sets of decussate and whorled 
organs is also found in flowers of Tasmannia. There 
are two distinct variations in this basic pattern of 
floral organ arrangement within flowers of Tasman- 
nia. In Tasmannia lanceolata a lateral pair of se- 
pals is initiated followed by a medial pair. A te- 
tramerous whorl of petal primordia is then initiated, 
with the positions of the four petals alternating with 
those of the four sepals. Further petals, stamens 
male flowers), or a carpel (female flowers) are then 


жм, 


initiated. In Tasmannia xerophila two вера! рп- 
mordia are initiated medially, followed by two petal 
primordia initiated laterally. A whorl of four organs 
(which may be petals, stamens [male flowers], or 
carpels [female flowers]) are then initiated alter- 
nating with the “whorl” comprised of the pair of 
sepals and the first pair of petals. More organs can 
also be initiated. The similarity in floral organ ar- 
rangement between the flowers of these two species 
of Tasmannia is that in both, two sets of decussate 


Volume 87, Number 3 
2000 


Doust 377 


Comparative Floral Ontogeny 


organs are followed by a tetramerous whorl of or- 
gans. The differences are in the position of the first 
pair of organs (lateral sepals in Т. lanceolata vs. 
medial sepals in T. xerophila) and in the identity of 
the second pair of organs (medial sepals in T. lan- 
ceolata vs. lateral petals in T. xerophila). All other 
species of Тазтапта examined conform to the pat- 
tern shown by T. xerophila. 

The initiation of the perianth in Pseudowintera 
is consistent with the general pattern described 
above. There is usually at least one pair of decus- 
sate organs, the laterally positioned sepals, but 
there may also be one or two decussate pairs o 
petals alternate with the sepal tips. Vink (1970) and 
Е. B. Sampson (pers. comm.) reported that three 
sepal tips may also occur, along with trimerous 
whorls of petals and stamens, as illustrated by Vink 
(1970, fig. 3e). A transition to spiral arrangements 
of the stamens and carpels starting with the inner- 
most petals also has been observed. 

e floral arrangement in Drimys winteri 1s un- 
like that in the other genera because there appears 
to be no relationship between position of the lateral 
sepals or calyx and the succeeding petals. The lack 


~ 


of relationship in lateral flowers may be because 
the delay between initiation of the two organ types 
may deprive the petals of positional information 
from the sepals. There is, of course, no positional 
information given by the calyx in terminal flowers, 
as the calyx is initiated as an annular primordium 
encircling the floral meristem. There is also a va- 
riety of whorled and spiral floral organ arrange- 
ments in D. winteri, implying that petal, stamen, 
and carpel arrangement is not constrained by sepal 
position. Irregularities in the position of the pri- 
mordia have been correlated with asymmetry of the 
floral meristem (unpublished results; Erbar & 
Leins, 1983) 


AFFINITIES OF TAKHTAJANIA 


Takhtajania perrieri was originally described as 
a species of Bubbia by Capuron (1963), and its 
floral architecture is similar to that of Bubbia how- 
eana. Both species share a decussate pattern of the 
sepals and first two pairs of petals, followed by a 
whorl or whorls of petals and several whorls of sta- 
mens in fours or fives. However, the basic pattern 
of decussate pairs of organs followed by whorls of 
organs has been shown to be common throughout 
the family, with exceptions being found only in the 
relationship between the sepals and the first pair 
of petals in Zygogynum (parallel to each other), the 
terminal flowers of Drimys winteri (no decussate or- 
gans), the lateral flowers of D. winteri (opposite se- 


pals followed by various arrangements of petals, 
stamens, and carpels), and some flowers of Pseu- 
dowintera spp. (opposite petals alternating with the 
sepals but further petals either in whorls or spirals). 
A basic pattern in the family of four petals alter- 
nating with the sepal tips was also postulated by 
Vink (1988). Thus the similar patterns of floral or- 
gan arrangement in Takhtajania perrieri and Bubbia 
howeana show the general relationship in the fam- 
ily rather than being evidence of a special shared 
relationship. 

Both Zygogynum bailloni and Takhtajania per- 
rieri have fused outer petals. Such fusion does not 
generally occur in the corolla of the flowers of the 
other genera in the family and may provide evi- 
dence of a shared relationship. The method of fu- 
sion of outer petals in Z. bailloni is both through 
connation in the basal regions as well as variable 
amounts of post-genital fusion in the otherwise free 
apical regions. In the species of Zygogynum that 
display basally connate petals the mode of conna- 
tion may be the result of coalescence of the margins 
of the petal primordia as they grow, whereupon the 
epidermis in the region of connation disappears. 
However, in T. perrieri the petals cohere only 
through post-genital fusion of the cuticles, the fu- 
sion occurring by the interdigitation of the cuticles 
in the overlapping petal areas (as documented by 
Endress et al., 2000). Interdigitation of the cuticles 
was also observed to occur between the bases of 
the outer petals and the basal portions of the next 
inner whorl of petals. Post-genital fusion as it oc- 
curs in T. perrieri may be likened to a cohesion of 
the petals rather than a true connation as found in 
the basal portion of the outer petals of Zygogynum. 
This makes it possible that the morphological sim- 
ilarity between the fused outer petals of the two 
species is not homologous. 


CONCLUSIONS 


The study of the ontogeny of the flowers of Win- 
teraceae reveals both unifying patterns and unex- 
pected complexity. The underlying pattern of floral 
organ arrangement is one of sets of decussate and 
whorled organs, occasionally turning to spirals in 
Drimys and Pseudowintera. Differences within the 
basic pattern include the number and position of 
sepals and the position of the petals relative to the 
sepals. In all species irregularities in floral apex 
symmetry and in the size of organs in a whorl may 
perturb the regular whorled or spiral pattern, lead- 
ing to flowers with more irregular floral organ ar- 
rangements. There is a clear morphological division 
in the family between those taxa that retain the 


378 


Annals of the 
Missouri Botanical Garden 


calycine calyptra until anthesis (Тазтапта and 
Drimys) vs. those where the calyptra is ruptured 
early in development (Bubbia, Zygogynum, Exos- 
permum, Pseudowintera, and Takhtajania). How- 
ever, differences in the position and number of se- 
pals between Tasmannia lanceolata, T. xerophila, 
and Drimys winteri suggest that the two genera 
should not be combined into one. Differences in 
the position of the first pair of petals relative to the 
sepals suggest that Zygogynum and Exospermum 
are distinct from Bubbia. The decussate and 
whorled arrangement of the floral organs in Takh- 
tajania perrieri is closest to that of species of Bub- 
bia, but this pattern is common throughout the fam- 
ily and does not serve as evidence of a special 
relationship between the two taxa. The manner of 
fusion of the outer petals in 7. perrieri is strictl 
post-genital and by interdigitation of the cuticles, 
whereas that in Z. bailloni is primarily by conna- 
tion of the basal region of the petals. These differ- 
ences do not support a close relationship between 
Takhtajania and Zygogynum. These observations 
are congruent with the molecular phylogeny of Ka- 
rol et al. (2000), where Takhtajania is basal, and 
its unique syncarpellate gynoecium autapomorphic. 

The ontogenetic data presented in this paper are 
a step toward understanding the evolution of floral 
form in Winteraceae. On the basis of robust inter- 
pretations of phylogenetic relationships within the 
family (other papers, this issue), an understanding 
of the evolution of diverse and variable floral forms 
within such a well-defined monophyletic group will 
also advance our understanding of the evolution of 
the flower in basal angiosperms. 


Literature Cited 

Bailey, 1. W. & С. С. Nast. 1943. The comparative mor- 
phology of “ з= eae. П. Carpels. J. Arnold Ar- 
bor. 24: 472 


& . The bes iui morphology 
of the Wihtteraceac. ve Summary and conclusions. J. 
Arnold Arbor. 26: 37-4 
Bessey, C. E. 1915. The „РЕЖИМ taxonomy x flow- 
ering plants. Ann. Missouri Bot. Gard. 2: 109-1 
Burtt, B. L. 1936. Bubbia ет В. L. Burtt, = 
aceae. Hooker's Icon. Pl. 34: t. 3315, 1-3. 
Capuron, R. о à Madagascar d'un 
nouveau représentant (Bubbia R. Capuron) de la famille 
gee Adansonia 3: 373-378. 
The Evolution and Classification of 
Flowering кыз 2nd ed. The New York Botanical Gar- 


, Bronx. 
Doust, A. N. L. 1997. Variability and pattern in the flow- 
ers of the Winteraceae (Magnoliidae). Amer. J. Bot. 84: 
40. arias 
& A. 


N. Drinnan. 1999. Floral form and onto- 

E enetic process in Winteraceae. XVI International Bo- 
ical Congress 917. [Abstract.] 

Ehrendorfer, F., F. Krendl, E. Habeler & W. Sauer. 1968. 


Chromosome numbers © evolution in primitive angio- 
sperms. Taxon 17: 33 
1986. 


Endress, P. K. жек ИИИ structures and phy- 
logenetic significance of extant primitive angiosperms. 
ү 152: 1-28. 


A. Igersheim, F. B. Sampson & С. E. Schatz. 
2000. Floral: structure of о and its systematic 
уел in Winteraceae . Missouri Bot. Gard. 87 
347-365. 
а С. & P. Leins. 1983. Zur Sequenz von В] епог- 
ganen bei einigen Magnoliiden. Bot. Jahrb. Syst. 103: 
3-449 


43 

Frame-Purguy. D. 1996. Carpel development in Tasman- 
nia insipida Winteraceae. Int. J. Plant Sci. 157: 698— 
702. 

› К. Endress. 


шереп, А.& 1997. Gynoecium diver- 


sity and Vier of the Magnoliales and winteroids. 
Bot. 1 Linn. 5 oe 213-271. 
Karol, K. G., G. Е. Schatz & Е. А. Zimmer. 2000. 


Molecular наши (ог the phylogenetic position of 
Takhtajania in the Winteraceae: Inference from nuclear 
Anar and eg cu gene spacer sequences. Ann. 
souri Bot. 14—432. 
| einfellner, W. Do Wie sind die Winteraceen-Karpelle 
tatsüchlich — I. Die о von Lie Sektion 
Tasmannia. s Bot. Z. 112: 5 
dnd die nate een-Karpelle tat- 
süchlic h gebaut? m Die Karpelle von Bubbia, Belliol- 
um, Pseudowintera, Exospermum und Zygogynum. Oes- 
terr. Bot. Z. 113: 245-264. 
Leroy, ]-.Е. 1977. A compound ovary with open carpels 
in Winteraceae cs noliales): Evolutionary implica- 
78. 


tions. E ience 196: 977— 

Nandi, O. L, M. W. Chase & P. K. Endress. 1998. A 
d cladistic analysis of angiosperms using rbcL 
and non-molecular data sets. Ann. Missouri Bot. Gard. 
85: 137-212 

Qiu, Y-L., M. W. Chase, D. H. Les & C. R. Parks. 1993. 
Molec ular di ако ө of the Magnoliidae: Cladistic 
analyses of nucleotide sequences of the plastid gene 
rbcL. Ann. Missouri Bot. Gard. 80: 5 587. 606. 

Затрзоп, Е. В. 1963. Тће Йога] ii d of Pseudow- 
intera, the New Zealand member o ie ao Win- 
teraceae. ш. _ 403 

—— Williams & P. р e 1988. The 

orphology and taxonomic dus of Tasmannia glau- 
cifolia (Winteraceae), a new Australian species. Austral. 


Schatz, G. E & A. Ramisamihantanirina. 
1998. Takhtajania perrieri rediscovered. Nature 391: 
133-134. 

Smith, A. С. 1943а. The American species of Drimys. J. 
Arnold eua 24: 1-33. 

= пон notes on the Old World spe- 
cies of Winterac ‘eae. J. Arnold Arbor. 24: 119-164. 

————. 1969. A rec тн of the genus Tasmannia 
(Winteraceae) Taxon 

Suh, Y., L. B. Thien, H. E. Reeve & E. A. Zimmer. 1993. 
Molec is ва and phylogenetic implications of 
internal transer 


Tucker, S 1959. Ontogeny of the inflorescence and 
flower in Drimys winteri var. chilensis. "Uni Calif. Publ. 
336. 


The Winteraceae of the Old World I. 
ys—Morphology and taxonomy. 


1970. 
Pseudowintera de m 
Blumea 18: 225 


Volume 87, Number 3 
2000 


Doust 
Comparative Floral Ontogeny 


1977. The Winteraceae of the Old World П. 
Zygogynum—Morphology and taxonomy. Blumea 23: 
219-250. 


1983. The ететш of the Old World un The 
TOE species of Bubbia. Blumea 28: 311 
—— ———. 1985. The тонне of the Old "dl v D x- 
ospermum links Bubbia to Zygogynum. Blumea 3 
55. 
1988. Taxonomy in Winteraceae. Taxon 37: 


691-698. 


T 
Ј. С. Rohwer & V. Bittrich P 
Cin of Vascular Plants, 
о Марпојпа, Наш melid and oo 
milies. Springer-Verlag, Berlin, Heidelber 
к С. А. & W. J. Harvey. 1982. 

patterns in the Winteraceae. cen 21: 
327. 


s), The Families 


993. Winteraceae. Pp. 630—638 in K. Kubitzki, 


and 


ү 2, Flowering Plants, Di- 


ява 


и flavonoid 


329- 


THE POLLEN OF 
TAKHTAJANIA PERRIERI 
(WINTERACEAE)! 


F. B. Sampson? 


ABSTRACT 


Pollen ne and ultrastructure of d qe (Capuron) Baranova & J.-F. I 


of the Winteraceae, were studied an 


Pollen is in 


permanent tetrahedral ra whic а аге йе ere in the family. As in 


eroy, the Malagasy member 
n of other members of this primitive angiosperm family. 
mys and Pseudowintera, the 
pollen tubes. 


t is concluded that previous reports of colpate e tric hotomoc colpate 
(tric бокалдан sate) apertures, based on the study of dried material from the type specimen, are incorr 


' exine 15 


reticulate and tectate-columellate and most columellae have characteristic expanded bases where bc: join E ^d 


layer. Beneath the foot layer is an endex 
teraceae. As in some other 
consisting of tangentially aligned lamellae. The se 
plugged 


but in contrast to Belliolum, Bubbia, Exosperm 


a 
tal exine is highly reduc a and s 
with endexine, suggesting that pollen initogis is asynchrono 
‚ Tasmannia, and Zygogynum, in which the 


e and an intine, which is two-layered in apertural areas, as in othe 
e tus is nig 


with an Sic pli. thickened MERE 

gaps in the ectexine are 
as in н Pseudowintera and Drimys, 
e gaps remain or are 


ous within a tetrad, 


blocked by the deposition of intine after pollen: ieee. On the basis of its pollen morphology, Takhtajania seems most 


closely related to Drimys and Pseudowintera. 


Key words: 


Bubbia perrieri, flora of Madagascar, Magnoliidae, pollen morphology, Takhtajania, tetrad pollen, Winteraceae. 


The rediscovery of Takhtajania perrieri (Schatz 
et al., 
liquid-preserved polliniferous material for the first 
time. Although there have been three quite detailed 
studies of the pollen of this plant (Straka, 1963; 
Lobreau-Callen, 1977; Praglowski, 1979), they 
were based on dried material from the type speci- 
men, collected nearly 90 years ago. These three 


1998) has provided the opportunity to study 


authors disagreed on the interpretation of some as- 
pects of the pollen of Takhtajania. 

Takhtajania pollen is in permanent tetrahedral 
tetrads, a feature that occurs in all other members 
of the ее with the exception of four зре- 
cies of ур which have monads (Sampson, 
1974; Belek. 1979; Vink, 1993). Monad pol- 
len in these Zygogynum species seems to have 
evolved from the tetrad condition (Sampson, 1981; 
Doyle et al., 1990a, b). Previous studies on the pol- 
len of Takhtajania indicated that its pollen tetrads 
are larger than those of other Winteraceae. It has 
been claimed, too, that a few apertures of Takhta- 
jania are of the trichotomosulcate type, with a 
three-slit sulcoid aperture, in contrast to the in- 
variably round or oval apertures of other members 
of the family. 


The present study was undertaken, therefore, to 
provide further information and illustrations of the 
pollen of Takhtajania based on liquid-preserved 
material. This will enable further comparisons to 
be made with the pollen of other genera of Winter- 
aceae, which were studied in detail by Praglowski 
(1979). It should be noted that the material used 
in the present study was obtained about 150 km 
southeast of the original type collection. This raises 
the slight possibility that some differences between 
pollen in previous and present studies may be the 
result, not only of dried versus preserved material, 
but also of differences between the two populations 
of Takhtajania. For as Smith (1943), Vink (1970), 
Ehrendorfer et al. (1979), and others have demon- 
strated, there can be considerable morphological 
variability between populations of some species in 
the Winteraceae. However, such differences have 
not been demonstrated for their pollen grains. Ge- 
neric names used follow those of dicas (1979), 
except that as recommended by Smith (1969) Tas 
mannia is substituted for Drimys sect. inan 
The merging of Bubbia, Belliolum, and Exosper- 
mum into Zygogynum (Vink, 1985) is therefore not 
ollowed in the present paper. 


I thank Peter Raven and George Schatz for pe о of Takhtajania Viii in bs. study. Special thanks 


are ы 'Ч to Barry Martin and Karen Reader (ог as 


with the electron micros . I wish to thank Dallas 


aet ја assistance with acetolysis of the pollen and Joka Dawson and Ilse Bi e for translating papers in 


French an 
financial assistance for the electron microscope 
? School of Biological Sciences, 


rman, respectively. The School of d 'al Sciences, Victoria University of Wellington, kindly provided 


studie 
Victoria University of Wellington, P.O. Box 600, Wellington, New Zealand. 


ANN. Missouni Bor. GARD. 87: 380—388. 2000. 


Volume 87, Number 3 Sampson 381 
2000 Pollen of Takhtajania perrieri 


Figures 1-6.  Takhtajania perrieri ee micrographs). —1. Pollen tetrad showing Bd of the ide E 
T ые pee (scale bar — 10 jum). —2. Pollen tetrad with the uppermost pollen gra n polar view and all fo 
apertural papillae visible (scale жа = 10 jum). —3. Apertural region of a pollen grain in рене view. 5 геріоп 


indicated by asterisk; white arrow shows expanded base of а columella (scale bar = 5 шт). —4. Pollen tetrad with a 
sterile pollen grain ЗИ which lacks а protuberant papilla, with the annulus surrounding the aperture in polar 
view (scale bar = 10 jum). —5. Pollen tetrad with a sterile pollen grain (lowermost) in which the aperture has become 
orn into a colpus-like SRi (scale bar = 10 шт). —6. Center of a pollen tetrad with portions of three pollen grains 
visible (scale bar = 5 ш 


382 Annals of the 
Missouri Botanical Garden 


Rise 
а ‘as 
2 ге et № oe г 
Figures 7-10. — 7. Takhtajania perrieri (SEM): a pollen tetrad with apertural regions of two pollen grains (lower 
left and upper right) in semi-lateral view (scale Баг = 10 рт). —8. Pseudowintera colorata (SEM): a pollen tetrad with 
approximately similar orientation to the previous figure (scale bar = 10 jum). —9. Takhtajania perrieri (TEM): section 
of part of a tetrad showing most of a pollen grain in non-median section passing though the external fringes of the 
annulus, with white arrows indicating the endexine, a black arrow indicating the intine, and an isk showing the 
cytoplasm of the generative cell, which has a central nucleus (scale bar = 5 рт). —10. Takhtajania perrieri (TEM): 


indicate gaps in the extexine, which have been filled with endexine (scale bar = 1 qum). 


MATERIAL AND METHODS Mature and nearly mature pollen for scanning 
- . . electron microscope (SEM) study was dehydrated 

Flowers and flower buds of Takhtajania perrieri in ап ethyl alcohol/acetone series, critical-point 
(Capuron) Baranova & J.-F. Leroy were collected dried, sputter-coated with a thin layer of gold, and 
(P. J. Rakotomalaza et al. 1342) on 13 June 1997 examined with a Philips 505 SEM. Pollen for trans- 
from the Anjanaharibe-Sud Special Reserve, An- mission electron microscope (TEM) study was post- 
tsiranana, Madagascar, and fixed in FAA. fixed in 1% OsO,, dehydrated, embedded in Spurr's 


Моште 87, Митбег 3 
2000 


Затрзоп 383 
Pollen of Takhtajania perrieri 


resin, sectioned, and stained with uranyl acetate 
and lead citrate. Some mature pollen for light mi- 
croscopy (LM) was acetolyzed and some was de- 
hydrated in a TBA series, embedded in Paraplast, 
sectioned with a rotary microtome at 10 jum, and 
stained with haematoxylin, safranin, and fast green. 
Measurements of external pollen features were 
based on LM using acetolyzed material to enable 
comparison with other genera of Winteraceae de- 
scribed in the pollen monograph of Praglowski 
(1979); 50 tetrads were sampled. 
or comparative purposes, pollen tetrads of 
Pseudowintera colorata (Raoul) Dandy, obtained 
from native forest, Brown Cow Ridge, N.W. Nelson, 
New Zealand (voucher WELTU 12949), were ex- 
amined under SEM, with treatment and fixation as 
for Takhtajania. 


RESULTS 
EXTERNAL MORPHOLOGY 


The most striking feature of the pollen tetrads of 
Takhtajania, which is shared by some other mem- 
bers of the family, is an expansion of the aperture, 
suggesting premature commencement of growth of 
the pollen tube (Figs. 1, 2). There is minor variation 
in the size of these “protuberant papillae” (Bailey 
& Nast, 1943) even between grains of the same 
tetrad, which may reflect differences in the degree 
of hydration of individual pollen grains. The surface 
of the aperture membrane is relatively smooth, in 
contrast to a surrounding verrucate (warty) annulus 
(Fig. 3). A few tetrads were observed in which one 
or more of the pollen grains did not show a bulging 
of the aperture (Fig. 4). Sections of tetrads revealed 
that such grains were sterile. Sometimes the aper- 
ture membranes of these pollen grains were torn 
(Fig. 5), with the ruptured part varying in shape. 

The tetrads are of the acalymmate type (Van 
Campo & Guinet, 1961), “in which each individual 
pollen grain is encompassed by its own ectexine 
which is usually incompletely developed in the sep- 
tal part” (Praglowski, 1979: 4), as in other Winter- 
aceae. Pollen grains are in tetrahedral tetrads, ra- 
dially symmetrical, heteropolar, semitectate, and 
rounded to rounded-triangular in polar view (Fig. 
2), uniaperturate with distal polar apertures, mostly 
round (ulcerate), although a few were oval. In lat- 
eral view, each pollen grain has a hemispherical 
distal (exposed) face (Fig. 1), excluding the shape 
of the apertural protrusion, and a proximal region, 
where the grain is joined to the other three mem- 
bers of the tetrad, consisting of three flattened in- 
clined rather triangular surfaces converging near 
the “geometrical centre” of the tetrad. Praglowski 


(1979: 4) defined this as “the point at the centre of 
the free space between four coalescing pollen 
grains of a tetrahedral tetrad.” 

Diameter of tetrads, excluding apertural protru- 
sions, which are destroyed by acetolysis, 65—83 p.m 
(mean 74 jum); polar axis of individual pollen 
grains 23-33 рт (28 pm); largest diameter per- 
pendicular to the polar axis 50-60 jum (55 pm). 
More than 9046 of the apertures examined were al- 
most exactly circular with a diameter (including the 
surrounding annulus) of 13-25 рт (20 jum); mean 
diameter of apertures, excluding annulus, 12.5 jum. 
The few oval apertures observed ranged from 6 X 
10 jum to 8 X 11 pm. Width of annulus са. 2.5-6 
pm (Fig. 4). 

The reticulum on exposed surfaces of the tetrads 
has mostly medium-sized lumina with shapes rang- 
ing from square, rectangular, or polygonal to oval, 
circular, and irregularly curved sinuous outlines 
(Figs. 1, 2, 4—7). Muri forming the reticulum are, 
therefore, straight or curved and not invariably 
winding, as Praglowski (1979) found in pollen from 
herbarium material. As Praglowski (1979) noted, 
there are about 25 to 30 lumina in a pollen grain 
seen in lateral view (Fig. 1). Some lumina are 
smaller near the borders between members of the 
tetrad (Figs. 1, 6, 7) and, as in all members of the 
family with tetrad pollen, the reticulum (or perfo- 
rate tectum in Exospermum) is completely inter- 
rupted near where individual grains are fused (Figs. 
1, 6, 7). Only unbranched columellae were ob- 
served supporting the muri. Lobreau-Callen (1977) 
noted that rarely the columellae are duplicolumel- 
late, although Praglowski (1979) commented this 
was not clearly confirmed. Columellae are cylin- 
drical, not quite as wide as the muri, and they are 
usually splayed out at their proximal ends forming 
a rounded flattened cushion, which joins onto the 
foot layer (Figs. 3, 6). The outer surface of the foot 
layer is mostly smooth, but at intervals there are 
small, mostly spherical, ornamentations of varying 
size (Figs. 1-7). Thickness of the sexine was 4—6 
um and the nexine up to 1.5 jum, excluding thinner 
parts near the aperture of each pollen grain. 


ULTRASTRUCTURE 


Extraseptal ectexine is 5—7.0 jum thick, exclud- 
ing thinner regions near apertures. Muri are ellip- 
tical to obovate in cross section and generally about 
1.5 рт wide and 1.2-2.2 jum high, with a height 
width ratio of 1:1 to 3:2. Columellae ca. 2-4 шт 
in height, i.e., as high as or higher than muri. In 
extraseptal parts, the foot layer is continuous except 
in apertural regions of the tetrads and is thickest 


Annals of the 
Missouri Botanical Garden 


Figures 11-14. 


Takhtajania perrieri (TEM mic коргар). — 11. Section through the center of а tetrad, with the 
1 pm). 


interseptal zone of fusion forming a space near th nter (scale bar = — 12. Median section of most of a 
ollen grain in a tetrad passing through the center of an apertural papilla (scale > Баг = 5 wm). —13. Section d 
part of the annulus Шеп grain with an asterisk indicating the center of the lamellated endexine (scale ba 


— 14. Section through part of the wall of an apertural papilla with underlying peripheral cytoplasm (scale bir = 


1 wm). 


near the exposed fringes of the pollen grains, where 
it is 0.5-1.0 jum thick and of similar thickness to 
the columellae and tectum (Fig. 9). It decreases in 
thickness toward the distal aperture of each pollen 
grain (Fig. 9). In septal parts (where the members 
of a tetrad are contiguous), the foot layer is thinner 
and measures ca. 0.1–0.4 jum. The inner surface 
of the foot layer is smooth to undulate (Figs. 9-11). 
Coherence between members of a tetrad is achieved 
by fusion of short columella-like elements extend- 


ing from the foot layers of adjacent pollen grains 
(Figs. 9-11). In a few places there are small breaks 
in the foot layer in septal regions, which are 
"plugged" by endexine in mature tetrads (Fig. 10). 
Beneath the foot layer is an endexine that is con- 
siderably thinner than the foot layer, except near 
the aperture (Figs. 9, 12). In septal regions, it is 
approximately one-third the thickness (0.1—0.2 р, 
of the foot layer and even at low magnifications is 
readily distinguishable by its greater electron den- 


Моште 87, Митбег 3 
2000 


Затрзоп 385 
Pollen of Takhtajania perrieri 


sity (Figs. 9-11). In extraseptal regions, excluding 
the annulus, it is ca. pam in thickness. It 
reaches a comparatively massive thickness of up to 
3 jum in the annulus region, and here it is made 
up of compressed lamellar elements more or less 
tangentially aligned (Fig. 13). Near the external 
fringes of the annulus and somewhat beyond this, 
the endexine has an outer, more homogeneous part 
and an inner region consisting of irregular globules, 
presumably sporopolleninous, separated by elec- 
tron-transparent spaces giving it a spongy appear- 
ance (Figs. 9, 

The intine, which underlies the endexine, is of 
similar thickness to the latter in septal regions 
(0.05—0.2 рт) and has a homogeneous appearance. 
It is usually readily distinguishable from the end- 
exine, even at low magnifications (Fig. 9), because 
of its lower electron density. As Figure 12 illus- 
trates, extraseptal intine increases dramatically in 
thickness toward the apertures, where it reaches up 
to 1.8 jum, and as commonly occurs in other Win- 
teraceae (Praglowski, 1979) it loses its homoge- 
neous appearance. There is a further increase in 
thickness of the intine in the apertural papilla re- 
gion to up to 2.4 jum, where the wall appears to 
consist entirely of intine (Fig. 14). It can be seen 
that in these thicker regions, outer parts of the in- 
tine have inclusions of a vesicular-like nature, 
which is common in similar regions of many angio- 
sperms. Only the innermost part of the intine ap- 
pears more or less homogeneous (Fig. 14). 


DISCUSSION 


It would seem that artifactual splits in the ap- 
erture in dried pollen, which had been obtained 
from the type specimen, similar to that shown in 
preserved pollen in Figure 5, led Straka (1963, 
1975) to describe the pollen of Takhtajania as col- 
pate and sometimes trichotomocolpate. Figure lc 
in Straka (1963: 357) is a photomicrograph showing 
a triradiate tear in the aperture membrane, which 
is labeled as a “trichotomocolpate” aperture. Sim- 
ilarly, Lobreau-Callen (1977: 447) illustrated what 
was termed a colpate (slit-shaped) aperture in a 
photomicrograph (plate 1, fig. 4), which is very sim- 
ilar to the torn aperture shown in Figure 5 of the 
present paper. Praglowski (1979: 19) also noted ap- 
ertures were different from other Winteraceae in 
their great variation in shape "ranging from pori to 
colpi." It is concluded, therefore, that previous re- 
ports of these varied types of apertures in Takhta- 
jania are incorrect, and that it resembles other 
members of the family in possessing round or some- 
what oval apertures. The study by Lobreau-Callen 


(1977) was the only previous one to illustrate pollen 
of Takhtajania under SEM, and two of his illustra- 
tions (plate 2, figs. 1 and 4) do show a bulging 
apertural membrane, despite the fact that pollen 
was obtained from dried herbarium material. It is 
unfortunate that it has become well established in 
the literature that Takhtajania pollen is sometimes 
trichotomocolpate (trichotomosulcate), but one can 
appreciate the limitations imposed on previous 
workers when studying pollen from a type specimen 
collected in 1909. On the other hand, true tricho- 
tomocolpate apertures are well defined. For exam- 
ple, the recently discovered genus Anacostia, a new 
basal angiosperm from the early Cretaceous of 
North America and Portugal, has some pollen 
grains with clearly delimited and attractively or- 
namented trichotomocolpate apertures (Егиз et al., 
1997). 

The present investigation supports previous stud- 
ies that Takhtajania has considerably larger tetrads 
than other members of the family whose pollen has 
been examined. Endress et al. (2000 this issue) 
suggested that the presence of these large tetrads 
may indicate polyploidy, as indicated for some oth- 
er Winteraceae by Hotchkiss (1955). Straka (1963) 
stated that the acetolyzed pollen tetrads were 60 
um in diameter; Lobreau-Callen (1977) noted their 
mean diameter was 57.4 рт, without citing the 
range of sizes; and Praglowski (1979), who gave 
mean and size range for the pollen of most other 
Winteraceae examined, gave a single size of 65 шт 
diameter for the acetolyzed tetrads, presumably be- 
cause of the limited material available. The range 
of tetrad size obtained in the current study (65—83 
um) extends the upper limits of the size range, but 
no tetrads were found as small as those measured 
by Straka (1963) and Lobreau-Callen (1977). The 
mean (74 рт) is considerably higher than that giv- 
en by Lobreau-Callen (1977). It should be empha- 
sized that acetolysis can increase the size of pollen 
units by 3096 or more, and the extent of the in- 
crease may depend on methods of acetolysis used 
(D. C. Mildenhall, pers. comm. 1999). In fact, tet- 
rads of Takhtajania, which had been dried by the 
critical point method for SEM (Figs. 1, 2, 4, 5, 7) 
had a diameter (excluding the apertural papillae) 
averaging ca. 47 рт. It is highly probable that the 
tetrads of other Winteraceae undergo a similar in- 
crease in size with acetolysis. Thus, Pseudowintera 
colorata, which had tetrads with a mean diameter 
of 50 jum following acetolysis (Praglowski, 1979), 
measured ca. 35 jum under SEM, when material has 
been dried by the critical point method (unpub- 
lished pers. obs.). 

Lobreau-Callen (1977) and Praglowski (1979) 


386 


Annals of the 
Missouri Botanical Garden 


made detailed comparisons between the pollen of 
Takhtajania and other genera in the family. Lob- 
reau-Callen (1977) concluded that based on its 
structure and sculpture, its pollen is closest to that 
of Belliolum, from which Takhtajania can be dis- 
tinguished by the difference in size of perforations 
in the internal walls (septae), which are large in 
Belliolum. It was subsequently found for several 
genera in the family that if these gaps are still open 
at the time of pollen grain mitosis and therefore the 
four members of a tetrad share a common cyto- 
plasm, division into tube and generative cells with- 
in each pollen grain is synchronous within a tetrad, 
e.g., in Belliolum (Sampson, 1981). It is probable 
that pollen grain mitosis is asynchronous within 
each tetrad of Takhtajania, because the small and 
infrequent gaps in the ectexine forming the septae 
are plugged with endexine, which is formed before 
division—at least in those species that have been 
investigated. On the other hand, species with syn- 
chronous division have, in older tetrads, the septal 
gaps plugged with intine, which is formed subse- 
quent to ai mitosis ее 1981), or large 
gaps may re n mature tetrads, as illus- 
trated by Praglowski (1979) for Bubbia howeana (F. 
Muell.) Tiegh. Assuming that Takhtajania has asyn- 
chronous pollen mitosis, it shares this feature with 
Pseudowintera and Drimys. Whether or not this in- 
dicates a relationship between these three genera 
depends on whether asynchronous mitosis, the 
adaptive significance of which is not obvious, has 
evolved independently in these taxa. Furthermore, 
this feature has not been investigated in all species 
in the family, and future studies may demonstrate 
that it is not consistent within a genus. For exam- 
ple, although asynchronous division occurs in all 
three species of Pseudowintera, only one species of 
Drimys has been examined (Sampson, 1981). It has 
been suggested that the asynchronous type has 
evolved from the synchronous (Sampson, 1981). 
In contrast to other Winteraceae, Drimys, Takh- 
tajania, and Pseudowintera share protuberant ap- 
ertural papillae, which may or may not indicate a 
relationship between the genera, depending on 
whether or not this characteristic evolved indepen- 
dently in these three genera. Bailey and Nast 
(1943) noted that van Tieghem (1900), who studied 
pollen of all genera (excluding Takhtajania), al- 
though he provided no illustrations, found that the 
pollen of the three New World species of Drimys 
he examined formed protuberant papillae when 
moistened, in contrast to Tasmannia (Drimys sect. 
Tasmannia), Pseudowintera, Bubbia, Belliolum, Ex- 
ospermum, and Zygogynum. Bailey and Nast (1943) 
confirmed these results. However, fresh and pre- 


served pollen of all three species of Pseudowintera 
have the papillae (Fig. 8). Both Takhtajania and 
Pseudowintera possess an annulus that is verrucate 
Figs. 3, 8) with smaller verrucae in Takhtajania, 
in contrast to the comparatively smooth-surfaced 
annuli illustrated for species of Drimys (sensu 
Smith, 1969) by Lobreau-Callen (1977) and Prag- 
lowski (1979). Tetrads of Pseudowintera differ from 
those of Takhtajania in generally lacking expanded 
tips to their columellae where they touch the foot 
layer and in possessing numerous small perfora- 
tions in the tectum near their external boundaries 
Fig. 8). 

Praglowski (1979), while agreeing that Takhta- 
јата pollen differs considerably from that of Bub- 
bia, the genus in which Takhtajania perrieri had 
first been placed (Capuron, 1963), disagreed with 
the conclusion of Lobreau-Callen (1977) that it was 
closest to pollen of Belliolum. Praglowski (1979) 
questioned the value of the ratio between the width 
of muri and the diameter of bacula (columellae) 
used by Lobreau-Callen (1977) for intergeneric 
segregation and concluded its pollen was closest to 
that of Drimys (syn. Drimys sect. Drimys). Lobreau- 
Callen (1977) also had noted similarities between 
Takhtajania and Drimys pollen. Coetzee and Prag- 
lowski (1988) described two types of fossil Winter- 
aceae pollen resembling Тазтапта and Bubbia 
from the Miocene of South Africa. This discovery 
and the Early Cretaceous occurrence of Wintera- 
ceae pollen in Israel (Walker et al., 1983) led them 
to suggest that the Winteraceae had a West Gond- 
wanan origin and an early differentiation, possibly 
in Africa, before migration to Australasia and Mad- 
agascar. They suggested furthermore that a dis- 
persal route from the south could probably explain 
the close resemblance between the pollen of Takh- 
tajania and Drimys of the New World. Doyle et al. 
(1990a) noted that extinct relatives of the Winter- 
aceae (Afropollis) were an important component of 
Early Cretaceous tropical floras and extended into 


. — 


— 


Laurasia, and commented that their present austral 
temperate distribution was attained later. The win- 
teraceous affinity of Afropollis has, however, been 
questioned by Friis et al. (1999 

Ultrastructural detail in pollen of the Wintera- 
ceae (e.g., intine and endexine structure, endexine 
fine structure) does not seem to differ significantly 
between genera (Praglowski, 1979, and unpub- 
lished pers. obs.) However, further studies are 
needed to confirm this, utilizing freshly collected 
pollen and TEM fixation techniques. 

Table 1, which includes information from Prag- 
lowski (1979), the present study, and unpublished 
personal observations, summarizes a number of pol- 


387 


Pollen of Takhtajania perrieri 


Sampson 


Volume 87, Number 3 


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388 


Annals of the 
Missouri Botanical Garden 


len characters of the genera. Assuming that some 
or all similar characters are not the result of con- 
vergent evolution, the pollen of Takhtajania most 
closely resembles that of Pseudowintera and 

mys. Endress et al. (2000) reached somewhat dif. 


Takhtajania fits best in the Pseudowintera—Zygo- 
gynum (including Bubbia, Belliolum, and Exosper- 
mum) clade, “which is sister to the Tasmannia/Dri- 
mys clade.” Hopefully, further studies, including 
those using molecular data, will provide further in- 
formation on the relationships between Takhtajania 
and other Winteraceae. 


Literature Cited 


Bailey, I. W. & C. С. Nast. 1943. The comparative mor- 

phology of the Winteraceae. I. Pollen and stamens. J 
Arnold Arbor. 24: 340-346. 
Meer M. Van & Ph. Guinet. 1961. Les pollens com- 
ées.—Lexemple des Mimosacées. Pollen Spores 3: 

200-218. 

Capuron, R. 1963. Présence à Madagascar d'un nouveau 
représentant (Bubbi perrieri R. Capuron) de la famille 

ansonia, n.s., 3: 373—378 

rii J. A. & J. Praglowski. 1988. Winteraceae pollen 
from the Miocene of the southwestern Cape (South Af- 
rea). Grana 27: 27-37 

Doyle, „СТ. Hotton < J. V. Ward. 1990а. Early 
Cretac eous tetrads, zonasulc e Е and Wintera- 
сеае. I. Taxonomy, morphology, and ultrastructure. 
Amer. J. Bot. 77: 1544-1557. 

. 1990b. Early Cretaceous tet- 

rads, жишш ulate pollen, and a - Cladis- 

tic analysis and implications. Amer. J. Bot. 77: 1558- 

1568. 


r F., 1. Silberbauer-Gottsberger & G. Gottsber- 

. Variation on the population, racial and spe- 

cies мү in the primitive relie angiosperm Пети Dri- 

mys | in South America. Pl. . Evol. 
132: 53-83. 

Endress, P. K., A. Igersheim, F. B. Sampson & G. 
Schatz. 2000. Floral structure of я апа из 
systematic position in Winteraceae. Ann. Missouri Bot. 
Gard. 87: 347-365. 


Friis, E. М., P. В. Crane & К. В. Pedersen. 1997. Ana- 


costia, a new basal angiosperm from the Early Creta- 
ceous of North America and Portugal with trichtomo- 
colpate/monocolpate pollen. Grana 36: 225-244. 

‚ К. R. Pedersen & P. Н. Crane. 1999, Early an- 
giosperm diversification: The diversity of pollen asso- 
ciated with angiosperm reproductive structures in Early 
Cretaceous Lp from Portugal. Ann. Missouri Bot. 
Gard. 86: 259-296. 

Hotchkiss, A. T 1955. Chromosome numbers and pollen 
tetrad size in the Winteraceae. Proc. Linn. Soc. New 


R. Cap. Rapports ен avec les autres genres 


8 5. 
НЕ 


e. Pp. 1-38 т S. Nilsson 
(editor), Wu Enn and “жн Flora 8. Almqvist & 
Wiksell, Stockholm. 

Sampson, Е В. 1974. A new pollen type in the Winter- 
aceae. Grana 14: 

1981. B ашына: Ж versus asynchronous mitosis 

1 permanent pollen tetrads of the Winteraceae. 

а 20: 19– a 


en 


Schatz, G., Lowry II & A. || 
aem uaa uu rediscovered. e 391: 
IX, 


Smith, Pu r 156 The American species of Drimys. J. 
Arnold bug 24: 1-33 

. À rec vonsiderátion of s genus Tasmannia 

(Winteracenc) Taxon 18: 286–2% 

Straka, H. 1963. Über die migliche phylogenetische Be- 

deutung der Pollen Morphologie der madagassischen 

Bubbia perrieri R. Cap. А Grana Palynol. 


4: 3 


1975. Pollen- und Sporenkunde in Grundbe- 
griffe = modernen Biologie 13: 1-238, Gustav Fischer 
ed., ga 

пе E van. 1900. Sur les dicotylédones du groupe 
des Homoxylées. J. pn ndn) E о 334 A 

Vink, W. 1970. The Winteraceae e Old World. 
и апа Drm Е рне and Medi 
Blun | 22 5-354. 

e Winteraceae of the Old World V. Ex- 

ospermum links Bubbia to Zygogynum. Blumea 31: 39- 

55. 

& H. S. Mac ЕЯ 


Winteraceae. /n: Р. Morat 


——— 199 
(editors), P de la Nouvelle-Calédonie ba 90-17 


Muséum National d'Histoire Naturelle, P. Pa 
Walker, J. W., С. J. Brenner & А. 6 Walker. 1983. Win- 
teraceous pollen in the Lower Cretace eous of Israel: Ear- 
ly evidence of a Magnolialean angiosperm family. Sci- 


ence 220: 1273-1275. 


EMBRYOLOGY OF 
TAKHTAJANIA 
(WINTERACEAE) AND A 
SUMMARY STATEMENT OF 
EMBRYOLOGICAL FEATURES 
FOR THE FAMILY! 


Hiroshi Tobe? and Bruce Sampson? 


ABSTRACT 


We present the first report on the embryology of Takhtajania. By adding its data to those already known from other 
Winteraceae (Drimys, Pseudowintera, Tasmannia, and Zygogynum s.l.), we summarize embryological features of the 
proposed basal position of Takhtajania in the family and a рне sister-group а 
. Comparisons within 
between the мааа showed that Takhtajania agrees well with other Winteraceae Ра Па НА having по distin 
s basal position in the family. Although Winteraceae and Canellaceae share a number of basic 
embryological iba including an exotestal seed coat, Winteraceae are clearly distinct by virtue of the following 
features: the outermost one of the middle layers and even part of the connective tissue in the anther developing fibrous 
thickenings at anthesis; ovules anatropous (not р as іп СапеПасеае); the писгорује formed by the inner 
integument alone (rather than by two integuments as in Canellaceae); an exostome formed after fertilization; a persistent 
micropylar part of the tegmen composed of variously ЕРЕ thick-walled cells; and the exotesta palisadal. Winter- 
aceae are thus a well-defined group аи and, despite their modern widespread distribution, genera show 
little diversification in embryological character 


words: Canellaceae, embryology, ы anatomy, Takhtajania, Winteraceae. 


The Winteraceae are a relatively small woody di- 
cotyledonous family, comprising about 65 species 
assigned to Drimys, Pseudowintera, Takhtajania, 
Tasmannia, and Zygogynum sensu lato а. 
Belliolum, Bubbia, and Exospermum). They 
broadly distributed from the Philippines to Tas 
mania and New Zealand, South and Central Amer- 
ica, and Madagascar (Vink, 1993). Because of its 
relict Gondwanan distribution as well as its dis- 
tinctive primitive-type floral morphologies, the fam- 
ily has long attracted the attention of researchers 
of plant evolution and morphology. Within the fam- 
ily, Takhtajania, comprising the only extant species 
T. perrieri (Capuron) Baranova & J.-F. Leroy in the 

rica/Madagascar region, has been considered 
most isolated, and in fact it differs from the three 
other genera in having flowers with two carpels 
united to a paracarpous gynoecium, instead of free 
carpels or a number of carpels united to a eusyn- 
carpous gynoecium. Takhtajania thus has been 
placed in a subfamily of its own, Takhtajanioideae, 
and the other genera in another subfamily Winter- 


oideae (Leroy, 1978; Takhtajan, 1997). 


Takhtajania has been very poorly understood in 
general with respect to its morphological and evo- 
lutionary traits because only a single collection 
made in 1909 had been available for research until 
recent times (Leroy, 1980). A number of trees were, 
however, rediscovered at the Anjahanaribe-Sud 
Special Reserve (14^45'S, 49°29’E) in Madagascar 
in 1994 (see Schatz et al., 1998; Schatz, 2000 this 
issue). An extensive collection was subsequently 
made for morphological, anatomical, cytological, 
and molecular studies (for many results, see other 
articles in this Annals issue). 

In this paper we present the embryology of Takh- 
tajania, which has not been studied before. Taking 
the data from this study together with those from 
the four other genera (Drimys, Pseudowintera, Tas- 
тапта, and Zygogynum s.l.), which have already 
been studied relatively well, we will summarize em- 
bryological features of the whole family Wintera- 
ceae. Molecular evidence based on combined se- 
quence data from mitochondrial atpl and matR, 
plastid atpB and rbcL, and nuclear 185 rDNA has 


revealed that Winteraceae are sister to Canellaceae 


! We are а to Porter P. Lowry П and George E. Schatz for their efforts я T the materials used in this 


sti, and to Pet 
? Department of Bota 


r H. Raven for his support and encouragement to complete this 
ny, Graduate School of Science, Kyoto University, Kyoto 606- "8502, 


3 School of Biological Sciences, Victoria University of Wellington, P.O. Box 600, Wellington, New Zealand. 
ANN. Missouni Bor. GARD. 87: 389—397. 2000. 


390 


Annals of the 
Missouri Botanical Garden 


with support of 100% bootstrap value, and that 
Takhtajania has a basal position within the Win- 
teraceae (Qiu et al., 1999). We will discuss whether 
or not the sister-group relationship between Win- 
teraceae and Canellaceae is supported, whether or 
not the basal position of Takhtajania in Wintera- 
ceae is supported, and how Winteraceae are a co- 
herent group distinct from the Canellaceae. 


MATERIALS AND METHODS 


Flower buds and fruits of Takhtajania perrieri in 
various stages of development were collected at the 
Anjahanaribe-Sud Special Reserve in Madag 
(vouchers: Rakotomalaza et al. 1342, Schatz 2748, 
Ravelonarivo s.n. in 1998; all at MO) and fixed with 
FAA (five parts stock formalin; five parts glacial 
acetic acid; 90 parts 50% ethanol). Twenty-eight 
flower buds, four open flowers, and 17 fruits were 
dehydrated through a t-butyl alcohol series, em- 
bedded in Paraplast (melting point 57—58?C), and 
sectioned using a rotary microtome following stan- 
dard paraffin methods. Sections cut at about 6–10 
pam thickness were stained with Heidenhain's hae- 
matoxylin, Safranin-O, and FastGreen FCF, and 
mounted with Entellan. To examine their fine struc- 
ture, several anthers were embedded in Spurr's res- 
in after dehydration through an acetone series and 
sectioned with glass knives using an ultramicro- 
tome. Sections cut at about 1-2 рт thickness were 
stained with Toluidine blue O. 


OBSERVATIONS 
ANTHERS AND MICROSPORES 


The small anther is supported by a more or less 
laminar filament (Fig. 1), and is tetrasporangiate 
(Figs. 2, 3). Prior to maturation the anther com- 
prises 5 to 6 cell-layers: an epidermis, an endothe- 
cium, 2 to 3 middle layers, and a tapetum (Fig. 7). 
Because of the lack of younger flower buds, it is 
not certain how the anther wall developed. The ta- 
petum is glandular, and its cells are 2-nucleate 
(Fig. 4). During maturation, the epidermal cells are 
enlarged to some extent but nearly collapsed even- 
tually, and the endothecium develops fibrous thick- 
enings (Figs. 8, 9). The middle layers mostly de- 
generate, but like the endothecium, the outermost 
layer usually develops fibrous thickenings (Figs. 8, 
9). Part of the connective tissue may also develop 
fibrous thickenings (Fig. 8). Anther dehiscence 
takes place by longitudinal slits, with each slit 
common to two microsporangia of a theca (Fig. 9). 

Meiosis in the microspore mother cell is accom- 
panied by simultaneous cytokinesis (Fig. 5). The 


shape of resultant tetrads is tetrahedral. Unfortu- 
nately, no buds could be sectioned that would have 
showed stages in the division of the 1-сеПеа mi- 
crospores leading to the formation of vegetative 
(tube) and generative cells. Two different types of 
division have been found. In the synchronous type, 
which occurs in Tasmannia and Zygogynum s.]., 
the nuclei of all four pollen grains within a tetrad 
are at exactly the same stage of mitotic division 
because there are cytoplasmic connections through 
aps in their internal walls. However, in the asyn- 
chronous type, which occurs in Drimys and Pseu- 
dowintera, division of the microspore nuclei is 
asynchronous because there are no gaps in the in- 
ternal walls of each tetrad at this time (Sampson, 
1981; Prakash et al., 1992). It has been suggested 
that the asynchronous type has evolved from the 
synchronous, and in view of the basal position of 
Takhtajania it would therefore be of interest to 
know if it too has the synchronous type of pollen 
mitosis. 
Pollen grains are 2-celled at the time of shedding 
(Fig. 6 


OVULES, NUCELLUS, AND MEGAGAMETOPHYTE 


Early in development six to seven ovules with a 
long funicle are pendulous in an ovarian locule 
(Figs. 1, 2; see also Endress et al., 2000 this issue). 
They are anatropous at maturity (Fig. 15) and cras- 
sinucellate. The archesporium is 1-celled (Fig. 10). 
The archesporial cell divides periclinally to form 
the primary parietal cell above and the primary 
sporogenous cell below (Fig. 11). While the primary 
parietal cell further divides periclinally into more 
cells, the primary sporogenous cell develops into a 
megaspore mother cell. Thus ovules have one en- 
larged megaspore mother cell below a longitudinal 
row of five to six parietal cells. The megaspore 
mother cell undergoes meiosis to produce a linear 
tetrad of megaspores (Fig. 12). In the megaspore 
tetrad the chalazal megaspore functions (Fig. 12) 
and develops successively into a 2-, 4- (Fig. 13), 
and 8-nucleate embryo sac. Thus the mode of the 
embryo sac development is of the Polygonum type. 
An organized embryo sac has eight nuclei in seven 
cells: an egg cell, two synergids, two polar nuclei, 
and three antipodal cells (Fig. 14). The organized 
embryo sac is ellipsoid in shape and positioned at 
the upper region of the nucellus (Fig. 15). The an- 
tipodals degenerate before or soon after fertilization 
Fig. 14) 

During megasporogenesis and megagametogene- 
sis, some apical epidermal cells of the nucellus di- 
vide periclinally to form a 2-cell-layered nucellar 


— 


Volume 87, Number 3 Tobe & Sampso 391 
2000 Embryology of Takhtajania 


Figures 1—9. poe of anther and microspores in Takhtajania. —1. E maar section (LS) of flower 
showing a small anther. —2. Transverse section (TS) of flower showing dehiscing anthers. . TS of anther. —4. TS 
of po showin ng 2-nucleate glandular tapetal cells. —5. Meiosis in mic пароша т showing simultaneous 
cytokinesis. —6. Microtome sections of 2-celled mature pollen grains in tetrad. —7. Microtome section of young anther 
showing wall structure. —8. TS of older anther showing that fibrous thickenings develop in the outermost one of middle 
layers and even in part of the connective tissue as well as in the endothecium. —9. Microtome section of dehiscing 
anther. bed ne an, anther; co, connective tissue; end, endothecium; ep, epidermis; mc, microspore mother cell; 
ml, middle layer; ov, ovule; t, tapetum. Scale bar equals 1 mm in Figs. 1 and 2, 100 jum in Figs. 3, 8, and 9, and 20 
um in Figs. 4—7 


392 Annals of the 
Missouri Botanical Garden 


Figures 10-18. Development of nucellus, embryo sac, and integuments in Takhtajania. —10. Longitudinal section 
(LS) of оуше primordium. —11. LS of young ovule with a a megaspore mother cell. —12. LS of young ovule showing a 
linear sir of megas spores, in bis ch the chalazal megaspore is functional and the other megaspores are degenerating. 

of older ovule with a 4- мака embryo sac, showing only two nuclei in photo. —14. LS of mature оуше 
with an e embryo sac, sho owing o e synergid, (fused) polar nuclei, and one antipodal cell in photo. —15. LS 
of nearly — ovule showing the mic E formed by the inner integument alone. —16. Transverse section of mature 
ovule. —17. LS of mature ovule showing the micropylar part. —18. LS of mature ovule showing the thickness “ 


integuments. ле ons: ant, antipodal cell; arc, archesporial cell; dm, degenerating megaspore; es, embryo sac; 
fm, functional megaspore; ii, inner integument; mi, micropyle; mmc, megaspore mother cell; n, nucleus of embryo sac; 
nuc, nuc т оЬ, ит к oi, outer integument; р, parietal cell; po, polar nucleus; sy, synergid. Scale bar equals 500 
pam in Fig. 18, 100 um in Figs 15-17, and 20 jum in Figs. 10-14. 


Volume 87, Number 3 Tobe & Sampso 393 
2000 Embryology of Takhiajania 


Figures 19-27. Development of seeds and seed coats in Takhtajania. —19. Longitudinal pected (LS) of seed. 


—20. Micropylar part in LS of older seed. —21. LS of older seed showing a globular p osperm. 
— 22. Nearly mature fruit opened yi ially. —23. Transverse section (TS) of young seed. € LS of y young seed 
showing cellular DEN —25. LS of young seed coat. —26. LS of nearly mature seed coat. —27. TS of nearly 

ature seed coat. Abbreviations: em, о еп, endosp эегт; ents, endotesta; exts, exotesta; mts, mesotesta; nuc, 


nucellus; se, seed; tg, tegmen; ts, testa. Scale bar m A mm in Fig. 19, 3 mm in Fig. 22, 200 um in Figs. 20 and 
21, 100 jum in Figs. 23, 26, and 27, and 50 jum in Fig. 2 


394 Annals of the 
Missouri Botanical Garden 


Table 1. Comparisons of Takhtajania with other Winteraceae and Canellaceae in embryological characters. 


Characters Canellaceae Takhtajania Other Winteraceae® 


Anthers and microspores 


Number of sporangia 1 4 4 

Thickness of anther wall 5 cell-layered 5 or 6 cell-layered 5 or 6 cell-layered 

Mode of wall formation ? ? asic 

Anther epidermis Persistent Collapsed Collapsed 

Endothecium Fibrous Fibrous Fibrous 

Middle layers Crushed Crushed except fibrous Crushed except fibrous 
outer cell-layer outer cell-layer 

Tapetum Glandular Glandular Glandular 

Number of nuclei in tapetal — 2-4, fused 2 2 (or more), fused 

ell 

Cytokinesis in meiosis Simultaneous Simultaneous Simultaneous 

Shape of microspore tetrads Tetiahedis 1 Tetrahedral Tetrahedral 

Mature pollen 2-celled 2-celled 2-celled 


Ovules, nucellus, and megagametophyte 


Ovule orientation Campylotropous! Anatropous Anatropous 
Hypostase Not formed Not formed Not formed 
Number of archesporial cells 1 1 
Nature of nucellus Crassinucellate Crassinucellate Crassinucellate 
Thickness of parietal tissue — ? 5—6 cells thick 2-5 cells thick 
Mode of embryo sac forma- Polygonum Polygonum Polygonum 

tion 
Antipodal cells Ephemeral Ephemeral Ephemeral 
Nucellar cap Not formed 2 cells thick 2-3 cells thick 
Nucellar tissue in mature Present Present Present 

ovule 
Obturator ? Present Rarely present? 

Integuments 

Number of integuments 2 2 2 
Thickness of ii (early stage) ? 3(-4) cell-layered 2(-3) cell-layered 
Thickness of ii (late stage) 3 cell-layered 3(—4) cell-layered 2(-3) cell-layered 
Thickness of oi (early stage) — 2 4—5 cell-layered 3(-4) cell-layered 
Thickness of oi (late stage) 4-8 cell-layered 4—5 cell-layered 3(—4) cell-layered 
Vascular bundles ? Absent Absent 
Micropyle formation By п and oi By ii By ii 
Endothelium Not formed Not formed Not formed 


Endosperm and embryo 
Mode of endosperm formation ? ab initio Cellular ab initio Cellular 
Туре of embyrogeny ? ? Unknown, irregular type 
(Drimys winteri) 


Seed and seed coat 


Cells of endostome after fer- | Unspecialized Persistent, thick-walled; Persistent, thick-walled; 
tilization outermost cells enlarged outermost cells enlarged 
Pachychalazy No ous митова): No No 
Aril/wing Not forn Not formed Not formed 
Endosperm in mature seed Copious, ruminate* Copious Copious 
Type of seed coat Exotestal® je tal Exotestal 
Thickness of testa ? 5 cell-layers thick 4—6 cell-layers thick 
Cells of exotesta Lignified® quim palisadal Lignified, palisadal 
Cells of mesotesta Unspecialized or oily idio- Rectangular, tanniniferous Little specialized 
blasts 
Cells of endotesta Unspecialized Rectangular, tanniniferous | Unspecialized, crushed 
Cells of tegmen Crushed Unspecialized rushed, or rarely tangen- 


tially elongate, thick- 
walled 


Volume 87, Number 3 
2000 


Tobe & 


Sampso 395 
Embryology of Takhtajania 


Table 1. 


Continued. 


Characters Canellaceae 


Takhtajania Other Winteraceae® 


References 
an (1961, 1962) 


Corner (1976), Parmeswar- Present study 


Bhandari авина Bhandari 
& Venkat 
(1968), ups (1976), 
De Boer & Bouman 
(1974), Prakash et al. 
(1992), Sampson 

963), Smissen 7 

Swamy (1952), У 
(1970, 1977) 


= 


PIETS ii, inner integument; oi, outer integum 
! Ovules of Canellaceae are describe 


d as ana Boe "(Netolitzky 1926; later authors). However, a drawi 


of г 
longitudinal section of an ovule of Warburgia stuhlmannii Engl. (Parameswaran, 1962, fig. 25 on p. 173) une that 


the ovule is campylotropous rather than anatropous 
? According to Corner (1976: 
appears to be pachychalazal, but this must be confirmed. 


86), the description at Parameswaran (1961) suggests that the large seed of Cinnamosma 


* According to Corner (1976: 86), “Baillon described a vestigal aril round the hilum of Canella, but later authors 


have not reporte 


* According to Jaera (1961), Cinnamosma (C. macrocarpa and C. madagascariensis) has ruminate pue esses 
n 


in peed coats, which a 
Based on earlier fe. diene Corner (1976: 8 
ии, and 


6) noted that the exotesta is com 
that the seeds appear exotestal. The structure of the seed coat d попа has been studied very 


e not known in Canella, Cinnamodendron (including ap ap ), and № 


of breed ded (except 


poorly, and more detailed study of the seed coat is needed for critical compariso 
h 


following ta 
ча than Takhtajania: 


species of Тазтапта (Prakash et al., 


1992), Zygozynum bailloni Tiegh. (Swamy, 1952 


xa have been studied for some or part of the bubo, dn РРА of the genera of Winteraceae 
“Belliolum haplopus” (B. L. Burtt) A 

k. f. (Corner, 1976), D. winteri (Bhandari & Venkataraman, oe 
(Raoul) Dandy (Bhandari, 1963; Corner, 1976), P. axillaris (J. R. Forst 


mith (Corner 1976), Drimys (Tasmannia) piperita 
kohera oer & и 1974), | 
. & G. Forst.) Dandy 


Саа 1963), те 


d Z. stipitatum Baill. (syn. 


ја 
Exospermum stipitatum) (Smissen, 1993). Unless otherwise stated, embryological features presented above are common 


to all the taxa studied. 


e presence of an a is reported in all three species of Tasmannia studied by Prakash et al. (1992) but is 


not clear in other gene 


8 According to ca (1976: 282) in “Belliolum haplopus” 
242), who s 5 е 

r, there is no clear и ы for this, and а! о 
ceae indio inte that the micropyle is formed by the inner integument alone 


s is reiterated by Johri et al. к 
' in the formation of the micropyle. Н 
sources available for this character in Winter 


m mic pr is formed by the endostome and the 


ate that “1 eguments are 


and is covered by the outer integument (testa) after fertilization. 


cap. Cells of the nucellus remain around the mature 
embryo sac (Fig. 15). An obuturator is formed near 
the micropyle (Fig. 15). A hypostase is not formed. 


INTEGUMENTS 


The ovule is bitegmic, having an inner and an 
outer integument (Figs. 15-18). Neither the inner 
nor outer integument are multiplicative. The inner 
integument is 3-cell-layered (Fig. 18) from the be- 
ginning to older stages of development except at 
the apical part where it is about 5- to 7-cell-layers 
thick (Fig. 15). Some cells of the inner epidermis 
of the inner integument may, however, divide in 
places to increase the thickness, so that the inner 
integument is 4-cell-layered in places. The outer 
integument is 4- to 5-cell-layered from the begin- 
ning to older stages of ovules (Fig. 18). 

By the fertilization stage the inner integument 
always develops beyond the outer integument to 


cover the tip of the nucellus, so that the micropyle 
is formed by the inner integument alone (Figs. 15, 

7). However, after fertilization, the testa (devel- 
oped outer integument) develops further and reach- 
es the tip of the tegmen (developed inner integu- 
ment) (Figs. 19, 20). Therefore, when looked at in 
seeds, the micropyle appears to have been formed 
by both the inner and the outer integument. 


ENDOSPERM AND EMBRYO 


Fertilization is porogamous. Endosperm forma- 
tion ab initio is of the Cellular type. No free en- 
dosperm nuclei have been observed in young seeds. 
Compared to the whole size of the nucellus, which 
is enlarged after fertilization, the embryo sac com- 
posed of cellular endosperm is small and narrow 
(Figs. 20, 21). In nearly mature or the oldest seeds 
available, the endosperm is positioned in the center 


396 


Annals of the 
peer Botanical Garden 


of the seed and is surrounded by a massive nucellar 
tissue, which appears to be degenerating (Fig. 19). 
We did not examine embryogenesis in detail in 
this study, but fragmentary observations of early 
and late embryogenesis indicate that it proceeds 
normally to form a globular embryo (Fig. 21). In the 
nearly mature or oldest seeds available, we could 
not observe a dicotyledonous embryo. The seeds 
seem to be released from a fruit before the globular 
embryo develops into a dicotyledonous stage. 


SEED AND SEED COAT 


Nearly mature seeds are ellipsoid and somewhat 
angular (Figs. 19, 22) without any appendages such 
as an aril or a wing. The seed coat is formed by 
both the tegmen and the testa. Both the tegmen and 
the testa do not increase their thickness after fer- 
tilization. The tegmen is 3- to 4-cell-layers thick, 
and the testa is 4- to 5-cell-layers thick (Fig. 25). 
As the seed develops, cells of the exotesta radially 
elongate to assume a palisadal structure, while 3 to 
4 underlying cell-layers of the mesotesta and the 
endotesta are rectangular in shape and tanninifer- 
ous (Figs. 25-27). On the other hand, попе of the 
cell-layers of the tegmen exhibit any particular spe- 
cialization and probably degenerate or collapse ex- 
cept at the micropylar part. Cells of the micropylar 
part of the tegmen are variously enlarged and thick- 
walled to form a persistent cap-like structure (Fig. 
20). The seed coat is thus exotestal (for terminology 
of seed coat types, see Corner, 1976; Schmid, 
1986) 


DISCUSSION 


In Table 1 embryological features of Takhtajania 
are summarized in comparison with those of the 
other Winteraceae as well as of the Canellaceae. 
Embryologically Takhtajania agrees well with the 
four other genera of Winteraceae (Drimys, Pseu- 
dowintera, Tasmannia, and Zygogynum s.1.). No 
particular characters suggest the distinctness of 
Takhtajania from the other Winteraceae and its 
basal position within the family. The embryological 
features of the family are summarized as follows. 

Anther tetrasporangiate; anther wall develop- 
ment of the Basic type (unknown in Takhtajania); 
the anther epidermis becomes collapsed; the en- 
dothecium, the outermost cell-layer of the middle 
layers, and occasionally part of the connective tssue 
developing fibrous thickenings; other middle layers 
degenerating, and tapetum glandular; tapetal cells 
2-nucleate; cytokinesis in meiosis simultaneous; 
the predominant shape of microspore tetrads tet- 
rahedral; mature pollen grains 2-celled. 


Ovule anatropous, bitegmic, and crassinucellate; 
archesporium l-celled; parietal tissue 5-6 cells 
thick, tetrads of megaspores linear or T-shaped; 
mode of embryo sac formation of the Polygonum 
type; antipodal cells ephemeral; the mature embryo 
ellipsoid and positioned at the upper region of an 
enlarged nucellus; 2-cell-layered nucellar cap 
formed; hypostase not differentiated; an obturator 
formed; ovule or seed not pachychalazal; inner in- 
tegument 3(—4) cells thick, and outer integument 
4—5 cells thick; both integuments not multiplica- 
tive; vasculature absent in both the integuments; 
endothelium not formed; micropyle formed by the 
inner integument alone, but covered by an exosto- 
me after fertilization. 

Fertilization porogamous; endosperm formation 
of ab initio Cellular type; mode of embryogeny of 
an undetermined type (in Drimys winteri J. 
Forst. & G. Forst.). Mature seeds albuminous with 
a very small embryo; seed coat exotestal, having 
lignified palisadal exotestal cells to form the most 
specialized mechanical structure; cells of mesotesta 
and endotesta little specialized or crushed but rect- 

lar and tanniniferous in Takhtajania; tegmen 
usually crushed 


COMPARISIONS WITH CANELLACEAE 


Based on embryological data that is available for 
Canellaceae, the Winteraceae resemble Canella- 
ceae in general, including having an exotestal seed 
coat (Table 1). Based on the presence of lignified 
exotestal cells (except in Cinnamosma), Corner 
(1976: 86) stated that the seeds of Canellaceae ap- 
pear exotestal and indicate alliance with the Win- 
teraceae. In addition, according to Huber (pers. 
comm. in Kubitzki, 1993), seeds of Canella alba 
Nees are exotestal, with a collapsed tegmen and 
nucellus, abundant and thin-walled endosperm free 
of starch, and a small and slender embryo, and they 
agree with those of Winteraceae (and llliciales). 
However, information on the seed coat structure of 
Canellaceae is still extremely limited for critical 
comparison. Although the seed coat structure of 
several species of Canella and Cinnamosma has 
been briefly summarized (Corner, 1976), its devel- 
opment has never been documented. Therefore the 
exact structure of any species of the Canellaceae is 
not clear enough to verify how the seed coat struc- 
ture of the Winteraceae resembles or is different 
from that of Canellaceae. Nor can we clarify wheth- 
er or not the two families share exactly distinct seed 
coat structure from that of related taxa such as Pi- 
perales sensu lato (Qiu et al., 1999). Canellaceae 
need further detailed studies of these features. 


Volume 87, Number 3 
2000 


Tobe & Sampson 
Embryology of Takhtajania 


Despite the limited data available for the Canel- 
laceae, the whole family Winteraceae is clearly dis- 
tinct from it, and all the five genera of the family 
share a few characteristic features that are not 
found in Canellaceae. For instance, in Winteraceae 
fibrous thickenings occur in the outermost one of 
the middle layers (and occasionally even in part of 
the connective tissue) as well as in the endotheci- 
um, while they occur only in the endothecium in 
Canellaceae; the micropyle is formed by the inner 
integument alone in Winteraceae, but is formed by 

oth the inner and outer integuments in Canella- 
ceae; the exostome is formed after fertilization in 
Winteraceae and not by the fertilization stage as in 
Canellaceae; ovules are anatropous in Winteraceae 
but campylotropous in Canellaceae; seeds of Win- 
teraceae have a persistent micropylar part of the 
tegmen composed of variously enlarged, thick- 
walled cells, which is not reported in Canellaceae; 
the exotesta is palisadal and formed by radially en- 
larged, lignified exotestal cells in Winteraceae but 
not palisadal in Canellaceae. These distinctive em- 
bryological features support the coherence of the 
Winteraceae including Takhtajania and highlight 
the distinctness of the Winteraceae from the Ca- 
nellaceae. 

As discussed above, there is no embryological 
evidence to distinguish Takhtajania from the re- 
mainder of the family. In other words, Winteraceae 
are a well-defined group = Ancestral 
Winteraceae date back to the Barremian stage о 
the Early Cretaceous in ae gaa Gond- 
wana, and their descendents migrated from there to 
the southern temperate zone and eventually to the 
current distribution areas (Doyle, 2000 this issue). 
However, despite its modern widespread distribu- 
tion, the family has been little diversified in em- 
bryological characters since it diverged from a com- 
mon ancestor with Canellaceae. 


Literature Cited 


Bhandari, N. N. 1963. Embryology of Pseudowintera co- 
lorata. A vesselless dicotyledon. Phytomorphology 13: 
pP 


. Venkataraman. 1968. vg of Dri- 
mys winteri. J. Arnold Arbor. 49: 509—52 

Corner, E. J. H. 1976. The Seeds of Deco edm 2 vols. 
Cambridge fax Press, Cambridge. 

De Boer, R. & F. Bouman. 1974. Integumentary studies 
in the Polycarpicae. III. е winteri (Winteraceae). 
Acta Bot. Neerl. 23: 19– 

Doyle, J. A. 2000. ned relationships, and geo- 
graphic history of Winteraceae. Ann. Missouri Bot 
Gard. 87: 303- ree 

Endress, P. K., rer m heim, F. B. Sampson G. 
Schatz. 2000. Floral dtum of Takhtajania m its 


J. G. ids CER Bitt 


systematic hs in Winteraceae. Ann. Missouri Bot. 
Gard. 87: 347-365. 


Johri, B. M., K. B. Ambegaokar & P. S. € 1992. 


Comparative Baba er и Angiosperms. 2 vols. 
p Berlin. 

Kubitzki, K. 1 Canellaceae. Pp. 200-203 in K. Ku- 
bitzki et al. pe The Families and Genera of Vas- 
cular eget ‚ Vol. 2. Springer-Verlag, Berlin 

Leroy, J.-F. 1978. Une sous-famille кшй de Win- 


teraceae endémique а Madagascar: Takhtajanioideae. 
о 17: 385-395. 

— velles remarques sur le genre Takh- 
йшй и eae—Takhtajanioideae). Adansonia 20: 
9-20. 


Шеге: К. 1926. Anatomie der Angiospermern Затеп. 
K. Linsbauer Handbuch der Pflanzenanatomie Vol. 10. 
ae ide er Borntraeger, Berlin. 

Parameswaran, N. 1961. Ruminate endosperm in the Ca- 
nellaceae. Curr. Sci. 30: 3 

1962. Floral Сабин Бена and embryology in 
some des of the Canellaceae. Proc. Indian Acad. Sci. 


A. L. Lim & Е. B. Sampson. 1992. Anther 
and mul War s in Tasmannia (Winteraceae). 
Austral. J. Bot. 40: 877—885. 

. Lee, ч Bernasconi-Quadroni, D. Е. Soltis, 
P. S. Soltis, M. Zanis, E. A. Zinmmer, Z. Chen, V. Sav- 
alainen & M. W. 1999, The ай angio- 
sperms: Evidence from mitochondrial, plastid and nu- 
clear genomes. Зе 402: 404—407. 

ч F. B. . The floral morphology of на 

ега: The New pes member of vesselless Win 
aceae. гое 13: 402—423. 

. Synchronous versus asynchronous mitosis 

within d pollen tetrads of the Winteraceae. 
Grana 20: to 

Schatz, G. E. . The rediscovery of a Malagasy en- 
demic: Takhtajania T- (Winteraceae). Ann. Mis- 
souri pis Ain 87: 297-302. 


ПЗА . Ramisamihantanirina. 1998. 
unn pie Bison. Nature 391: 133- 


ЖЕ], К. 1986. On Cornerian and other terminology of 
angiospermous and gymnospermous seed coats: Histor- 
ical perspective and terminological recommendations. 

1. 


3. Some aspects of the embryology, mor- 
phology and anatomy of Exospermum stipitatum. B. Sc. 
(Hons) Project, Victoria University of Wellington, New 
Zealand. 

d B. G. L. 1952. Some aspects in the embryology 

X Zygogynum ар; У. Tiegh. Proc. Natl. Inst. Sci. 
: 399-4 


ering Plants. crate Univ. Press, New Yor 
Vink, W. 1970. The v duda сре of the Old World l. 
Pseudowintera and Drimys—Morphology and taxonomy. 


mea 18: 225-354 
—— —. 19 


11. The Winteraceae of the Old World 2. Zyg- 
ogynum—Morphology and taxonomy. Blumea 23: 219— 


. Winteraceae. Pp. 630—638 т К. Kubitzki, 
rich (editors), The Families and 
Genera of Vascular Plants, Vol. 2. Springer- Verlag, Ber- 
lin. 


NOTES ON THE VASCULAR 
ANATOMY OF THE FRUIT OF 
TAKHTAJANIA 
(WINTERACEAE) AND ITS 
INTERPRETATION! 


Thierry Deroin? 


ABSTRACT 


During fruit set the vase cular skeleton of the и gynoecium did not alter равни but there is a transfer 


Key words: 


s lc meridian- nce an 


y lateral ones, in accordance 
enere e ire some phloem fibers) 
E characte is genus among the 


with wider morphogenetic ра Ак: syncarpy and likely dehiscence. 
ovary, кые ыда т paracarpy, Takhtajania, winteroids. 


About 20 years ago, the description of bicarpel- 
late syncarpy in Takhtajania perrieri (Capuron) Bar- 
anova & J.-F. Leroy from Madagascar (Leroy, 1977, 
1978, 1980; Vink, 1978) catalyzed a phase of re- 
search and analysis regarding the evolution of the 
gynoecium of Winteraceae and of Magnoliales in 
general. More recently, Leroy (1993) presented a 
brief synthesis of the debate resulting from this dis- 
covery, which was all the more interesting since the 
information available at the time gave him every 
reason to believe that the genus Takhtajania was 
extinct. However, after considerable effort, Mala- 
gasy foresters and their colleagues from the Mis- 
souri Botanical Garden found living plants of T. 
perrieri in 1997, and thereby made it possible to 
undertake a study of the vascular anatomy of the 
fruits of this species. The results presented here 
thus represent an unexpected continuation of an 
earlier study of the gynoecium of this plant (Deroin 
& Leroy, 1993). 


MATERIALS AND METHODS 


The fruits examined in this study were collected 
by C. Birkinshaw (483, MO) in the Anjanaharibe- 
Sud reserve. They were fixed in the field in FAA 
and then transferred in the lab to a glycerol: etha- 
nol: water solution for storage. One fruit was de- 
hydrated using tertiary butanol and embedded in 


paraffin (melting point = 60?C) using standard pro- 


cedures (Gerlach, 1984), and then cut into 12 рт 
transverse sections, which were stained successive- 
ly with 0.5% aqueous Astrablue and 10% Ziehl's 
fuchsin, and then dehydrated using acetone and 
mounted in Eukitt. 

A similar protocol was used for an older flower 
from the type material (Perrier de la Báthie 10158, 

P) after restoration using 10% aqueous ammonia, 
postfixation in FAA, and the preparation of 8 jum 
sections (Deroin & Leroy, 1993). Comparison of the 
material from these two sources was thus facilitat- 
ed. Photographs were prepared using a Zeiss pho- 
tomicroscope with an orange filter and Ilford Pan F 
Plus 50 ASA film. 


OBSERVATIONS 


The material available contained only the later 
stages of fruit development (Fig. 1). The series of 
fruits showed that: (1) no growth, either in length 
or thickness, occurs outside the gynoecium; (2) a 
basal stipe(s) observed in the fruit must have 

ormed during an earlier stage, as little if any fur- 
ther elongation can be seen in the late series stud- 
ied here, and the stipe is not yet differentiated at 
anthesis (Vink, 1978), although its vascularization 
can already be detected (Deroin & Leroy, 1993); 
(3) the ovary locule (1) does, however, grow consid- 
erably, and is asymmetrical in the plane of the two 
carpels, one of which elongates more than the other. 


ess my gratitude to Pete Lowry (MO) for his English translation of the original draft, Dawn Frame (Montpellier, 


ex 
fae e) ‘Tor her ~~ help in literature and discussions, as we 


and constructive 


as the anonymous reviewer for his careful corrections 


2 Laboratoire de Phare А Muséum National d’Histoire Naturelle, 16, rue Buffon, F-75005 Paris, France. de- 


roin@mnhn.fr 


ANN. Missouri Bor. GARD. 87: 398—406. 2000. 


Моште 87, Митбег 3 
2000 


Deroin 399 
Fruit Vascular Anatomy 


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e 1. Four developmental stages of Takhtajania fruit. In the sample studied, dehiscence (C) can take place 


ur 
before maximum size is reached. a: androecial zone; 
s: stipe. 


This differential growth could explain the rather ir- 
regular dehiscence that proceeds downward from 
the stigmatic scars 

The anatomy of ilia pedicel and receptacle of the 


flower (Fig. 2). The highly elliptic stele, with 15 
vascular strands (Fig. 2A), gives rise to 2 median 
T we 2B, C), and then their lateral bundles 
(Fig. 20-С). The stele, which then becomes cir- 
cular see broadly medullate in cross section (Fig. 
2H), gives rise farther up (Fig. 21, J) to all of the 
separate traces feeding the petals and stamens (ca. 
27 in all). The histological changes that take place 
following anthesis are minimal: lignification of the 
xylem (vessels stainable with fuchsin) and forma- 
tion of subero-phellodermal strata at the level of 
the corolla and the androecium. No secondary phlo- 
em-xylem structures form. 

At the extreme base of the stipe (Fig. 2K) the 
gynoecium appears to lack cork, and the stele di- 
vides (Fig. 2L—O) in connection with the dorsal su- 
tures of the carpels. Higher in the stipe (Fig. 2P- 
S) the stele condenses in the plane of the sutures, 
forming two symmetrical lines of bundles. Two di- 
vided median carpellary bundles can thus be rec- 
ognized, as often seen in Winteraceae, which are 
fused to the meridian-lateral bundles to form me- 
dian complexes (mc) and the two synlateral bundles 


c: calyx and corolla zone; |: area of the ovary locules: p: pedicel: 


(sl), which are relatively narrower at this level. The 
stele then becomes more complex (Fig. 2T) by giv- 
ing rise to meridian-lateral traces, whereas the su- 
tures extend far to the interior by a well-differen- 
tiated epidermis (Fig. 4C). The center is obliterated 
by large-celled parenchyma, although at anthesis 
this level is hollow (Deroin & Leroy, 1993, figs. 1, 


At the base of the locule in the fruit (Fig. 2U) 
the lateral and median-lateral vascularizations are 
much more developed than the median carpellary 
complexes, whereas the inverse is true at anthesis. 
Up to the mid-level of the locule, these divided 
median complexes give rise on each side to 3 me- 
dian-lateral bundles (1, 2, and 3, Fig. 3A-C), 
whereas the lateral carpellary bundles are generally 
separate (Fig. 3A—E, 11, and 12). 

ruit dehiscence proceeds from the top down as 
the wall is torn either between the separated lat- 
erals or next to the synlateral. No specialized tis- 
sues can be seen at this level (Fig. 4F). The vas- 
cularization at the top of the fruit is formally 
identical to that observed in the ovary (Fig. 5A, C, 
E): meridian-lateral traces 1 and 2 fuse back with 
the laterals to form the placental bundles (pl, Fig. 
5B, D). It should be noted, however, that pl2 is 
comprised only of the median-lateral traces 1 and 
2 of carpel 2. The median-lateral traces 3 terminate 


400 Annals of the 
Missouri Botanical Garden 


5mm 
шини шыш 


Figure 2. Ascending sections of Takhtajania fruit. —A. Pedicel. —B-G. Perianth. —H-J. Androecium 


peripheral cork). —K-T. Stipe. —U. Extreme base of the ovary locule. |: lateral bundle; mc: median complex; sl: 
synlateral; 1, 1', 2, 2', median-laterals. 


note the 


Моште 87, Митбег 3 Deroin 401 
2000 Fruit Vascular Anatomy 


К рез e 3. Ascending sections of Takhtajania fruit. —A-F. Lower half e 2t Ae —G-—J. Upper half of the ovary. 
, a2: apical vascular arcs; m: carpellary median; pl: plac d bundle; 1, 1', 2, 2', 3, 3', meridian-laterals. 


402 


Annals of the 
Missouri Botanical Garden 


$ тањи ^ бода 
ево @° 


s 
> 


+. 


cf. Fig. 3E). 


| 500 um 


ia. — А. Synlateral bundle (sl') ( — В. Meridian-lateral 


Figure 4. Histology of the fruit wall of Takhtajan 
bundle 2' (cf. Fig. ЗЕ above). —C. Transverse section of the wall at the level of the median bundle m1. — 
section of the wall at the level of the insertion of the seeds. ansverse seclion of the wall at the level of the 
(sl) (cf. 


meridian-lateral (middle of the ovary locule). —F. Transverse section of the wall at the level of the synlateral 


‘ig. 3F); note the unspecialized dehiscence cleft, with no epidermal boundaries. 


Volume 87, Number 3 Deroin 403 
Fruit Vascular Anatomy 


~ а K 


A^ 
` 


à > -E7 ` Ps. чИ nm y. 
4t; У = ры. МА m. Af 


5 | ~ D 4 NA. а T 7 a 

igure 5. Placental vascularization of Takhtajania. —A, C, E. Ascending sections of one of the two placentas at 
anthesis. —B, D. Ascending sections of half a placenta at fruit stage (cf. 1', pl 1’, Fig. ЗН, I), dehiscence cleft visible 
on right. —F. Apical vascular arc a2 (cf. Fig. 3, I). 


Annals of the 
Missouri Botanical Garden 


in the wall, and the placental bundles fuse back 
with the median bundles to form vascular arcs al 
and a2 (Figs. 31, J, SF). The vascularization con- 
nected to the stigmas naturally disappears along 
with the latter. 

The histological changes of the gynoecium that 
take place from anthesis to formation of fruit appear 
to be minor. The ovary wall (Fig. 4E) shows a rather 
ordinary structure at mid-height: a thin epidermis 
that is highly fuchsin-stainable throughout the loc- 
ule and covered with a fairly thick hypodermis that 
is almost palisadal in places; and lacunar paren- 
chyma ca. 15 cells thick, including some fuchsin- 
stainable secretory cells as well as cells with cal- 
cium oxylate crystals, but lacking sclerified cells. 
The vascular bundles are surrounded by a crown 
of highly fuchsin-stainable cells and are largely 
comprised of metaxylem and metaphloem that are 
separated by 3 to 5 layers of cambial cells. The 
protoxylem is crushed and the protophloem is 
transformed into sclerified fibers with a lumen that 
is sometimes narrow (Fig. 4A, B). No secondary 
formations are present in the gynoecium. 

At all levels the external epidermis clearly ex- 
tends to the center of the divided medians (Fig. 4C, 
D). 


DISCUSSION 


On the basis of this study, five important features 
of the fruit of Takhtajania can be identified: 

(1) Winteraceae appear to be characterized in 
general by floral vascularization that lacks a corti- 
cal system, and in which secondary vascular struc- 
tures are absent (Nast, 1944; Deroin & Frame, 
1998). By contrast, lignification of the xylem and 
the appearance of phloem fibers are clearly delayed 
in Takhtajania, as compared, for example, to Ex- 
ospermum or Zygogynum (Deroin & Frame, 1998), 
in which differentiation occurs at anthesis. The to- 
tal absence of sclerenchyma in Takhtajania, espe- 
cially in the fruit wall, appears to be unique within 
the family, based on the work of Harvey (1982: 
164). This type of unspecialized histology is also 
found in the syncarpous gynoecia of Annonaceae— 
Monodoroideae (Deroin, 1997), although at anthe- 


(2) The evolution of a stipe, as in most species 
of Bubbia (Winteraceae) (Harvey, 1982: 169), 
makes it easier to interpret the vascularization at 
the base of the gynoecium and thus its morphology. 
In the lower half of the gynoecium the stele divides 
to form two closed steles (Fig. 2K—P), which in fact 
clearly represent an incomplete fusion of the pel- 
tate bases of the two carpels. This peltation was not 


visible at anthesis (Deroin & Leroy, 1993) and only 

appears later in the development of Takhtajania, 

whereas it is rather frequent and probably primitive 

in other Winteraceae (Leinfellner, 1965, 1966a, 

1966b; Endress, 1994; Igersheim & Endress, 
997). 


— 


Several very small, isolated bundles are found in 
the medullary parenchyma (Fig. 20–Р) of the fruit 
that are not yet present at anthesis, but which are 
likely homologous with the basal plexus of the gy- 
noecia in Annonaceae and even in Magnoliaceae 
(Deroin, 1999). Higher up, the steles open once 
again toward one another while folding back around 
the median complexes, which divide, as is gener- 
ally the case in Winteraceae. 

(3) From a strictly vascular perspective, the syn- 
carpy found in Takhtajania is not particularly ad- 
vanced, as confirmed by the organization of the 
stipe and the synlateral bundles, which are often 
disjointed (Fig. 20, I1', and I2'; Fig. ЗЕ, Il, and 
12). In this respect the situation is similar to that 
found in Monodora brevipes Benth. (Annonaceae) 
(Deroin, 1997). 

(4) Fruit dehiscence in the material studied is 
initially difficult to interpret, as the splitting could 
have resulted from the dehydrating action of the 
fixative, which would account for the irregularity of 
the opening. Nevertheless, the simple fact that this 
phenomenon can occur is of interest, as no other 
syncarpous member of Magnoliales has dehiscent 
fruits. Moreover, the possible occurrence of dehis- 
cence in the mericarps of apocarpous Winteraceae 
is still poorly understood (Harvey, 1982: 167), and 
was not reported for members of the family in New 
Caledonia (Vink, 1993). 

On the contrary, differentiation of the epidermis 
between the halves of the divided median bundles 
suggests primitive dehiscence at this level, which 
is in fact still functional in many Magnoliaceae and 
even some archaic Annonaceae such as Anaxago- 
rea and Xylopia. The presence of apical vascular 
arcs (Fig. 31, J, al, and a2) linking the placental 
zones, which are present in all Winteraceae studied 
to date (Bailey & Nast, 1943; Nast, 1944; Ueda, 
1978; Harvey, 1982) except Drimys (Tucker & Gif- 
ford, 1964; Tucker, 1975; Doweld, 1996), does not, 
however, favor dorsal dehiscence. 

(5) The vascular rearrangements during fruit de- 
velopment (Fig. 6) confirm that the nutritive role is 
transferred from the median carpellary bundle (A) 
to the lateral bundles (B), a phenomenon that ap- 
pears to be very generalized among Annonaceae 
(Deroin, 1997) and Magnoliaceae (Deroin, 1999). 
The strengthening in the fruit of the meridian-lat- 
eral bundles of the first and second orders (Fig. 6, 


Моште 87, Митбег 3 
2000 


Deroin 
Fruit Vascular Anatomy 


4 
pI---- А ^--pl 
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l' mc | A 
‘igure 6. 
(A) to mature fru 
and 3: median- Wer bundles. 


І, 1’, 2, 25, which then contribute in large part to 
the apical placental bundles (Fig. 6, pl', pl), leads 
to a vascular pattern that is functionally compara- 
le to that of the carpel in Ambavia gerrardü 
(Baill.) Le Thomas, a rather primitive Malagasy An- 
nonaceae, in which the ovules are fed by the me- 
ridian-lateral traces (Deroin & Le Thomas, 1989). 
In the case of Takhtajania (Fig. 6B), eight func- 
tional meridian-lateral bundles (four per carpel) 
feed the eight seeds, with an additional role being 
played by the synlateral traces. The remaining four 
meridian-lateral traces, of the third order, most 
likely represent the relictual vascularizations of 
four additional ovules. While the closely related ge- 
nus Bubbia has an average of 12 ovules per carpel 
(Harvey, 1982: 172), the genus Takhtajania appears 
to have undergone a reduction in the number of 


me | B 


ruo Sella schemes of transfer of vascular functionality in the gynoecium of Takhtajania from pan 
it (B). a: apical vascular arc; |, lateral; m, median; mc, median complex; pl, placental bundle; 1, 2 


ovules (to 4 per carpel in the material studied), 
perhaps following fusion of the carpels. This re- 
duction does not appear to have occurred in the 
syncarps of Annonaceae—Monodoroideae (Deroin, 
1991), and instead results from the distinctive pla- 
centation of Winteraceae. 


CONCLUSIONS 


Study of the fruit of Takhtajania shows that only 
modest changes have taken place in the organiza- 
tion of the ovary, but that they are nevertheless very 
instructive. The latent peltation of the carpels and 
the formation of a very short stipe resemble the 
tricarpellate apocarpous gynoecium of Anaxagorea 
luzonensis А. Gray, a primitive Annonaceae (De- 
roin, 1988), and thus suggest the early appearance 


405 


406 


Annals of the 
Missouri Botanical Garden 


of syncarpy as in Annonaceae—Monodoroideae (De- 
roin, 1991), which preserves in some way the prim- 
itively open carpel structure. The vascularization of 
the seeds shows a delayed pattern which parallels 
that of the ovules of the syncarpous gynoecium of 
Canellaceae (Leinfellner, 1967); the gynoecium of 
Takhtajania is thus paedomorpic, with a carpel pat- 
tern very near that of Bubbia, a rather few-carpelled 
and thus archaic genus (1 to 11 carpels after Har- 
vey, 1982: 148). In these two genera the dehiscence 
was at first dorsal, as demonstrated by the epider- 
mal differentiation at the median trace level, the 
ventral dehiscence being restored in Takhtajania 
because of the imperfection in vascular syncarpy. 
By comparison, in Annonaceae—Monodoroideae, all 
with indehiscent fruits, however, vascular syncarpy 
is yet more advanced in /solona than in Monodora, 
sometimes even with an ultimate reduction and dis- 
appearance of synlateral traces (Deroin, 1997). 
When considering the continuum from the flower 
to the fruit, this morphological and functional con- 
vergence among Gondwanan Magnoliales appears 
to highlight their shared dispersal syndrome, likely 
due to common paleoenvironmental constraints. 
As a final remark, it is noticeable that the main 
vascular rearrangements during the fruit set are lo- 
cated in the winteraceous gynoecium only, while 
the regular receptacle vasculature too is subject to 
disruptions in Magnoliaceae and Annonaceae, in 
close relation with seed nutrition (Deroin, 1999). 


Literature Cited 


Bailey, 1. W. & С. С. Nast. 1943. The comparative mor- 
phology of the S eae. II—Carpels. J. Arnold Ar- 
bor. 24: 472-481. 

ез = 1988. Aspects anatomiques et biologiques de 
le r des Annonacées. Unpublished Thesis, Univer- 

sity of Paris 11, Orsay, 590. 

199 Distribution of stigmatic plate patterns 
and valuton i in Eos naceae. Compt. Rend. Acad. Sci. 
Paris, Sér. = Vie 312: 561—566. 

———. 1997. Confirmation and origin of the paracarpy 

e on some methodological 


— 


т VE den with co 
aspects. Candollea 32:4 

1999. e се ү papel of the vascular archi- 
ure of flowers in Anno naceae and Magnoliaceae, 


tectu 


Flower vasculature in Mag- 
onaceae and Winteraceae: Lessons in 
The First пон Ѕут- 


‚ РВ. 


поћасеае, "Anno 
evolutionary constraints. 
posium on the Family Magnoliaceae, Guanzho 
China, Abstracts: 26. 

J.-F. Leroy. 1993. On the interpretation of ova- 
ry vascularization in Takhtajania (Winteraceae). Some 


anatomical characters related to the u par- 
acarpy. в Rend. Acad. Sci. Paris, Sér. 3, Sci. Ме 
316: 725— 


L К Тһотаз. 1989. Оп KE per and ev- 
olutive potentialities of Annonaceae: С; mbavia 
gerrardii (Baill.) Le Thomas, an endemic в. зре- 
cies. Compt. Rend. Acad. Sci. Paris, Sér. 3, Sci. Ме 


–65 
. 1996. On the origin of the carpel as evi- 

denced by its vascular skeleton. Phytomorphology 46: 

-394. 

Endress, P. K. 1994. Diversity and Evolutionary Biology 
of Tropical Flowers. Cambridge Univ. Press, Cambridge. 

Gerlach, D. 1984. Botanische Mikrotechnik, ed. 3, Thie- 
me, Stuttgart. 

Harvey, W. J. 1982. Systematic Studies in Bubbia Van 
Tieghem and Related Genera. Unpublished Thesis, 
University of Reading. 

Igersheim, A. & P. K. Endress. 1997. Gynoecium diver- 
sity and systematics of the Magnoliales and winteroids. 

t. J. Linn. Soc. 124: 213-271. 

I einfellner W. 1965. Wie sind die Winteraceen-Karpelle 
tatsächlich gebaut? I—Die Karpelle d кчы Sek- 
tion Tasmannia. Osterr. Вог. Z. 112: 55 

1966a. Wie sind die Winteraceen- јарка pelle tat- 
sche h gebaut? II—Über das Vorkommen einer ring- 
fórmigen Plazenta in den Karpellen von Drimys, Sektion 

Wintera. n Bot. Z. 113: 84—95. 

1966b. Wie sind die Winteraceen-Karpelle tat- 

siichlic h a шї? III—Die Karpelle von Bubbia, Bel- 

liolum, Pseudowintera, Exospermum und Zygogynum. 

Osterr. В. . Z. ПЗ: “ase -204. 

967. Uber die Karpelle verschiedener е 

i: * Pleodendron (Canellac eae). Österr. Bot 

—507. 


1977. А compound ovary with open carpels 


Leroy, J.-F. 
Evolutionary implica- 


in Winteraceae (Magnoliales): 
tions. Science 196: 977-978. 

9 Une sous- Fea ает де Win- 
"s eae endémique a Майар l'akhtajanioi- 
deae. Adansonia, ser. 2, 17: 383-305, 

1980. Nouvelles remarques sur le genre Takh- 
tajania (Winteraceae-Takhtajanioideae). Adansonia, 
ser. 2, 20: 9— 20. 


993. Histoire des travaux sur le Takhtajania. 
Pp. 109-1 19 in Origine et Évolution des Plantes à 
Fleurs. Masson, 
The comparative ааа of the 
—Vascular anatomy of the flowering 
5 


Tucker, 5. С. 1975. үнө ү vasculature к the ovular 
supply in Drimys. Amer. J. Bot 
— & Е. M. Gifford. 1964 у а a vasc a айы: of 
"d Jawceolata. Phytomorphology 14: 197-203. 
Ueda, 978. Vasculature in the carpels of Belliolum 
Bob (Winteraceae). Acta Phytotax. Geobot. 29: 
—125. 
Vink, W. 1978. The Winteraceae of the Old World. Ш. 
Notes on the ovary of Takhtajania. Blumea 24: 521 
525. 


Winteraceae. /n: P. Morat & H. S. Mac d 
dins. cum de la Me Calédonie 19: 90-17 
Muséum National d'Histoire Naturelle, Paris 


CHROMOSOMES OF 
TAKHTAJANIA, OTHER 
WINTERACEAE, AND 
CANELLACEAE: 
PHYLOGENETIC 
IMPLICATIONS! 


F. Ehrendorfer? and M. Lambrou? 


ABSTRACT 


The somatic chromosome number of Takhtajania perrieri, the only re 
2n 6. The possibly paleotetraploid karyotype , Е. arie die Chromosome size and 


is documented as 
havior, as well as sat of interphase 


nuclei, correspond to 
noliid assemblage. Chromosome numbers available for all other naive and the а Can ella 
ight of the postulated Wd od Use oed of the two families, 
ава ог and parallel d 
extinction can explain the karyological Porta and distribution of the 


ит 


espread among the 
ceae (Winterales) 


ochromosome type, w 


d series coupled with much 


w survivin Еа 


Key words: СапеПасеае, chromosome number, diploidization, кечан paleonolsbloidy: Takhtajania, Winteraceae. 


Takhtajania perrieri (Capuron) Baranova & J.-F. 
Leroy was discovered in 1909 by Perrier de la Báth- 
ie in the Massif du Manongarivo of northwestern 
Madagascar, where the population is now extirpated. 
Only in 1963 was it described as Bubbia perrieri by 
Capuron. After 85 years, its rediscovery in another 
locality of northwestern Madagascar, 150 km to the 
east, in the Anjanaharibe-Sud Special Reserve 
(Schatz et al., 1998) has made it possible to collect 
material for a first karyological study of this fasci- 
nating relict plant. After several failures with fixa- 
tions of untreated roots and flower buds (smallest 
about 2 mm diam.), already too advanced for the 
study of microsporocyte meioses or pollen mitoses, 
preliminary results were finally obtained on mitotic 
chromosome number and morphology, as well as in- 
terphase nuclear structure from pretreated root tips. 

This breakthrough has stimulated a critical com- 
pilation of all chromosome counts (and additional 
karyological observations) published so far on the 
families of Winteraceae and Canellaceae. That 
these families are related and somewhat isolated as 
Winterales within the Magnoliid assemblage (sister 
to Piperales sensu lato) has been well supported 
recently by plastid, mitochondrial, and nuclear 
DNA sequences (Soltis et al., 1999; Qiu et al., 
1999). The aim of the present report is to summa- 


rize the still quite incomplete karyological data on 
all taxa of Winterales (Table 1) and to compare 
them with available information from DNA data as 
well as other evidence (see Vink, 1970, 1985, 
1993; Kubitzki, 1993). Thus, we hope to contribute 
to a more unified picture of the evolution and phy- 
logenetic history of the two Winterales families. 


MATERIALS AND METHODS 


The material used for our study was collected by 
C. Birkinshaw, 7-11 September 1997, at the An- 
janaharibe-Sud Special Reserve, 14^45'S, 49°29’E, 
at an altitude of 1100—1300 m. Juvenile plants less 
than 20 cm tall but exhibiting adult leaf morphol- 
ogy were observed growing adjacent to reproductive 
adults. Ten of these were transplanted into contain- 
ers and transported to Antananarivo. This popula- 
tion is represented by the fruiting voucher C. Bir- 
kinshaw et al. 483. At the time that root tips were 
fixed for the present study (June 1998) only two of 
the ten plants remained alive; both of these have 
subsequently died. 

Root tips were pretreated in a 0.05% solution of 


озан, and stored in а deep-freezer until required. 


! We are very grateful to P. Raven for his interest and support of this study and to G. Schatz in particular for collec "ing 


the material for our karyological analysis, i nes root tips 
my of Sc 


14, Austri 


made Vind the te 


s, С 
rtment of Higher Plant ‚екан апа са ни of Botany, University of Vienna, А-1030, Rennweg 


ANN. Missouni Bor. GARD. 87: 407—413. 2000. 


Annals of the 


408 


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409 


Ehrendorfer & Lambrou 


Volume 87, Number 3 


2000 


Chromosomes of Winteraceae and 


Canellaceae 


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410 


Annals of the 
Missouri Botanical Garden 


Continued. 


Table 1. 


Published name Provenance n/2n Reference 


Taxon 


Canellaceae 


Goldblatt in Raven (1975), Gold- 


= 28 


Canella alba Murr. USA, Florida 


Canella winteriana (L.) Gaertn. 


blatt (1977) 
Occhioni (1948) 


Brazil, Rio de Janeiro 


same 


Cinnamodendron axillare (Nees 


& Mart.) Endl. 
Capsicodendron dinisii (Schw.) 


Occhioni (1945) 


= 26 


Brazil, cult. 


Capsicodendron Dinissii 


Occhioni? 


! Obviously erroneous for 2n 


26 (see Raven & Kyhos, 1965). 


? Possibly a triploid plant (Raven & Kyhos, 1965) or rather a wrong determination; no voucher! 


? Obviously an autotetraploid plant. 


(now limited to populations from central and southern Chile and adjacent Argentina), the exact determination of 


4 [n view of the former wide circumscription of Drimys winteri 


cultivated material remains doubtful 


43 or 2n = 86, respectively. 


? The genera Bubbia, Belliolum, Exospermum, and Zygogynum s. str. are united by Vink (1985, 1993) under Zygogynum sensu lato. 


8 To be incorporated into Cinnamodendron (Kubitzki, 1993)? 


€ Partly with one additional fragment. 


5 Obviously erroneous for n 


Chromosome preparation by the air-drying meth- 
od was performed according to Geber and Schweiz- 
er (1988), except that the root apices were softened 
in an enzymatic psi (2% cellulase plus 20% 
pectinase) at 37°C 3 hours. e slides were 
stained in a DAPI ушы -2 phenylindole) 
solution of 2 pg/ml in Мс Ilvain's buffer p 
20 minutes and mounted in Mc Ilvain's buffer pH7 
plus glycerol 2:1 (see Schweizer & Ambros, 1994). 
Chromosome fluorescence was observed with a 
Leizt Orthoplan fluorescence microscope with filter 
block A for DAPI epiluminescence. Microphoto- 
graphs were taken on Kodak TMAX 400 film. 


RESULTS 


Well spread earlier and later mitoses and inter- 
phase nuclei from pretreated root tips of Takhtajania 
perrieri are shown in Figure 1. The nuclei belong to 
the prochromosome type: Their intensively stained 
elements correspond to the proximal and evidently 
heterochromatic segments of the chromosomes, 
which remain = condensed during interphase and 
form about 36 larger and smaller chromocenters 
(Fig. 1b, c). These chromocenters exhibit a certain 
tendency toward agglomeration (particularly obvious 
in those of larger size) and are embedded in a very 
weakly stained granular-fibrous matrix. During mi- 
totic prophase and early metaphase condensation of 
chromosome segments in a proximal — distal direc- 
tion becomes apparent with the distal regions first 
visible as fibrous attachments (Fig. 1a). This process 
is concluded toward the end of metaphase (Fig. 1b), 
just before the onset of anaphase. 

From the careful inspection of about 20 mitotic 
plates the somatic chromosome number 2n — 36 
can be ascertaine hese chromosomes are of 
quite different sizes, E largest ones (about 4 to 5 
pairs) reach a length of 2.0-2.5 jum in our pretreat- 
ed material, the smallest (one pair), not always eas- 
ily recognizable (see arrow heads in Fig. 1а and b), 
about um. We are not yet able to present a 
Iu oca for Takhtajania, but there are certainly 
not four structurally = identical chromosome sets 
and therefore no clear karyological indications for 
tetraploidy. 


DISCUSSION 


The interphase nuclei, chromosome size, and mi- 
totic condensation pattern found in Takhtajania per- 
rieri correspond well with the prochromosome type 
described by Tanaka (1971), Okada (1975: type C), 
and Morawetz (1986a: Gyrocarpus type) for many 
Magnoliid orders such as Magnoliales (Magnoli- 
aceae, Annonaceae, etc.) and Laurales d 
Calycanthaceae, etc.). Within Winteraceae and 


Volume 87, Number 3 
2000 


Ehrendorfer & Lambrou 
Chromosomes of Winteraceae and 
Canellaceae 


411 


Figure 1. 


DAPI = chromosomes (a, b) and interphase nuclei (c, 
tips. In early metaphase 


chromosomes fully condensed only in proximal s 


d) of у леду perrieri from pretreated root 
zments, distally still fibrous, in lat 


metaphase (b) completely ene Arrows mark the pair of smallest EE ue sa Bowie nuclei (c, d) are of 


the prochromosome type. The bar corresponds to 10 jum 


terales comparable karyological data are still limited, 
but Drimys s. str., in spite of its much higher chro- 
mosome number, exhibits quite similar karyological 
1984). Thus 


there is no inconsistency in this respect to the place- 


" 


features to Takhtajania (Morawetz, 


ment of Takhtajania into the Winteraceae, and of 
Winterales, together with Magnoliales and Laurales, 
into the Magnoliid assemblage. Nevertheless, these 
karyological features may vary quite considerably 
within orders and families (e.g., Cassytha within 

urales: Okada, 1975; Uvariopsis and Tetrameran- 
thus within Annonaceae: Morawetz, 1986a, b, 1988). 
This also applies to Winteraceae because prelimi- 
nary and unpublished observations indicate that Tas- 
mannia not only deviates from all other members of 
the family by its low chromosome number (x — 13), 
but also by much "ager chromosomes and inter- 
phase nuclei that correspond rather to type B of 
Okada (1975). Together with the molecular evidence 


(Suh et al., 1993; Karol et al., 2000 this issue; Fig. 
2) this argues strongly against the combining of Tas- 
mannia and Drimys into a single genus as advocated 
by Vink (1970, 1988, 1993). Nevertheless, much 
more detailed and comparative studies on karyo- 
types, banding patterns, and DNA quantities in the 
members of Winterales and other Magnoliids are 
necessary before we can better understand the kar- 
yoevolution of these basal Angiosperms. 

What are the phylogenetic implications of the 
chromosome number 2n = 36 established for Takh- 
tajania relative to other members of Winterales? 
The main evidence compiled in Table 1 is com- 
pared in Figure 2 with our present knowledge about 
the evolutionary divergence of the order. This 


shows that from four extant clades three (1—3) have 


— 


remained оп a low ploidy level: (1) Takhtajania (п 
= 18) and (2) Tasmannia (n = 13) within Winter- 
aceae, and (3) Canellaceae (n = 14, 13, 11). Clade 


Annals of the 
Missouri Botanical Garden 


412 
Zygogynum (2) 86 
Zygogynum (1) 43 
Belliolum 43 ө 
© 
Bubbia 43 s 
Pseudowintera 43 а 
Drimys 43 = 
Tasmannia 13 
Takhtajania 18 
Cinnamodendron 11 g 
н Capsicodendron 13 5 
— Canella 14 Я 
Figure 2. Molecular tree (ITS and trnL) of Wintera- 


ceae = Canellaceae, adapted from Karol et al. (2000) 

h genera for which chromosome numbers are 2 
haploid на (п) inserted. Zygogynum (1) = Z. bail- 
lonii, (2) = Z. balansae. See Table 1. 


(4) has become stabilized on a higher polyploid lev- 
el (n = 43, rarely with additional polyploidy: n = 
86) and includes all remaining Winteraceae, i.e., 
Drimys, Pseudowintera, and the Zygogynum s.l. 
group of closely related genera. It is obvious that 
considerable evolutionary changes and numerous 
extinction events during the well documented and 
extremely long geological history of Winterales 
(since the Lower Cretaceous: see Doyle, 2000 this 
issue) must have resulted in the present-day highly 
relictual pattern of the Winterales. 

There is some evidence that polyploids have be- 
come diploidized by considerable restructuring of 
their karyotypes (Drimys: Morawetz, 1984; Takh- 
tajania: see above) and now appear as “paleopo- 


Robertsonian changes of base numbers at different 
ploidy levels. In Annonaceae and Eupomatiaceae 
this pattern of chromosomal evolution is still better 
preserved and more easily interpreted (Morawetz 
1986a: fig. 16; 1988). It exhibits dysploid differ- 
entiation on the diploid level with extant taxa on n 
= 7 & 8 > 9 9 10 as well as 4x, бх, and higher 
polyploid series on each of these base numbers. 
This pattern can serve as a model for the evolu- 
tionarily much more “eroded” Winterales where 
such a reconstruction suffers from many extinction 
gaps. Nevertheless, Figure 3 illustrates possible 
links between chromosome numbers of extant rep- 
resentatives (Grant, 1982). 

According to the ITS and trnL tree (Suh et al., 
1993; Karol et al., 2000), Takhtajania with n = 18 
and Canella with n = 14 are the most basal 
branches of Winteraceae and Canellaceae, respec- 
tively. If paleotetraploidy is дй for these two 
extant relicts, then n = Tasmannia (Winter- 
aceae) as well as n = 13 and n = 11 in the re- 
maining Canellaceae could have originated by de- 
scending dysploidy. The origin of the highly 
polyploid Winteraceae clade (4) with n = 43 (pa- 
leo-12x?) was evidently linked with a remarkable 
duplication event in the ITS region (Suh et al., 
1993; Karol et al., 2000). The present diversity 
center of this clade is in Australasia, i.e., former 
eastern Gondwanaland, from where it could have 
reached New Zealand with Pseudowintera and also 
South America via Antarctica (?) with Drimys. 

In retrospect, Winterales correspond in their evo- 
lutionary pattern to other tropical, woody and “bas- 
al” Angiosperm groups (Ehrendorfer, 1976, 1987, 
1995). They exhibit features of stasigenesis (evo- 
lutionary “erosion,” much extinction), but also show 
signs of very active and recent eco-geographical ra- 


lyploids." In addition, chromosomal rearrange-  diation (e.g., in Drimys: Ehrendorfer et al., 1979; 
ments must have led to  dysploid and/or Таѕтапта: Vink, 1 
86 
36 37 38 39 40 41 42 43 44 45 46 47 48 .. 12x |5 
© 

18 19 20 21 22 23 24 6x & 

11 12 13 14 15 16 17 18 4x ri 
7 8 9 2x 
< > 
dysploidy 


re 3. Possible origins of haploid chromosome numbers (n) of extant Winteraceae and Canellaceae (in bold 


igu 
print: 11, 13, 14, 18, 43, 86 


numbers in regular print). 


) as a result of polyploid and dysploid changes in hypothetical ancestral taxa (chromosome 


Volume 87, Number 3 
2000 


Ehrendorfer & Lambrou 
Chromosomes of Winteraceae and 
Canellaceae 


Literature Cited 

Borgmann, E. 1964. Anteil der Polyploiden in der Flora 
des Bismarckgebirges von Ostneuguinea. Zeitschr. Bot. 
52: 118-172 

Carr, G. D. & G. McPherson. 1986. Chromosome num- 
bers of New Caledonian plants. Ann. Missouri Bot. 
Gard. 73: 486—489 

Darlington, C. D. & A. P. Wylie. 1955. Chromosome на 
of ее e George Allen & Unwin, 

Doyle, J. A. 2000. Paleobotany, relationships. i geo- 
graphic hio of Winteraceae. Ann. Missouri Bot. 
Gard. 87: 303-316. 

Ehrendorfer, F. 1976. Evolutionary — of chro- 
mosomal differentiation wien in Gymn 
primitive Angiosperms 
itor), Origin and Early Evolution of Angiosperms. Co- 


lumbia Univ. Press, New York. 
987. Differentiation trends in tropical woody 
Angiosperms. Pp. 227-237 in K. M. Urbanska (editor), 


Differentiation funus in Higher Plants. Academic 
Press, London, New York. 

1995. Evolution und Lebensraum tropischer 
Holzpflanzen. Bayer. Akad. Wissensch., Rundgesprüche 
d. Komm. f. Okologie 10: 127-136. Verlag Dr. Pfeil, 
München. 

, F. Krendl, E. Habeler & W. Sauer. 1968. Chro- 
mosome numbers and evolution in primitive Angio- 
кш Тахоп 17: 337-353. 

I. Silberbauer-Gottsberger & G. Gottsberger. 1979. 
Vocatus on the Бени нана and species level іп the 
nus Drimys (Winteraceae) in 

53-83. 


Cytochemical hetero- 
Е diffeeentiation in Sikapi alba (Cruciferae) 
using a simple air-drying technique for е сћго- 
mosome spreads. Pl. Syst. Evol. 158: 

Goldblatt, P. 1977. New or notewort n p M 
counts in the Angiosperms. Ann. Missouri Bot. Gard. 
63: 889—895. 


Grant, V. 1982. Chromosome number patterns in primi- 
РА 94. 


966. Contribution to a 
chromosome atlas of the New Zealand flora. New Zea- 
land J. Bot. 4: 255-266 

Hotchkiss, A. T. 1955. Chromosome numbers and pollen 
tetrad size in the Winteraceae. Proc. Linn. Soc. New 
South Wales 79/80: 47—53. 

Karol, K. G., Y. Suh, G. E. Schatz & E. A. Zimmer. 2000. 
Molecular evidence for the phylogenetic position of 
Takhtajania in the Winteraceae: Inferenc uclear 
ribosomal and chloroplast gene spacer sequences. Ann. 
Missouri Bot. Gard. 87: 414—432. 

Kubitzki, K. 1993. Canellaceae. Pp –200 in K. Ku- 
bitzki, J. G. Rohwer & V. Bittrich (editors), The Fami- 
lies and Genera of Vascular Plants, Vol. 2. Flowering 
Plants, Dicotyledons, Magnoliid, Hamamelid and Car- 
yophyllid Families. Springer-Verlag, Berlin, Heidelberg. 

Morawetz, W. 1984. How stable are genomes of tropical 
woody plants? Heterozygosity in C-banded karyotypes 
of Porcelia as compared with Annona (Anonnaceae) and 
Drimys (Winteraceae) Pl. Syst. Evol. 145: 29-39. 

——. marks on karyological d ees 
EPUA in pea woody plants. Pl. Syst. Evol. 
49-100. 

1986b. Systematics and karyoevolution in Mag- 
naliidae: Tetrameranthus as compared with other Ann- 


nonaceae genera of the same chromosome number. Pl. 

Syst. Evol. 154: 147-173 

88. Karyosystematics and evolution of Aus- 

tralian Annonaceae as compared with Eupomatiaceae, 
acca фр and аме РІ. Syst. 
Evol. 159 

Occhioni, Р. on О iia de cromósomios em “Cap- 
sicodendron". Rodriguesia 9: 37-42. 

1948. Contribuição ao estudo da família “Са- 

nellaceae", estudo citológico. Arq. Jar. Bot. Rio de Ja- 
neiro 8: 97-103. 

Okada, H. 1975. Karyomorphological studies x er 
Polycarpicae. J. Sci. Hiroshima Univ. Ser PA 
Bot. 15: 115-200. 

Qiu, Y.-L., J. Lee, F. Bernasconi-Quadroni, D. E. Soltis, 
P. S. Soltis, M. Zanis, E. A. Zimmer Z. Chen, V. Savo- 
lainen & M. W. Chase. 1999. The earliest angiosperms: 
ван from mitochondrial, plastid and nuclear ge- 

omes. Nature 402: 404—407. 

быш. J. А. & С. Milne. 1973. Chromosome numbers of 
some ec Angiosperms. Notes Roy. Bot. Gard. Ed- 
inburgh 32: 423-428. 

Haven, P H. УА The bases of н рћујорепу: 

Cytology. Ann. PS Bot. Gard. 6 4—764. 

& р. W. 1965. New pe es concernin 
the original basic diese: number of Angiosperms. 
Evolution 19: 244—248 

Schatz, G. E., P. P. Low & A. Ramisamihantanirina. 
1998. Takhtajania perrieri rediscovered. Nature 391: 

‚ 133-13 

onrad D. n 

ing, stain combinat 


| F. Paara 1994. Chromosome band- 
ons for specific regions. Pp. 97-1 12 
nalysis Protocols. Human 
ress, Totowa, New 

Soltis, P. S., D. E. So ltis ‹ & M. W. Chase. 1999. Angio- 

sperm ze пе вв imis prs genes as a tool 
or comparative ogy. Nature 402: 402-404. 
чоо, G. L. 1958. yi the he hybrid origin of the angio- 
sperms. Nature 402: 40 
Strasbureer, E. 1905. Die E adis von Drimys Win- 
und die Endospermbildung bei Angiospermen. Flo- 
ra "05: 215-231. 

Suh, Y., L. B. Thien, H. E. Reeve & E. A. Zimmer. 1993. 
Molecular evolution and phylogenetic implications of 
internal transcribed spacer sequences of ribosomal 
DNA in Winteraceae. Amer. J. Bot. 80: 1042-1055. 

Sun, B. Y., T. Е Stuessy & D. J. Crawford. 1990. Chro- 
mosome counts from the flora of the Juan Fernández 
Islands, Chile. III. Pacific Sci. 4: 258-264. 

Tanaka, R. 1971. Types of resting nuclei in Orchidaceae. 
Bot. Mag. (Tokyo) 84: 118-112. 

Vink, W. 1970. The Winteraceae of the Old World, I. 
Pseudowintera and Drimys—Morphology and taxonomy. 
Blumea 28: 225-354. 

—— ———. 1985. The Winteraceae of the Old World, А, 
Lion links Bubbia to Zygogynum. Blumea 31: 
39-55. 


—— —— —. 1988. Taxonomy in Winteraceae. Taxon 37: 
691-698. 

————. 1993. Winteraceae. Pp. 630—638 т К. Kubitzki, 
J. G. Rohwer & V. Bittrich (editors), The Families and 


Genera of Vascular Plants, Vol. 2 


er and relation- 


ship in the Magnoliales. J. Arnold ur 14: 376-385. 


MOLECULAR EVIDENCE FOR Kenneth С. Karol,?? Youngbae Suh,‘ 
THE PHYLOGENETIC о о D 
POSITION OF TAKHTAJANIA 

IN THE WINTERACEAE: 

INFERENCE FROM NUCLEAR 

RIBOSOMAL AND 

CHLOROPLAST GENE 

SPACER SEQUENCES! 


ABSTRACT 


The nucleotide sequences of the internal transcribed spacers (ITS 1 and ITS 2) and 5.85 ue region of nuclear 
ribosomal DNA, as well as the non-coding trnL-trnF spacer regions of the chloroplast DNA, were determined and 
analyzed to estimate the phylogenetic position of Takhtajania perrieri (Capuron) Baranova & J. n Leroy within the 
Winteraceae. Using representatives of each genus of Canellaceae as — (Canella, Capsicodendron, Cinnamoden- 
dron, Cinnamosma, Pleodendron, and Warburgia), both maximum parsimony and maximum likelihood analyses place 
Takhtajania in a basal position sister to the remainder of the кыш Although the overall topology within the 
Winteraceae was mostly congruent between nuclear and chloroplast data sets, the trnL-trnF data resulted in lower 
support values in comparison to the ITS data, and failed to resolve basal relationships in the family, yielding alternative 
equally parsimonious solutions. The combined nuclear/chloroplast data set resulted in a single tree identical to that 


xt branch within the family. Potentially conflicting signals from nuclear 
and chloroplast data indicate that further taxon sampling or additional sequence data may be required to infer infra- 


familial phylogenetic relationships for Canellaceae 
1 


words: Canellaceae, combined data, cpDNA, ITS, Likelihood, phylogeny, Takhtajania, Winteraceae. 


Known until recently from only a single collec- 
tion made in 1909, Takhtajania perrieri has long 
been an enigma among the basal angiosperm family 
Winteraceae (Leroy, 1978). Its rediscovery in 1994. 
(Schatz et al., 1998) has provided the opportunity 
to reevaluate those morphological features anoma- 
lous within the family, as well as to explore the 
phylogenetic relationship of this isolated, relictual 
Malagasy endemic. Based upon morphological 
characters, two alternative hypotheses have been 
proposed regarding the position of Takhtajania 
within the Winteraceae: (1) Takhtajania is a mem- 
ber of a clade also comprising Pseudowintera, Bel- 
liolum, Bubbia, Exospermum, and Zygogynum (the 


latter 4 genera sometimes combined into a single 
genus Zygogynum s.l.) (Vink, 1988; Endress et al., 
2000 this issue); or (2) Takhtajania represents the 
sister taxon to the remainder of the Winteraceae 
(Leroy, 1978; Vink, 1988). The aim of the present 
study is to infer the phylogenetic position of Takh- 
tajania based upon analyses of molecular sequence 


А previous molecular phylogenetic study (Suh et 
al., 1993) utilized the internal transcribed spacer 
(ITS) region of the nuclear ribosomal DNA (nrDNA) 
to address questions of generic relationships in the 
Winteraceae. Tissue of Takhtajania, however, was 
unavailable. In addition to presenting a phylogeny 


! We thank Peter Raven for his enthusiastic encouragement | this E me his help in obtaining samples of 


Canellaceae. hank Leonard Thien fo 
thorough critical reviews of the manuscript. This 
and by a Smithsonian/Mellon Foundation award for ou 


or Winteraceae materia and 
work was As funded. i gam KOSEF 941-0500-002-2 to YS 


r project on basal angio 


and Jam James Reveal for providing 


rm dris This paper was 


submitted by KGK in partial fulfillment of the requirements for PBIO450 at the Un esit of Mary 
? Laboratory a Molecular Systematics, National Museum of Natural History, Smithsonian dereud Washington, 


D.C. 20560 


3 Current pias Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Mary- 


land 20742, U.S.A. 


* Natural Products Research Institute, Seoul National University, Seoul, 110-460, Korea. 
5 Missouri Botanical Garden, P.O. Box 299, St. Louis, Missouri, 63166-0299, U.S.A 


ANN. Missouni Bor. GARD. 87: 414—432. 2000. 


Моште 87, Митбег 3 
2000 


Karol et a 
Molecular Evidence 


for the family solely through ingroup analysis, Suh 
et al. (1993) identified a single gene duplication 
event in the Zygogynum s.l. clade, a possible mo- 
lecular marker for the inclusion (or exclusion) of 
Takhtajania within the Belliolum/Bubbia/Exosper- 
mum/Zygog ynum assemblage. 

If an ITS duplication event either was not iden- 
tified in Takhtajania or occurred independently, 
then outgroup analysis would become critical for 
investigating its relationship to the remainder of the 
family. Both morphological and molecular (rbcL) 
cladistic studies, which examined relationships 
among basal angiosperms, have identified Canel- 
laceae (represented in these studies by Canella 
alone) as a putative sister taxon to Winteraceae 
(Donoghue & Doyle, 1989; Qiu et al., 1993; Chase 
et al., 1993). Canella also appeared to be closely 
related to Winteraceae on the basis of secondary 
metabolites (Gottlieb et al., 1989). Canellaceae are 
comprised of six genera found in tropical Africa 
(Warburgia), Madagascar (Cinnamosma), and trop- 
ical America (Capsicodendron and Cinnamoden- 
dron in South America, Canella and Pleodendron 
in the Caribbean) (Cronquist, 1981). Although Suh 
et al. (1993) had encountered difficulties in the 
alignment of ITS sequences between Canella and 
Winteraceae, it was decided to reattempt alignment 
after sequencing additional representatives of Ca- 
nellaceae. 

This paper reports on the implications of new 
ITS sequence data for Takhtajania and the five oth- 
er genera of Canellaceae; Canella ITS sequence 
had been previously determined by Suh et al. 
(1992). In addition, sequence data have been gen- 
erated for two tandem chloroplast encoded (cp- 
DNA) spacer regions, and analyzed both separately 
and in combination with the ITS sequences. The 
first spacer is a group I intron and is located be- 
tween the conserved trnL (UAA) 3' exon and trnL 
(UAA) 5' exon. The second, an intergenic spacer, 
is located between the trnL (UAA) 5' exon and trnF 
(GAA) (Taberlet et al., 1991). Restriction fragment 
length polymorphisms of this molecule have been 
used to address species-level relationships within 
Mexican pines (Perez de la Rosa et al., 1995). Se- 
quence data generated from the trnL-F region also 
have been utilized to resolve phylogenetic relation- 
ships at both the species level (Mes & Hart, 1994 
and the generic level (Compton et al., 1998; 
McDade & Moody, 1999). Several studies (слећу 
et al., 1996; Molvray et al., 1999) have demonstrat- 
ed the utility of combining the ITS and trnL-F spac- 
er sequences for phylogenetic reconstruction. Thus, 
a complete survey of both ITS and trnL-F regions 
for all genera of both Winteraceae and Canellaceae 


м 


s been conducted to estimate the phylogenetic 
S 506 of Takhtajania. 


MATERIALS AND METHODS 


SPECIES SAMPLED AND SOURCES OF PLANT 
MATERIAL 


The species included in this study are presented 
in Table 1, along with voucher information, litera- 
ture citation, and GenBank accession numbers for 
both the ITS and trnL-F spacer sequences. This list 
includes at least one representative species from all 
genera in both the Winteraceae and Canellaceae. 
In the instances where ITS has been determined 
previously (all Winteraceae [Suh et al., 1993] ex- 
cept Takhtajania, plus Canella [Suh et al., 1992]), 
identical DNA extracts were used to amplify and 
sequence the trnL-F spacers. 


DNA EXTRACTION, AMPLIFICATION, AND 
SEQUENCING 


Standard CTAB methods of DNA isolation (Doyle 
& Doyle, 1987) were used to obtain total genomic 
DNA for polymerase chain reaction (PCR) ampli- 
fication. To generate ITS amplicons, PCR was per- 
formed using plant specific primer ITS5 (Suh et al., 
1993) and universal primer ITS4 (White et al., 
1990) under the conditions described in Suh et al. 
(1993). To generate the trnL-F spacer amplicons, 
PCR was performed using universal primers “c” 
and “f” (Taberlet et al., 1991), also under the con- 
ditions described in Suh et al. (1993). 

Sequence data were generated from PEG (poly- 
ethylene glycol) purified double-stranded PCR 
products (Morgan & Soltis, 1993) with the dye-ter- 
minator cycle-sequencing protocol for the ABI 
373A Sequencer (Applied Biosystems, Inc.). The 
entire forward and reverse strand of ITS was deter- 
mined for each species using the amplification 
primers (see above) and primers ITS3 (White et al., 
1990) and С5.85 (Suh et al., 1993). To determine 
the sequence for both strands of the trnL-F spacers, 
the amplification primers were used as well as 
primers “d” and “e” (Taberlet et al., 1991). The 
resulting bsc, were edited with Se- 
quencher version 3.1 (Gene Codes, Inc.); regions 
corresponding to the amplification primers were de- 
leted. The final edited consensus sequences were 
exported for alignment. 


SEQUENCE ALIGNMENT 


All sequences were aligned manually with the 
aid of Se-Al version 1.0al (Rambaut, 1996) mul- 


tiple sequence editor. 


Annals of the 
Missouri Botanical Garden 


a List of specimens used in this study with locality and voucher information, ITS literature citation for 
previously published sequences, and GenBank accession numbers for both ITS and trnL-F. 


Species Voucher 


Genbank 


ITS citation Genbank ITS trnL-F 


Winteraceae 

Belliolum pancheri (Baill.) 
Vink (= Zygogynum panch- 
eri (Baill.) Vink) 

Bubbia comptonii (Baker f.) 
Dandy (= Zygogynum 
comptonii (Baker f.) Vink) 

Drimys winteri J.R. Forst. & G. 
Forst. 

Exospermum stipitatum (Baill.) 
Tie ay ex Morot (= 


LBT 205, NO 


Zygo- 202, NO 
m stipitatum Baill.) 
Рон ae axillaris (J.R. 
“orst. . Forst.) Dandy 
Pseudowintera colorata (Raoul) 


Dandy 
Takhtajania perrieri (Capuron) 
Baranova & J.-F. Leroy 
aza et al. 1342, 
Tasmannia insipida R.Br. ex 
DC. ton, LB , 
Tasmannia lanceolata (Poir.) 
A.C. Smith 
Zygogynum acsmithii Vink 


Zygogynum — Tiegh. 
(= Zygogynum a LBT 
Baill. ue balans 
(Tie 


203, NO 


New Caledonia, Mt. Koghis, 


New Caledonia, Prokoméo, NW 
of Canala, LBT 204, 


South America, Chile, (Berke- 
ley B. G.) 45.307, NO 
New Caledonia, Mt. Panié, LBT 


New Zealand, N. Island, Aka- 
tarawa, LBT 300, NO 

New Zealand, N. Island, Aka- 
tarawa, LBT 301, NO 

Madagascar, Anjahanaribe-Sud 
Special Reserve, Rakotomal- 


Australia, Queensland, Ather- 
NO 
Australia, (Berkeley B. G.) 
60. 
New Caledonia, Mts. near Lac 


Enhuit, LBT 201, NO 
New Caledonia, Dzumac Mts., 


gh.) Vink) 
Zygogynum bicolor Tiegh. 


Canellaceae 


Canella winterana (L.) Gaertn. 
Capsicodendron dinisii 
(Schwacke) Occhioni 


Cinnamodendron ekmanii Sleu- 
mer 


Cinnamosma madagascariensis 


Dan 

icin macranthum 
(Baill.) Tie 

iii salutari (G. Bertol.) 
Chiov. 


New Caledonia, Plateau de 
Dogny, LBT 200, NO 


South America, LBT 124, NO 

South America, Butzke et al. 
11521, US 

Dominican Republic, Samaná 
Penninsula, García & Veloz 


6866, SD 
Madagascar, Lowry 4991, MO 
s Rico, Axelrod 10783, 


es Africa, Mpumalanga, 
Goldblatt 11314, MO 


Suh et al., 1993 AY004119 AY004135 
Suh et al., 1993 AY004123 AY004140 
NO 
Suh et al., 1993 AY004126 AY004143 
Suh et al., 1993 AY004121 AY004138 
Suh et al., 1993 AY004124 AY004141 
Suh et al., 1993 AY004125 AY004142 
This paper AY004129 AY004146 
Suh et al., 1993 AY004127 AY004144 
Suh et al., 1993 AY004128 AY004145 
Suh et al., 1993 AY004122 AY004139 
Suh et al., 1993 AY004120 AY004137 
Suh et al., 1993 AY004118 AY004135 
Suh et al., 1992 L03844 AY004152 
This paper AY004132 AY004149 
This paper AY004133 AY004150 
This paper AY004131 AY004148 
This paper AY004134 AY004151 
This paper AY004130 AY004147 


Nuclear ITS. The alignment of Winteraceae 
ITS sequences of Suh et al. (1993) was used as a 
si to easily wile the Takhtajania ITS se- 

The ralogous copies (GenBank 
AY004111-AYO04117) of the ITS region as deter- 
mined by Buckler et al. (1997) were not included 
in the alignment. 


Alignment of ITS sequences within Canellaceae 
also was easily determined by eye. d of 
Canellaceae with Winteraceae seque into a 
single data set required additional чи шан ал to 
generate a plausible alignment. One region of ITS 
1 and another in ITS 2 were unalignable across 
both families, though these regions were alignable 


Моште 87, Митбег 3 


Karol et al. 
Molecular Evidence 


among families. These regions were treated as non- 
overlapping insertion/deletion (indel) events in the 
final data set. Inferred indel regions were included 
in all phylogenetic analyses with the resulting gaps 
treated as missing. 

Putative secondary structures of ITS 2 were de- 
termined for each taxon employing the minimum 
free-energy program mFOLD (Zuker, 1989) using 
constraints previously described by Hershkovitz 
and Zimmer (1996). In their study, the putative sec- 
ondary structure of Canella ITS 2 was determined, 
and this structure was used as a guide to determine 
structures for all remaining Canellaceae. 

Chloroplast trnL-F. The trnL-F spacer regions 
were easily aligned by eye within and between both 
families, with the resulting indels treated identi- 
cally to those in the ITS data set. 

Combined data set. The aligned ITS and trnL- 
F spacer sequences were then combined into a sin- 
gle data set. Two partitions were defined (ITS and 
trnL-F) and subjected to the data partition-homo- 
geneity test (Farris et al., 1995). A thousand rep- 
licates were performed and the resulting P-value 
was used to determine if using the combined data 
set for phylogenetic reconstruction would be appro- 
priate. 

In summary, three data sets were examined: (1) 
the ITS data set consisting of the ITS 1, 5.85, and 
ITS 2 regions; (2) the trnL-F data set consisting of 
a group I intron, the 5'-trnL exon, a non-coding 
intergenic spacer region, and partial sequence of 
trnF; and (3) a combined data set consisting of both 
the ITS and trnL-F regions. 


PHYLOGENETIC ANALYSES 


All phylogenetic reconstruction analyses were 

conducted using version 4.0Ь1 or 4.052 of PAUP* 
(Swofford, 1998). 

Maximum parsimony. For each data set, phy- 
logenetic reconstruction under maximum parsimo- 
ny (MP) was conducted by utilizing the Branch and 
Bound search option in PAUP* with TBR branch- 
swapping, MULPARS, and ACCTRAN options ac- 
tive. Characters were assigned equal weights at all 
nucleotide positions (Fitch, 1971). Robustness of 
cladistic linkages was evaluated with 1000 boot- 
strap replicates (Felsenstein, 1985; Sanderson, 

989). Decay values (Bremer, 1988; Donoghue et 
al., 1992) for each node were calculated using the 
TOPOLOGICAL CONSTRAINTS option in PAUP*. 

Maximum likelihood. The strategy employed 
utilized the likelihood-ratio test statistic (Felsen- 
stein, 1981; Goldman, 1993; Yang et al., 1995) to 
determine the best model of DNA substitution for 


each of the three data sets described above. The 
models considered include the general time-revers- 
ible model (GTR, equals REV of Yang, 1994a), 
Hasegawa et al. (1985; denoted HKY85), Kimura 
(1980; denoted K2P), and Jukes and Cantor (1969; 
denoted JC69). Equations for calculating likeli- 
hoods under these DNA substitution models are 
given in Swofford et al. (1996). An iterative pro- 
cedure to first evaluate models and optimize model 
parameters was used on an initial set of trees gen- 
erated from the maximum parsimony analysis. 

For each model of DNA substitution, models that 
incorporate rate variation across sites were ex- 
plored. These included equal rates assumed across 
all sites, a proportion of sites assumed to be in- 
variable with equal rates assumed across variable 
sites (“I” [Hasegawa et al., 1985]), all sites as- 
sumed to follow a discrete approximation of the 
gamma distribution (“Г” [Yang, 1994b]), and some 
sites assumed to be invariable with gamma distrib- 
uted rates at variable sites (“I + Г” [Gu et al., 
1995; Waddell & Penny, 1996]. 

For the combined data set, a set of models was 
tested which explored the possibility that the rel- 
ative rates between the two molecules differed sig- 
nificantly. This class of models assigns sites to clas- 
ses and then estimates the relative rate for each 
class separately (Felsenstein, 1991; Goldman & 
Yang, 1994). Sites were assigned to two classes (ITS 
and trnL-F). This model is denoted as “+ SS." 

For each data set, a maximum likelihood heuris- 
tic search with TBR branch swapping was then per- 
formed using parameters estimated from the tree 
with the best likelihood score under the best model. 
If the resulting tree was different in topology from 
that of the original tree, the resulting tree was used 
to further optimize the model parameters. With pa- 
rameters re-optimized, a heuristic search of ten rep- 
etitions of random taxon addition and TBR branch 
swapping was performed using fully defined model 
parameters. Bootstrap methods (Felsenstein, 1985; 
Sanderson, 1989) with 1000 replicates were per- 
formed to estimate robustness of nodal support. 


RESULTS 


The combined data set used in this study can be 
found at TreeBASE (http://www.herbaria.harvard. 
edu/treebase/). 


SEQUENCE ALIGNMENT 


Nuclear ITS. ITS sequence alignment proved to 
be a relatively simple task within both Winteraceae 
and Canellaceae. With the exception of two regions, 


one in ITS 1 (bp 1143-1163) and the other in ITS 


418 Annals of the 
Missouri Botanical Garden 


Table 2. List of the insertion/deletion (indel) events for the trnL-F/ITS combined data set. Character пате 

for trnL-F; 1-115 for ITS), ded information (character number in the combined data set), indel length (bp), dose 
type (I = parsimony-informative, U = uninformative, H = homoplastic when mapped on Figs. 1 and 2a), and the 
species where indels were ^e are noted. Bp — Belliolum pancheri, Bc — Bubbia comptonii, Dw — Drimys winteri, 
Es — Exospermum stipitatum, Pa — Pseudowintera axillaris, Pc — Pseudowintera colorata, Tp — Takhtajania perrieri, 
Ti hid: insipida, Tl = Tasmannia lanceolata, Za = Zy um acsmithii, Zba — Zygogynum balansae, Zbi 
— Zygogynum bicolor, Cw — Canella winterana, Cd — а dinisii, Се = Cinnamodendron ekmanii, Ст = 
Cinnamosma madagascariensis, Pm = Pleodendron macranthum, Ws = Warburgia salutaris. 


Position in combined 


Character data set Length (bp) ИОН Clade 
trnL-F 

A 114 1 | Winteraceae/Canellaceae 
B 179-181 3 I Winteraceae/Canellaceae 
C 207-212 6 U Tp 

D 247-255 9 U Cw 

E 252-256 5 I Winteraceae/Canellaceae 
F 7—2 8 U 8 

G 282-286 4 U Cm 

H 7 1 U Cw 

I 319-331 13 U Cw 

J 323-331 9 U Tp 

K (a, b, c) 359-364. 1 Н a = (Ti, Tl, Tp), b = (TI, Tp), 

с = TI 
I 379-384 6 | Dw, Pc, Pa, Bc, Bp, Es, Za, 
Zba, Zbi 
M 385-388 4 U C 
414—418 5 I Winteraceae/Canellaceae 

O 586-591 6 H Run of Adenines (see text) 
P 590—694 105 I Winteraceae/Canellaceae 
Q 633 1 U Dw 

R 634 1 U Tp 

5 706 1 I Winteraceae/Canellaceae 
T 710 1 I Winteraceae/Canellaceae 
U 717-718 2 I Winteraceae/Canellaceae 
V 140—750 11 U Ws 

W 712 1 U Ti 

X 897 || Н Са, Cm, Ws 

ITS-1 

l 993 1 1 Winteraceae/Canellaceae 
2 997 1 I Be, Bp, Es, Za, Zba, Zbi 
3 998—999 2 I Cd, Ce, Cm, Pm, W 

4 1000 1 U Cd 

5 1003-1013 11 U Cd 

6 1002 ] I Pa, Pc 

7 1003 || U Tp 

8 1008 1 | Winteraceae/Canellaceae 
9 1009 1 Џ IW 

10 1027-1028 2 I Winteraceae/Canellaceae 
11 1040 1 Џ м 

12 1041 1 | Winteraceae/Canellaceae 
13 1042 || H eras 

14 1045 1 | d, Cm, 

15 1048-1049 2 H Canellacea 

16 1050-1051 2 I о сене банане 
17 1052 1 [ Cd, Ст, Ws 

18 1053-1054. 2 U Ws 

19 1058 1 Н Canellaceae, Dw 

20 1059 1 Н 


Volume 87, Number 3 
2000 


а!. 
Емдепсе 


419 


Table 2. Continued. 


Position in combined 


Character Length (bp) I/U/H Clade 
21 1060-1061 2 U Dw 
22 1068 1 U Cw 
23 1076-1077 2 U Cw 
24 1081-1082 2 I Tl, Ti 
25 1083-1084 2 I Winteraceae/Canellaceae 
26 1090 || Н Winteraceae, Ст, Рт 
27 1091 1 Н Се, Cw, Pm 
28 1094-1106 13 U Cd 
29 1194—1097 4 U Ws 
30 1198—1104 7 Н Се, См, Pm, Ws 
31 1105-1106 2 U Pm 
32 1112-1113 2 U Dw 
33 1114 1 || Winteraceae, Cw 
34 1115 1 | Winteraceae/Canellaceae 
35 1116-1118 3 I Winteraceae, Cw 
36 1119-1122 4 I Winteraceae/Canellaceae 
37 1128 1 U w 
38 1135 || 1 Winteraceae/Canellaceae 
39 1141-1142 2 H Winteraceae, Cd, Cw 
40 1143-1146 4 I Winteraceae/Canellaceae 
41 1147-1163 17 I Winteraceae/Canellaceae 
42 1148-1149 2 U Cd 
43 1158 1 1 Ст, Ws 
44 1176 1 U Cw 
45 1185-1188 4 I Winteraceae/Canellaceae 
46 199 || U w 
47 1206-1225 20 I Winteraceae/Canellaceae 
48 1209-1212 4 I e, Pm 
49 1223-1225 3 I Ce, Pm 
50 234 1 | Winteraceae/Canellaceae 
51 1244. 1 U Cw 
52 1245 1 | Winteraceae/Canellaceae 
53 1262-1277 16 I Winteraceae/Canellaceae 
54. 1262-1263 2 I Ce, Pm 
55 1272 1 1 Cd, Ст, Ws 
56 1273 | 1 Се, Рт 
57 1283-1284 2 H Ce, Cw, Pm 
58 1287 1 Н Cd, Cm, Cw, Ws 
59 1292-1293 2 I Bc, Bp, Es, Za, Zba, Zbi 
60 1299 || H m, Pm 
61 1308-1326 19 I Winteraceae/Canellaceae 
62 1311 1 I Bp, Es, Za, Zba, Zbi 
63 1312-1313 2 H Bc, Pa, Ti, Tl, Tp 
64 1314-1316 3 U p 
65 1325 1 H Winteraceae, Ws 
66 1326 1 H Cd, Cm, Cw 
67 1331 1 U Cw 

5.88 
68 1487 1 U Cw 
ITS-2 

69 1498 1 Н Pa, Pc, Bc, Es, Za, Zba, Zbi 
70 1504. 1 I Winteraceae/Canellaceae 
71 1511-1513 3 I Winteraceae/Canellaceae 
72 1514 1 Н Cd, Се, Ст, Pm 
73 1523 1 Н Са, Се, Cw, Pm, Ws 


420 


Annals of the 
Missouri Botanical Garden 


Table 2. Continued. 


Position in combined 
data set 


Character Length (bp) ИОН Clade 
74. 1524—1525 2 Н Bc, Bp, Cm, Es, Ti, Tl, Za, 
Zba, Zbi 
75 1526 1 I c, Bp, Cm, Es, Za, Zba, Zbi 
76 1527 1 I Winterac se nella eds 
77 1534 1 Џ Dw 
78 1535-1539 5 I Winteraceae/Canellaceae 
79 1540 || H Canellaceae, Bc 
80 1543-1544. 2 U lw 
81 1545 1 1 Winteraceae/Canellaceae 
82 1553-1554. 2 I а, Ре 
83 1573-1575 3 I Winteraceae/Canellaceae 
84. 1595-1596 2 I 
85 1603 1 H Ба Bp, Се, Ст, Es, Pm, Ti, ТІ, 
Za, Zba, Zbi 
86 1604 ] I Bc, Bp, Cm, Es, Ti, Tl, Za, 
Zba, Zbi 
87 1612-1618 7 U Cw 
88 1651 1 U p 
89 1652-1653 2 I Winteraceae/Canellaceae 
90 1659 1 1 Winteraceae/Canellaceae 
91 1662 || U Pm 
92 1663 1 I Winteraceae/Canellaceae 
93 1672 1 Н Winteraceae, Ст 
94. 1676 1 1 id, С 
95 1683-1688 6 U Ce 
96 1690-1691 1 I Ра, Ре 
97 1692-1706 15 I Winteraceae/Canellaceae 
98 1692-1695 4 U Tl 
99 1700-1701 2 U Tp 
100 1707-1722 16 I Winteraceae/Canellaceae 
101 1709 1 U Ce 
102 1713-1716 4 U Cw 
103 1717-1718 2 H Cw, Pm 
104 1719–1720 2 U Ce 
105 1726-1727 2 U TI 
106 1730 1 | Winteraceae/Canellaceae 
107 1737 1 U Pe 
108 1738 || Џ Ра 
109 1736-1748 10 U с 
110 1750-1759 11 I Winteraceae/Canellaceae 
111 1757-1758 2 U p 
112 1763-1764 2 H Cw, Cd, Pm 
113 1765-1774 10 I Cd, Cm, Ws 
114 1775 1 Н Са, ст 
115 1776-1778 3 U р 


2 (bp 1690-1722), the alignment between families 
was also relatively straightforward. The two prob- 
lematic regions were partitioned equally between 
the two families as non-overlapping separate indels 
(e.g., positions 1143-1153 were coded as gaps in 
Winteraceae and positions 1154—1163 were coded 
as gaps in Canellaceae). The effect of this was to 
reduce the probability of incorrect homology as- 


sessment between families while retaining potential 
phylogenetic information within each family. 
Putative secondary structures for ITS 2 were de- 
termined for each sequence, and these structures 
were generally consistent with the final alignment 
(data not shown). An apparent contradiction be- 
tween secondary structure and DNA alignment was 
found in the v5 region (see Hershkovitz & Zimmer 


Моште 87, Митбег 3 
2000 


Karol et al. 
Molecular Evidence 


[1996] for details) and corresponds to bp 1704— 
1734. Although this region consistently formed a 
bulge in secondary structure analyses across all in- 
dividuals, the base composition and sequence 
length clearly differed between families (11-15 bp 

wit 0% G/C in Winteraceae and 7-16 bp 
with 75-85% С/С in Canellaceae). This result fur- 
ther supports treatment of this region as non-over- 
lapping indel events. No known secondary structure 
exists for ITS 1; consequently, structural evidence 
supporting the alignment of ITS 1 could not be gen- 
erated. 

A total of 115 indels were identified in the ITS 
alignment, of which 53 (46%) were parsimony-in- 
formative, 38 (33%) were autapomorphic, and the 
remaining 24 (21%) were homoplastic. Table 2 pre- 
sents these indels along with position information 
(character number in the data set), indel length, 
and the species where indels were found. 

Chloroplast trnL-F. The trnL-F spacer regions 
were easily aligned by eye within and between fam- 
ilies. Overall, a total of 24 indels (Table 2: A-X) 
were identified in the alignment. Of them, 9 
(37.5%) were parsimony-informative, 12 (50.0%) 
were autapomorphic, and the remaining 3 (12.5% 
were homoplastic. Especially noteworthy was indel 
P (Table 2) with 105 bases present in the Winter- 
aceae, including Takhtajania, but this indel was not 
present in the Canellaceae. 

Combined data set. Two partitions of the com- 
bined alignment were defined, (1) trnL-F (bp 1– 
987) and (2) ITS (bp 988-1778). These partitions 
were tested for congruence with the data partition- 
homogeneity test (1000 replicates) and yielded a P 

0.367. This result fails to reject the null 
hypothesis of congruence between the two parti- 
tions. Therefore, all phylogenetic analyses were 
performed on both the individual and combined 
data sets. 


— 


value o 


PATTERNS OF SEQUENCE EVOLUTION 


The aligned combined data set was 1778 char- 
acters long. Positions 1 to 987 represented the 
trnL-F chloroplast sequence region; positions 988 
to 1778 represented the ITS nuclear ribosomal se- 
quence region including the 5.8S coding region. 
Unlike the situation for certain genera of Wintera- 
ceae (Belliolum, Bubbia, Exospermum, and Zygo- 
gynum), no polymorphisms were detected in the 
ITS sequence for Takhtajania or any member of the 
Canellaceae. 

The ITS 1 sequences in Winteraceae ranged 
from 238 bp in Takhtajania to 252 bp in Drimys. 


In Canellaceae, ITS 1 sequences ranged from 244 


bp in Capsicodendron to 274 bp in Pleodendron. 
The 5.85 coding region was 164 bp in all individ- 
uals of both families, although it was only 163 bp 
long in Canella. ITS 2 sequences in Winteraceae 
ranged from 215 bp in Pseudowintera colorata to 
228 bp in Belliolum, with Takhtajania having a 222 
bp sequence. Canellaceae ITS 2 sequences ranged 
from 187 bp in Cinnamodendron to 211 bp in Ca- 
nella. 

The Winteraceae chloroplast group I intron se- 
quences ranged from 490 bp in Tasmannia insipida 
to 508 bp in Takhtajania perrieri. In Canellaceae, 
these sequences ranged from 477 bp in Canella 
winterana to 495 bp in Cinnamosma madagascar- 
iensis. The 5' trnL exon sequences were 50 bp in 
all individuals of both families. The intergenic 
spacer sequences in Winteraceae ranged from 360 
bp in Drimys to 368 bp in both Tasmannia insipida 
and Takhtajania perrieri. All Canellaceae sequenc- 
es were markedly shorter, ranging from 247 bp in 
Warburgia salutaris to 259 bp in Canella winter- 
ana, Cinnamodendron ekmanii, and Pleodendron 
macranthum. The 40 bp partial trnF exon sequenc- 
es were identical in length in all individuals of both 
families. 


PHYLOGENETIC ANALYSES 
Maximum parsimony 


Nuclear ITS. Using the Branch and Bound 
search procedure in PAUP*, with equal weighting 
of positions and Canellaceae selected as the out- 
group, a single most parsimonious tree of 489 steps 
(366 steps excluding uninformative characters) was 
obtained with the ITS data set. The consistency in- 
dex (Kluge & Farris, 1969) was 0.8221 (0.7623 
excluding uninformative о and retention 
index (Farris, 1989) was 0.8793. Overall, 317 var- 
iable nucleotide кин were counted; of these, 
206 (or 26% of the 791 total characters) provided 
parsimony-informative changes. 

о assess robustness of these results, the bootstrap 
procedure (1000 replicates) and decay analyses were 
run in PAUP* (TOPOLOGICAL CONSTRAINTS 
were used to determine decay values for each node). 
These results are shown in Figure 1. 

A polytomy including Belliolum, Exospermum, 
and Zygogynum (Zygogynum s.l.) was supported as 
monophyletic and sister to Bubbia by a 9796 boot- 
strap value and decay index of + 4. The Bubbia + 
Zygogynum s.]. clade was monophyletic and sister 
to Pseudowintera (represented by two species, 
monophyletic in 9996 of the bootstrap iterations 
[decay = + 5]) supported by a 10046 bootstrap 
value (decay = + 10). Drimys was sister to this 


422 Annals of the 
Missouri Botanical Garden 


Zygogynum bicolor 


Belliolum pancheri 


Zygogynum balansae 


Exospermum stipitatum 


Zygogynum acsmithii 


Bubbia comptonii 


p^ Pseudowintera axillaris 


99/100 | 3/3 
Pseudowintera colorata 


| 1111 24/31 


LT 111 Drimys winteri 
9^0 Tasmannia insipida 
95/128 p 1118 | 
00/100 ' 99/100 7/9 
61/+91 +8/+13 Tasmannia lanceolata 


|||] | 28/40 
1 Ји и 


Takhtajania perrieri 


21/23 


Warburgia salutaris 


Cinnamosma madagascariensis 


Capsicodendron denisii 


Cinnamodendron ekmanii 


+25/+27 Pleodendron macranthum 


51/56 
— -HHHHHH-HH-H— —- canella winterana 


Figure l. Single most о tree generated both from the ITS data set alone and the combined ITS/trnL-F 
data set. Tree length for ITS = 489 steps (366 steps excluding uninformative characters), consistency index = 


steps (434 steps excludin ng uninformative characters), consistency index = 0.8381 (0.7788 excluding uninformative 
characters), and retention index = 0.8994. Numbers to the left of the slash represent those generated from ITS data 
alone, while numbers to the right of the slash represent those generated from the combined data set. Branch lengths 
shown above branches, bootstrap values (1000 replicates) and decay values shown below branches, respectively. Dark 
bars represent non-homoplastic insertion/deletion events inferred from the ITS alignment (see Table 2). 


Volume 87, Number 3 
2000 


Karol et al. 423 


Molecular Evidence 


assemblage in 95% of the bootstrap iterations (de- 
cay = + 3). Tasmannia (represented by two spe- 
cies) was monophyletic with 99% bootstrap support 
(decay = + 8) and sister to the Drimys + Pseu- 
dowintera + Bubbia + Zygogynum s.l. clade in 
86% of the bootstrap iterations (decay = + 3). 
Takhtajania was found sister to the remainder of 
the Winteraceae with a bootstrap value of 94% (de- 
cay = + 8). 

In the Canellaceae, Cinnamosma and Warburgia 
formed a clade sister to Capsicodendron in 73% of 
the bootstrap replicates, with a decay index of + 
3. Cinnamodendron and Pleodendron formed a 
well-supported clade (100% of the bootstrap rep- 
licates, decay = + 25) sister to the Capsicodendron 
+ Cinnamosma + Warburgia clade in 88% of the 
replicates (decay = + 7). Finally, Canella was 
found sister to the remainder of the family with 
84% bootstrap support (decay = + 4). 

Chloroplast trnL-F. Using the Branch and 
Bound search procedure in PAUP*, with equal 
weighting of positions and Canellaceae selected as 
the outgroup, two most parsimonious trees of 103 
steps (67 steps excluding uninformative characters) 
were obtained with the trnL-F data set. The con- 
sistency index was 0.9223 (0.8806 excluding un- 
informative characters), and the retention index was 
0.9657. Overall, 90 variable nucleotide positions 
were counted; of these, 55 (or 5.6% of the 987 total 
characters) provided parsimony-informative chang- 
es. These two trees are presented in Figure 2 along 
with bootstrap values and decay indices. 

With the exception of alternative equally parsi- 
monious basal relationships, the topology generated 
by the trnL-F sequence data for Winteraceae was 
not in conflict with the tree generated by the ITS 
data, albeit there were both lower resolution and 
lower bootstrap support values. The polytomy of 
Belliolum, Exospermum, and Zygogynum (Zygo- 
gynum s.l.) expanded to include Bubbia (78% boot- 
strap and decay = + 2). The two species of Pseu- 
dowintera were unresolved with respect to each 
other and to the Bubbia + Zygogynum s.l. роју- 
tomy, and together these were only weakly sup- 
ported as monophyletic and sister to Drimys (69% 
bootstrap and decay = + 1). The two species of 
Tasmannia were strongly supported as sister taxa 
in 9896 of the bootstrap iterations (decay = + 5). 
Finally, the trnL-F sequence data could not resolve 
basal relationships within the family; both Takhta- 
jania (Fig. 2a) and Tasmannia (Fig. 2b) as sister to 
the remaining Winteraceae were equally parsimo- 
nious. 

Within Canellaceae, except for a strong sister re- 
lationship between Cinnamodendron and Pleoden- 


dron (8896 bootstrap and decay = + 2), the topol- 
ogy generated by trnL-F data resulted in much 
lower levels of bootstrap support, and, moreover, 
suggested a fundamental split between Old World 
and New World lineages. Thus, rather than being 
sister to the remaining Canellaceae as suggested by 
the ITS-based phylogeny, Canella was nested with- 
in a weakly supported clade of New World genera. 
To force such a division of the family into Old 
World and New World clades requires an additional 
11 steps in the ITS-based phylogeny. 

Combined data set. Using the Branch and 
Bound search procedure in PAUP*, with equal 
weighting of positions and all Canellaceae genera 
selected as outgroups, a single most parsimonious 
tree of 593 steps (434 steps excluding uninforma- 
tive characters) was obtained with the ITS/trnL-F 
combined data set. The consistency index was 
0.8381 (0.7788 excluding uninformative charac- 
ters) and the retention index was 0.8994. The to- 
pology derived from this combined data set was 
identical to that generated from the ITS data alone. 
The branch lengths, bootstrap values, and decay 
indices are presented in Figure 1 

Overall, with the addition of the trnL-F data, 
bootstrap values either remained similar or in- 
creased in comparison with the ITS-based phylog- 
eny. Within the Winteraceae, the most notable in- 
creases resulted in stronger support for basal 
relationships in the family. The position of Takh- 
tajania as basal and sister to the rest of the family 
was further strengthened (ITS — 9446 bootstrap and 
+ 8 decay index vs. combined data = 99% boot- 
strap and + 11 decay index), as was the position 
of Tasmannia as the next branch within the family. 
The combined data set provided especially strong 
support for the inclusion of Drimys in the clade 
consisting of all other members of the family and 
sister to Tasmannia (ITS = 86% bootstrap and + 
3 decay index vs. combined data = 97% bootstrap 
and + 6 decay index). Within the Canellaceae, 
bootstrap support also remained similar or in- 
creased only slightly with the addition of the trnL- 
F data (Fig. 1). 


Maximum likelihood 


Nuclear ITS. For the ITS data set, the best 
model of DNA substitution was the general time- 
reversible model with a proportion of sites assumed 
to be invariable and equal rates assumed across 
variable sites (СТЕ + I). With this model, param- 
eters were fully optimized using the tree generated 
from the parsimony search (Fig. 1). With the model 
parameters defined, a heuristic search was con- 


424 Annals of the 
Missouri Botanical Garden 


a 2 Zygogynum bicolor 
Belliolum pancheri 
Zygogynum balansae 


78/+2 
—_ Exospermum stipitatum 


Zygogynum acsmithii 


69/+1 — — — Bubbia comptonii 


Pseudowintera axillaris 


3 
79/+2 


Pseudowintera colorata 


46/+0 1 Drimys winteri 
1 eee 
Tasmannia insipida 
33 7 


100/+31 98/+5 | 2 
Tasmannia lanceolata 


+ rt Takhtajania perrieri 
Cinnamodendron ekmanii 


Pleodendron macranthum 


Canella winterana 


Capsicodendron denisii 


[H^— Warburgia salutaris 
1 
49/41 6 
Lys Cinnamosma madagascariensis 


Figure 2a, b. Two most parsimonious trees келерде using trL- F data. Tree length = = 103 steps (67 steps excluding 
uninformative cha ын саа index = 0.9223 (( aracters), and retention index 
= 0.9657. Branch lengths shown above branches, bootstrap values (1000 replicates) а decay values shown below 
branches, respectively. s bars (Fig. 2a) represent non-homoplastic insertion/deletion events inferred from the trnL- 
F alignment (see Table 2). 


Volume 87, Number 3 Karol et al. 425 
2000 Molecular Evidence 


b _ Zygogynum bicolor 


Belliolum pancheri 


— — Zygogynum balansae 


78/+2 


Exospermum stipitatum 


Zygogynum acsmithii 


1 
69/+1 — — —— Bubbia comptonii 


Pseudowintera axillaris 


79/+2 
0 у 
Pseudowintera colorata 


Drimys winteri 


47/+0 


Takhtajania perrieri 


0 "eem 
Tasmannia insipida 
8 
98/+5 | 3 
Tasmannia lanceolata 


Cinnamodendron ekmanii 


33 
100/+31 


Pleodendron macranthum 


Canella winterana 


С. icodend denisii 


2 ; : 
mE Warburgia salutaris 
1 
49/+1 | 6 
Cinnamosma madagascariensis 


Figure 2. Continued. 


426 Annals of the 
Missouri Botanical Garden 


Zygogynum bicolor 
2 
63 | Belliolum pancheri 


— Zygogynum balansae 


wo 
o 


r Exospermum stipitatum 
100 
— Zygogynum acsmithii 


89 — Bubbia comptonii 


Pseudowintera axillaris 


Pseudowintera colorata 


Drimys winteri 


0 Таѕтаппіа insipida 


Tasmannia lanceolata 


Takhtajania perrieri 


Warburgia salutaris 


80 Cinnamosma madagascariensis 


Capsicodendron denisii 


Canella winterana 


[ Cinnamodendron ekmanii 


Е Pleodendron macranthum 
0.05 substitutions/site 


of four most es! phylograms generated with ITS data (—In = 3371.16) with bootstrap values 
ie ыы е branches 


Моште 87, Митбег 3 
2000 


Karol et al. 
Molecular Evidence 


427 


Zygogynum bicolor 
Belliolum pancheri 
| Zygogynum balansae 


— Exospermum stipitatum 


|Zygogynum acsmithii 


—— Bubbia comptonii 


— Pseudowintera axillaris 


'Pseudowintera colorata 
Drimys winteri 


| Tasmannia insipida 


98 
M Tasmannia lanceolata 
Takhtajania perrieri 
~ Warburgia salutaris 


^^ я Ј 


denisii 


— — 0.005 substitutions/site 


Figure 
(1000 ть above branches. Figure 3c is оп pag 


ducted that resulted in three trees, one of which 
was identical in topology to the most parsimonious 
tree (Fig. 1). These three trees were used to further 
optimize model parameters. The resulting parame- 
ter values did not differ from those generated by 
using the single maximum parsimony tree. Thus, 


ingle most likely phylogram ке а trnL-F data (—1п 


Cinnamosma madagascariensis 


Cinnamodendron ekmanii 
87 | 
Pleodendron macranthum 


Canella winterana 


1996.3473) with bootstrap values 


ten heuristic searches (with random taxon addition, 
multrees, and steepest descent active) were con- 
ducted using the parameters defined in the initial 
search. These ten random taxon addition searches 
generated a total of four equally likely trees (—In = 
3371.16), one of which is presented in Figure 3a. 


428 Annals of the 
Missouri Botanical Garden 
C Zygogynum bicolor 
64 
e4|- Belliolum pancheri 
Zygogynum balansae 
98 


Г Exospermum stipitatum 


— Zygogynum acsmithii 


— Bubbia comptonii 


=| Pseudowintera axillaris 
99 


Pseudowintera colorata 


e 


Dr imys winteri 


Tasmannia insipida 


ed 


Tasmannia lanceolata 


Takhtajania perrieri 
Warburgia salutaris 


Cinnamosma madagascariensis 


denisii 


[ Стпатодепагоп ekmanii 


—— 0.01 substitutions/site 


Figure 3c. 
eani values (1000 replicates) above branc 


In all four trees the relationships within Winter- 
aceae were identical to those generated by the MP 
search, with Takhtajania sister to the remainder of 
the Winteraceae. The four trees differed in the po- 
sition of Canella in the Canellaceae. Either Canella 
was sister to the Warburgia + Cinnamosma + Сар- 
sicodendron clade, or sister to the Cinnamodendron 
+ Pleodendron clade, or sister to the remainder of 


Single most likely phylogram oo with combined ITS/trnL-F data set (In 
hes 


L Pleodendron macranthum 
Canella winterana 


= —5508.26165) with 


the family, or, finally, in an unresolved polytomy 
with the three above-mentioned clades. Although 
Canella is included within a monophyletic Canel- 
laceae in 100% of the bootstrap iterations, boot- 
strap values do not support a resolved phylogenetic 
position of Canella within the family. 

Chloroplast trnL-F. For the trnL-F data set, the 
best model of DNA substitution was the HKY85 


Моште 87, Митбег 3 
2000 


Karol et al. 
Molecular Evidence 


model with a proportion of sites assumed to be in- 
variable and equal rates assumed across variable 
sites (HKY85 + 1). With this model, parameters 
were fully optimized using the two trees generated 
from the parsimony search (Fig. 2a, b). With the 
model parameters defined, a heuristic search was 
conducted that resulted in a single tree identical in 
topology to one of the four most likely trees gen- 
erated from the maximum likelihood ITS search 
(Fig. 3a). Using this tree, parameters were re-esti- 
mated and the resulting optimized values were used 
for ten heuristic searches (random taxon addition, 
multrees, and steepest descent active). The result 
of these searches generated a single most likely tree 
(—In = 1996.3473), which is presented in Figure 
3b along with bootstrap values for each node. 

The single tree from the maximum likelihood 
search was identical to one of the two trees gen- 
erated by the maximum parsimony analysis using 
trnL-F with one exception. The placement of Ca- 
nella sister to Pleodendron and Cinnamodendron 
was not supported. Rather, Capsicodendron, Cin- 
namosma, and Warburgia were supported as an un- 
resolved trichotomy in 69% of the bootstrap itera- 
tions. Cinnamodendron and Pleodendron were 
supported as sister taxa in 8796 of the bootstrap 
iterations, and finally Canella was found in an un- 
resolved trichotomy with the two above-mentioned 
clades in a monophyletic Canellaceae in 100% o 
the bootstrap replications. Within the Winteraceae 
an unresolved polytomy including Belliolum, Bub- 
bia, Exospermum, and Zygogynum was found to be 
sister to the two species of Pseudowintera in 71% 
of the bootstrap replicates. In 7846 of the bootstrap 
iterations, Drimys joined the above in a monophy- 
letic clade, but was only weakly supported by a 
5496 bootstrap value as sister to them. There was 
less than 5096 bootstrap support for the basal po- 
sition of Takhtajania sister to the remainder of Win- 
teraceae. 

Combined data set. For the combined data set, 
the best model of DNA substitution was the general 
time-reversible model. Among site rate variation 
was accommodated for by dividing the data into two 
partitions (ITS and trnL-F), thus allowing among 
site rate variation to be estimated for each molecule 
separately (GTR + SS). With this model, parame- 
ters were optimized using the tree generated from 
the maximum parsimony search (Fig. 1). With the 
model parameters defined, a heuristic search was 
conducted that resulted in a single most likely tree 
identical in topology to the most parsimonious tree 
using the same data set (Fig. 1). This tree was used 
to further optimize model parameters. The resulting 
parameter values did not differ, and thus ten heu- 


ristic searches (random taxon addition, multrees, 
and steepest descent active) were conducted using 
the parameters defined in the initial search. These 
ten random taxon addition searches generated a 
single most likely tree (In = —5508.26165). The 
bootstrap values are presented in Figure 3c. 

In general, with the exception of support for Ca- 
nella as basal within Canellaceae, the bootstrap 
values were consistent with those generated by the 
parsimony analyses of the combined data set. In 
the parsimony analysis, Canella was sister to the 
remainder of the Canellaceae in 85% of bootstrap 
iterations, whereas the likelihood analyses recov- 
ered this relationship in only 53% of the bootstrap 
replicates. 


DISCUSSION 


In the initial molecular phylogenetic study of 
Winteraceae, Suh et al. (1993) found it difficult to 
align the ITS region with certainty for several ex- 
emplar outgroup species. The present study in- 
creased generic sampling in Canellaceae from a 
single exemplar (Canella) to a representative of all 
genera, which greatly facilitated homology assess- 
ment between the two families. In addition, the 
more slowly evolving trnL-F chloroplast regions, for 
which no difficulty in alignment was encountered, 
were investigated. With Canellaceae as the out- 
group, the rooting of Winteraceae is no longer am- 
biguous. Phylogenetic analyses of ITS and com- 
bined ITS/trnL-F DNA sequences provided a 
well-resolved molecular phylogeny for the Winter- 
aceae that placed Takhtajania sister to the remain- 
ing genera in the family. Analysis of trnL-F se- 
quence data alone failed to resolve the basal 
topology within the family, with either Takhtajania 
or Tasmannia equally parsimonious as the basal 
branch. Nevertheless, the addition of trnL-F se- 
quences to a combined ITS/trnL-F analysis resulted 
in increased support values for the same topolo 
derived from ITS sequences alone. Maximum like- 
lihood analyses of each of the three data sets mir- 
rored those obtained through maximum parsimony. 

ITS and trnL-F sequences provided generally 
weaker support for phylogenetic relationships with- 
in Canellaceae. Although parsimony analysis of ITS 
data revealed moderate support for a basal position 
of Canella sister to the remaining members of the 
family, parsimony analysis of trnL-F data weakly 
suggested a fundamental New World/Old World 
split in Canellaceae. Parsimony analysis of the 
combined molecular data set resulted in a topology 
identical to that generated by ITS data alone, with 
only slight increases in bootstrap support and iden- 


430 


Annals of the 
Missouri Botanical Garden 


tical decay values. Maximum likelihood analyses of 
each of the three data sets further demonstrated the 
poor resolution these molecules provide for esti- 
mating basal relationships in Canellaceae. Never- 
theless, ITS, combined ITS/trnL-F, and all maxi- 
mum likelihood analyses strongly suggest that the 
southeastern Brazilian endemic Capsicodendron 
forms a clade with the African/Malagasy genera 
Warburgia and Cinnamosma. 

The alignment of ITS and trnL-F sequences for 
the 12 species of Winteraceae and 6 species of Ca- 
nellaceae revealed numerous probable insertion- 
deletion events. When mapped on the trees, these 
indels furnished additional support for certain 
clades. A total of 37 indels (29 in ITS [Fig. 1] and 
8 in trnL-F [Fig. гај) support the separation of Win- 
teraceae from Canellaceae. Three ITS indels sup- 
port the monophyly of Pseudowintera, and three dif- 
ferent ITS indels support the monophyly of the 
Zygogynum s.l. (Bubbia, Belliolum, Exospermum, 
and Zygogynum) assemblage. One indel each in 

oth nuclear and chloroplast sequences (Figs. 1 
and 2a, respectively) support inclusion of Drimys 
in the Zygogynum s.l. + Pseudowintera clade, and 
thus provide additional evidence of its distinctness 
from Tasmannia. Within Canellaceae, four ITS in- 
dels support the sister relationship between Cin- 
namodendron and Pleodendron, whereas a single 
ITS indel supports a sister relationship between Af- 
rican Warburgia and Malagasy Cinnamosma. The 
basal position of Canella in the family is further 
suggested by three ITS indels that support the 
monophyly of the remaining genera of Canellaceae. 
None of the indels found in the trnL-F Canellaceae 
sequences were phylogenetically informative. In to- 
tal, 139 indels were scored overall and 114 (82%) 
were consistent with the trees generated from the 
sequence data. The remaining 25 were homoplastic 
and in some cases difficult to score. For example, 
in trnL-F sequences, long adenine runs occurred at 
varying numbers in closely related taxa. The length 
of these runs was not consistent when mapped on 
the tree, and delimiting homology was difficult 
when scoring this region. No indels were detected 
in either ITS or trnL-F that bear on the position of 
Takhtajania as sister to the remainder of Wintera- 
ceae. 

Zygogynum s.l., Pseudowintera, and Takhtajania 
exhibit a short, early-rupturing involucrum (the 
congenitally fused two, or rarely three, outermost 
perianth parts, which could as well be considered 
the “calyx”) that is persistent in fruit vs. the con- 
dition in both Drimys and Tasmannia where the 
ultimately caducous, late-rupturing involucrum be- 
comes as long as the fully developed next inner 


tepal whorl, and therefore completely encloses the 
flower just prior to anthesis (Doust, 2000 this is- 
sue). А persistent, short calyx occurs in Canella- 
ceae that is similar to the ruptured, persistent in- 
volucrum of basal Takhtajania, suggesting that 
Pseudowintera and Zygogynum s.l. have retained 
the ancestral plesiomorphic condition of an early- 
rupturing involucrum. In conjunction with the hy- 
pothesized phylogenetic positions of Drimys and 
Tasmannia, the differing ontogenetic patterns of te- 
pal initiation and development exhibited by these 
two genera (Doust, 2000) imply that the late-rup- 
turing, caducous involucrum has evolved twice in- 
dependently. Similarly, deep red tepal color, pre- 
sent in Canella and other Canellaceae, as well as 
Takhtajania and тап 
may be the ancestral condition, with the evolution 
of white tepals through the loss of pigmentation 
and/or the acquisition of an optical tapetum (En- 
dress et al., 2000 т issue) occurring a number of 
times ашан 

The historical pm implications of 
Takhtajania in a basal position sister to the re- 
maining genera of the Winteraceae have been dis- 
cussed in the context of the distribution of fossil 
Winteraceae (Doyle, 2000 this issue) and ecophys- 
iological constraints (Feild et al., 2000 this issue). 
It is reasonable to assume that the basal branch 
leading to Takhtajania became isolated in Mada- 
gascar after reaching there from continental Africa, 
and also that Madagascar may have played a prom- 
inent role in the migration of the family to Aus- 
tralasia and South America via Antarctica during 
the mid-Cretaceous. The presence of two different 
winteraceous pollen types in the Miocene of the 
southwestern Cape possibly referable to Tasmannia 


Zygogynum s.l. species, 


and the most advanced Zygogynum s.l. clade is 
more puzzling and problematic (Coetzee & Prag- 
lowski, 1988). With South Africa relatively isolated 
from both South America and Antarctica from the 
mid-Cretaceous onward (Smith et al., 1994), either 
all of the currently extant clades of Winteraceae 
had already evolved prior to that point in time, or 
Madagascar may have continued to serve as a con- 
duit for migration of more advanced Winteraceae 
back into South Africa from Australasia. In com- 
parison to the Southern Gondwanan pattern of ra- 
diation in Winteraceae, the preliminary phyloge- 
netic hypothesis of infrafamilial relationships 
within Canellaceae suggests an "inverted" Northern 
Gondwanan biogeographical history. Basal clades of 
Canellaceae are centered in northern South Amer- 
ica and the West Indies, with a more advanced 
clade exhibiting a southeastern Brazil/African-Mal- 
agasy split related to the middle Cretaceous sepa- 


Volume 87, Number 3 


Karol et al. 431 
Molecular Evidence 


ration of South America and Africa (Goldblatt, 
1993) and a subsequent dispersal event to Mada- 
gascar. Thus, the modern-day sympatric occurrence 
of the two Malagasy endemic genera Cinnamosma 
(Canellaceae) and Takhtajania (Winteraceae) may 
well be the end result of vastly divergent biogeo- 
graphic histories. 


Literature Cited 


Bremer, K. 1988. The limits of amino acid sequence data 
in angiosperm phylogenetic reconstruction. Evolution 
42: 795-803. 

Buckler, E. S., IV, A. Ippolito & T. P. Holtsford. 1997. 
The evolu&on of ribosomal DNA: Divergent paralogues 

and op aue implications. Genetics 147: 821—832. 

,D [0] 


Сћаве, 7. Soltis, В. Imstead, D. Morgan, 
D es, "B D. Mishler, M Duvall, . Price, 
H. G. Hills, Y.-L. Qi ron, J. H. Re 


Y. Xiang, G. M kett, P. S. Soltis, S. M. Swensen, 
S. E. Williams, P. A. Gadek, C. J. Quinn, L. E. Eguiarte, 
E. Golenberg, G. H. Learn, Jr., S h 


: An айа al nuclenlde se 
quences from the nd gene rbcL. Ann. Missouri Bot. 
Gard. 80: 528-58 

Coetzee, J. A. & J. Куана. 1988. Winteraceae pollen 
from the Miocene of the southwestern Cape (South Af- 
rica): Relationship to modern taxa and phytogeograph- 
ical significance. Grana 27: 27—37. 

n, J. A., A. Culham & J. L. Jury. 1998. Reclas- 

on of Actaea to include Cimicifuga and Souliea 

a uneulaceac} Phylogeny inferred from morphology, 
nrDNA ITS, and cpDNA trnL-F sequence variation. 
Taxon 47: Eo rdi 

Cronquist, A. 1981. An Integrated System of Classifica- 
tion of Flowering Plants. Columbia Univ. Press, New 

k 


York. 

Donoghue, M. J. & J. A. Doyle. 1989. Phylogenetic anal- 
ysis of angiosperms and the relationships of Hamame- 
lidae. Pp. 17-45 in P. R. Crane & S. Blackmore (edi- 
tors), Evolution, pura. and Fossil History of the 
Hamamelidae, -= a rendon hoan Oxford. 

, R. G. Olm ‚ ]. Е5 & J. D. Palmer. 

1992. К ee Bs | Dipsacales base 

on rbcL sequences. Ann. Missouri Bot. Gard. 79: 333— 


Doust, A. N. 2000. Comparative floral ontogeny in Win- 
teraceae. Апп. Missouri Bot. Gard. 87: 366-379 

Doyle, J. A. 2000. Paleobotany, relationships, and g 
graphic history of Winteraceae. Ann. Missouri Bot. 
Gard. 87: 303-316. 

Doyle, J. J. & J. L. Doyle. 1987. A rapid DNA isolation 
procedure for EE Ab rai of fresh leaf tissue. Phy- 
tochem. Bull. 19: 11-15. 

Endress, P. K., A. ы heim, F. В. Sampson & С. 
Schatz. 2000. Floral adie of Takhtajania and its 
systematic position in Winteraceae. Ann. Missouri Bot. 


9. The retention index and the rescaled 
consistency index. Cladistics 5: 417—419. 


, M. Kallersjé, A. С. Kluge & С. Bult. 1995. Test- 
ing significance і in congruence. Cladistics 10: 315-319. 
i M. Holbrook. 2000. 


5 
т 
un 
$ 
É 
D | 
о 
© 
2. 
e 
e 
у= | 
с. 
со 
-l 
w 
| 


ive. А 

Felsenstein, J. 1981. Evolutionary trees from DNA se- 
quences: A maximum likelihood approach. J. Molec. 
Evol. Е. 368-376 

985. Cond ikio limits on E | ар- 
ues | the bootstrap. Evolution 39: 783— 

——.. in a пина beris лы 
version 24 | Seat 

Fitch, W. M. 1971. p defining the course of evo- 

for a specific tree topology. 


. Yuan, P. Kupfer & P. Taberlet. 
Phylogenetic use of noncoding regions in the 
Gentiana L.: Chloroplast a (UAA) intron versus nu- 
"im ribosomal internal зрее spacer sequences. 

c. Phylogenet. Evol. 5: 460-466 

& P. (editor). 1993. Biological Relationships Be- 
tween Africa and South America. Yale Univ. Press, New 
Haven and London. 

Goldman, N. 1993. Il. ip a Е of DNA 
substitution. J. Molec. : 182 

ang. 1994. Ў D Suit em of nucle- 

otide substation for protein-coding DNA sequences. 
Molec. Biol. Evol. 11: 725-736. 

Gottlieb, O. R., M. A. C. Kaplan, K. Kubitzki & T. Barros. 
1989. Chemical dichotomies in the Magnolialean com- 
plex. Nordic J. Bot. 8: 437-444. 

Gu, X., Y.-H. Fu & W.-H. Li. 1995. Maximum likelihood 
estimation of the heterogeneity of substitution rate 
among nucleotide sites. Molec. Biol. Evol. 12: 546—557. 

Hasegawa, M., H. Kishino & T. Yano. 1985. Dating the 
human-ape split by a angl clock of mitochondrial 

. J. Molec. Evol. 22: 

Воено M. A. & E. A. т. 1996. Conservation 
patterns in angiosperm rDNA 1752 sequences. Nucl. 
Acids Res. 24: 2857-2867. 

Jukes, T. H. & C. R. Cantor. 1969. Evolution of protein 


lutionary rate of base substitutions through i њи 
studies ч nucleotide sequences. J. Molec. Evol. 
111-12 

Kluge, A. с & Ј. 5. Farris. 1969. Mer RM 
and the evolution of айнаш. pu 1 

Leroy, J.-F. 1978. Une зоиз-Е x 
teraceae endémique à Madagascar: ins Pu 
deae. Paw x 2, 17: 383-395. 

McDade, L. A L. Moody. 1999. Phylogenetic re- 
lationships PER WA LUE Evidence from noncod- 
ing trnL-trnF chloroplast DNA sequences. Amer. J. Bot. 

: 70—80 


Mes, T. H. M. & H. T. Hart. 1994. Sedum surculosum 
and S. jaccardianum (Crassulaceae) share a unique 70 
bp deletion in the chloroplast DNA trnL (UAA)—trnF 
(GAA) E PE spacer. Pl. Syst. Evol. 193: 213-221. 

Molvray, M., P. J. Kores & M. W. Chase. 1999. Phylo- 
genetic relationships within Korthalsella РЕН 
based on nuclear ITS and plastid trnL-F sequence data 
Amer. J. Bot. 86: 249—260. 

Morgan, D. R. & D. E. Soltis. 1993. Phylogenetic ie 

lat 


tionships among пне of Saxifragaceae sensu 


432 


Annals of the 
Missouri Botanical Garden 


based on rbcL sequence data. Ann. Missouri Bot. Gard. 
80: 631-660. 
Peres ж. la Вова, J., S. A. Harris & А. Farjon. 1995. 
t DNA gii in Mexican pines. 
Theor A Gen 91: 1101-11 

u, Y.-L., e, D. H. с С. Ң. Parks. 1993. 
p es ы: женен of the Magnoliidae: Cladistic 


analyses of nucleotide sequences of the plastid gene 
rbcL. Ann. Missouri Bot. Gard. 80: 587 

Rambaut, Se-Al Sequence Alignment Editor 
Version 1.0 1. Departm 


alpha ent of Zoology, University 

of Oxford, Sot Parks Road, Oxford OX1 4JD, U.K. 
Sanderson, M. J. ;onfidence limits on phylogenies: 
The bootstrap visited Cladistics 5: 
Schatz, С. Е., Р.Р. |, П&А ть 
1998. тбл р perriert rediscovered. Nature 391: 


и С. Smith & В. М. Funnell. 1994. Atlas 
of Меха and Cenozoic coastlines. Cambridge Univ. 
Press, Cambridge 

Suh, Y., L. B. Thien & E. A. Zimmer. 1992. Nucleotide 

sequences of the internal transcribed spacers and 5.8S 

МА gene in Canella winterana (Magnoliales; Canel- 
laceae). Nucl. Acids Res. 20: 6101—6102. 

. Thien, H. E. Reeve & E. A. Zimmer. 1993. 


‚ L. 
Molecular evolution and сше о. и 


ot. 80: 

Swofford, D. L. 1998. PAUP*- Phylogenetic Analysis Us- 
ing Parsimony (*and Other Methods), version 4.0. Sin- 
auer, Sunderland, Massachusetts. 


— ———, С. J. Olsen, P. J. Waddell & D. M. Hillis. 1996. 
Phylogeny Е: Pp. 407—514 in D. M. Hillis, 
C. ире & В. Mable (editors), Molecular System- 
atics, ed. 2. — Sunderland, Massachusetts. 

Taberlet. P., L. Gielly, G. Pautou & J. Bouvet. 1991. Uni- 
versal primers for amplification of three non-coding re- 
s of chloroplast DNA. Pl. Molec. Biol. 17: 1105- 

109. 

Vink b. 1988. Taxonomy in Winteraceae. Taxon 37: 

691-698. 

ире} Р. J. & D. Penny. 1996. Evolutionary trees of 
apes and humans from DNA sequences. Pp. 53-73 in 

C. R. Peters (editors), Handbook of Sym- 
bolic Evolution. Clarendon Press, Oxford. 

White, T. J., T. Birns, S. Lee & J. Taylor. 1990. Ampli- 
fication and direct sequencing of fungal ribosomal RNA 
genes for phylogenetics. Pp. 315-322 in M. Innis 
Sel nshy & T. White (editors), PCR Proto- 
cols: A Guide to Methods and Applications. Academic 
Press, San Die 

Yang, Z. 1994a. JN the nn ч nucleotide sub- 
stitution. J. Molec. Evol. 39: 105-11 

————. 1994b. Maximum likelihood r^s esti- 
mation from DNA sequences with variable rates 
sites: Approximate methods J. Molec. Evol. 42: 204 
307. 


—., №. Goldman & A. Friday. 1995. Maximum like- 
lihood trees from DNA p ew A peculiar statistical 
и. problem. Syst. Biol. 4 399. 

Zuker, 1989. On finding all ina foldings of an 
RN A prn Science 244: 4. 


BOOK REVIEW 


Ribeiro, J. 5. da 5. et al. 1999. Flora da Reserva 
Ducke. Guia de indentificagáo das plantas vas- 
culares de uma floresta de terra-firme na Ama- 
zónia Central. 800 pp. Softcover. ISBN 85-211- 
0011-76. INPA (Brazil) and DFID (U.K.). Retail 
price: $50 U.S. (through http://www.balogh.com). 


I am not normally one to gush praises on a new 
flora book, but this one deserves a score of 12 on 
a scale of 0 to 10 (I admit to giving extra credit on 
my exams). Why such a high rating? Because this 
book excels in a number of important criteria, such 
as high information content, richness of graphics, 
full-color pages, high degree of innovation, reason- 
able price, and all this for a small reserve near 
Manaus in central Amazonian Brazil that shares a 
good number of its species with the much broader 
Amazon basin and neighboring countries. The only 
downfall for some will be the Portuguese text, but 
the book is so graphics-oriented and self-guiding 
that it is still very useable by readers not familiar 
with that language. Still, an English translation of 
this book would be welcomed by many. 

The book is a field guide—in the true sense— 
something you can actually bring into the field with 
you and use to identify forest plants. In fact, it con- 
veniently comes with a transparent, thick plastic 
jacket that wraps around the open face of the book 
and secures to the rear dust cover with a Velcro 
strip. Neat! It is designed for ease of use, that is, 
non-technical and focused mainly on vegetative 
characters, since so often the plants (especially the 
trees) one encounters in tropical forests lack flowers 
or fruits at the time of one's visit. The flora covers 
2175 species of vascular plants, each one illustrat- 
ed by color photographs that are diagnostic of that 
particular group. For instance, nearly all trees have 
small photographs of the outer bark, a bark slash 
(to show exudates or inner cortex characters), an 
entire leaf, a detail of leaf venation, and often some 
other distinguishing feature such as a leaf gland or 
pulvinus. The species entries themselves do not in- 
clude images of flowers or fruits, but at the begin- 
ning of each family, there are composite plates 
showing a large part of the diversity of flowers and 
fruits in the family members of the reserve (take a 
look at pages 152 to 155 to be amazed by the close- 
up photos of Lauraceae flowers and fruits!). 

The Ducke Reserve is a 10 X 10 km forest pre- 
serve situated on the outskirts of Manaus. It has 
been the site of many research projects and silvi- 


cultural experiments over the past decades, but has 
now become a virtual island of forest amid the sub- 
urban sprawl of the capital of Brazilian Amazonia. 
The much broader geographical utility of the flora 
derives from the habitat diversity that is present in 
the reserve. There are four main vegetation types, 
associated with different soils and drainage pat- 
terns: (1) hilltop plateau forest, with clayey soils 
and good drainage; (2) slope forest, actually a gra- 
dient from the hilltop forest down to more sandy 
and poorly drained soils lower down; (3) “campi- 
narana,” or what might also be called “Amazonian 
caatinga," lower forests on white-sand spodosol 
soils; and (4) “baixio,” or alluvial plains along 
streams that have poor drainage and are often wa- 
terlogged. Only small streams traverse the reserve, 
so it lacks the seasonally inundated areas known 
locally as “varzea” or “igap6.” 

I particularly recommend the illustrated glossa- 
ries, which cover pages 24 to 95. They are the best 
I have seen anywhere and include collages of small 
photographs of particular characters, for instance 
leaf glands, exudates, stilt and tabular root varia- 
tions, leaf types and shapes, among many others. 
Identification works by a technique of nested dia- 
gram boxes with the characters written in or some- 
times illustrated, leading to the actual species pag- 
es with further diagram boxes that lead to groups 
of species that can be perused and compared (there 
are short diagnostic text descriptions to accompany 
the graphics). Another endearing aspect of the book 
is feature boxes, or vignettes, interspersed in par- 
ticular families. For instance, under Clusiaceae, a 
feature box (p. 253) illustrates and narrates the re- 
cently published story of parakeet pollination in 
Moronobea coccinea. There is also a valiant attempt 
at a "rapid key" that uses little colored markings 
on the right-hand margin of species pages to narrow 
down a series of criteria listed on pages 94 and 95. 
It is a great concept, though somewhat awkward in 
practice. 

Let me cite a personal example of how useful 
this book is as a field guide. During a recent trip 
to San Carlos de Rio Negro, Venezuela, some 1000 
km to the north, Gerardo Aymard and I collected a 
sterile canopy tree with bipinnate leaves, clearly a 
legume and to our eyes likely an Abarema species, 
but one we had never seen with such huge, “dixie- 
cup"-like petiolar glands. Returning to Caracas, I 
searched the manuscript of the Mimosaceae for the 


ANN. Missouni Bor. GARD. 87: 433—434. 2000. 


434 


Annals of the 
Missouri Botanical Garden 


Flora of the Venezuelan Guayana and then looked 
through the entire family at the National Herbarium 
(VEN), to no avail. It took me a mere five minutes 
to follow the keys through the Ducke Reserve book 
and come across a very likely candidate, Abarema 
adenophora. Upon 
Grimes’s (1996) superb monograph of the genus, I 
confirmed the identification, the first report for Ven- 


consulting Barneby and 


ezuela. 

My only real quibble with the book is in the 
introductory section, where the authors briefly dis- 
cuss the diversity and phytogeography of the re- 
serve. As one of the causes explaining the high 
diversity of the area, they discuss and actually de- 
pict a hypothetical Pleistocene “Lago Amazonas” 
reaching from the base of the Andes to nearly the 
mouth of the Amazon and draining through the pre- 
sent-day Orinoco River to the Caribbean rather 
than into the Atlantic. The original paper postulat- 
ing this lake (Frailey et al., 1988) presented some 
evidence of the existence of a lake in the upper 
Amazon (in Acre State, close to the border of Peru 
and Bolivia), but their extrapolation of the extent 
of the lake to the lower Amazon and draining out 
through the Orinoco was and continues to be highly 
speculative. However, a recent paper by Oliveira 
and Daly (1999), whose interpretation the Ducke 
book repeats, takes this shaky concept and makes 
it appear as widely accepted dogma. Likewise, the 
same section of the book steps into the “refugia” 
quagmire, asserting that the area of Manaus is a 
species refugium characterized by a high degree of 
local endemism with elements from many different 


phytogeographical regions. It is well known (Nelson 
et al., that Manaus is the epicenter of plant 
collecting efforts in the Amazon, and that other 
parts of the basin are so woefully undercollected or 
in fact totally unsampled that any such conclusions 
stand on very slippery ground. 

These small details aside, this book is a truly 
masterful work, many years in the making, and de- 
serving of all the praise and wide distribution it can 
get. Mike Hopkins, one of the 14 authors listed on 
the title page, merits special credit for managing 
the project and overseeing the team of very capable 
Brazilian botanists who carried out most of the 
work, both in the field and in the office. —Paul E 
Berry, Department of Botany, University of Wiscon- 
sin—Madison, 132 Birge Hall, 430 Lincoln Drive, 
Madison, Wisconsin 53706, U.S.A. 


Literature Cited 


Barneby, R J. W. Grimes. 1996. Silk tree, Guana- 
caste, Monkey’s Earring. A generic system for the syn- 
p us Mimosaceae of the 


Americas. Part 1. Abare- 

oo and allies. Mem. New York Bot. Gard. 74: 
1 

в. i D., . Lavina, A. Rancy & J. P. de Souza 
Filho. 1988. a piene Pleistocene тепе lake т 
the Amazon basin and its significance to Amazonian 
ш and тонги Acta Ain. 1834) 119– 
14 


"Ead В. W., C. is Ferreira, M. F. da Silva & M. L. 
Kawasaki. 1990. Endemism centres, refugia and bo- 
tanical Ет Данай | in Brazilian Amazonia. Nature 
345: 7 6. 

Oliveira, rà A. de & D. C. Daly. 1999. Geographic dis- 
tribution of tree species occurring in the region of Ma- 
naus, Brazil: Implications for regional diversity and 
conservation. Biodiversity and Conservation 8: 1245- 
1259. 


Volume 87, Number 3, pp. 297—434 of the ANNALS OF THE MISSOURI BOTANICAL GARDEN 
was published on October 10, 2000. 


| 


Missouri Botanical Garden Press М 


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species, about one-fourth the total number in the 
genus. Each species is represented by text, 
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СОМТЕМТ5 


Investigations into the Systematic Botany and Phylogenetic Relationships of Takhtajania perrieri 
apuron) Baranova & J.-F. Leroy (Winteraceae) 

The Rediscovery of a Malagasy Endemic: Takhtajania perrieri (Winteraceae) ~- 
George E. Schatz 297 
Paleobotany, Relationships, and Geographic History of Winteraceae _ James A. Doyle 303 
Wood and Bark Anatomy of Takhtajania (Winteraceae); Phylogenetic and Ecological 

Implications Sherwin Carlquist 317 


Winteraceae Evolution: An Ecophysiological Perspective 

Taylor S. Feild, Maciej A. Zwieniecki & М. M. Holbrook 323 

Anatomy of the Young Vegetative Shoot of Takhtajania perrieri Me ee ыы 

hard С. Keating 335 

_ Floral Structure of Takhtajania and Из Systematic Position in ede ULL TET 
Peter К. Endress, Anton Igersheim, Е В. Sampson & George E. Schatz 347 


Comparative Floral Ontogeny in Winteraceae Andrew N. Doust 366 
The Pollen of Мов porte (Winteraceae) Е B. Sampson 380 
ryology of Te ) and a Summary Statement of Embryological 
Features for the F UR Hiroshi Tobe & Bruce Sampson 389 
Notes on the Vascular Anatomy of the Fruit of Takhtajania (Winteraceae) and Its Inter- 
pretation Thierry Deroin 398 
incer of Takhtajania, Other Winteraceae, and Canellaceae: Phylogenetic Impli- 
cat Е Ehrendorfer & M. Lambrou 407 


| Malecular Р Ба for the Phylogenetic Position of Takhtajania in the Winteraceae: 
Inference from Nuclear Ribosomal and Chloroplast Gene Spacer Sequences __- 

____- Kenneth б. Karol, Youngbae Suh, George E. Schatz & Elizabeth A. Zimmer 414 

Book Review. Flora da Reserva Ducke by J. S. da S. Ribeiro et al., reviewed by Раш E. Berry 433 


Cases Боа Crateva simplicifolia J. S. Miller, drawn by Barbara Alongi, from the book- 
let for the 45th Annual Systematics хана Missouri Botanical Garden. First published in 
_ Novon 8: 167-169 (1998). 


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Volume 87 Annals 
Number 4 of the 
2000 Missouri 


Botanical 


Garden 


PHYLOGENETIC Lucinda A. McDade,? Thomas F. Daniel,? 
RELATIONSHIPS WITHIN Ha ie qoc im 

THE TRIBE JUSTICIEAE 

(ACANTHACEAE): EVIDENCE 

FROM MOLECULAR 

SEQUENCES, MORPHOLOC Y, 

AND CYTOLOGY' 


ABSTRACT 


We used molecular sequence data from the nuclear ribosomal internal transcribed spacers and from the intron and 
spacer of the trnL-trnF chloroplast region to study phylogenetic relationships within the large (ca. 2000 species), wide- 
ranging, and taxonomically difficult tribe pe ieae (Acanthaceae). The partition homogeneity test indicated that the 
data sets for the two loci were congruent, and separate analyses of the two m similar results. Analysis of the т 
data set provides a highly resolved hypothesis of relationships, much of it strongly supported. Justicieae are strc 
supported as monophyletic; within the tribe, five lineages and one ар и grade are related as follows: jad 
anthemum lineage (Isoglossinae (атара ит lineage [multiple clades of Old World “justicioids” (Diclipterinae + New 
World “justicioids”)]})]. Many aspects of this phylogenetic hypothesis are supported by data from morphology and 
cytology, and some conform to earlier classific ‘ations of the group. There аге, however, a number of novel aspects. 
Notably, the large genus Justicia (ca. 700 species) is not ао 18 пс; the Old World members form а grade and the 
New World members are monophyletic only if a number of other genera are included. The very strongly supported 
р relationship between Diclipterinae and the New World “justicioid” lineage i is novel, and we cannot identify 

molecular synapomorphies to confirm this 1 re pres Rhinacanthus, а “justicioid” (Justicia and morphologically 
ка и genera) by all but ane criteria, rongly supported as a basal member of Diclipterinae, and cytological 
evidence confirms this placement. The Pse и ‚тит lineage is only modestly supported as monophyletic and may, 
in fact, represent a series of d lineages. These plants are marked by having four staminal elements (four stamens 
or two stamens plus two staminodes), a plesiomorphic condition for all Аса 'eae. Additional evidence (both taxa 
and characters) will be necessary to resolve this uncertainty, as well as to determine the phylogenetic status of Old 
World *justicioids." Our analysis does provide considerable resolution of enia within monophyletic lineages, 
and Mec relationships are discussed in the context of non-molecular evidence and previous classifications 
y words: Acanthaceae, cp trnL-trnE, cytology, Justicieae, Lamiales, molecular sequences, morphology, nr ITS, 
Ка сивих 


= 


' For help in acquiring plant materials we thank K. Balkwill, M.-J. Balkwill, M. Butterwick, A. Faivre, M. Foote, W. 
Haber, P. Jenkins, J. MacDougal, R. Olmstead, Ornduff, R. Scotland, D. Shindelman, B. Tankersley, M. Turner, T. 
Van Devender, and M. Zjhra; ARIZ, CAS, and MO; C. Marcopolus and the staff of the San Francisco Conservatory of 
Flowers, and the staffs of the Duke University iis nd Strybing Arboretum and Botanical Gardens, Mildred E. 
Mathias Botanical Garden, Waimea Arboretum and Botanical Garden, and Witwatersrand National Botanic Garden. We 
thank D. Swofford for salio available test versions of PAUP*, D. Maddison for making available test versions of 


ANN. Missouni Вот. GARD. 87: 435—458. 2000. 


436 


Annals of the 
Missouri Botanical Garden 


“I will not say that ... [my] classification of the 
Justicieae . . . is perfect. On the contrary, І am con- 
vinced that it is no more than a tentative effort" 
(Bremekamp, 1965: 30). 

The tribe Justicieae is the largest and arguably 
most systematically challenging lineage in the plant 
family Acanthaceae. Justicieae comprise about 

000 species, and their nearly worldwide distri- 
bution essentially matches that of the family as a 
whole. As documented by Lindau (in 1895) and 
supplemented by numerous other contributions 
since that time, the macromorphological (especially 
habit and floral) and palynological diversity within 
the tribe is considerable. Composition, infratribal 
relationships, and affinities of Justicieae have been 
controversial ever since Lindau's (1895) compre- 
hensive infrafamilial classification of Acanthaceae. 
Lindau grouped Justicia L. and several of its pre- 
sumed close relatives into a tribe characterized by 
having an androecium of two stamens and *Knótch- 
enpollen" (2- or 3-aperturate pollen with 1 to 3 
rows of insulae on each side of the apertures). The 
fact that several taxa of Lindau's Justicieae do not 
have this type of pollen whereas several genera with 
two stamens and *Knótchenpollen" were classified 
in other tribes by Lindau is indicative of problems 
with his classification. 

Tribe Justicieae was classified by Lindau as part 
of "Imbricatae," a supratribal group that was de- 
fined by imbricate corolla aestivation. This pattern 
of aestivation is likely primitive for Acanthaceae, 
and Imbricatae are a heterogeneous group, as has 
been recognized for some time (Bremekamp, 1965) 
and confirmed by recent phylogenetic work (Hed- 
rén et al., 1995; Scotland et al., 1995; McDade & 
Moody, 1999; McDade et al., 2000). Further, Scot- 
land et al. (1994) showed that there is more than 
one variant of imbricate aestivation among Acan- 
thaceae. Other authors have demonstrated that aes- 
tivation varies within lineages (Manktelow et al., in 
press) and even within genera (Schónenberger & 
Endress, 1998) such that its use as a defining char- 
acter may unwarranted. Moreover, Lindau's 
(1895) tribes and subtribes of Imbricatae, including 
Justicieae as described above, are not clearly cir- 


cumscribed morphologically, and many are dispa- 
rate assemblages. The character basis for the clas- 
sification is inconsistent in that characters used by 
Lindau in his keys sometimes conflict with those 
employed in descriptions of tribes or genera in- 
cluded therein. For example, Isoglossinae are char- 
acterized as having two stamens, but plants of some 
genera placed in this subtribe by Lindau have four. 
Many of Lindau’s taxa contain a core of genera that 
undoubtedly belong together, along with an odd as- 
semblage of others. For example, his Diclipterinae 
include Dicliptera Juss., Hypoestes Sol. ex R. Br., 
Periestes Baill. (= Hypoestes), and Peristrophe Nees, 
a group readily recognized as cohesive by essen- 
tially all students of Acanthaceae. To this subtribe, 
however, Lindau added Tetramerium Nees. Plants 
of this last genus are morphologically more similar 
to Carlowrightia A. Gray and Anisacanthus Nees, 
which were placed by Lindau in Graptophylleae, 
than they are to other Diclipterinae. These prob- 
lems have made it difficult for subsequent шш 
of Acanthaceae to classify newly described genera 
with any о such that Lindau’s classifica- 
tion is not widely used. 

Bremekamp (1965) outlined a revised infrafam- 
ilial classification of Acanthaceae, but did not al- 
ways specify the generic composition of his tribes 
and subtribes. In a major realignment of taxa, he 
included six of the tribes placed by Lindau in Im- 
bricatae (Asystasieae, Graptophylleae, Isoglosseae, 
Justicieae, Odontonemeae, Pseuderanthemeae) into 
an expanded Justicieae with three subtribes (Jus- 
ticiinae, Rhytiglossinae, and Odontoneminae). Jus- 
ticiinae and Rhytiglossinae (a substitute name for 
Lindau's Isoglossinae) were distinguished from 
Odontoneminae on the basis of an androecium of 
two stamens with no staminodes. Justiciinae were 
further distinguished by presence of a rugula (a 
channel in the upper lip of the corolla in which the 
style rests during anthesis), whereas plants with 
lenticular biporate pollen were assigned to Rhytig- 
lossinae (— Isoglossinae). Most of the remaining 
Justicieae were placed in Odontoneminae. Plants 
belonging to this last group retain the plesiomorph- 
ic state of one or more of the characters that marked 


MacClade, and two anonymous reviewers for pedum input on an earlier version of the manuscript. This research was 
U.S 


partially supported by grants from the 


National Science Foundation to LAM (DEB BSR-8507517. DEB 


BSR- 


9707693), and from ie University of Мө small grants program. TFD's collecting activities were partially ~~ 


by the In-house Research Fund a 
Mu Made the American Philosophical Society, and the 
Research Training Group in the A 


ан of Arizona’s Undergraduate Biological Кезеагс 


Analysis of Biological о. 


nd Lindsay Field Researc à Fund of the California Academy of Sciences, the Or. 


ristensen Research Institute. The University of Ari 
(NSF DIR-9113362, BIR-9602246) кеў е 
Participation program support < 


rted К 
artments of Ecology and Evolutionary Biology, and Plant Sciences, University of Arizona, Tucson, Arizona 
S.A. 


3 Department of Botany, California Academy of Sciences, Golden Gate Park, San Francisco, California 94118, U.S 


Volume 87, Number 4 
2000 


McDade et al. 
Phylogeny of Tribe Justicieae 


Bremekamp’s other two subtribes, including ab- 
sence of a rugula, androecium with four staminal 
elements (four fertile stamens or two stamens plus 
two staminodes), and pollen with colpoid streaks (= 
pseudocolpi). It is mostly feasible to assign newly 
described genera to Bremekamp’s subtribes; how- 
ever, Justiciinae and Odontoneminae, in particular, 
are very large groups of plants such that little is 
really accomplished by classifying to this level. 
Further, it is clear that although Justiciinae and 
Rhytiglossinae (= Isoglossinae) are likely marked 
by synapomorphies and may be monophyletic, 
Odontoneminae are not and are thus likely not 
monophyletic. 

Problems in classifying Justicieae sensu Bre- 
mekamp (and as used herein) do not end at the 
subtribal level. The tribe includes a number of gen- 
era that are not clearly diagnosed as distinct from 
related genera (including Justicia with ca. 700 spe- 
cies; see Graham, 1988, who noted a conservative 
estimate of some 600 species at that time). This 
often makes routine identification difficult at best. 
In sum, there is a great deal to be learned about 
relationships in this richly diverse lineage of Acan- 
thaceae. 

Recent progress has affirmed the monophyletic 
status of Acanthaceae and advanced our under- 
standing of relationships between Justicieae and 
the three other main lineages of Acanthaceae 
(McDade et al., 2000). Justicieae and Ruellieae 
sensu lato (s.l, i.e., sensu Manktelow et al., in 
press) are sister taxa. Barlerieae (including Whit- 
fieldieae, see Manktelow et al., in press) are sister 
to these two together, and Acanthoideae (sensu 
Bremekamp, 1965) are sister to the first three to- 
gether. The monophyly of each of these, as well as 
the pattern of relationships among them, is sup- 
ported by molecular sequence data and, in some 
cases, by morphological synapomorphies (McDade 
& Moody, 1999; Manktelow et al., in press). There 
has, however, been little progress in understanding 
relationships within Justicieae. Based on admitted- 
ly sparse sampling, particularly of Old World 
plants, McDade and Moody (1999) and McDade et 
al. (2000) proposed at least five sublineages of Jus- 
ticieae. Other authors, notably Daniel (1986), Dan- 
iel and Chuang (1993), and Daniel et al. (1984, 
1990), have informally proposed that certain groups 
of genera are closely related, but these proposals 
have not been in the context of phylogenetic anal- 
yses nor have they been formalized. 

Here we present a phylogenetic analysis based 
on molecular sequence data from two regions (the 
nuclear ribosomal ITS and chloroplast trnL-trnF 
spacer and intron) for a reasonably representative 


sample of Justicieae. We discuss these results in 
the context of evidence from other sources includ- 
ing morphology, chromosome numbers, and geo- 
graphic distribution. 


MATERIALS AND METHODS 
TAXON SAMPLING 


We obtained sequences for species representing 
all of Lindau's (1895) suprageneric taxa of Imbri- 
catae that were included by Bremekamp (1965) in 
his Justicieae. In addition, we attempted to sample 
two or more species of the larger genera of Justi- 
cieae, with emphasis on genera that occur in both 
the Old and New Worlds, e.g., Justicia, Dicliptera. 
From the very large genus Justicia, we included 
representatives of more than half of the sections 
recognized by Graham (1988). To root our hypoth- 
esis of relationships among Justicieae, we used two 
members of each of the other main lineages of 
Acanthaceae s. str. as outgroups: Ruellieae s.l. (one 
species each of Ruellia and Sanchezia), Barlerieae 
(one species each of Barleria and Lepidagathis), 
and Acanthoideae (one species each of Aphelandra 
and Stenandrium) (Appendix 1). We included the 
most distantly related groups within these lineages 
for which we had material and were able to obtain 
sequences. This should have the effect of *break- 
ing" long branches (sensu Felsenstein, 1985) and 
thus increase confidence in our results (see Gray- 
beal, 1998) 


MOLECULAR METHODS 


For most samples, DNA was extracted from fresh 
leaf material or material dried in silica gel; recently 
collected herbarium specimens were the source of 
DNA for the remaining species (Appendix 1). Total 
genomic DNA was extracted using the modified 
CTAB method of Doyle and Doyle (1987). Some 
acanths have pigmented compounds that apparently 
complex to DNA; this was dealt with by js ie 
the DNA, as described by McDade et al. (2000 

For the nr ITS sequences, a fragment comprising 
itsl, the 5.8s gene, and из2 (Baldwin, 1992; Bald- 
win et al., 1995) was amplified. Early in this pro- 
ject, we used the “universal” primers “154” and 
“its5” (Baldwin, 1992). As noted by McDade et al. 
2000), some samples amplified with these primers 
yielded fungal contaminants. Using primers 
“С26А” and “N-nc18S10” designed for plants (Wen 
& Zimmer, 1996) effectively ended this problem. 
Optimal polymerase chain reaction (PCR) condi- 
tions to amplify double-stranded DNA varied some- 
what among taxa, and we used a "touchdown" tem- 


— 


438 


Annals of the 
Missouri Botanical Garden 


perature cycling profile to circumvent the process 
of optimizing PCR conditions for each taxon (see 
McDade et al., 2000). For the trnL-trnF sequences, 
a fragment comprising the trnL intron, the 3'trnL 
exon, and the intergenic spacer between this exon 
and the trnF gene of the chloroplast genome (Ta- 
berlet et al., 1991) was amplified using the “с” and 
"f" primers designed by these same subs Stan- 
dard PCR techniques were used to amplify double- 
stranded DNA 

Sequences were generated on ABI automated se- 
quencers at the University of Arizona DNA se- 
quencing facility using initially the same primers 
as in amplification. This yielded high quality se- 
quences for essentially all of the cp trnL-trnF PCR 
templates and for about two-thirds of the nr ITS 
samples. However, the nr ITS region is extremely 
G-C rich in many Acanthaceae (see McDade et al., 
2000) and poly-C or -G strings > 5 bp 


present in some taxa. DNA polymerase frequently 


long are 


was unable to read through these long repeats of 


Gs or Cs such that incomplete sequences were ob- 
tained. When only a partial its1 sequence was ob- 
tained using primer “N-nc18S10” (anchored in the 
18s rDNA gene), we attempted to complete the se- 
quence using primer “C26A” (anchored in the 26s 
rDNA gene). For some templates, neither the se- 
quencing reaction primed with "N-nc18S10" nor 
that primed with A" yielded a complete se- 
quence. When this occurred, we sequenced using 
internal primers “its2” and “its3” (Baldwin, 1992), 
which are anchored in the 5.8s gene and yield se- 
quence for its] and its2, respectively. Both strands 
were sequenced except when sequencing with one 
primer yielded a complete sequence with no am- 
biguities (ca. % of the cp trnL-trnF templates and 
м of nr ITS templates) or when sequencing with all 
available primers did not yield completely overlap- 
ping forward and reverse sequences (five of the nr 
ITS templates). 

Electropherograms of all sequences were proof- 
read manually. Overlapping portions were recon- 
ciled by reverse-complementing one, aligning the 
two, and double-checking any inconsistencies 
against the electropherograms; mismatches were 
coded as uncertain. 


ALIGNMENT AND ANALYSES 


Sequences were aligned separately by eye in 
SeqApp (Gilbert, 1994), and then moved into 
MacClade version 4.0411 (Maddison & Maddison, 
1999). These are available on request from LAM. 
As documented by McDade et al. (2000), the nr 


ITS region is quite divergent among Acanthaceae. 


Although alignments were straightforward within 
Justicieae, one region of its] (aligned positions 65— 
204; ca. 1596 of the total aligned length) could not 
be aligned with confidence between Justicieae and 
the six taxa used as outgroups. The six outgroup 
taxa were thus scored as missing for those nucle- 
otide positions. Most of the total 3.596 missing data 
for the ingroup are in the highly conserved 5.8s 
gene. Most of the remaining missing data are in the 
its] sequences for three taxa (Fittonia Coem., Asys- 
гама gangetica, and Spathacanthus parviflorus); 
these sequences could not be completed despite 
attempts using all available primers. Alignment of 
the nr ITS sequences required introduction of many 
gaps, most of which were one or two bp in length, 
restricted to a single taxon, and in highly variable 
and/or G-C rich regions of the sequences. However, 
14 indels were shared by two or more taxa, had 
concordant 5’ and 3’ termini, and did not overlap 
gaps in the sequences for other taxa. Information 
on these indels was added to the matrix as pres- 
ence/absence characters. 

The cp sequences were easily aligned across all 
taxa, including outgroups, despite the fact that, as 
noted by McDade and Moody (1999), this region is 
prone to length mutations. Seventeen parsimony in- 
formative indels were added to the matrix as pres- 
ence/absence data. For the cp locus, 3.296 of data 
are missing; almost all of the missing data are in 
the jd conserved 5' end of the intron. 

Data matrices were analyzed separately using 
PAUP* 4 Ob2 (Swofford, 1999). parsimony 
analyses were conducted using rigorous heuristic 
searches, i.e., 20 random addition sequences (all 
analyses bond a single island sensu Maddison, 
1991) and TBR swapping; gaps were treated as 
missing data. Multiple most parsimonious (MP) 
trees were combined as strict consensus trees. 

In addition to standard measures of fit of char- 
acters to the resultant trees (consistency index, re- 
tention index), the strength of support for individual 
nches was estimated using bootstrap values 


> 
ы 
~ 


(Felsenstein, 1985) and decay or Bremer indices 
(Bremer, 1988; Donoghue et al., 1992). For the nr 
ITS matrix, bootstrap (BS) values reported are from 
200 full heuristic replicates with 20 random addi- 
tion sequences and TBR branch swapping. The cp 
data provide poor resolution in some distal portions 
of the phylogeny such that most bootstrap replicates 
generated enough MP trees to swamp computer 
memory; it was thus not possible to use rigorous 
branch swapping methods. Instead, BS values are 
from 1000 replications with 500 random addition 
sequences each and no branch swapping. For both 
data sets, decay values for each branch were de- 


Volume 87, Number 4 
2000 


McDade et 


al. 439 
Phylogeny of Tribe Justicieae 


Table 1. 


Characteristics of the nuclear ribosomal ITS region (59 taxa) and chloroplast trnL-trnF (52 taxa) in Jus- 


ticieae (outgroup taxa not included in these calculations). Reporting of variable and parsimony informative sites includes 


within 


sites 


hereas sites within gaps were excluded for calculation of pairwise distance 


8. ! Includes 25 and 28 


bp of the 18s and 26s ribosomal genes, respectively, that flank its] and its2, plus the 5.8s gene. ? Includes the 3' trnF 
exon and 40 bp of the trnF gene that flanks the trnL-trnF spacer. 


nr ITS Intron + 
itsl its2 region trnL3' intron Spacer Spacer 
Raw length 193-278 211-234 410—512 347-521 224—332 710—824 
Aligned length 356 288 644(860) 6 73 103 (11912) 
Variable sites (proportion) 209 (0.59) 176 (0.61) 421 (0.49)' 165 (0.26) 160 (0.34) 347 (0.31)? 
Parsimony informative sites 175 (0.49) 114 (0.40) 308 (0.36)! 73 (0.12) 78 (0.16) 167 (0.15)? 
Pairwise distances (range, %) 0.8-31.5% 0.5-33.2% 0.5-23.8% 0-9.6% 0-15.9%  0.4–10.9%2 
GC content (range) 0.66–0.77 0.66–0.77 0.67-0.76 0.33-0.38 0.36-0.42 0.35-0.38? 
Scored indels, number 12 14 7 10 17 


termined by first using MacClade to prepare a set 
of trees each with a single branch resolved. These 
trees were then loaded into PAUP as constraint 
trees, and the program was asked to find the short- 
est trees inconsistent with the constraint tree. The 
difference between the length of these trees and the 
globally shortest trees is the decay index (DI) for 
the branch in question. 

We obtained sequences for only one of the two 
loci for 13 taxa (see Appendix 1). Four of these 
(Justicia brandegeana, J. spicigera, J. comata, An- 
isacanthus puberulus) were sequenced for nr ITS 
but not the cp locus because sequences obtained 
earlier permitted us to judge that the more slowly 
evolving cp locus would be essentially invariant in 
these compared to close relatives (see McDade et 
al., 2000). DNA of the other taxa could not be am- 
plified for the missing locus or more than М of the 
sequence was missing even after attempts to se- 
quence using all available primers. These taxa were 
included in the analysis of the locus for which com- 
plete (or nearly complete) data were available but 
were pruned fi 
The data sets thus pruned to include complete se- 
quences for the same set of 55 taxa (49 Justicieae 
+ 6 outgroups) were combined into a single N 
US file using the file editing capabilities of PAUP*. 
The nr ITS and cp trnL-trnF data sets were tested 
for congruence using Farris et al.’s (1995) Incon- 
gruence Length Difference test (implemented in 
PAUP* as the partition homogeneity test). Phylo- 
genetic analyses of the combined data sets were 
conducted as described above; bootstrap values 
(200 replicates with 5 random addition sequences 
each) and decay indices (as previously described) 
were generated for each branch. 


rom the data sets before combining. 


Alternative phylogenetic hypotheses were eval- 
uated by using MacClade to prepare trees that re- 
flect the relationships of interest. These were load- 


ed into PAUP* as constraint trees, and the program 
was asked to find the shortest trees consistent with 
the topology in question. The difference between 
the length of these trees and the globally shortest 
trees provides an indication of the parsimony cost 
(in terms of additional evolutionary transitions) in- 
volved in accepting the alternative hypothesis. 

To compare patterns of molecular evolution be- 
tween loci and among lineages, matrices of pairwise 
HKY85 distances for both loci and for the com- 
bined sequences were output from PAUP* and 
moved into JMP (Sall & Lehman, 1996) for analy- 
sis. The relationship between pairwise distances for 
the two loci was examined using correlation. Rates 
of evolution were compared between selected sister 
lineages using a modified relative rates test (Sarich 
& Wilson, 1973). Distances between members of 
two sister lineages and their closest relative were 
tabulated, and t-tests were used to compare the 
means of these pairwise distances. 


RESULTS 
MOLECULAR EVOLUTION 


Within the nr ITS region, its] and its2 are rough- 
ly similar in variability except that its] has more 
informative indels than its2 (Table 1). The cp trnL- 
trnF spacer is considerably more variable than the 
trnL3' intron, and the spacer is likewise more 
prone to indels although this difference is not 
. Whether considered in terms of overall 
variable sites or parsimony informative sites, the nr 
ITS region is twice as variable among these taxa as 
the cp trnL-trnF region (Table 1). Similarly, maxi- 
mum pairwise distances are more than twice as 
great for the nr ITS sequences as for the trnL-trnF 
data. Across all taxa, pairwise distances for the two 
loci are positively correlated (r — 0.669, N — 1176, 
P < 0.0001), suggesting that although the two loci 


Annals of the 
Missouri Botanical Garden 


are evolving at different rates, those rates are fairly 
consistent across Justicieae. Variation in rates of 
evolution among lineages will be discussed below 
in the context of phylogenetic relationships. 


PHYLOGENETIC RELATIONSHIPS 


Results of the partition homogeneity test indicate 
that the two data sets are not incongruent (P = 
0.34). Further, except as regards taxa for which se- 
quence data from only one locus were available, 
the topology obtained from the combined data set 
(Fig. 1) differs from the trees obtained from the 
separate data sets (not shown) only in degree of 
resolution or in weakly supported portions. For ex- 
ample, the nr ITS data do not resolve Ptyssiglottis 
T. Anderson as part of Isoglossinae, but do not sup- 
port any other placement of this taxon. Similarly, 
the cp trnL-trnF data do not resolve relationships 
among most species of Old World Justicia, whereas 
the nr ITS data provided a fully (but weakly) sup- 
ported hypothesis of relationships among these 
taxa. As expected, given congruence of the data 
sets and increased number of characters in the 
combined data set [308 and 218 parsimony infor- 
mative characters in the nr ITS and cp trnL-trnF 
data sets, respectively, for a total of 526 in the com- 
bined data set (note that these tallies of parsimony 
informative characters are from the data sets 
pruned to include taxa for which both sequences 
were available) branch support increases mark- 
edly in the combined topology. For all of these rea- 
sons, discussion of relationships is based on the 
outcome of the combined analysis. 

Figure 1 presents the strict consensus of most 
parsimonious (MP) trees produced by analysis of 
the combined data sets. Note that only major line- 
ages within Justicieae are labeled to emphasize 
higher level patterns of relationship; Figures 2 and 
3 provide detail on relationships within lineages. In 
our discussion of relationships, when lineages iden- 
tified here largely conform to previously recognized 
taxa, we use names of these established taxa; when 
lineages have not been previously recognized or 
named, we use informal names 

The combined analysis — strong support 

$ = 


= 17). Within Justicieae, five lineages аге moder- 
ately, e.g., the Pseuderanthemum lineage, BS = 75, 
DI = 2, to very strongly, e.g., the Tetramerium lin- 
eage, BS = 100, DI = 28, supported as monophy- 
letic. The Pseuderanthemum lineage is sister to all 
other Justicieae, wares are strongly supported as 
monophyletic (BS = 
are moderately well шы as monophyletic (BS 
= 82, = 3), and as sister to all Justicieae ex- 
cluding the Pseuderanthemum lineage (BS = 89, 
DI = 3). The Tetramerium lineage is very strongly 
supported as monophyletic; this group is sister to a 
monophyletic lineage that includes Diclipterinae 
and Justicia and close relatives. These taxa, i.e., all 
Justicieae above the Tetramerium lineage exclusive 
of Diclipterinae and inclusive of both Old and New 
World plants, will be referred to subsequently as 
“justicioids.” The labels New World “justicioids” 
and Old World “justicioids” will be used to refer to 
geographically delimited assemblages. The “justi- 
cioids” and Diclipterinae gu is strongly sup- 
ported as monophyletic (BS — 97, CI — 6). There 
is strong support for сед of the New World 
“justicioid” lineage (BS = 100, DI = 12) and of 
Diclipterinae (BS = 100, DI = 9), and for the sis- 
ter-group relationship of these lineages (BS = 94, 
DI owever, Old “justicioids” 
placed as a paraphyletic assemblage below New 
World “justicioids” + Diclipterinae. 

Figures 2 and 3 are strict consensus trees show- 
ing placement of all included taxa, and bootstrap 
and decay support for all branches. Taxa for which 
sequence data for only one locus was available 
have been added to Figures 2 and 3 using stylistic 
conventions, i.e., 


). Isoglossinae 


are 


branches are angled and taxon 
labels and support values are indicated in smaller 
type, to signal that the result is based on partial 
data. One randomly chosen MP tree is presented 
as Figure 4 to illustrate branch lengths. 

There is only mod- 
est support for monophyly of this lineage from the 
combined analysis (BS = 75, DI = 2). Nr ITS data 
indicate that Spathacanthus Baill. belongs here, 


Pseuderanthemum lineage. 


and that the two included species are each other's 
closest relatives with strong support (Fig. 2; BS = 


for monophyly of Justicieae (Fig. 1, B 00, РГ 89, DI = 4). Our data do not resolve relationships 
> 
Figure 1. Major у of relationships among Justicieae from ae analysis p е пг ITS апа ср 


ai ae sequence data. Strict consensus о 


and support for these). Note that all labeled li 
фе є ехс a of ‘Old World “justicioids,” 
text. 


most parsimonious trees (ler 
ott and s 'ay indic 'es are presented for major lineages only (see 

ine ineages are monophyletic (indicated by solid v 
which are a grade (indicated by dashed vertical line), as dise assed i in the 


gth = 2455, 0.557, RI = 0.690). 
Figs. 2 ы 3 for jd of n within 
rtical lines) with 


Volume 87, Number 4 
2000 


McDade et al. 441 
Phylogeny of Tribe Justicieae 


100 
EN 
a 
BS 
0! 
94 
97 
6 gm 
a 
89 
3 
100 
1 
X 100 
"| 28 
100 82 
17 3 
Out- 15 
Groups 2 


New World 
"justicioids" 


Diclipterinae 


| 

| 

| Old World 
| "justicioids" 
| 
l 


Tetramerium 
Lineage 


Isoglossinae 


Pseuderanthemum 
Lineage 


442 Annals of the 
Missouri Botanical Garden 


| Anisacanthus 


Tetramerium 
Carlowrightia 
Gypsacanthus 
Chalarothyrsus 
Henrya 
Fittonia 
Pachystachys 


Streblacanthus 


әдеәш” штагшраја J 


Ecbolium 
Schaueria 


|Stenostephanus 


[sos] 


Razisea 


Isoglossa grandiflora 
Ptyssiglottis 


c 
© 
С, 
© 
wu 
cl 
& 
~ 
© 
м. 
S 
> 
3601550 


Out- 


Oplonia 
Groups 


Herpetacanthus 
| Spathacanthus 


2 
© 
= 
S 
S 
= 
= 
© 
5 
= 
5 
pm шпшәцјирләрпәѕ 


Figure 2. Relationships among members of the Pseuderanthemum us age, Isoglossinae, and the oe lin- 
eage. Vertical lines link congeneric species when these are monophyletic. Taxa with bold, large typeface > labe n thick 
branches are in the combined analysis (nr ITS + cp trnL-trnF); taxa for which data for only the nr ITS w НЫЕ 
are added on thinner, angled branches with smaller typeface labels. Data for both loci were obtained for ут и 
thurberi and Stre Hd cordatus, whereas only nr ITS data were obtained for a second member of each gen 
puberulus and S. roseus; see Appendix 1). Bold, large typeface bootstrap and decay values are from the ae! 
analysis; those in smaller typeface are from the analysis of the nr ITS data alone. 


Volume 87, Number 4 McDade et al. 443 
2000 eii i of Tribe Justicieae 


J. spicigera nr-its 
"s. > J. brandegeana 


Poikilacanthus 
Megaskepasma 
Justicia caudata 
J. longii 
Harpochilus 


cp, trnL-trnF ] 
D. magaliesbergensis 
D. extenta 


4 Sprornsnf,, 
ро M MƏN 


72 Dicliptera sp. 9194 
100 D. suberecta = 
. e 
D. resupinata = 
BS p Hypoestes aristata > 
- 
DI H. phyllostachya 5 
Peristrophe - 
Rhinacanthus 
«50 "E" | 
Justicia betonica | 
68 Justicia sp. 9024 І 
4 јр“ Justicia sp. 9010 |: 
4 Justicia sp. Zjhra 983 1 A = 
J. extensa EE- 
| = 
J. adhatoda 1 >. 2 
• Lnd 
Anisotes za 
Fig. 2 Duvernoia m 
94 p Кипра | 
6 bk Metarungia i 
Figure 3. Relat ое а у World “justicioids,” Diclipterinae, and the New World “justicioid” lineage. Taxa 
with bold, large typeface labels on thick brane hes are in the combined analysis (i.e., nr ITS + ep trnL-trnF); taxa for 
which data for only one pen was available are added on thinner, angled branches or to the upper left. Bold, large 
typeface Sig and decay values are from a combined analysis; those in smaller typeface are from separate analyses 
of either the nr ITS or cp trnL-trnF data set. The positions of Justicia brandegeana (nr ITS) and ја (cp trnL- 
trnF) аге Bite and strongly supported, whereas J. comata and J. spicigera (nr ITS), Dicliptera extenta and D. 
magaliesbergensis (cp trnL-trnF) are placed in polytomies with other New World Justicia and with Dicliptera, respec- 
vely. 


among basal groups in the lineage [i.e., Spathacan- species also share two unique indels, one in each 
thus (nr ITS only), Herpetacanthus Nees, Asystasia genic region. Further, there is considerable molec- 
Blume]. The two representatives of Asystasia are, ular divergence between the two Asystasia species, 
however, strongly supported as monophyletic; these and between Asystasia and other members of the 


Annals of the 
Missouri Botanical Garden 


444 


15 Justicia caudata 
T = = Megaskepasma erythrochlamys 
ran 
27 y T rahe и тасгапіћиѕ 


6 20 треш neesiana 
21 24 = Dicliptera а 
m Г). suber 
i 12 Dicliptera » 9194 
uidi phyllostachya 
— |] ровове aristata 
Rhi онш gracilis 
Tustin betoni. 


11 22 


Justicia ха Daniel 9010 
30 Justicia exten 
235... Justicia adhatoda | 
22. Anisotes madagascariensis 
30 12. Duvernoia acon е 
23 5 к klos 
Meta rungia ain 
19 НОТА specio 
_Anisacanthus thurberi 
"mt = Carlow рн arizonica 
13 25 Eom nervosum 
12 umen id non 
121 [lo » Chalarothyrsus amplexicaulis 
44 Hen nrya insular: 


56 Echo 
17 Isoglossa poise) flora аш 
Isoglossa? "m Daniel 9106 
sf Razisea spic 
Stenostephanus silvaticus 


24 
17 


2 = pen 
Piyssiglottis pubisepala 
1 ои апрепса 
11 e = а sp. Daniel 9129 
B stenophsli us 
22 || 13 Маскауа bella 
Za Chileranthemum a 
| 10 Odontonema tubaeform 


26 


8 
1 Daniel 6737cv 


Figure 4. One (randomly chosen) of the most parsimonious trees from analysis of the nr ITS and ср trnL-trnF 
combined sequence data for all taxa for which data for both loci were available. Branch lengths are proportional to 
number of changes using ACCTRAN optimization; numbers on branches report branch length. 


Pseuderanthemum lineage (note branch lengths in aya (BS = 100, DI = 12). This last group includes 


Fig. 4). Above this unresolved group of basal gen- 
era, there is weak support for Mackaya Harv. as 
sister to all others (BS — 68, DI — 2), and strong 
support for monophyly of the lineage above Mack- 


one Malagasy species (Daniel 6737cv) that corre- 
sponds to specimens at P that were annotated as 
“Justicia petiti Benoist." It does not, however, ap- 
pear that Benoist ever formally described this tax- 


Volume 87, Number 4 
2000 


McDade et al. 
Phylogeny of Tribe Justicieae 


on, and it remains either unnamed or described 
with another name. There is little molecular diver- 
gence within the lineage above Mackaya (note short 
branch lengths, Fig. 4), and only the sister taxa 
Ruspolia Lindau + Ruttya Harv. are resolved with 
strong support (BS = 99, DI = 5). 

Isoglossinae. А lineage that largely conforms to 
Lindau's (1895) Isoglossinae is identified by the 
combined analysis as monophyletic with modest 
support (BS = 82, DI = 
above Ptyssiglottis is extremely strongly supported 
as monophyletic (BS = 100, DI = 15). The Old 
World genus /soglossa Oerst. may not be monophy- 
Јенс: a putative species of Isoglossa (i.e., Daniel 
9106) is more closely related to Siaip hani 
Nees, Razisea Oerst., and Brachystephanus Nees 
(BS = 100, DI = 9) than to I. grandiflora. The nr 
ITS data place Old World Brachystephanus with 
New World Razisea + Stenostephanus, with strong 
support (BS = 99, DI = 7; note that these support 
values are from the nr ITS analysis alone). The two 


3). The group of genera 


species of Stenostephanus are placed together, but 
with only modest support (BS = 80, DI = 1). There 
is, however, strong support for monophyly of Razi- 
sea + Stenostephanus (BS = 100, DI = 12). Short 
branch lengths among Razisea and Stenostephanus 
(Fig. 4) indicate that there is little molecular di- 
vergence among these three species. 

The Tetramerium lineage 
is extremely н supported as monophyletic 
(BS = = 28). Nr ITS data indicate that 
Schaueria сва is part of this lineage but do not 
resolve its relationships. Further, the nr ITS data 
place the two sampled species of Streblacanthus 
Kuntze as sister taxa, and also place the two sam- 


Tetramerium lineage. 


pled species of Anisacanthus together (cp data not 
available for S. roseus and A. puberulus). There is 
strong support for monophyly of a lineage above 
Ecbolium Kurz (and Schaueria, nr ITS data only) 
(BS = 91, DI = 5), and moderate support for the 
lineage above Hoverdenia Nees (and Mirandea 
Rzed., nr ITS data only) (BS = 75, DI = 4). The 
combined data set provides strong support for sis- 
ter-group relationships of Streblacanthus + Pach- 
ystachys Nees (BS = 95, DI = 5) and Chalaroth- 
yrsus Lindau + Henrya Nees ex Benth. (BS = 94, 
DI = 4), and nr ITS data provide moderate support 
for Hoverdenia + Mirandea (BS = 78, DI = 3; note 
that these values are from the nr ITS data alone). 
Although there is weak support for the internal 
nodes of the phylogeny distal to Pachystachys + 
Streblacanthus, monophyly of a lineage including 
Anisacanthus, Tetramerium, Carlowrightia, and 
Gypsacanthus E. J. Lott, V. Jaram. & Rzed. seems 
highly likely based on a shared deletion in the 


spacer region of trnL-trnF that is ca. 150 bp in 
ngth. 
* Justicioids" and Diclipterinae. 
support from the combined analysis (Fig. 3) for a 
clade comprised of much of Lindau's (1895) Di- 
clipterinae plus all included “justicioids” (i.e., spe- 
cies of Justicia and the allied genera Rungia Nees, 
Metarungia Baden, Duvernoia E. Mey. ex Nees, Ап- 
isotes Nees from the Old World; Harpochilus Nees, 
Megaskepasma Lindau, and Poikilacanthus Lindau 
from the New World) (BS = 97, DI = 6). Within 
this lineage, the representatives of Justicia are not 
monophyletic. Instead, the Old World species are a 
paraphyletic assemblage that is basal to the Diclip- 
terinae (as here circumscribed, see below) + the 
New World “justicioids” lineage. New World Jus- 
ticia are monophyletic only if Poikilacanthus and 
Megaskepasma are included. Old World genera tra- 
ditionally aligned with Justicia are placed as a se- 
ries of lineages basal to Old World Justicia, with 
strong support for Rungia + Metarungia (BS = 94, 
DI = 6), for monophyly of all others above this pair 
(BS = 94, DI = 5), and then weak support for 
precise placement of Duvernoia, Anisotes, and the 
Old World representatives of Justicia. 

There is strong support for monophyly of the Di- 
clipterinae + New World “justicioids” lineage (BS 

= 94, DI = 7), and very strong support for mono- 
е of each of these lineages (Fig. 3). The lineage 
referred to here as Diclipterinae largely conforms 
to the core of Lindau’s taxon of that name. Rhina- 
canthus Nees (not part of Lindau’s Diclipterinae) is 


— 
[=] 


There is strong 


strongly supported as a basal member of the line- 
age, with very strong support for monophyly of all 
included taxa above Rhinacanthus (BS = 100, DI 
= 32). Hypoestes is monophyletic with strong sup- 
port (BS = 93, DI = 6), and the cp trnL-trnF data 
place Peristrophe as sister to Hypoestes. The genus 
Dicliptera is monophyletic, including both Old and 
New World members. Especially above Rhinacan- 
thus, branch lengths are notably longer in Diclip- 
terinae than in adjacent groups (Fig. 4). Using Jus- 
ticia betonica as the outgroup, a relative rates test 
indicates that sequence divergence in Diclipterinae 
is higher than in its sister group (the New World 
*justicioid" lineage). This is true for the loci con- 
sidered separately (nr ITS: t — 3.229, 13 df, P — 
0.0066; cp trnL-trnF: t = 3.89, 12 df, P = 0.0021) 
as well as for the combined data (t — 3.838, 9 df, 
Р = 0.004). 

New World “justicioids” (i.e., representatives of 
Justicia plus Harpochilus, Megaskepasma, and Po- 
ikilacanthus) are together monophyletic, with strong 
support (Fig. 3, BS — 100, DI — 12). ITS se- 


quence data place J. brandegeana as sister to Po- 


446 Annals of the 
Missouri Botanical Garden 
ikilacanthus with strong support (BS = 87, DI = plants of the other two lineages, with just a few 


4, nr ITS data alone), and place J. comata ты Ј. 
spicigera as part of the clade that includes all other 
New World Justicia, but without resolution. F igure 
4 indicates that branch lengths are quite short 
among members of the New World “justicioid” lin- 


eage, and a relative rates test shows lower levels of 


sequence divergence in this clade compared to its 
sister group, Diclipterinae. 


DISCUSSION 


Justicieae are strongly supported as monophylet- 
ic in our analysis, and this has been confirmed in 
other analyses with richer samples of other Acan- 
thaceae (McDade & Moody, 1999; McDade et al., 
2000). This result is supported by nucleotide sub- 
stitutions as well as by indels: all Justicieae in- 
cluded here share three indels in the cp locus. Un- 
usual tricolporate hexapseudocolpate pollen grains 
(see figs. 7-10 in Daniel, 1998a) occur in all lin- 
eages of Justicieae as here delimited (although 
shifts to other types of grains are synapomorphies 
for some clades as discussed below). Pollen grains 
of this type are not known among Acanthaceae out- 
side of Justicieae and, to our knowledge, are not 
known in other angiosperms. We here propose this 
character as а synapomorphy for Justicieae. Bre- 
mekamp (1965) proposed that this pollen type char- 
acterizes plants of his subtribe Odontoneminae; it 
is thus not surprising that his taxon is not mono- 
phyletic but instead assembles a heterogeneous 
group of Justicieae that lack more derived pollen 
types. 

Lack of hygroscopic trichomes on the seeds char- 
acterizes Justicieae but may not be a synapomor- 
phy. Seeds of plants of Ruellieae s.l., the sister 
group of Justicieae (see McDade et al., 2000), have 
hygroscopic trichomes (exceptions are genera pre- 
viously included in Louteridieae and Trichanther- 
eae, which have apparently lost these structures). 
Seeds of plants of Barlerieae + Whitfieldieae (these 
together comprise the sister group of Justicieae + 
Ruellieae s.l.; see Manktelow et al., in press) also 
have hygroscopic trichomes, again with a few ex- 
ceptions in which they have apparently been lost 
secondarily. However, there is evidence that the hy- 
groscopic trichomes differ between Ruellieae s.l. 
and Barlerieae + Whitfieldieae such that they are 
not likely homologous (Grubert, 1974; Scotland et 
al., 1995; Manktelow, 1996). If this is the case, 
then the common ancestor of these lineages would 
have lacked hygroscopic trichomes. Regardless of 
the phylogenetic status of the character, lack of hy- 
groscopic trichomes distinguishes Justicieae from 


exceptions. Acanthoideae also lack hygroscopic tri- 
chomes on the seeds, but these plants lack also the 
synapomorphies that link the other three lineages 
of Acanthaceae s. str., i.e., Ruellieae s.l., Barler- 
ieae, Justicieae (e.g., ставе: articulated stems, 
colporate or porate pollen) and are thus unlikely to 
be confused with them. 

Pseuderanthemum lineage. The position of 
these taxa as basal within Justicieae is clear from 
our data, but there is only modest support for their 
monophyly. Further, we know of no morphological 
synapomorphies for the entire lineage. Compared to 
other Justicieae, these plants have an androecium 
of four staminal elements (all four may be fertile or 
two may be reduced to staminodes). However, this 
is no doubt plesiomorphic for Justicieae: it is hy- 
pothesized to be a synapomorphy for all Lamiales 
s.l. (i.e., sensu Olmstead et al., 1993), and the other 
major lineages of Acanthaceae include taxa with 
this trait. Additional data will be necessary to test 
whether the lineage is indeed monophyletic or in- 
stead a series of lineages at the base of Justicieae. 
Most aspects of relationships within the Pseuder- 
anthemum lineage are not resolved with confidence 
by our data. There is little divergence among most 
of these taxa for the loci examined here, and it will 
be necessary to work with morphological characters 
or more rapidly evolving loci to make progress in 
unraveling these phylogenetic relationships. 

There is some corroborating evidence for those 
aspects of relationships within the Pseuderanthe- 
mum lineage that do emerge from our analysis. 
Whereas plants placed in unresolved fashion at the 
base of the lineage have four fertile stamens, two 
of these are reduced to staminodes in plants of the 
Mackaya-Ruttya clade. Further, although chromo- 
some numbers have not been determined for Chil- 
eranthemum Oerst. or for the unidentified Malagasy 
taxon represented by Daniel 6737cv, all other gen- 
era in the Mackaya-Ruttya clade appear to have a 
base chromosome number of x — 21 (Daniel & 
Chuang, 1989; Daniel et al., 
Chuang, 1993, 8). The only chromosome num- 
ber known for Mackaya (Old World, 2 species), n 
— 42, is from a single count by Daniel and Chuang 
(1989); n — 42 has not been recorded for any other 
genus in the lineage. А chromosome complement 
of n — 21 has been infrequently reported in genera 
representing various other lineages within the fam- 
ily, but does not seem to characterize other large, 
suprageneric groups. Below the Mackaya-Ruttya 
clade in the Pseuderanthemum lineage, chromo- 
some counts are not available for Herpetacanthus 
(New World, ca. 10 species) or Spathacanthus (Neo- 


Моште 87, Митбег 4 
2000 


McDade 
piedi 4 Tribe Justicieae 


447 


tropical, 3 species), and x = 21 may thus be a 
synapomorphy for a more inclusive lineage. Inter- 
estingly, Asystasia (Old World, ca. 50 species) has 
x — 13 and includes species with polyploid deriv- 
atives of that number (Daniel, 2000). Several au- 
thors have suggested that x — 7 is symplesiom- 
orphic for Acanthaceae (Grant, 1955; Raven, 1975; 
Piovano & Bernardello, 1991; Daniel & Chuang, 
1993). If so, x — 13 has evolved via both polyploidy 
and dysploidy, and x = 21 may represent a ћеха- 
ploid derivative of this base number. 

There is very strong molecular support for the 
Chileranthemum-Ruttya clade but, within that 
clade, the only strongly supported relationship is 
that of Ruspolia and Ruttya as sister taxa. These 
two genera differ from others in the Chileranthe- 
mum-Ruttya clade in having monothecous sta- 
mens. Ruspolia (Africa and Madagascar, 4 species) 
and Ruttya (Africa, 3 species) are clearly closely 
related based on both molecular and morphological 
evidence, and a natural intergeneric hybrid, X Rut- 
tyruspolia A. Meeuse & de Wet, is known between 
them 

Heterostyly has been reported or observed by 

D or LAM in all genera of the Chileranthemum— 
Ruttya clade except Ruttya (based on the limited 
material available, it is not possible to determine 
whether Daniel 6737cv was collected from a het- 
erostylous population). Given that this trait has only 
recently been observed in Ruspolia (LAM, pers. 
obs.), its absence in Ruttya may reflect lack of 
study rather than absence of the character. Heter- 
ostyly is otherwise unknown among Acanthaceae 
[the stylar polymorphism reported by Long (1971) 
for Ruellia caroliniensis (Walter) Steud. is related 
to corolla size but not to anther position]. As noted 
by Daniel (1995a), distinctions among Chileranthe- 
mum (New World, 3 species), Odontonema Nees 
(New World, ca. 30 species), Oplonia Raf. (Amer- 
ican tropics and Madagascar, 14 species), and 
Pseuderanthemum Radlk. (pantropical, ca. 60 spe- 
cies) pertain primarily to differences in form of the 
corolla, which likely represent adaptations for dif- 
ferent pollinators. These traits are well known to 
vary at low taxonomic level in Acanthaceae (e.g., 
Ezcurra, 1993, for Ruellia; Graham, 1988, for Jus- 
ticia). We know of no diagnostic morphological fea- 
tures for these genera, and the monophyly of each 
should be tested. 

Based on both molecular and morphological 
data, it seems clear that Daniel 6737cv should not 
be placed in Justicia; like other plants in the Pseu- 
deranthemum lineage, it has an androecium of four 
staminal elements (two stamens plus two stami- 
nodes in this case, in contrast to Justicia, which 


lacks staminodes), and tricolporate hexapseudocol- 
pate pollen (which is rare in Justicia). Like many 
Malagasy plants, its taxonomic placement requires 
additional study; our results do not indicate a clear 
generic assignment for the species. 

The Pseuderanthemum lineage is a taxonomically 
heterogeneous group, including representatives of 
Lindau's (1895) tribes Asystasieae (Asystasia, 
thacanthus), Graptophylleae (Ruspolia), Isoglosseae 
(Herpetacanthus), Odontonemeae [Chileranthemum, 
Mackaya, Odontonema, Ruttya, and Oplonia (as 
Anthacanthus Nees)], and Pseuderanthemeae 
(Pseuderanthemum). To complete our understand- 
ing of this lineage and to test its monophyletic sta- 
tus, it will be important to include representatives 
of all Justicieae that have four fertile stamens or 
two stamens and two staminodes. As will be clear 


Spa- 


from his classification of the genera included here, 
Lindau (1895) distributed plants with four staminal 
elements in a number of tribes including Asysta- 
sieae (e.g., Asystasiella Lindau, Chameranthemum 
Nees, and Thomandersia Baill. in addition to those 
already included here), Graptophylleae (Grapto- 
phyllum Nees), Isoglosseae (Podorungia Baill., 
Chlamydacanthus Lindau, Forcipella Baill.), Odon- 
tonemeae (Ballochia Balf. f., Phialacanthus Benth., 
Filetia Miq.), and Pisulecntbemeae (Codonacan- 
thus Nees). Many of these genera have not been 
studied in the century since Lindau's (1895) clas- 
sification was proposed. More recently described 
genera with four staminal elements include Pran- 
ceacanthus Wassh. and Pulchranthus V. M. Baum, 
Reveal & Nowicke. Based on our results we predict 
that plants in most of these genera will be placed 
either as part of a monophyletic Pseuderanthemum 
lineage or as a series of basal lineages within Jus- 
ticieae. It would also be useful to test monophyly 
of some of the larger genera in this lineage, espe- 
cially those that are pantropical in distribution 
(Pseuderanthemum) or that have disjunct ranges 
(Oplonia, with 5 Malagasy and 9 New World spe- 
cies). Given the uncertainties noted above regard- 
ing exact taxonomic composition, we estimate that 
the Pseuderanthemum lineage comprises about 200 
species. 

As noted above, whether or not the Pseuderan- 
themum lineage is monophyletic, there is very 
strong support from our analysis for monophyly of 
all other Justicieae. These plants have an androe- 
cium of two stamens; that is, with only two excep- 
tions of which we are aware, they have lost even 
staminodial remnants of the other pair. Remarkably, 
plants of Chalarothyrsus (here placed as a member 
of the Tetramerium lineage, see below) have four 


dithecous stamens. Two species of Ptyssiglottis 


448 


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(here placed as the basal member of Isoglossinae, 
see below) are reported to have small or remnant 
staminodes (Hansen, 1992). 

Isoglossinae. Lindau’s (1895) key to tribes and 
subtribes distinguishes Isoglossinae based on the 
presence of two stamens with mono- or dithecous 
anthers and “Giirtelpollen” (i.e., girdled pollen). 
However, in describing the subtribe and in assign- 
ing genera to it, he included also some plants that 
lack this pollen type and have four stamens (e.g., 
Herpetacanthus, Populina Baill.). As discussed be- 
low, our results suggest that Lindau was correct in 
recognizing a group marked by girdled pollen. He 
was also correct in assigning additional genera that 
lack this synapomorphy to Isoglossinae, but erred 
in at least some assignments, e.g., Herpetacanthus 
is a member of the Pseuderanthemum lineage. 

In our analysis, Ptyssiglottis (southeastern Asia 
to Papuasia, 33 species) is placed in Isoglossinae 
with moderate support. These plants have dithecous 
anthers and a diversity of pollen types including 
the basic type for Justicieae (tricolporate hexapseu- 
docolpate pollen), but not the girdled biporate pol- 
len characteristic of “core Isoglossinae” (Hansen, 
1992; see below). Ptyssiglottis was placed by Lin- 
dau (1895) in Pseuderanthemeae, probably on the 
basis of its pollen. As delimited by Hansen (1992 
plants of Ptyssiglottis lack staminodia with the two 
exceptions noted above of small or remnant sta- 
minodia. We are, however, unable to point to clear, 
non-molecular synapomorphies linking this genus 
to other Isoglossinae. Hansen (1992) treated Ptys- 
siglottis as a member of Isoglossinae but did not 
describe synapomorphies supporting the relation- 
ship. It would be well to test the present hypothesis 
that Ptyssiglottis is part of Isogléssinae by addition- 
al data (DNA sequence or otherwise). If this is the 
case, then the genus can be viewed as transitional 
between more typical Justicieae and “core Isoglos- 
sinae,” which are marked by distinctive pollen 
morphological synapomorphies. 

“Core” Isoglossinae (i.e., Isoglossa through Sten- 
ostephanus) are extremely strongly supported as 
monophyletic in our analysis. In addition to se- 
quence data, this clade is marked by three unique 
and unreversed indels (two in the nr ITS locus). 
These plants also appear to share Lindau’s “Giir- 
telpollen,” which is typically biporate with pores 
that are surrounded by a more or less circular re- 
gion. The two circular regions are separated from 
one another by a peripheral band (continuous or 
interrupted) of varying width. This type of pollen is 
described and figured by Daniel (1999) for Sten- 
ostephanus and Razisea. Similar pollen has been 
described in Brachystephanus (Figueiredo & Keith- 


М 


Lucas, 1996) and /soglossa (Muller et al., 1989; 
Raj, 1961, as Rhytiglossa Nees ex Lindl.), although 
at least the former genus shows more diversity in 
features such as aperture number. The identity of 
Daniel 9106 from Madagascar has yet to be deter- 
mined, but preliminary examination of its pollen 
reveals biporate pollen like that of other *core" Is- 
oglossinae, and its macromorphological character- 
istics suggest its placement in /soglossa. If it indeed 
represents a species of this genus, then the two 
species of /soglossa included in our analysis do not 
form a monophyletic group. Instead, Daniel 9106 
is more closely related to Stenostephanus, Razisea, 
and Brachystephanus than to 1. grandiflora. Inter- 
estingly, in addition to sequence support for this 
relationship, Daniel 9106 also shares two indels in 
the trnL-trnF region with Stenostephanus + Razisea 
that /. grandiflora lacks. The nr ITS data support 
monophyly of Brachystephanus (tropical Africa and 
Madagascar, 13 species) and Neotropical Stenoste- 
phanus (ca. 75 species) and Razisea (4 species). 
These three genera share monothecous anthers as 
a morphological synapomorphy. The New World 
members of this sublineage have been divided into 
a series of small genera reflecting remarkable floral 
diversity (e.g., Cylindrosolenium Lindau, Kalbrey- 
eracanthus Wassh., Kalbreyeriella Lindau). Wood 
(1988) treated most of these as congeneric with Ha- 
bracanthus Nees, and Daniel (1995b, 1999) further 
combined Habracanthus with Stenostephanus. No- 
menclatural changes are gradually being made to 
reflect this recasting of generic limits, e.g., Was- 
shausen (1999). Our results indicate that there is 
considerable merit to this approach and confirm 
that Razisea is part of this group as well (Daniel, 
1999). However, investigation of relationships 
among the New World species using molecular data 
will require a locus evolving more quickly than 
those we have studied (note short branch lengths 
in Fig. 4) 

Isoglossinae, as here defined, are particularly 
poorly known cytologically. Among genera studied 
by us, chromosome counts have been determined 
for only five species. It is noteworthy that the Neo- 
tropical genera Stenostephanus and Razisea appear 
to share a common chromosome number of n = 18 
(Daniel, 1999). No chromosome numbers have been 
reported for the Old World genera Brachystephanus 
or Ptyssiglottis, and only a single count (n = 17) 
has been reported for the Old World genus Isoglos- 
sa (widespread in Old World tropics and subtropics, 
ca. 60 species) (Daniel & Chuang, 1998). 

To delimit Isoglossinae clearly, it would be well 
to include representatives of other genera that lack 
the synapomorphies that mark “core” Isoglossinae. 


Volume 87, Number 4 


McDade et al. 
ЕЯ of Tribe Justicieae 


449 


Genera that were included in this subtribe by Lin- 
dau (1895, 1897) and that would appear to be part 
of “core” Isoglossinae as defined here include Cy- 
lindrosolenium, Populina, and Oreacanthus Benth. 
These, and other more recently described genera 
with two dithecous stamens and “Giirtelpollen” 

e.g, Conocalyx Benoist, Sphacanthus Benoist), 
should be added. Acknowledging uncertainty about 
placement of some of these smaller genera, we es- 
timate that Isoglossinae include about 200 species. 

Monophyly of the larger genera of Isoglossinae 
should be tested, including Ptyssiglottis, Isoglossa, 
and Stenostephanus. Monophyly of Brachystephanus 
should be tested, and the usefulness of maintaining 
this genus separate from Stenostephanus also merits 
evaluation. Broad-scale studies of a number of taxa 
have demonstrated that morphologically similar 
genera in widely separated geographic regions are 
often congeneric (e.g., Stenandrium Nees—Stenan- 
driopsis S. Moore, Vollesen, 1992; Oplonia—Forsy- 
thiopsis Baker, Stearn, 1971; Mendoncia Vell. ex 
Vand., Afromendoncia Gilg ex Lindau, and Mon- 
achochlamys Baker, Benoist, 1925) 

Monophyly of all Justicieae above these first two 
lineages (i.e., Pseuderanthemum lineage and Iso- 
p Fig. 1) is moderately supported (BS = 

, DI = 3), but we know of no non-molecular 
аа that mark this lineage. 

Tetramerium lineage. Sequence data strongly 
support monophyly of this lineage; indeed, it is one 
of the most strongly supported clades in all of Jus- 
ticieae. In addition to sequence data, all of the 
plants included in our analysis uniquely share five 
indels in the nr ITS region. The genera sampled 
here were placed by Lindau (1895) in diverse 
tribes: Asystasieae (Chalarothyrsus, part of Henrya 
[as Solenoruellia Baill.]), Graptophylleae (Anisacan- 
thus, Carlowrightia, Pachystachys), Isoglosseae 
(Fittonia), and Odontonemeae [Ecbolium, Hover- 
denia, Schaueria, Streblacanthus, Tetramerium (in- 
cluding Непгуа)ј. Despite Lindau’s treatment and 
the fact that we can identify no clear non-molecular 
synapomorphies for the lineage, a group corre- 
sponding closely to that delimited here has been 
consistently identified based on monographic and 
cytological studies (Daniel, 1986, 1990; Daniel & 
Chuang, 1993; Daniel et al., 1984). A chromosome 
number of n — 18 seems to characterize the entire 
group, with additional diversity in some genera 


— ]4 (one count), 18 (six counts), 
(one count); chromosome data are lacking for Hov- 
erdenia and Schaueria]. Because n — 18 also oc- 
curs in Isoglossinae and in a few species of Justicia, 
it is difficult to determine whether it is a synapo- 


morphy for the Tetramerium lineage or for a more 
inclusive group. These plants share a number of 
traits that are symplesiomorphic at this level within 
Justicieae: tricolporate hexapseudocolpate pollen (a 
synapomorphy for all Justicieae); an androecium of 
two bithecous stamens and no staminodes (a syn- 
apomorphy for Justicieae above the Pseuderanthe- 
mum lineage); and thecae that are parallel, inserted 
at more or less the same height on the filament, 
and unappendaged (these thecal characteristics are 
variously modified in many members of the “justi- 
cioids" and Diclipterinae lineage). 

The placement of Chalarothyrsus within the Te- 
tramerium lineage is surprising in that these plants 
have four bithecous stamens, a trait known to occur 
among Justicieae only in the Pseuderanthemum lin- 
eage, as here defined. However, Chalarothyrsus has 
a chromosome complement of n — 18 (contra n — 
21, which characterizes the Pseuderanthemum lin- 
eage, see above). We can only conclude that there 
was a reversal to four functional stamens in the 
evolutionary history of plants of this unispecific ge- 
nus from western Mexico. Given that occasional 
flowers with atypical androecia have been observed 
in a number of species of Acanthaceae (TFD & 
LAM, pers. obs.), such a reversal does not seem 
especially improbable. Further, trees placing Chal- 
arothyrsus with the Pseuderanthemum lineage are 
68 steps (396) longer than the MP trees. 

Within the Tetramerium lineage, most aspects of 
relationships are not strongly supported. Monophy- 
y of all included genera above Ecbolium (Old 
World, 22 species) [+ Schaueria (New World, ca. 
10 species), nr ITS data only] is strongly supported, 
and that of Anisacanthus through Streblacanthus 1s 
moderately well supported. We are not able to iden- 


— 


tify clear non-molecular synapomorphies for these 
lineages. There is moderate to strong support for 
sister relationships between Hoverdenia (Mexico, 1 
species) + Mirandea (Mexico, 4 species) (nr ITS 
only), Pachystachys (West Indies and South Amer- 
ica, 12 species) + Streblacanthus (Central and 
South America, 4 species), and Chalarothyrsus + 
Henrya (North and Central America, 2 species) but, 
again, non-molecular evidence is lacking. A clade 
comprised of Anisacanthus through Henrya (Fig. 2) 
is not especially strongly supported by sequence 
data (BS = 66, DI = 2), but is marked by a unique 
22 bp deletion in the nr ITS region. Similarly, a 
clade composed of the New World genera Anisa- 
canthus (ca. 20 species), Tetramerium (28 species), 
Carlowrightia (24 species), and Gypsacanthus (1 
species) is only weakly supported by sequence data 
but, as noted above, these plants share a > 150bp 
deletion in the trnL-trnF spacer. This shared de- 


450 


Annals of the 
Missouri Botanical Garden 


letion argues strongly for their monophyly and also 
explains the weak support from sequence data: the 
cp region that is absent in these four genera is the 
most variable of the cp locus (i.e., the most likely 
source of nucleotide substitutions among close rel- 
atives). The paucity of morphological evidence for 
relationships in the Tetramerium lineage reflects 
Daniel and Chuang’s (1993) statement regarding 
problematic generic delimitations among these 
plants. Generic boundaries are difficult at best and 
are mostly based on differences in floral morphol- 
ogy that reflect adaptation to pollinators (e.g., bees 
and flies in Carlowrightia and Henrya; humming- 
birds in Anisacanthus). Among genera in this lin- 
eage, only Henrya and Chalarothyrsus have unam- 
biguous apomorphies (i.e., fused bracteoles and 
pollen traits in the former, seeds fused to the cap- 
sule valves in the latter) that are not likely related 
to recent selection by pollinators. 

Interestingly, this Tetramerium lineage is almost 
exclusively New World in geographic distribution. 
Mexico is especially rich in its members and sev- 
eral genera are endemic there [i.e., Chalarothyrsus, 
Gypsacanthus, Hoverdenia, Mirandea, Aphanosper- 
ma T. F. Daniel; (the last is not included in our 
sample but undoubtedly belongs here on the basis 
of macromorphological, palynological, and cytolog- 
ical similarities; Daniel, 1988, 
m their greatest c centu of species in Mex- 

o (e.g., Anisacanthus, Carlowrightia, Henrya, Te- 
а Ecbolium is the only Old World mem- 
ber in the present sample, but its placement here 
is not surprising: Vollesen (as cited in Balkwill & 
Balkwill, 1998) and Daniel (1998b) have suggested 
that these plants are congeneric with the North 
American Yeatesia Small, a genus not included in 


and others 


our analysis but which clearly belongs in this lin- 
eage. The Old World genera Megalochlamys Lin- 
dau, Angkalanthus Balf. f., and Calycacanthus K. 
Schum. likely also belong here and should be in- 
cluded in future work (the same is true of New 
World Yeatesia and Aphanosperma, as noted above). 


Assuming that we are correct about placement of 


these genera in the Tetramerium lineage, the group 
includes about 150 species. 

“Justicioids” and Diclipterinae lineage. Mono- 
phyly of the “justicioids” and Diclipterinae line- 
age is strongly supported in the combined analysis 
(recall that “justicioids” is used here to include 
Justicia and allied genera, both Old and New World 
in distribution). Among these plants, there is a 
marked tendency for increased complexity in an- 
ther morphology and ornamentation compared to 
other Justicieae. Thus, the thecae are usually in- 
serted at different heights on the filament and not 


perfectly parallel; basal appendages of various 
shapes and sizes occur on the thecae in many spe- 
cies. This lineage is also marked by evolution of a 
rugula: a channel-like structure on the internal sur- 
face of the upper lip formed by parallel ridges of 
corolla tissue. During anthesis, the style lies in this 
rugula, sometimes fitting so snugly that force is re- 
quired to dislodge it. If we are correct in placing 
the evolution of the rugula here, then this structure 
is lost in Diclipterinae (as here delimited) above 
Rhinacanthus (see below). The genera not included 
in Diclipterinae are placed either as part of a non- 
monophyletic grade that includes Old World “jus- 
ticioids” or as part of a clade, the New World “jus- 
As will be clear from the 
following discussion, our analysis confirms that ge- 
neric delimitations are problematic among “justi- 
cioids,” adding the additional complication that 
neither “justicioids” as a whole nor the genus Jus- 
e are monophyletic 

Old World ^ usticioids," The taxa represented 
in our analysis are not monophyletic, nor do the 
species of Old World Justicia that we have included 
form a monophyletic lineage. However, it is inter- 
esting that all other Old World "justicioids" are 
basal to the six Old World species of Justicia that 
are included. The sister-group relationship between 
Rungia (Paleotropics, ca. 20 species) and Metarun- 
gia (Africa, 3 species) is strongly supported in our 


ticioids" lineage. 


analysis. These plants share a placenta that rises 
elastically from the base of the capsule at maturity 
(this trait is also found in Dicliptera, see below). In 
plants of both genera, the bracts have a distinctive 
hyaline or colored border. There is strong support 
for monophyly of the lineage above Rungia + Me- 
tarungia, but we know of no non-molecular evi- 
dence for this relationship. Relationships among 
other Old World “justicioids” are not strongly sup- 
ported by our analysis, although it is interesting 
that the three Malagasy Justicia are moderately well 
supported as monophyletic (BS — 68, DI — 
Conclusions regarding relationships among Old 
World “justicioids” are unwarranted based on our 
limited sample (10 of at least 300 species) and the 
inconclusive pattern of relationships among these. 
Further, all Justicia species from the Old World are 
monophyletic in trees that are only four steps 
(0.296) longer than the MP trees. Similarly, con- 
straining all Old World “justicioids” to monophyly 
requires only eight additional steps (0.3%) com- 
pared to the shortest trees. On the other hand, ev- 
idence from indels suggests that the present hy- 
pothesis is correct in placing some Old World 
“justicioids” closer to the Diclipterinae + New 
World “justicioid” lineage than to other Old World 


Volume 87, Number 4 
2000 


McDade et al. 
Phylogeny of Tribe Justicieae 


"justicioids." The three species of Malagasy Justi- 
cia and J. betonica share a 3 bp indel in the cp 
locus with the Diclipterinae + New World “justi- 
cioids" lineage, and these plus J. extensa share a 
7 bp indel, also in the cp locus, with the Diclip- 
terinae + New World “justicioids” lineage. 

Students of Acanthaceae have differing opinions 
regarding the validity of a number of genera that 
are clearly closely related to Old World Justicia 
(e.g., Monechma Hochst., Adhatoda Mill., Aulojus- 
ticia Lindau, Old World Siphonoglossa Oerst., As- 
cotheca Heine, Trichocalyx Balf.f., Chlamydocardia 
Lindau, and Sarojusticia Bremek.). Adding repre- 
sentatives of these groups will both expand our 
sample of Old World “justicioids” and test their 
validity as genera. The geographic range of Old 
World “justicioids” is extensive (Africa through 
west Asia to southeast Asia and Australia) and un- 
der-sampled here (we have included only African, 
Malagasy, and south Asian species). These plants 
present a series of intriguing biogeographic patterns 
that can be usefully addressed once we have a well 
resolved phylogeny for a denser and geographically 
broader sample. The phylogenetic status of Justicia 
is discussed further under “Тће Justicia Problem," 
below. 

Strong support for monophyly of the Diclipteri- 
nae + New World “justicioid” lineage is perhaps 
the most surprising component of our results. To 
our knowledge, this relationship has not previously 
been suggested, and we cannot identify any clear 
non-molecular synapomorphies to corroborate the 
strong support from molecular sequence data. Cer- 
tainly this hypothesis of relationships should be 
tested with additional data. 

Diclipterinae. There is strong support from our 
analysis for a monophyletic Diclipterinae, including 
Rhinacanthus. This lineage conforms to the core of 
Lindau's (1895) Diclipterinae in which he placed 
Peristrophe, Hypoestes, and Periestes (= Hypoestes), 
in addition to Dicliptera. However, Lindau included 
also Tetramerium and Rungia here; we have shown 
that these genera have relationships elsewhere in 
Justicieae. Further, he placed Rhinacanthus in 
Odontoneminae. In fact, Rhinacanthus (Old World, 
ca. 20 species) shares a rugula with “justicioids” 
and lacks a number of the morphological synapo- 
morphies that otherwise characterize Diclipterinae. 
Nonetheless, molecular data (including two unique 
and unreversed indels in the cp locus) strongly sup- 
port Rhinacanthus as a member of this lineage, and 
cytological data corroborate this placement. Diclip- 
terinae including Rhinacanthus seem to share a 
base chromosome complement of x — 15. Although 
chromosome numbers of n = 15 and n = 30 аге 


not uncommon elsewhere in the family, among Jus- 
ticieae п = 15 is known only іп a few species of 
Justicia. Among Rhinacanthus, Peristrophe, and 
Hypoestes, п = 15 and 30 are the most commonly 
reported numbers. Many Old World Dicliptera have 

— 13, but n — 15 is known from both African 
and Malagasy species (Kaur, 1970; Daniel, unpub- 
lished data). We thus suggest that n — 13 repre- 
sents dysploid evolution from n — 15. Interestingly, 
all New World species of Dicliptera for which 
counts have been obtained have n — 40 and thus 
appear to be ancient polyploids (Daniel, 2000; 
Daniel & Chuang, 1993). 

Diclipterinae above Rhinacanthus are one of the 
most strongly supported lineages in our analysis; 
these plants are also well marked by morphological 
synapomorphies. Corollas of these plants lack the 
rugula that is otherwise characteristic of the “јиз- 
ticioids" and Diclipterinae lineage. Hypoestes, Per- 
istrophe, and Dicliptera all share a specialized type 
of compound inflorescence (Balkwill & Getliffe 
Norris, 1988). Additionally, in these three genera 
the corolla is resupinate through 180 degrees of 
rotation. This trait has apparently been lost sec- 
ondarily in a number of New World Dicliptera. In- 
terestingly, in some species of New World Diclip- 
tera, the corolla is resupinate through a full 360 
degrees: in these plants, the corolla appears to be 
normally oriented, but this is not achieved in the 
normal way (Daniel, 1995c). 

The two species of Hypoestes are strongly sup- 
ported as each other's closest relative, and cp data 
place Peristrophe sister to these two, again with 
strong support (Fig. 3; these taxa also share an 11 
bp indel in the cp locus). Hypoestes (Old World, ca. 
150 species) differs from Peristrophe (Old World, 
ca. 25 species) and Dicliptera (Old and New World, 
ca. 80 species) by having monothecous anthers. We 
know of no non-molecular synapomorphies that 
mark the Hypoestes + Peristrophe clade. 

Chloroplast data (including three indels, one ca. 
50 bp long) place all five species of Dicliptera to- 
gether, and the combined data set provides strong 
support for monophyly of the three species (includ- 
ing two from the New World and one from the Old 
World) for which we have sequence data for both 
genic regions. Species of Dicliptera have fruits with 
elastic dehiscence of the placentae; this distin- 
guishes them from other Justicieae except Rungia 
+ Metarungia, which also have this trait, as noted 
above. Our results agree with the conclusions of 
Balkwill and Getliffe Norris (1988) in indicating 
that elastic placentae evolved separately in these 
two groups; subtle differences in the trait are to be 
sought. In fact, fruits of a number of other genera 


452 


Annals of the 
Missouri Botanical Garden 


including Tetramerium and Henrya in Justicieae 
have placentae that separate from the fruit wall, but 
are not considered elastic (Daniel, 1986, 1990). 
These characters merit further comparative study 
across Acanthaceae to identify homologies and 
analogies. 

Diclipterinae, with an estimated diversity of 300 
species, have not suffered from the proliferation of 
small genera observed in other lineages of Justi- 
cieae. Dicliptera and Hypoestes are large genera 
whose monophyly should be tested. Especially in- 
teresting is that the biogeographic range of Diclip- 
tera has a cytological correlate, as noted above; 
phylogenetic work within this genus is certainly 
warranted. 

Our results 
suggest the existence of a monophyletic group that 
includes all New World Justicia and members of 
related genera. In addition to sequence support for 
this relationship, all taxa share two indels in the cp 
locus. Generic limits among New World “justi- 
cioids” have been unsettled for some time. For ex- 
ample, among species included here, Л. brande- 
geana was described originally in Beloperone and 


ew World “Justicioid” lineage. 


subsequently moved to its own genus, Calliaspidia 
Bremek., by Bremekamp (1948). Justicia longii was 
treated in Siphonoglossa until this group was stud- 
ied by Hilsenbeck (1990). Our analysis indicates 
that submerging these genera in Justicia was war- 
ranted. In her infrageneric classification of Justicia, 
Graham (1988) included these and many other gen- 
era in Justicia. Daniel (1991) noted that Poikila- 
canthus (ca. 14 species) has but little to distinguish 
it from New World Justicia except pollen morphol- 
ogy, as was also noted by Raj (1961) and Breme- 
kamp (1965). In fact, among species of New World 
Justicia there is already a remarkably rich diversity 
of pollen morphology (Daniel, 1998a). Megaskepas- 
ma, with a single Neotropical species, is a plant 
with spectacular magenta bracts, long white corol- 
las, and pollen that is similar to at least one species 
of Poikilacanthus (Daniel, 1991, 1998a). Finally, 
Harpochilus (3 species) is a poorly known genus 
endemic to Brazil with highly specialized corollas. 
Morphological distinctions between this genus and 
Justicia have not been fully investigated. 

We estimate that the New World “justicioid” lin- 
eage includes about 400 species, with much still 
remaining to be discovered about species-level di- 
versity. In this context, our sample of eight species 
(including the three for which we have only nr ITS 
data) is extremely sparse. On the other hand, our 
sample is taxonomically diverse, as described 
above, such that we regard the hypothesis that the 
entire group is monophyletic as robust. However, 


species representing other genera, both currently 
recognized as well as those already synonymized 
with Justicia, should be included (e.g., Sebastiano- 
Schaueria Nees, Clistax Mart., Chaetochlamys Lin- 
dau, Chaetothylax Nees, Neohallia Hemsl., Tabas- 
cina Baill.). On the other hand, as indicated by 
branch lengths in Figure 4, there is remarkably lit- 
tle molecular diversity among the sampled species. 
For both the nr ITS and trnL-trnF sequences, New 
World “justicioids” have lower rates of divergence 
than their sister group (Diclipterinae). These low 
levels of molecular divergence contrast with species 
diversity (the New World “justicioid” lineage is as 
species-rich as any other lineage of Justicieae), as 
well as with morphological and cytological dispar- 
ity. Plants of this lineage range from prostrate herbs 
to trees, have corollas from a few mm to at least 
8.5 cm in length, and have a startling range of di- 
versity in pollen morphology (Daniel, 1998a). At 
least 11 different chromosome numbers, ranging 
11 to л = 31, have been reported for 
New World species of Justicia alone. It is remark- 


from n = 


able that this explosion of morphological and cy- 
tological diversity is not reflected by molecular di- 
vergence, at least at these loci. 

The Justicia problem. Our results indicate that 
Justicia is not monophyletic, that New World Jus- 
ticia is monophyletic only if a number of other “јиз- 
ticioids" are included, and that Old World species 
of Justicia are unlikely to form a monophyletic 
group whether or not other "justicioids" are includ- 
ed. Clearly, a great deal remains to be learned 
about phylogenetic relationships of plants de- 
scribed in this genus. Even species-level diversity 
remains poorly documented among “justicioids”: 
Index Kewensis (Davies, 1991, 1996) reports 81 
new species described in Justicia alone between 
1986 and 1995. 

The results of our analysis point to a strong phy- 
logenetic distinction between Old and New World 
“justicioids,” despite the fact that they share many 
macromorphological characteristics, have similar 
chromosome number patterns, and the same wide 
range of pollen types. Constraining New + Old 
World “justicioids” to monophyly requires 32 ад- 
ditional steps (1.596) compared to the shortest 
trees. There are no established benchmarks against 
which to evaluate an increase of 1.596 in tree 
length, but note that decay indices (Figs. 2, 3) in- 
dicate that only Diclipterinae above Rhinacanthus 
would be resolved in trees 30 steps longer than the 
MP trees. That is, trees in which all *justicioids" 
are monophyletic would lack resolution among es- 
sentially all other Justicieae. 

n her infrageneric classification. of Justicia 


Volume 87, Number 4 
2000 


McDade et a 
Phylogeny of Tribe Justicieae 


based on study of 295 species of the genus, Graham 
(1988) recognized 16 sections. Interestingly, Gra- 
ham’s work seems to have presaged our results in 
that none of her sections have species in both hemi- 
spheres (7 are restricted to the Old World, 9 to the 
New World). However, the characters used to dis- 
tinguish the sections often seem extremely fine 
[e.g., the distinctions between the Old World sect. 
Justicia and the New World sect. Drejerella are 
based on corolla color (cream versus red) and size 
(less than 25 mm versus more than 35 mm)]. We 
have thus tended to view her work as a monumental 
step toward characterizing variation among species 
of Justicia, but still preliminary. Our sample of 
улеме“ does not permit evaluation of Gra- 
ham’s (1988) sections, but she seems to have been 
correct in separating New and Old World members. 
Clearly, our own contribution can be viewed as pre- 
liminary as well in that we are still far from a full 
understanding of relationships among all “justi- 
cioids.” 


CONCLUSIONS 


The morphological diversity among Justicieae as 
well as the sheer size of the group in terms of num- 
ber of species has made coming to terms with in- 
fratribal relationships quite challenging. In such di- 
verse groups, the simplicity of molecular sequence 
data simplifies comparison. More importantly, plac- 
ing comparative data in a phylogenetic context per- 
mits distinguishing shared derived characters that 
argue for relationships from shared primitive char- 
acters that do not 

It will be clear from the foregoing discussion that 
Lindau (1895) correctly diagnosed Diclipterinae 
and Isoglossinae (although he placed in both of 
these subtribes some genera that we demonstrate 
belong elsewhere) but otherwise shuffled things up 
rather thoroughly. Bremekamp (1965 
correct in recognizing Isoglossinae (his Rhytiglos- 
sinae) and Justiciinae. The fact that he did not in- 
clude Diclipterinae in the latter is mirrored by our 
inability to point to non-molecular synapomorphies 
that support the embedding of Diclipterinae in Bre- 


was likewise 


mekamp's Justiciinae (our “justicioids”). Breme- 
kamp's Odontoneminae thus included Diclipterinae 
(except Rhinacanthus), the Tetramerium lineage, 
and the Pseuderanthemum lineage, a non-mono- 
phyletic group of plants whose morphological basis 
lies in retention of primitive characters. More re- 
cently, Balkwill and Getliffe Norris (1988) present- 
ed a classification for southern African Acanthaceae 
that improves upon Bremekamp's classification by 
removing Diclipterinae (again, as here defined ex- 


cept that Rhinacanthus is placed with their Justi- 
сппае) from Odontoneminae. These authors also 
recognize Isoglossinae, Justiciinae (our Old World 
"justicioids" plus Rhinacanthus), and Odontone- 
minae. This latter group includes elements of our 
Pseuderanthemum and Tetramerium lineages. Fi- 
nally, our results corroborate the findings of Mc- 
Dade and Moody (1999) and McDade et al. (2000) 
regarding clades within Justicieae, while expanding 
the sample of Justicieae considerably. In particular, 
these earlier analyses included no Old World “jus- 
ticioids. 

As discussed throughout, our results are limited 
in a number of ways. First, although much of our 
phylogenetic hypothesis is remarkably strongly 
supported, a few key aspects are not. 
tablishing the p status of the Pseuder- 
anthemum lineage with confidence requires addi- 
tional research ad Second, in some lineages 
(notably most of the Pseuderanthemum lineage and 
New World “justicioid” lineage), the genic regions 
examined here do not provide sufficient variation 
to elucidate relationships with confidence. It will 
be necessary to add another, more rapidly evolving 


otably, es- 


locus to our molecular tools in order to unravel re- 
lationships in these groups and at lower taxonomic 
levels than explored here. Third, sampling remains 
insufficiently dense to address all of the interesting 
phylogenetic problems in Justicieae, notably with 
Old World “justicioids.” Stating this 
shortcoming more positively, our results provide a 
framework to which additional taxa can be readily 
added to address specific questions regarding phy- 
logenetic relationships. In addition to taxa whose 
characters permit predictions about their placement 
into this framework, a few present character com- 


regard to 


binations that defy predictions (e.g., Leandriella 
Benoist with biporate pollen similar to “core” Iso- 
glossinae and four stamens characteristic of basal 
Justicieae). Fourth, in many cases, our knowledge 
of morphology, palynology, and cytology among Jus- 
ticieae remains inadequate for the sort of large- 
scale, comparative project that we have undertak- 
en. We anticipate addressing some of the 
limitations of the present study in continuing col- 
laborative efforts. 


Literature Cited 


Baldwin, B. G. 1992. Phylogenetic utility of the internal 
transcribed spacers of nuclear ribosomal DNA in 
plants: An RCM from the Compositae. Molec. Phy- 
logenet. Evol. 1: 3-16 

„Б crue. J. M. Porter, M. F. Wojciechows- 
ki, C. 5. Campbell & M. J. Boule. 1995. The ITS 
region of nuclear ribosomal DNA: A valuable source of 


454 


Annals of the 
Missouri Botanical Garden 


cvidence on angiosperm phylogeny. Ann. Missouri Bot. 
7-277. 


, K. & F. Getliffe Norris. 1988. Classification of 
the Ас anthaceae: A southern African perspective. Моп- 
орг. Syst. Bot. Missouri Bot. Gard. 25: 503-516. 

Balkwill, M.-J. & К. Balkwill. 1998. A preliminary anal- 
ysis of LE patterns in a large, pantropical ge- 
nus, Barleria L. (Acanthaceae). J. Biogeogr. 25: 95- 

10 


Benoist; R. 1925. — 'ées de Madagascar. Bull. Mus. 
atl. Hist. Nat. 31: 386—388. 
кр C. E. B. pu^ Notes on the Acanthaceae of 
a. Verh. Kon. Lc Akad. Wetensch., Afd. Natuurk., 
Tweede Sect. 45: 1-78. 
. 1965. Delimitation and puede of the Acan- 
Жаса ае. Bull. Bot. Surv. India 7 30. 

Bremer, K. 1988. The e of amino acid sequence data 
in angiosperm phylogenetic reconstruction. Evolution 
42: 795-803. 

Mac T. F. 1986. Systematics of Tetramerium (Acantha- 
ceae). Syst. Bot. Monogr. 12: 1-134. 

1988. Aphanosperma, a new genus of Acantha- 
ceae yug КЕ 9 with unusual diaspores. Amer. J. Bot. 
5—550. 


75: 


: ios Systematics of Henrya (Acanthaceae). 
Contr. nd Michigan Herb. 17: 99-131. 

— А synopsis of о Papin 
— in о Bull. Torrey Bot. Club 1 51-458. 

1995a. Revision ог Odontonema aie eae) 
in Mexico. Сой Univ. Michigan Herb. 20: 147-171. 

. 1995b. New and Јак Mexican Acantha- 
сеае. | | Proc. Calif. Acad. Sei. 48: 253–284. 
а Pp. 1-158 т D. Е. Breed- 
love a Flora о die m 4. California Acad- 
emy of Sciences, San Francis 

. 1998. “Pollen Muri of Mexican Acantha- 
ystematic significance. Proc. Calif. 
156. 


ceae: dm ands 
Acad. Sci. 50: 217— 
98b. Barleria (Acanthaceae) in the New World: 
BI. qe systematic implications. Amer. J. 
Bot. (suppl.) 85: 12 
= 1999 gsm of Stenostephanus Sy anthaceae) 
in Мей о. Conte Univ. Michigan Herb. 2 
. 2000. Additional chromosome е ars of Amer- 
ican Acanthaceae. Syst. Bot. 25: 15-25 
& T. I. Chuang. 1989. SDN numbers of 
some cultivated Ac о Bailey 93. 
& 19¢ “Chromosome so of New 
World Ac ва HEN sy Bot. 18 89. 
& . 199. 


DE Me m of culti- 


7-93 


vated Асап Tee Бет systematic implications. Pp. 
309-330 in P. Mathew & M. Sivadasan (editors), Di- 
versity and Taxonomy of Tropical Flowering Plants. 
Mentor ye | 
‚В. . Baker. 1984. Chromosome 
numbers ud their pane implications in some 
North American Acanthaceae. Syst. Bot. 9: 346-355. 
‚ T. I. Chuang & M. A. Baker. 1990. Chromosome 
numbers of American Acanthaceae. Syst. Bot. 15: 13 
25. 


Davies, R. A. (editor). 1991. Index Kewensis, Supplement 
Nineteen. Clarendon Press, Oxford. 
1996. Index m Supplement Twenty. Roy- 


al Botanic сетан 
Donoghue, М. J., К. Oln istead, J. F. Smith & J. 
Palmer. 1992. а relationships of Dipsac a 


based on rbcL sequences. Ann. Missouri Bot. Gard. 79: 


Doyle, J. J. . Doyle. 1987. A rapid isolation pro- 
cedure for di quantities of fresh leaf tissue. Phyto- 
chem. Bull. Bot . Amer, 19: 11-15 

1993. b nnn s of Ruellia (Ac 'anthaceae) in 

Ann. Missouri Bot. Gard. 8 


Ezcurra, C. 
southern South America. 
787-845 

Farris, J. S. Kallersjé, А. С. Kluge & С. Bult. 1995. 
ws энне. of incongruence. Cladistics 10: 

319. 


15- 
Felsenstein, J. 1985. Confidence limits on phylogenies: 
An approach using the bootstrap. Evolution 39: 783- 


На ава: E. & M. Keith-Lucas. 1996. Pollen ен 
ogy of Brachystephanus (Acanthaceae—Justicieae). 
na Palynol. 35: 65-73. 
7. 1994. SeqApp: A biosequence editor and 
analysis application. Privately published by the author. 
A. W. 1988. Delimitation and infra-generic 
43: 


Graham, 
classification of Justicia (Acanthaceae). Kew Bull. 
551-624 

Grant, W. Е. 1955. A cytogenetic study in the Acantha- 
ceae. Brittonia 8: 121—149. 

Is it better to add taxa or characters 


Graybeal, A. 1998 
47: 9— 


to a difficult uhylugenatie problem? Syst. Biol. 
17. 

Grubert, М. 1974. Studies on the distribution of myxos- 
permy among seeds and fruits of Angiospermae and its 
ec e importance. Acta Biol. Venez. 8: 315—551. 

Hanse 1992. The genus Pty. тр (Асатћасеае). 
А кагы pU iei Opera Bot. 116: 1—58. 

Hedrén, M., M. e & К. С. DLE 1995. Re- 
lationships in the npe m 'eae and related families as 
suggested by iege gredi of rbcL nucleotide se- 

PI. Syst. Evol. 194: 93-109 

). Sy ystematics of Justicia sect. Pen- 

). Р Pl. Syst. Evol. P4 19-235. 


quences, 
Hilsenbeck, R. ч 19% 

taloba (Acanthaceae 
Holmgren, P. . H. Holmgren & . Barnett (edi- 

tors). 1990. index Herbariorum, Part 1: M ве of 


Sci. & Cult. 36: 103—106. 

Lindau, G. 1895. Acanthaceae. Pp. 274—354 in A. Engler 
& K. Prantl (editora) ж natiirlichen Pflanzenfamilien 
4(3b). Engelmann, Leipz 

1 . Ас: о americanae et азайсае no- 
уае vėl minus cognitae. Bull. Herb. Boissier 5: 643- 


= 


Long, R. W. 1971. Floral ројутогрћу and amphimictic 
breeding m in Ruellia caroliniensis (Acanthaceae). 


Amer. J. B о 

Maddison, D. e discovery and importance of 
multiple sane al m most-parsimonious trees. Syst. Zool. 
40: 315-328. 


& W. P. Maddison. 1999. MacClade: Analysis of 
Phylogeny and Character Evolution. Test version 
Оа11, to be ee b: Sinauer Associates, Sun- 
derland, Massachuse 
Manktelow, M. 1996. Phoulopsis (А thaceae) 
graph. wet seu Upsal. : 
ade, B. аш. C. A. Fu M.- 
J. E n. pres nigmatic s Whitfieldieae 
(Acanthaceae): Delimitation and phylogenetic relation- 
ships based on molecular and M за! data. Syst. 
Bo 
McDade, L. 


- 
— 


А mono- 


A. & M. L. Moody. 1999, Phylogenetic rela- 


Volume 87, Number 4 
2000 


McDade et al. 
Phylogeny of Tribe Justicieae 


455 


tionships among Acanthaceae: Evidence from noncod- 
ing trnL-trnF chloroplast DNA sequences. Amer. J. Bot. 
6: 70-80. 


, 9. E. . M. L. Moody & E. Waters. 2000. 
Pilone jena among Acanthaceae: Evi- 
dence from two genomes. Syst. Bot. 25: 105-120. 

Muller, J., M. Schuller, H. Straka & B. Friedrich. 1989. 
И Майка е! Мазсагетса. Кат. 182: 
Аса hp D Subtrop. d 67: 138-187. 

Olmstead, ., B. Bremer, К. M J. D. Palmer. 
1993. A parsimony analysis of ha diana: sensu lato 
based on rbcL sequences. Ann. Missouri Bot. Gard. 80: 
700-722. 

Piovano, M. A. & L. M. Bernardello. 1991. iip cds 
numbers in Argentinian Acanthaceae. Syst. Bot. 16: 

—97. 


Кај, В. 1961. Pollen —-— studies in the Acan- 
thaceae. е Рајуп 3–108 (+ 44 plates). 

Raven, P. H. 1975. The uis of Dem d pw 
и ra Missouri Bot. Gard. 62: 724— 

Sall, J. & A. Lehman. 1996. JMP Start а. ja Gilda 
to Statistical and Data Analysis Using JMP® and JMP 
IN® Software. Duxbury Press at Wadsworth Publishing, 
Belmont, California 

Sarich, V. M. & A. C. Wilson. 1973. Generation t time and 


genomic evolution in primates. Science 179: 1144— 
147 


Schönenberger, J. & P. K. Endress. 1998. Structure and 
development of the flowers in Mendoncia, Pseudocalyx, 
and Thunbergia d and their systematic im- 
plications. Int. J. P . 159: 446—465. 

Scotland, В. W., pe) & T. J. Lawrence. 1994. 
Corolla ontogeny E aestivation in the Acanthaceae. 
Bot. J. Linn. Soc. 114: 49–65. 

j Sweere, P. A. Reeves & R. G. Olmstead. 

1995. Higher-level systematics e Acanthaceae deter- 

mined 2 еле DNA sequences. Amer. J. Вог. 82: 


26 
Stearn 
ia And ы. (Acanthaceae). Bull. 
Hist.), Bot. 4: 259-323. 
Swofford, D. L. 1999. PAUP*: Phylogenetic Analysis Је 


ing Pars er Sinauer Associates, Sunderland, Mas- 


. A survey of the Pew p ra Oplon- 
. Mus. (Nat. 


sachus 
Taberlet, P. Т. Gielly, С. Pautou & J. Bouvet. 1991. Uni- 
versal primers for amplification of three non-coding re- 


gions of chloroplast DNA. Pl. Molec. Biol. 17: 1105— 


( 
у еек, К. 1992. The Old World species of Stenandrium 
(Aca се Acantheae). Kew Bull. 47: 202 
Wasshau р. С. e genus Stenostephanus 
Ка in Bolivia. Harvard Pap. Bot 279 
288. 


Wen, J. & E. A. Zimmer. 1996. бен, and biogeog- 
raphy of UT . (the ginseng genus, Araliaceae): In- 
ferences from ITS sequences a nuclear ribosomal DNA. 
Molec. ју sry Evol. 6: 167-177. 

Wood, J. R. 1. 1988. EE Acanthaceae—Some new 
Фасса and some reconsiderations. Kew Bull. 43: 
1-51 


APPENDIX 1. 


Taxa, Genbank accession numbers (nr ITS followed by 
trnL-trnF; NA = not available), and sources of plant ma- 
terials from which DNA was extracted for sequencing of 
the nuclear ribosomal internal transcribed spacer region 


= chloroplast trnL-trnF spacer and intron. Abbrevia- 

ns for herbaria follow Holmgren et al. (1990). Fresh 
acd or material placed directly into silica gel was 
used except as indicated (HS = 


5 
5 
-B 
= 
о 
© 
= 
® 
Р. 


alphabetically within lineage), a 
press) with regard to others. To facilitate phylogenetic 
placement of taxa of oe Appendix 2 lists Justicieae 
alphabetically and assigns each to a lineage (or grade in 
the case of Old World ' justicioids") 
Taxon; Genbank accession number (nr ITS, trn L-trnF); 
source and voucher information. 


Acanthoideae 
dde i-o pilosulum (S. F. Blake) T. F. Daniel; 
AF169758, AF061827; Mexico. Sonora: Yécora 
Municip El Kipor, Van Devender & Reina G. 97- 
== campanensis Durkee; AF169760, AF061829; 
па. San Blas: near Mandinga, Rfo Mandinga, 
Mc Dade 852 (DUKE). 

о 

Ruellieae s.l. 

Серен speciosa Leonard; AF169385, AF063113; Cul- 
tivated, Duke Pug d greenhouses, Durham, 
€ Carolina, U.S.A., Accession No. 66-462 (na- 

South Аніёйса), acm 1180 (ARIZ). 

Rue Ша ‘californica (Rose) I. M. Johnst.; AF167704, 
! 115; Cultivated, Univ. Arizona campus, Тис- 
son, d na, U.S.A., McDade 1157 (ARIZ) (native 
to a Mexico). 

Barlerie 

bie lupulina ма AF169751, AF289758; Culti- 
vated, San Fra o Conservatory of Flowers, San 
Franci isco, California, U.S.A., Daniel s.n. (CAS) (na- 

Mauritius). 

irren Hedrén; AF169752, AF063121; - 'ot- 
land et al. (1995) [DNA provided by R. Olmstead 
(шегине of Washington) and R. Scotland (Oxford 
en 

Justic 

кеу эн E "p i 

Asystasia gangeti (L. Anderson; AF289793, 

"289748; ке, "Ме ‘Ispruit, Mpumalanga, 

South Africa, Daniel & Balkwill 9386 (CAS) (wide- 

Spread i in Old World tropics and subtropics). 

; AF289794, AF289749; Madagascar. Fianarantsoa, 
лос ин National Park, Daniel 9129 (CAS 

eo ed pyramidatum (Lindau) F. Dan iel; 

89797, AF289752; (HS) Mexico. Chiapas: Mun- 

icipio La Trinitaria, Breedlove & Daniel 70767 


c. 
< 


=~ 


(CAS). 

Daniel 6737cv; AF289799, AF289754; (HS) Cultivated, 
n Francisco Conservatory of Flowers, San Francis- 
co, California, U.S.A., Daniel 6737cv (CAS) (native 

o Madagascar). 
ооа stenophyllus Gómez-Laur. & Grayum; 
AF289795, AF289750; (HS) Costa Rica. Limón: Re- 
е Hitoy Cerere, Herrera 3855 (ARIZ). 
Mackaya pu Harv.; AF289796, oo Cultivated, 
2 E g Arboretum, San Francisco, California, 
. Daniel s.n. (CAS) Leg to souhen Africa). 
Odontonema tubaeforme (Bertol.) K AF169748, 
AF063127; Cultivated, Duke Universi greenhous- 
es, bs North Carolina, U.S.A., Accession No. 


456 


Annals of the 
Missouri Botanical Garden 


66-153, иа 1182 (ARIZ) (native to Mexico and 


microph n (Lam.) Stearn; AF289798, 
9753; (HS) Cultivated, San Francisco Conser- 
of Flowers, San Franciso, California, U.S.A., 
Ornduff 7814cv (CAS) (native to the West Indies). 
МАРА alatum (Nees) Radlk.; AF169749, 
63130; Cultivated, Duke oe greenhous- 
es, т. North Carolina, U.S.A., Accession No. 
84-055, McDade 1183 (ARIZ) (native to North and 
Central America). 
Ruspolia ео (С. В. Clarke) Milne-Redh.; AF289800, 
755; (HS) Cultivated, Waimea Arboretum and 
eae Garden, Oahu, Hawaii, U.S.A., Daniel & 
Butterwick 6635 (CAS) (native to Africa). 
Ruttya fruticosa Lindau; AF289801, AF289756; Cultivat- 
Francisco por pM of Flowers, San 
Fiane "isco, California, U.S.A., Daniel s.n. (CAS) (na- 
tive to Africa). 

Spathacanthus dg Sa г AF289802, NA; (HS) 
Costa Rica. а erva Biológica Carara, Mo- 
rales 1347 (ARI 

S. parviflorus Secon “AF289803, AF289757; (HS) Mex- 


ico. Chiapas: El Triunfo Biosphere Reserve, Daniel 
) 


Isoglossinae 
— lyallii Nees; AF289790, NA; Madagas- 
car. Fianarantsoa, Ranomafana National Park, Dan- 
iel 9101 с AS 
Isoglossa grandiflora C. B. Clarke; AF289788, AF289745; 
(HS) Cultivated, San Francisco Conservatory of 
qn San Francisco, California, U.S.A., D 
s.n. (CAS) (native to eastern tropical Africa 
Isoglossa? sp. 9106; AF289789, AF289746; Madagasc ar. 
Fianarantsoa, Ranomafana National Park, Daniel 
9106 (CAS 
u— pubisepala, (1 йаш) В. Hansen; AF28978 
4; (HS) Papua New Guinea. Madang, ca. 8 
km $ cw of Madang, Daniel 6630 (CAS 
Razisea spicata Oerst.; AF169848, AF06 3131; Costa 
ica. Heredia: La Selva Biological Station, Hammel 
7974 (DUKE). 
Stenostephanus chiapensis У F. Daniel; AF289792, 
A — 9747; Cultivated, 1 Francisco, California, 
., Breedlove & ee 72688cv (CAS) (native to 
sout in ern Mexico 
S. silvaticus (Nees) T. Е Daniel; AF169747, AF063132; 
С 3 Costa Rica. San José: Parque Nacional Braulio 
rrillo, Maas 7800 (MO). 
па Lineage 
Anisacanthus puberulus (Torr. Henr. С ы J. 
289778, NA; Cultivated, Uni na campu 
à son, Arizona, U.S.A., Mc Dade 1179 ARIZ) a 
to southwestern U.S.A. and northern eis о). 
А. thurber us A. Gray; AF169846, AF063122; U.S.A. 
: Pima County, Tucson Mountains, Van De- 
ied 88. 150 (ARIZ) 
н arizonica А. Gray; AF169845, AF063123; 
U.S.A. Arizona: е County, Tucson Mountains, 
Jib 89-24 (ARI 


diced. diae NE 


~ 


Lott: 


Lindau; deep dn 
740; (HS) Cultivated, San Francisco Obse 
vato Flowers, California, U.S.A., Daniel & Bar- 
won 4842gh (CAS) (native to western Mexico). 
Ecbolium syringifolium (Vahl) Vollesen; AF289786, 
AF289743; (HS) dgio qns yu Toliara, Taolanaro, 
Daniel & Butterwick 6733 (C 


Fittonia VASE (Lindl. ex Veitch) Brummitt; omg un 
AF289741; Cultivated, Univ. Arizona, Tucson, Ari 
zona, үн S.A., McDade 1178 (ARIZ) a to a 


Americ 
орлов nelsonii E. J. Lott, V. Jaram. & Rzed.; 
AF289779, AF289739; т Puebla: Municipio 
nn Daniel 8357 (CAS 
и insularis Nees ex Benth.; AF169843, AF063125; 
Sonora: near Alamos, Jenkins 89-432 


Hoverdenia speciosa Nees; AF289777, AF289738; (HS) 
Mexico. Hidalgo: Barranca de Tolantongo, Daniel & 
AS). 


Mirandea grisea Rzed.; AF289783, NA; (HS) Mexico. San 
Luis Potosí: ca. 3 km NW of La Calzada, Daniel & 
Baker 3717 (CAS 

еа Ішеа Nees; AF 169844, AF063128; iei 

Duke aed с Durham 
Carolina = n No. 84-055 (eave | to 
eru), McDade "n (ARIZ 
nos calicotricha (Link & in Nees; 
NA; ‚ Florida, 


AF289782, 
U.S.A., 
Foote 
Srelacanus cordatus (sada: AF289784, AF289742; 
Colón: vicinity of Portobelo, Daniel et al. 


S. roseus is (Radik) B. L. Burtt; AF289785, NA; Cultivated, 
Francisco Conservatory of Flowers, San Francis- 
co, California, U.S.A., Daniel s.n. (CAS) (native to 

1). 


еп 

Tetramerium nervosum Nees; AF169847, AF063133; 
U. x rizona: Pima County, near Patagonia, 

e & Jenkins 1154 (ARIZ). 

Old a “justicioids” (note that results reported 
herein indicate that this is not a И group 
but rather a grade). 

madagascariensis Benoist; AF289772, 

AF28 9733; (HS) Madagascar. Toliara: ca. 20 km N 
oliara, Daniel & Butterwick 6736 (CAS 

M и A. Meeuse; AF289774, AF289735; 

ted, Roodepoort, Witwatersrand National Bo- 
tanic о, Gauteng, South Africa, Daniel et al. 
А (CAS) (native to southern Africa). 
Justicia adhatoda L.; AF289773, AF289734; ш 
niv. Arizona campus, Tucson, Arizona, U.S.A., Bar 
60-393 (ARIZ) (native to southern Asia). 

J. betonica L.; AF289770, AF289731; Culte: Johan- 
nesberg, Gauteng, South Africa, Daniel 9369 (CAS) 
(native to eastern and southern Africa and the Indian 
Subcontine 

J. extensa T. о ue AF289771, AF289732; Cultivat- 
ed, San Francisco Conservatory of Flowers, San 
Francisco, California, U.S.A. Daniel s.n. (CAS) (na- 
tive to eastern tropical Africa 


). 
J. sp. 9024, AF289768, AF289729; Madagascar. Fianar- 


antsoa, Ranomafana National Park, Daniel 9024 
(CAS). 
J. sp. 9010; AF289769, AF289730; Madagascar. Fianar- 


antso. ^» Ranomafana National Park, Daniel 9010 

(С 

Ј. зр. ou. AF289767, AF289728; Madagascar. Masoala 
Peninsula, Vokoanina Forest, Zjhra 983 (CAS 

Metarungia galpinii (C. Baden) C. Baden; 
AF289737; Cultivated, Johannesberg, Gaute 
мини Africa, Daniel 9322 (CAS) (native to South 
Afr 

Rungia Hossi S. Moore; AF289775, 


AF289736; (HS) 


Volume 87, Number 4 
2000 


McDade et al. 
Phylogeny of Tribe Justicieae 


Papua New Guinea. Madang, Bundi, Daniel et al. 
6561 (CAS) 
Diclipterinae 
Dicliptera extenta 
ohannesberg, Gauteng, ina Africa, McDade 1 
(J) (native to southern Afric 
D. ТО nsis K. Balkwill МА, AF289725; Wit- 
rand National Botanic Garden, Gauteng, South 
hes a, Daniel et al. 9357 (CAS). 
D. resupinata (Vahl) Juss.; AF169841, AF063124; U.S.A. 
rizona: Pima County, png Catalina Mountains, 
у an Devender 84-269 (А 
р. у ке (André) iiie "com AF289722; 
vated, Tucson Botanical Garden, Tucson, Ari- 


zona, USA. McDade 1176 (ARIZ) (native to Uru- 


guay). 

D. sp. ode Mr ini AF289723; Madagascar. Fian- 
ara . Ranomafana National Park, Daniel 9194 
(CAS). 

Hypoestes aristata R. Br.; AF289765, AF289726; (HS) 
Cultivated, Mildred E. Mathias Botanical Garden 
i шм California, U.S.A., Daniel s.n. (CAS) 

e to tropical and southern Africa). 

а реа Baker; AF169842, AF167703; 
Cultivated, Univ. Arizona, Tucson, Arizona, U.S.A., 
McDade 1232 (ARIZ) (native to Madagascar). 


dai c d (Burm. Bremek.; NA, 
129; Cultivated, Missouri Botanical Garden. 
= M ed Missouri, U.S.A., Accession No. 


S. Moore, NA, AF289724; Sunes 


oe MacDougal 5047 (MO) (native to In- 


Rhinacanihus gracilis D AF289766, AF289727; 
ted, San Francisco Conservatory of Flowers, 
San мен Бы California ‚ U.S.A., Daniel s.n. (CAS) 
(native to Africa). 
New World dina 
Harpochilus nee Mart. ex. Nees; AF289762, 
AF289721; (H9) 1 Brazil. Bahia Mn Luzia, Souza 
et al. 5413 (CA 
Justicia е Wassh. & L. B. : AF289759, 
ж Cultivated, Univ. Arizona мн Тисѕоп, Аг- 
а, U.S.A., Starr с.32 (ARIZ) (native to Mexico). 
J. ae A. doni AF169837, AF063134; Mexico. So- 
nora: near Alamos, Faivre 64 (ARIZ). 
J. comata (1, vlan AF289760, NA; Costa Rica. Heredia: 
Selva Biological Station, Vere A RIZ 
J. longii Hilsenb.; AF169839, AF063135; U.S.A. Form 


County, Tucson кейиш а Devender 87- 


307 (ARIZ). 
J. шен пичи Е, МА; нај Univ. Ar- 
своп, Arizona, McDade 
1158 (ARIZ) ae to Mexico and edt Ameri- 
Lindau; AF169840, 
126; Cultivated, Wilson Botanical Garden, 
MM e Rica, McDade 253 (DUKE) (na- 
outh America). 

Poikilacanthus асаби Lindau; AF169838, AFO7066; 
Costa Rica. Alajuela Province: Monteverde Re- 

serve, Haber 707 (MO) 


ca). 
M egaskepasma erythrochlamys 
AF063 


458 Annals of the 
Missouri Botanical Garden 


Appendix 2. Alphabetical list of Justicieae included in the present analysis; each taxon is assigned to a lineage 
(or grade in the case of Old World “justicioids”), reflecting the results presented herein. See Appendix 1 for authors 
of names, Genbank accession numbers, and vouchers. 


Aniscanthus puberulus равна ji 
A. thurberi Tetrameriur 
Anisotes i AINE Old World асс ic "E 
Asystasia gangetica Pseuderanthemum lineage 
Asystasia sp. Daniel 9129) м nthemum lineage 
Brachystephanus lyallii Isoglossinae 
Carlowrightia arizonica ре diee 
Chalarothyrsus amplexicaulis Tetramerium lin 
Chileranthemum pyramidatum Pseuderanthemum a ш 
Daniel 6737cv (Unidentified) Pseu шаа lineage 
Dicliptera ца Diclipterinae 
D. magaliesbergensis Diclipterinae 
D беен Diclipterinae 
D. suberecta Diclipterinae 
D. sp. (Dar siel 9194) Diclipterinae 
Duvernoia оя га Old World * ‘justic m 
Ecbolium syringifolium Tetramerium linea 
Fittonia albivenis Tetramerium lineage 
Gypsacanthus nelsonii Tetramerium lineage 
Harpochilus neesianus New World “justicioid” lineage 
Henrya insularis Tetramerium lineage 
Herperacanıhus згепорћ; yllus Pseuderanthemum lineage 
Hoverdenia specios Tetramerium 
oestes aristata Diclipteri 
H. phyllostachya Diclipterinae 
Isoglossa grandiflorc Isoglossinae 
Isogloss a (Daniel 9106) Isoglossinae 
Justicia adhate Old World “justicioids” 
J. betonica Old World “justicioids” 
J. brandegeana New World “justicioid” lineage 
J. caudata New World “justicioid” lineage 
J. comata New World “justicioid” lineage 
J. extensa Old World “justicioids” 
J. longii New er “justicioid” lineage 
J. spicigera orld “justicioid” lineage 
J. sp. (Daniel 9024) Old мү “justicioids” 
J. sp. (Daniel Ae, Old World “justicioids” 
J. sp. (Zjhra 983) Old World е 
е kaya bella Pseuderanthemum linea 
gaskepasma erythroc hlamys New World * justicioid" Ген 

ранага galpinii Old World “justicioids” 

irandea grisea Tetramerium | Ч 
Odontonema tubaeforme Pseuderanthemum lineage 
Oplonia microphylla Pseuderanthemum lineage 
Pachystachys lutea Tetramerium lineage 
Peristrophe hyssopifolia D ig erinae 
Poikilacanthus macranthus orld “justicioid” lineage 
Ране дина еј alatum Mud lineage 
Pty теше pubisepala Isoglossinae 

= spicata un 
Пас ићи gracilis Diclipterin 
Rungia klossii Old Wo dd "iusticioids" 
Ruspolia seticalyx Pseuderanthemum lin 
к fruticosa Pseuderanthemum lineage 
Schaueria calicotricha Tetramerium lineage 
Spathacanths hoffmannii Pseuderanthemum lineage 

S. parviflorus ои lineage 
Stenostephanus chiapensis чыш 

silvaticus Isoglos 

Sireblacanthus cordatus Tetramerium li ineage 
S. roseus Tetramerium eu 
Tetramerium nervosum Tetramerium line 


Unidentified (Daniel 6737cv) Cibeles пала tug 


A PHYLOGENETIC ANALYSIS 
OF DICOMA CASS. AND 
RELATED GENERA 
(ASTERACEAE: 
CICHORIOIDEAE: 
MUTISIEAE) BASED ON 
MORPHOLOGICAL AND 
ANATOMIC CHARACTERS! 


Santiago Ortiz? 


ABSTRACT 


This study s a phylogenetic analysis of the ind Dicoma and the related genera Achyrothalamus, Erythroce- 


phalum, Pasaccardoa, and Pleiotaxis (Mutisieae, Astera 
corolla, ray 


characters was used for the analysis. The genera 


flore t у жени anther appendages, and sena bra 


the resulting consensus c Dru suggests that the genus Dicoma appears paraphyletic. A clade including Dicoma 


e number of synapomorphies. These 


№ ilic geographical origin of Deana (probably Madagascar or southern Africa), and to the evolution of the different 


species groups, are briefly discussed. 
Ke 


ey words: 


Asteraceae, cladistics, Dicoma, Mutisieae, phylogeny, tropical Africa. 


The “Dicoma group” (Asteraceae, Mutisieae, 
Mutisinae) of Bremer (1994) is considered by this 
author to be one of the most difficult groups to eval- 
uate phylogenetically within the Mutisieae. Accord- 
ing to Bremer (1994), it is characterized by its co- 
rolla distinctly divided into a narrow tube and wide 
limb, its non-mutisioid ray floret epidermis pattern, 
its acuminate apical anther appendages, and из 
mostly subapically pilose style branches. It com- 
prises Dicoma itself, with 50 species occurring in 
tropical Africa, Madagascar, and South Africa, and 
two species reaching Asia (D. schimperi (DC.) Baill. 
ex O. Hoffm. is present in the Arabian Peninsula, 
and D. tomentosa Cass. is present in India and Pak- 
istan). Other genera of this group are Erythroce- 
phalum Benth. with about 12 species occurring 
throughout tropical Africa (particularly tropical 
East Africa), Achyrothalamus О. Н 
gle species from Kenya and Tanzania, Pleiotaxis 
Steetz with about 25 species occurring throughout 
tropical Africa, Pasaccardoa Kuntze with 4 species 


offm. with a sin- 


from tropical Africa, and Gladiopappus Humbert 
with a single species, possibly extinct, from Mad- 
agascar. 

Prior to Bremer's (1994) *Dicoma group," similar 
groupings had been proposed by Jeffrey (1967) and 
Grau (1980). Indeed, Jeffrey considered this group 
to be one of the most distinct within the Mutisieae. 
Hansen (1991) suggested that most of the genera 
of this group be moved from the Mutisieae to the 
tribe Cynareae (Cardueae). His support included (a) 
the cuticular ornamentation of corolla epidermal 
cells; (b) the corolla divided into a narrow tube and 
a broader limb; (c) bilabiate flowers with upper 
limb lobules short and uncoiled or absent (except 
in Gladiopappus, in which they are long and coiled; 
Humbert, 1963); and (d) style branches with sub- 
apical sweeping hairs. Karis et al.’s (1992) cladistic 
analysis of the subfamily Cichorioideae indicated 
that Dicoma, Erythrocephalum, and Pleiotaxis form 
a monophyletic group within the tribe Mutisieae, in 
accordance with traditional views (see Hoffmann, 


1893a; Cabrera, 1977) 


! My thanks to А. A. Anderberg, Р. О. Karis, and B. 


my stay at the Natural History "cni of Stockholm, to an anonymous reviewe 
the keepers of the herbaria mentioned for the loan of study material, to Alfredo López 
(Tokio) for the illustrations, and to Guy Norman for ше о translation: of this: maniaci ript. 

© 


provement of this article, to t 


ordenstam for their invaluable advice and assistance during 
r who contributed decisively to im- 


је Compostela, Galicia, 


2 Laboratorio de Botanica, Fac ultade de Farmacia, l 
Spain. 


ANN. Missouni Bor. GARD. 87: 459—481. 2000. 


Annals of the 


460 


Missouri Botanical Garden 


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Ortiz 461 


Volume 87, Number 4 


2000 


Dicoma and Related Genera 


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Annals of the 


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Ortiz 
Dicoma and Related Genera 


463 


Table 3. Characters and states used in phylogenetic 
analysis. 


1. Woody (0) / herbaceous (1) 
2. Plant erect (0) / spreading to prostrate (1). 
3. Stem with conical pluricellular trichomes (0) / without 


pluricellular trichomes 
4. Leaf ovate to lanceolate (0) / obovate to oblanceolate 
1 


5. Leaf herbaceous (0) / coriaceous (1). 
6. Leaf margin not revolute (0) / conspicuously revolute 


7. Leaf not conduplicate (0) / conduplicate (1). 

8. Stem without subtending leaves (0) / with subtending 
leaves (1). 

9. Capitulum > 20 mm wide (0) / capitulum = 20 mm 
wide (1). 

10. Involucre campanulate (0) / obconic (1). 

11. Outer phyllaries erect-patent to erect (0) / patent to 
reflexe 

12. Phyllary midrib inconspicuous (0) / conspicuous (1). 

13. Phyllaries without longitudinal dark stripes (0) / with 
dark stripes running along the phyllaries (1) 

14. Phyllary sclerenchymal fibers in a continuous tis- 
sue (0) / in distinct bundles surrounded by parenchy- 
mal tissue (1) / concentrated on the abaxial face (2) 
/ concentrated on the adaxial face 

15. Resin ducts in phyllaries at the eer face (0) / 
reduced or absent (1) (see Fig. 1) 

16. Phyllaries acute to acuminate-pungent (0) / subacute 


). 
17. Innermost phyllaries with scarious margins (0) / + 
entirely scarious (1). 
18. fons ost нм longer than the rest (0) / shorter 
than the contiguous outer serie 


s (1). 
19. Phyllary glabrescent to hairy (0 ) / completely glabrous 
1 


(1). 

20. Receptacle epaleate (0) / paleate (1). 
21. Florets in capitula actinomorphic (0) / marginal florets 
bilabiate and disc florets actinomorphic (1) / zygo- 
morphic (transitional between actinomorphic and lig- 
ulate) (2) / marginal florets “true-ray” (Karis et al.. 
1992) and disc florets actinomorphic (3) 
Marginal florets hermaphroditic (0) / neuter (1) 
23. Corollas = as long as or only slightly longer than the 

involucre (0) / much longer than the involucre (1). 
. Disc corolla tube gradually dilates into limb (0) / 


N 
N 


N 
~ 


abruptly dilates into limb (1). 
. Disc corolla lobes erect to erect-patent (0) / recurved 
at the apex (1). 
. Disc floret epidermal cell cuticle ornamentation: mu- 


N 
сл 


€ 


~ 
с 


tisioid (0) / slightly transversely ondulate-striate to 
nearly smooth (1) / conspicuously longitudinally stri- 
ate and тшш ondulate (*intestine-like") (2) / 
senecioid (: 


N 
“з 


. Disc un UR not sclerified (0) / sclerified (1). 
. Disc corolla without n twin glandular hairs (0) / 
). 


~ 
со 


with long twin glandular 
. Dise corolla without а dus (0) / with simple 
irs (1) 


~ 
јој 


Table 3. Continued. 


30. Star-shaped calcium oxalate crystals in disc corolla 


3l. 


Uc 
N 


сл 
Ф 


сл 
сл 


. Marginal vascular tissue in disc cor 


subepidermal cells absent (0) / present (1) (see Fig. 
2) 


Vascular tissue in disc corolla bifurcate at the lobe 
sinuses (0) / bifurcate well below the sinuses (1) / 
bifurcate at the corolla base (2) 


. Marginal vascular tissue in the disc corolla broad (0) 


/ narrow (1). 
|| | 


submarginal (see E 3-3 and 3-4 (0) / = mad 
(Fig. 3-1 and 3-2) (1 


. Accessory vascular strands in the disc corolla lobes 
absent Ay | present 
5 iX 


(1). 
e disc corolla lobes with thick-bundled 
veins 0 ) v» ye thick-bundled veins (1). 


. Stamen insertion in disc corolla: at the tube/throat 
(1) 


junction (0) / near the corolla 


. Star-shaped calcium oxalate crystals in stamen fila- 


ments absent (0) / present (1). 


. Stamen filament collar inconspicuous (0) / conspicu- 


ous and not swollen (1) / conspicuous and swollen at 


the apic 


cal part. 
. Stamen s collar « 0.8 mm long (0) / > 1.5 


mm long (1). 


. Anther not protruding beyond the corolla (0) / pro- 
) 


truding beyond the corolla (1 


. Anther appendage apiculate (0) / acute to slightly 


acuminate (1). 


2. Anther tails calcarate (0) / ecalcarate (1). 
3. Anther tails long tapering (0) / with subacute to sub- 


rounded apex (1). 


. Contiguous anther tails free (0) / joined (1). 
. Anther tail apices with antrorse ramifications (0) / 


without ramifications (1 


. Apex of the anther tail ramifications obtuse (0) (Fig 


5-3) / acute (1) (Figs. 5-1 and 5-2 


7. Sporoderm thick (0) / thin (1). 

8. Pollen smooth to hae) granulate (0) / echinate (1). 
. Style with two veins (0) / four veins (1). 

. Star- id cr MEN in styles absent (0) / present (1). 
. Style branches adjacent (0) / style branches separate 


(1). 


. Style branches straight (0) / conspicuously curved (1). 
3. Style branches short (0) (Figs. 6-1 and 6-2) / long (1) 
-4). 


(Figs. 6-3 and 6 


. Stylar sweeping hairs absent (0) / reaching or almost 
a 


reaching the bifurcation (1) (Figs. 6-1 and 6-2) / in 
19: p not reaching the bifurcation (2) (Figs. 6- 
4) / in a subapical tuft (3) (Fi 


5. Stylar sweeping hairs of similar length (0) (Fig. 6-1) / 


with a sub-basal group slightly longer than the rest 
(1) (Fig. 6-2) / with a sub-basal group conspicuously 
longer than the rest (2) (Figs. 6-3 and 6-4 


. Style branch vascular tissue narrow (0) / very thick 


. Cypsela carpopodium present (0) / absent (1). 


Disc floret cypsela ellipsoid (0) / obovoid to obconic 
(1) 


Annals of the 
Missouri Botanical Garden 


Table 3. 


Continued. 


59. Cypsela of marginal florets without rostrum (0) / with 


— 
~ 


rostrum ( 

60. Cypsela + smooth (0) / conspicuously ribbed (1). 

61. Star-shaped crystals in the cypsela pericarp absent (0) / 
present (1) 

62. Superficial cypsela glands absent (0) / in a continuous 
layer (1) / on the ribs (2) / in the intercostal grooves 
(3) 

63. Cypsela twin hairs simple (0) / twin hairs bifurcate 
(1) / absent (2). 

64. Twin hairs all around the cypsela (0) (Fig. 7-: 
tween ribs (1) (Fig. 7-1 and 7-3). 

65. Twin-hair bases not bulbous-glandular (0) / conspic- 
uously bulbous-glandular (1) 


2) / be- 


66. Twin hairs of the cypsela base similar to those of the 
other parts of the cypsela (0) / conspicuously enlarged 
l 


67. Cypsela with biseriate glands (0) / without biseriate 
glands (1). 

68. Immature testa with similar proportion of needle crys- 
tals and square to short-rectangular non-oriented 
crystals (0) / with long-rectangular crystals oriented 
in the same direction (1) 

69. Mature testa pattern: Gochnatia type (0) / Erythroce- 
phalum type (1) / Dicoma type (2) / Dicoma welwit- 
schii type (3). 

70. Testa epidermal cells with the lateral and basal walls 
тен (0) / only the basal walls strengthened 


71. Pappus of bristles (0) / scales (1) / bristles and scales 
(2) / absent (3) 


72. Pappus bristles scabrid to barbellate (0) / plumose 


73. Pappus element persistent (0) / caducous (1). 

74. Pappus elements of one row (0) / several rows (1) 

75. Pappus and involucre of the same length (0) / pappus 
overtopping the involucre (1) 

76. Pappus erect to erect-patent after fruiting (0) / patent 
after fruiting (1). 

77. All the pappi of the capitulum florets of similar length 
(0) / pappi of marginal florets much longer than those 
of disc florets (1) / internal pappus bristles of marginal 
florets as long as pappus of disc florets, and external 
pappus bristles of marginal florets much longer than 
pappus of disc florets (2) 


While studying African taxa of the Mutisieae 
(Asteraceae), particularly Dicoma Cass. (see Ortiz 
< Rodríguez-Oubifia, 1994, 1996, 1997; Ortiz et 
al., 1998; Rodríguez-Oubifia & Ortiz, 1995, 1997), 
I investigated phylogenetic relationships within Di- 
coma, and between this genus and related taxa. 

The principal aims of the work reported here 
were to investigate the relationships between Di- 
coma and other genera and the phylogenetic rela- 


tionships within Dicoma, with the aims of identi- 
fying generic and infrageneric subdivisions and of 
assessing the validity of the subdivisions previously 
proposed by Lessing (1830, 1832), De Candolle 
1838), Harvey (1865), Hoffmann (1893a), Wilson 
(1923), and Pope (1991) (see Table 1). Another ob- 
jective was to obtain preliminary information about 
the geographical origin and evolution of different 


“~ 


species groups within Dicoma. 


MATERIALS AND METHODS 


The phylogenetic analysis was based on morpho- 
logical and anatomical study of specimens from the 
BM, COI, K, MO, and S herbaria. A number of 
species of Dicoma were selected as representative 
of the various morphological types present within 
this heterogeneous genus as follows. First, the sev- 
en species of the group I denominate the “D. ses- 
siflora group” (see Results) are representative of a 
morphological type observed in a total of 19 spe- 
cies, including D. auriculata Hutch. & B. L. Burtt, 
D. elliptica G. V. Pope, and D. gossweileri S. Moore. 
Second, the species D. niccolifera Wild. and D. ca- 
pensis Less. are representative of a morphological 
type observed in 7 species, including D. macroce- 


Hoffm. Finally, D. anomala Sond., D. aethiopica S. 
Ortiz & Rodr. Oubifia, and D. montana Schwick. 
are representative of a morphological type observed 
in some 10 species not considered in the present 
analysis, including D. galpinii Wilson, D. popeana 
5. Ortiz & Rodr. Oubifia, and D. somalense 8. 
Moore. Та addition, and with the aim of covering as 
much morphological variability as possible, we in- 
cluded species from all the known sections of the 
genus (see Table 1) 

A total of 34 species were included in the study: 
the single species of Achyrothalamus, 24 species of 
Dicoma, 3 species of Erythrocephalum, 3 species 
of Pasaccardoa, and 3 species of Pleiotaxis (see 
Table 2). I also studied Dicoma anomala Sond., D. 
zeyheri Sond., and D. capensis Less. in the field in 
South Africa. Gladiopappus vernonioides Humbert 
was not included, since I was unable to obtain ma- 
terial of this taxon. 

A total of 77 morphological characters were stud- 
ied (see Table 3). For microscopic examination, flo- 
ral parts were first boiled in water with a surfactant, 
then mounted in Hoyer's solution (Anderson, 1954). 
Phyllary and achene sections were cut by hand with 
razor blades. Some characters initially considered 
were excluded from the analysis because they were 
the same in all species including outgroups, such 
as endothecial cell wall thickening organization 


Ortiz 465 


Volume 87, Number 4 


2000 


Dicoma and Related Genera 


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466 


Annals of the 
ier Botanical Garden 


2. Star-sha 
= rt, dj bun & Vollesen 7870 (K)). Seale bar = 0.01 m 


(Dormer, 1962), which was always polarized, and 
cypsela vein union (Karis et al., 1992), which was 
always at the base. Some of these characters re- 
quire explanatory comments, as follows: 

Character 8. Many of the species studied com- 
show more or less bractiform leaves, 
denominated "subtending leaves" (Pope, 1991, 

‚ at the base of the capitula. 

Character 13. 

have two dark stripes running along the phyllaries. 


monly 


> 


Many of the species studied 


Character 14. Examination of cross sections of 
the phyllaries allows four morphological types to be 
distinguished, most notably on the basis of the ar- 
rangement of sclerenchymal fibers. In the first type, 
the sclerenchymal fibers form a more or less con- 
tinuous mass occupying the inner part of the phyl- 
lary (Fig. 1-1). In the second type, the sclerenchy- 
mal fibers, though likewise situated in the central 
part of the phyllary, form clearly distinct groups 
separated by a very lax parenchyma, which appears 
to be aeriferous (Fig. 1-2). In the third type, the 
sclerenchymal fibers are concentrated toward the 
abaxial face of the phyllary; in the center, which is 
basically occupied by parenchyma, there are sev- 
eral vascular bundles reinforced with sclerenchy- 


»ed calcium oxalate crystals in the пне се of the disc corolla of Dicoma aethiopica 


mal fibers (Fig. 1-3, 1-4). In the fourth type, the 
majority of sclerenchymal fibers are either on the 
adaxial face or in the midrib of the abaxial face, 
where they protect the vascular bundles (Fig. 1-5). 

Character 21. 
er extent D. oleaefolia, have capitula in which all 
florets are zygomorphic, with one of the corolla 
lobes separated from the others by a much deeper 


Dicoma carbonaria, and to less- 


incision and without an expanded limb; this mor- 
phological type seems transitional between the ac- 
tinomorphic and the ligulate type and is similar to 
that seen in certain primitive genera of the subfam- 
ily Barnadesioideae, such as Chuquiraga Juss., Da- 
syphyllum Kunth, and Schlechtendahlia Less. 
(Hoffmann, 1983a; Cabrera, 1977; Bremer, 1994). 
“True ^ florets have no adaxial lobes and a 3- 


haracter 26. This character was о 
studied in ray florets (Baagge, 1977, ‚ but in 
my opinion is also informative for species |. ћауе 
disc florets only. Indeed, Hansen (1991) studied 
epidermal cell cuticle ornamentation in disc florets 
of species of the various genera considered in the 
present study (despite the fact that the study in 
question centers on ray florets). This character has 


шш [(y() = (p pue? 7) req ә 5. “wu pd 0 
= (© pue [) лед э[вэб '((М) zp97 uosiDagq) злэЧу jew Ацо: a19[os INOYIIM pue $]э$$эл ломој цим “шроң '0 пјродууори ошо] JO ejoo 
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Dicoma and Related Genera 


Ortiz 


"t 
Ф 
a 
= 
5 
= 
~ 
со 
Ф 
E 
= 
о 
> 


468 


Annals of the 
Missouri Botanical Garden 


Figure 4. 


veins. Scale bar = 0.05 mn 


likewise proved useful in the present study, in 
which most of the species considered lack ray flo- 


rets. “Intestine-like” ornamentation (Karis et al., 


, 


1992) is equivalent to Hansen’s “rugose pattern of 


longitudinal bands” (Hansen, 1991). In the char- 
acterization of Gochnatia and Achyrothalamus I 
have followed Hansen’s (1991) approach, classify- 


ing ornamentation in these two genera as “muti- 


1 2 


Figure 5. 
tail шше ‘ations is acute (Dicoma saligna, Gossweiler 11331 (COD). —2. эе oe 
"is the apex of the anther tail ramifications is acute (Dicoma tomentosa Cas 

ces have antr 


Е! 
| 


Apex of the ae corolla of Dicoma кени a (Gilbert, Ensermu & Vollesen 7870 (Ку) with thick-bundled 
b 


sioid" and “senecioid” respectively, though with 
some reservations, since the distinction between 
these two typologies was not clearly defined in our 
material. 

Character 28. Some species have long twin 
glandular hairs with a small apical gland on the 
corolla (see Karis et al., 1992: 418, fig. 4H). In the 


rest of the species considered the disc corolla hairs 


3 


Distal part of the anther tails. —1. Anther apices lack antrorse ramifications and the apex of the anther 


antrorse ier ations 


Nordenstam 2454 . Anther 


have 


orse ramifications and the apex of the anther tail ramifications i is obtuse (Dicoma nu (Drake) 


Humber Capuron 18643 (K)). AT: anther tail. B: ramifications. Scale bar = 0.05 


Volume 87, Number 4 
2000 


Ortiz 
Dicoma and Related Genera 


1 2 


Apical part of the style with sweeping hairs. 


Figure 6 
е геас hing the bifure sation ee виа Bayliss 4519 (5)). 
. these almost reac ning E bifurcation (Dicoma sessiliflora Harv. 
subsp. sessiliflora. Wingfield 4416 (K)). Bet uds E branches with sw 
basal group of sweeping hairs conspicuously longer than the rest (Dico ин Buscal. & M 
; with a long group of sweeping hairs not rea 


group of sweeping hairs sli „tl, longer the 


242 (K)). —4. Long style branches 


3 4 


—1. Short style branches with sweeping hairs of similar 
—2. Short style branches with a sub-basal 
hairs in a subapical eae 


ching the bifurcation, diese with a sub- 


basal group of sweeping hairs m ‘uously longer than the rest (Dicoma tomentosa, Nordenstam 2454 (S)). SH: 


sweeping hairs. Scale bar = 0.5 


are “short glandular hairs” with a large apical gland 
(see Karis et al., 1992: 418, fig. 41). Rarely (either 
in the first or second type) “twin hairs” are present 
that are basically similar to the non-myxogenic 
short ovoid twin hairs of Karis et al. (1992: 418, 
fig. 4B), which are very short and which lack a 
conspicuous apical gland. 

Character 29. Disc corolla simple hairs are 
long simple hairs with one or two basal cells (see 
Karis et al., 1992: 419, fig. 5E) 

Character 32. The marginal vascular tissue of 
the lobes of the corolla of disc florets is thick in 
some taxa, made up of many vessels and surround- 
ed by sclerenchymal fibers (Fig. 3-1 and 3-2). In 
the remaining taxa, the marginal veins are narrow, 
comprised of fewer vessels and without sclerenchy- 


species show accessory 
veins of variable length in the corolla lobes, run- 
ning more or less parallel to the marginal/submar- 
ginal veins. 

Character 35. In some species, the veins of the 
disc corolla lobes form a highly characteristic 
dense bundle (Fig. 4) (see Karis et al., 1992 

Character 36. In most of the species studied, 
the stamen filaments are inserted at the tube/throat 


junction. In D. carbonaria and D. oleaefolia, by 
contrast, the stamen filaments are inserted practi- 
cally at the base of the corolla. The presence of two 
clearly distinct states is not in agreement with Karis 
(1993), who reported that this character varies little 
within each of the tribes of the subfamily Astero- 
ideae (except in the tribe Astereae) and is thus of 
considerable taxonomic value at the tribal level. 

Character 44. Dicoma carbonaria and D. 
oleaefolia share an interesting characteristic, the 
tails of adjacent pairs of anthers being joined, with 
their ramifications or hairs interwoven. The anthers 
of the remaining species are clearly separated. 

Character 45. In many taxa the apices of the 
anther tails show characteristic antrorse ramifica- 
tions, while the ramifications of the rest of the an- 
ther tail are always retrorse (Fig. 5-2 and 5-3). In 
other taxa, however, such apical ramifications are 
absent (Fig. 5-1). 

Character 55. In several taxa the stylar sweep- 
ing hairs reach (or almost reach, or extend beyond) 
the point of bifurcation of the style branches. In 
most of the other species considered, the stylar 
sweeping hairs a short subapical tuft that 
Hansen (1991) and Bremer (1994) considered char- 
acteristic of the species of the “Dicoma group.” In 


Annals of the 


470 


Missouri Botanical Garden 


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Volume 87, Number 4 
2000 


Ortiz 471 
Dicoma and Related Genera 


Figure 8. 
bar = 0.1 mm 


Dicoma tomentosa and D. welwitschii, a third pat- 
tern is observed: the sweeping hairs extend from 
close to the apex toward the bifurcation, but do not 
reach it. 

Character 59. Cypselas of neuter marginal flo- 

rets are rudimentary and may or may not have a 
rostrum. 
Character 62. Many of the studied taxa have 
unicellular cypsela-surface glands containing a res- 
inous material. These may be located between the 
ribs on the rib walls (Fig. 7-3), between the ribs in 
the intercostal grooves, or forming a continuous lay- 
er around the cypsela (Fig. 7-1), as in the D. car- 
bonaria group. Dicoma fruticosa shows a morphol- 
ogy intermediate between types 1 and 2, and was 
coded as inapplicable for this character. 

Character 63. Simple twin hairs have the two 
cells joined, and bifurcate twin hairs have the two 
cells separated (Karis et al., 1992: 418, fig. 4A). 

aracter 65. In some species the cypsela twin 
hairs have a conspicuously bulbous glandular base 
that contains a resinous material (Fig. 7-2 and 8). 
In other species the base of the cypsela twin hairs 
may be somewhat bulbous but is never glandular. 

Character 66. In Pasaccardoa grantii and P. 
jeffreyi, the twin hairs of the lower part of the cyp- 
sela are thicker than the rest; in the remaining spe- 
cies all hairs of the cypsela are similar. 

Character 67. Biseriate glands are composed 
of two rows of cells and a collapsed apical head, 


Twin hairs with bulbous-glandular base on the cypsela of Dicoma saligna (Gossweiler 11331 (COI). Scale 


and occur in the cypselas of many of the taxa an- 
alyzed. In many cases these are not easy to see and 
often are situated between the superficial glands 
(see character 62). 

Character 68. Subepidermal calcium oxalate 
crystals are observed in the immature testa of all 
species. They are subsequently hidden from view 
by the epidermis of the mature testa. In some spe- 
cies needle-shaped crystals and square to short- 
rectangular crystals, without uniform orientation, 
are present in roughly equal proportion (Fig. 9-1). 
In others, by contrast, almost all the crystals are 
narrowly rectangular and oriented in the same di- 
rection (Fig. 9-2 

haracter 69. The first character state corre- 
sponds to the testa type defined by Grau (1980) for 
Gochnatia (testa with lateral and basal walls of the 
epidermal cells strengthened). Similar but not iden- 
tical morphologies are observed in Pasaccardoa 
grantii, P. jeffreyi, and some species of Dicoma: in 
these cases the testa morphology is similar to the 
Gochnatia type, but the cells of the epidermis are 
of irregular shape, not linear. The second character 
state is the Erythrocephalum type as defined by 
Grau (1980); in addition to Erythrocephalum, the 
species of Achyrothalamus and Pleiotaxis fall into 
this category (basal walls of the epidermal cells of 
the testa with reinforcements, which give to the 
cells a lacunose appearance in frontal view). The 
third character state is the Dicoma type as defined 


472 Annals of the 
Missouri Botanical Garden 
Figure 9. Calcium oxalate crystals in the immature testa (testa subepidermis). —1. Pig needle-shaped 


crystals and square to short-rectangular crystals in the t 


a of Dicoma zeyheri (Mogg 16306 (K)). —2. Long-rec ПО 


crystals oriented in the same direction in Dicoma montana (L. E. Codd 8689 (K)) (see text). Seale bar — 0.05 n 


by the same author (basal walls of the epidermal 
cells of the testa strengthened with ribs). The fourth 
in D. welwitschii and P. 
baumii, is very similar to that observed in the other 


character state, observe 


species of Pasaccardoa, except that the epidermal 
cells of the testa are traversed by thick bands. We 
have not been able to characterize testa morphology 
in the Oldenburgia material studied; similarly, Kar- 


is et al. (1992) were unable to determine whether 
or not the testa of the species of this genus is “col- 
The testas of the two species of the D. 
carbonaria group show a morphology similar to 
Grau's (1980) Perezia type (testa with epidermal 
cells not strengthened), though study of additional 
material is necessary to confirm this 

Character 70. In some species ilie lateral and 


lapsed." 


Ortiz 473 


Volume 87, Number 4 


2000 


Dicoma and Related Genera 


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Gochnatia 

Oldenburgia 

Pleiotaxis subpaniculata 
Pleiotaxis rugosa 

Pleiotaxis pulcherrima 
Achyrothalamus marginatus 
Erythrocephalum microcephalum 
Erythrocephalum zambesianum 
Erythrocephalum scabrifolium 
Dicoma welwitschii (Ps?/D) 
Pasaccardoa baumii 
Pasaccardoa jeffreyi 
Pasaccardoa grantii 

Dicoma elegans (S?/B) 
Dicoma aethiopica (Ps?/B?) 
Dicoma anomala (Ps/B) 
Dicoma montana (Ps?/B?) 
Dicoma dinteri (Ps?/B) 


Dicoma b lensis (E?/D) 


о 


Dicoma nachtigalii (B/B) 
Dicoma gillettii (Ps?/B?) 
Dicoma fruticosa (К?/В?) 
Dicoma picta (R/B) 
Dicoma niccolifera (S?/B?) 
Dicoma capensis (S/D) 
Dicoma schimperi (H/B?) 
Dicoma tomentosa (E/D) 
Dicoma cana (Ps/B) 
Dicoma spinosa (M/B) 
Dicoma grandidieri (Ps?/B?) 
Dicoma relhanioides (M/B) 
Dicoma sessiliflora (PUP) 
Dicoma zeyheri (PUP) 
Dicoma saligna (Pt?/P?) 
Dicoma carbonaria (Ps?/B?) 


Dicoma oleaefolia (Ps?/B?) 


Figure 10. Strict consensus tree of Dicoma and related genera, а on the six equally most parsimonious clad- 
ode are % jackknife support (before the 
after Dicoma species indicate the section to 
ongs. Sections proposed by Hoffman (1893a) (before the slash): S, Steirocoma; В, Rhigiothamnus; 
E, Eudicoma; M, Mac ledium; H, Hochstetteria; Ps, Psilocoma; B, Brachyachaenium; Pt, Pterocoma. Sections proposed 


ograms obtained from a 
h 


slash) : 


nalysis of the data in Ta 


ble 4 


Тће numbers a 


and % bootstrap support (after the slash). Letters in parenthe 
which each speci 


each n 


Volume 87, Number 4 
2000 


Ortiz 
Dicoma and Related Genera 


475 


basal walls of the epidermal cells of the testa are 
strengthened with a highly characteristic morphol- 
ogy, U-shaped in cross section and similar to that 
of Gun (1980) Gochnatia subgroup. In the re- 
maining species only the basal walls are hiked 
with ribs. 

Character 71. Pappus is only absent in Achy- 
rothalamus. In the remaining species it is formed 
by rather fine bristles, as in Pleiotaxis and in many 
species of Dicoma, by bristles and scales as in D. 
bangueolensis and D. tomentosa, or by scales only 
as in Erythrocephalum, Pasaccardoa, Dicoma wel- 
witschii, D. spinosa, and D. relhanioides. 

Character 73. Erythrocephalum has a pappus 
of narrow caducous scales, while in all other spe- 
cies considered the pappus is persistent. 

Character 77. In most of the species analyzed 
the pappi of the different flowers that make up the 
capitulum are all of similar length. In Dicoma nic- 
colifera and D. capensis, the pappi of the marginal 
florets are much longer than those of the disc flo- 
rets. In D. elegans, the internal pappus bristles of 
the marginal florets are as long as the pappus of 
the disc florets, while the external pappus bristles 
of the marginal florets are much longer than the 
pappus of the disc florets. 

Polarization of characters was determined by the 
outgroup comparison method (Stevens, 1980; Wa- 
trous & Wheeler, 1981; Maddison et al., 1984), us- 
ing Gochnatia Kunth and Oldenburgia Less. as out- 
groups. Both genera are of the tribe Mutisieae, and 
in Karis et al.’s (1992) analysis they are basal to 
the taxa included in the ingroup. Gochnatia is a 
heterogeneous genus with nearly 68 species (Bre- 
mer, 1994), mostly from the Americas, though also 
Asia. For outgroup comparison, I selected 5 species 
of Gochnatia, representatives of three of the five 
sections of the genus (G. amplexifolia (Gardner) Ca- 
brera, G. attenuata (Britton) Jervis & Alain, G. cor- 
data Less., G. microcephala (Griseb.) Jervis & 
Alain, and G. picardae (Urb.) Jiménez) with similar 
habit to those of the Dicoma group. Oldenburgia is 
a genus of four species endemic to the Cape area 
in South Africa, from which three species (О. gran- 
dis (Thunb.) Baill., O. papionum DC., and O. par- 
adoxa Less.) were selected. 

Characters with only two states of which one was 
autapomorphic were not included in the analysis. 

ner parsimony analysis of the data matrix 


Wag 
(Table 4) was performed on a PC with the aid of 


the program PAUP* 4.0 (Swofford, 1998). Clado- 
grams were generated using a heuristic search with 
the TBR (tree bisection-reconnection) branch- 
swapping algorithm with random additions (100 
replicates). Support values for each clade were ob- 
tained by jackknife analysis (100 replicates) (Farris 
et al., 1996) and bootstrap analysis (Felsenstein, 
1985). Successive weighting (Farris, 1969) was per- 
formed for generating cladograms where the rela- 
tive weight of homoplasious characters was ге- 
duced. All multistate characters were treated as 
nonadditive. Some characters were coded as poly- 
morphic for some terminals. In the data matrix, un- 
known character states were indicated with “777, 
and inapplicable character states were indicated 
with a dash 


RESULTS 


The cladistic analysis yielded six equally most 
parsimonious cladograms, each 186 steps long, 
with a consistency index (CI) of 0.513, and a re- 
tention index (RI) of 0.82, including in both cases 
the only uninformative character of the matrix. The 
six cladograms showed only minor differences. One 
of the six cladograms is shown in Figure 

Analysis with successive weighting gave two 
cladograms (Fig. 12), with identical major-clade to- 
pology to the six equally most parsimonious clad- 
ograms (and the corresponding strict consensus 
tree) obtained without successive weighting. How- 
ever, the two successive-weighting cladograms dif- 
fered from the no-weighting strict consensus tree in 
the internal topology of the major clades. 

In the strict consensus tree (Fig. 10), the first 
division split (a) the first clade, comprising the 
genera Pleiotaxis, Achyrothalamus, and Erythro- 
cephalum, from (b) the genera Dicoma and Pas- 
accardoa. The second division split (a) the Mad- 
agascan endemics D. 
oleaefolia (hereinafter referred to as the D. car- 
bonaria group) from (b) the remaining species of 


carbonaria and 


Dicoma and Pasaccardoa. The third division split 
(a) the seven species of the Dicoma sections Pter- 
ocoma DC., Macledium Less., and Psilocoma 
Harvey, hereinafter referred to as the D. sessiflora 
group, from (b) the remaining species of Dicoma 
plus the three species of Pasaccardoa. The fourth 
division gave the last two major clades, one com- 
prising Dicoma welwitschii and the Pasaccardoa 


€— 


by Wilson (1923) (after the slash): D, Dimorphae; В, Barbellatae; P, Plumosae. Question marks indicate that the species 
in question has not been assigned to any section before this paper (see Table 1). 


476 


Annals of the 
Missouri Botanical Garden 


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Gochnatia 

Oldenburgia 

Pleiotaxis subpaniculata 
Pleiotaxis rugosa 
Pleiotaxis pulcherrima 
Achyrothalamus marginatus 
Erythrocephalum microcephalum 
Erythrocephalum zambesianum 
Erythrocephalum scabrifolium 
Dicoma welwitschii 
Pasaccardoa baumii 
Pasaccardoa jeffreyi 
Pasaccardoa grantii 
Dicoma elegans 

Dicoma aethiopica 

Dicoma anomala 

Dicoma montana 

Dicoma dinteri 

Dicoma bangueolensis 
Dicoma nachtigalii 

Dicoma gillettii 

Dicoma fruticosa 

Dicoma picta 

Dicoma niccolifera 

Dicoma capensis 

Dicoma schimperi 

Dicoma tomentosa 

Dicoma grandidieri 

Dicoma relhanioides 
Dicoma spinosa 

Dicoma cana 

Dicoma sessiliflora 

Dicoma zeyheri 

Dicoma saligna 

Dicoma carbonaria 


Dicoma oleaefolia 


One of the six equally most parsimonious cladograms of Dicoma and related genera scar gta the 
` = 1). 0 


character distribution. Character numbering follows that shown in Tables 3 and 4. № synapomorphy (« 
synapomorphy (c < 1). О autapomorphy. = parallelism. X reversal. 


species, the other comprising the species I refer 


to as the D. tomentosa group 


In general, the five major ciues revealed by the 
analysis are adequately supported by jackknife and 


bootstrap values (see Fig. 10), and the results of 


the analysis can therefore be considered sufficiently 


reliable to provide a basis for elucidation of phy- 
logenetic relationships among the genera and spe- 


Volume 87, Number 4 
2000 


Ortiz 
Dicoma and Related Genera 


477 


A 
dr 


mE R 


Е: 
Е 


- 


Gochnatia 

Oldenburgia 

Pleiotaxis subpaniculata 
Pleiotaxis rugosa 

Pleiotaxis pulcherrima 
Achyrothalamus marginatus 
Erythrocephalum microcephalum 
Erythrocephalum zambesianum 
Erythrocephalum scabrifolium 
Dicoma welwitschii 
Pasaccardoa baumii 
Pasaccardoa jeffreyi 
Pasaccardoa grantii 

Dicoma elegans 


Dir thi. 
Ip 


Dicoma anomala 
Dicoma montana 
Dicoma dinteri 
Dicoma bangueolensis 
Dicoma nachtigalii 
Dicoma gillettii 
Dicoma fruticosa 
Dicoma picta 
Dicoma niccolifera 
Dicoma capensis 
Dicoma schimperi 
Dicoma tomentosa 
Dicoma cana 
Dicoma grandidieri 
Dicoma spinosa 
Dicoma relhanioides 
Dicoma sessiliflora 
Dicoma zeyheri 
Dicoma saligna 
Dicoma carbonaria 


Dicoma oleaefolia 


Figure 12. One of the two equally most parsimonious cladograms of Dicoma and related genera obtained in the 


8 
successive weighting analysis. 


478 


Annals of the 
Missouri Botanical Garden 


cies considered. Only the clade corresponding to 
the D. tomentosa group had low jackknife and boot- 
strap support. 


DISCUSSION 


The results of the analysis reported here contra- 
dict those of Karis et al. (1992), in that Erythro- 
cephalum and Pleiotaxis appear to be more phylo- 
genetically advanced than Dicoma. 

The analysis indicates that the genera Pleiotaxis, 
Achyrothalamus, and Erythrocephalum constitute a 
monophyletic group (see Fig. 10), the members of 
which share a number of apomorphic characters, 
some of which are exclusive (such as anther tails 
with subrounded to subacute apex, or style branch- 
es separated). Within this group, Erythrocephalum 
and Pleiotaxis appear to be paraphyletic. Neverthe- 
less, confirmation of this would require a cladistic 
analysis of these three genera alone, with consid- 
eration of all or most of the species included within 
them; the present study focused on the species of 
Dicoma and on the characters relevant to the tax- 
onomy of this genus. Such an analysis should try 
to identify other characters that although of little 
diagnostic value in the present study might be of 
value in an analysis of these three genera alone. 
Despite these reservations, it is of interest that 
Achyrothalamus and Erythrocephalum showed only 
minor differences (characters 16, 19, 26, 31, and 
71); of these characters, the absence of a pappus 
(character 71) is the most widely used to distin- 
guish between Achyrothalamus and Erythrocephal- 
um (which has a caducous pappus). In this con- 
nection, our observations show that bifid twin hairs 
are present on the cypsela of Achyrothalamus, and 
that these hairs are apparently identical to those 
previously considered to be diagnostic of Erythro- 
cephalum and Hyaloseris (Karis et al., 1992). The 
presence of these hairs in Achyrothalamus does not 
seem to have been taken into account when decid- 
ing to consider this taxon as a separate genus, pre- 
sumably because they have not previously been de- 
tected: there is no mention of them in the original 
description of the genus (Hoffmann, 1893b), and 
Bremer (1994) described the cypselas of Achyroth- 
alamus as glabrous. 

The clade comprising Dicoma carbonaria and D. 
oleaefolia (the *D. carbonaria group") (see Fig. 11 
is well defined and clearly separated from the re- 
maining species of Dicoma. Important apomorphic 
character states shared by these species include (a) 
capitulum of all florets zygomorphic [since the sub- 
family Barnadesioideae is one of the most primitive 
groups within the Asteraceae (Bremer, 1994), the 


ч” 


character state considered apomorphic, as observed 
in D. carbonaria and D. oleaefolia, should in a wid- 
er context probably be viewed as a reversion to a 
more primitive type; like Cabrera (1959), Bremer 
(1987), and Karis et al. (1992), I consider the most 
primitive state to be that in which all florets are 
actinomorphic] (character state 21.2), (b) disc co- 
rollas much longer than involucre (23.1), (c) disc 
corolla veins separated as far as the corolla base 
(31.2), (d) stamens inserted close to the base of the 
corolla [the marginal nerves of the corolla remain 
separated to the base of corolla, as in certain spe- 
cies of the genus Stenopadus, of the Guayana High- 
lands (Carlquist, 1957; Bremer, 1994), one of the 
most primitive genera or possibly the most primi- 
tive genus (Hansen, 1991; Karis et al., 1992; Bre- 
mer, 1993a, b, 1994) of the Mutisieae] (36.1), (e) 
anther projected beyond the corolla, (f) anther tails 
of contiguous anthers joined (44.1), (g) style with 
four veins (49.1), (h) 
posed in a continuous layer all around the cypsela 
(62.1), (1) pappus much longer than involucre 
(75.1), and (j) pappus bristles patent after fruiting 
(76.1). All of these synapomorphic characters, some 


superficial cypsela glands dis- 


relating to relevant aspects of floral morphology, to- 
gether with a series of plesiomorphic characters not 
observed in the remaining species of Dicoma (such 
as the presence of resin ducts, the absence of star- 
shaped calcium oxalate crystals in subepidermal 
cells of the corolla, and the presence of narrow style 
branch nerves), support the consideration of the D. 
carbonaria group as a clearly distinct clade, and 
raise the possibility that these two arborescent 
Madagascan endemics should be considered as a 
separate genus. This would entail resurrecting the 
genus Cloiselia S. Moore, described by this author 
(Moore, 1906) for С. carbonaria (D. carbonaria). 1 
am unaware of other species of Dicoma with floral 
characteristics like those observed in the D. car- 
bonaria group; certain of these characters, as men- 
tioned above, are clearly primitive (corolla ligulate 
but without expanded limb, stamens inserted close 
to the base of the corolla, abundant long simple 
hairs on the corolla surface). These characters sug- 
gest a relationship with the genera of the Barna- 
desioideae (notably Dasyphyllum, Chuquiraga, and 
Schlechtendahlia) and the likewise primitive South 
American Mutisieae genus Stenopadus (see Ca- 
brera, 1977; Bremer, 1994). This raises the possi- 
bility that the relationship between the African Mu- 
tisieae and the more primitive South American 
representatives of this tribe is perhaps closer than 
is currently admitted. This would be consistent with 
the hypothesis of Bremer (1993a, b, 1994), whereby 


the Asian species of Mutisieae were derived from 


Volume 87, Number 4 
2000 


Ortiz 
Dicoma and Related Genera 


the South American species as a result of westward 
spread across the Pacific. It is also possible that 
the primitive characteristics of the 
group reflect the isolation and protection from com- 
petition that has affected the evolution of many oth- 
er plant and animal lineages on Madagascar. Of the 
Dicoma species not considered in the present study, 
the only taxon showing some degree of external 
morphological similarity to the species of the D. 
carbonaria group is D. incana (Bak.) O. Hoffm., 
likewise arborescent and likewise endemic to Mad- 
agascar; note, however, that floral morphology char- 
acters in this species follow the normal pattern for 
the genus. 

The next clade (see Fig. 11) comprises two 
clades, one of them including the seven species 
that I refer to as the D. sessiliflora group. That this 
clade is genuinely representative of a phylogenetic 
entity is supported by a series of relevant non-ho- 
moplastic (CI = 1) synapomorphic characters, in- 
cluding innermost phyllaries more or less entirely 
scarious (17.1), and corolla with long twin glan- 
dular hairs (28.1). Other synapomorphic characters 
are homoplastic (CI < 1), due either to parallel 
evolution (e.g., phyllary anatomy, marginal veins of 
the disc corolla lobes in marginal or almost mar- 


. carbonaria 


ginal position, base of the twin hairs of the cypsela 
very conspicuously bulbous-glandular) or to rever- 
sions to the plesiomorphic state (e.g., innermost 
phyllaries shorter than the contiguous outer series, 
no antrorse branches at the apex of the anther tails 
Plesiomorphic characters shared by the members 
of this group include marginal veins of the disc 
corolla lobes broad (32.0), cypsela ribs absent or 
very slender (60.0), and twin hairs all around the 
cypsela (64.0). Confirmation that the D. sessiliflora 


group constitutes a distinct entity within Dicoma, 


. 


м 


ог possibly а separate genus, will require a more 
detailed analysis considering all the species in the 
group. It is worth noting that some of the species 
of this group are more adapted to woodland con- 
ditions than those of the D. tomentosa group (which, 
together with D. welwitschii and Pasaccardoa, con- 
stitutes its sister group); the species of the D. to- 
mentosa group, though sometimes occurring in 
woodland habitats, are generally better adapted to 
dry, open sites, and even sub-desert or desert hab- 
itats (see Lisowski, 1991; Pope, 1992). 

The members of this latter clade (see Fig. 11) 
share a number of apomorphic characters including 
(a) a conspicuous phyllary midrib (12.1), (b) disc 
corolla lobes recurved at apex [the corolla lobes 
have a recurved apex, with the apices of the anthers 
and the pistil exserted; this may reflect the adap- 
tation of most species of this group to desert or sub- 


desert environments in which pollinators are very 
scarce, so that there is selection in favor of at least 
partial anemophily (see Whitehead, 1969; Lane, 
1996)] (25.1), (c) cypsela ribs conspicuously strong 
(60.1), (d) cypsela with twin hairs between the ribs 
(as also observed in the D. carbonaria group, and 
presumably reflecting parallel evolution) (64.1), 
and (e) immature testa (subepidermal layer of the 
testa) with long-rectangular crystals oriented in the 
same direction (68.1). This group splits into two 
clades: one in which D. welwitschii appears as a 
sister group of the three species of Pasaccardoa, 
and another comprising the D. tomentosa group. 
The latter might be referred to as Dicoma s. str., 
since it contains D. tomentosa, the type species of 
the genus. 

In view of the topology of the cladogram (Figs. 
10-12), one possible approach would be to transfer 
the species of the genus Pasaccardoa to Dicoma. 
Alternatively, Pasaccardoa could be maintained, in 
view of its various synapomorphic characters, one 
non-homoplastic (cypsela of the marginal florets 
with rostrum) and others shared with other species 
but not with the members of the D. tomentosa group 
(disc corolla tube abruptly dilating into limb, mar- 
gin of the disc corolla lobes conspicuously scleri- 
fied, no antrorse branches at the apex of the anther 
tails, pappus of scales). Pasaccardoa also has a 
number of relevant plesiomorphies including pres- 
ence of a cylindrical disc floret cypsela, and Goch- 
natia-type testa. This view is supported by the fact 
that D. welwitschii, which the analysis indicated to 
be a sister group to Pasaccardoa, shows a number 
of character states different from those of the D. 
tomentosa group, some apomorphic (phyllary scler- 
enchymal fibers concentrated on the abaxial face, 
margin of the lobes of the disc corollas conspicu- 
ously sclerified, absence of antrorse branches at the 
apex of the anther tails, testa of D. welwitschii type, 
pappus of scales) and some plesiomorphic (capit- 
ulum wider than 20 mm, absence of dark stripes 
along the phyllaries, and lateral and basal walls of 
testal epidermal cells strengthened). Some of these 
characters are shared with the species of Pasac- 
cardoa. However, the inclusion of D. welwitschii 
within Pasaccardoa does not seem to be justifiable, 
in view of the marked differences with respect to 
the species of this genus, notably P. grantii and P. 
Jeffreyi. These two species, in addition to P. pro- 
cumbens (not included in the present analysis), ap- 
pear to form a highly homogeneous group, with ex- 
clusive synapomorphic characters (such as twin 
hairs of the cypsela base conspicuously enlarged, 
and cypsela surface glands positioned between the 
ribs in the intercostal grooves) and other apomorph- 


480 


Annals of the 
Missouri Botanical Garden 


ic characters that are not exclusive but that are not 
present in D. welwitschii (capitulum made up of 
“true-ray” marginal florets and actinomorphic disc 
florets, marginal florets neuter, and cypsela of the 
marginal florets with rostrum). By contrast, D. wel- 
witschii shows a series of apomorphic characters 
including apex of the disc corolla lobe without 
thick-bundled veins, disc floret cypsela obconic, 
and testa of D. welwitschii type. The cladogram may 
thus indicate that D. welwitschii (from the central 
plateaus of Angola, and also present in Zaire) is a 
distinct genus. Nevertheless, this possibility would 
have to be confirmed by an analysis including all, 
or nearly all, species of Dicoma. 

Of the previously proposed sections of the genus 
Dicoma, Dimorphae and Barbellatae of Wilson 
(1923) are clearly paraphyletic (see Table 1, Figs. 
10, 11). Considering the sections accepted by Hoff- 
mann (1893a), the results of the present analysis 
are more or less consistent with Brachyachaenium 
Baker (D. nachtigalii), Macledium (Cass.) DC. (D. 
spinosa and D. relhanioides), and Pterocoma DC. 
(sect. Plumosae Wilson) (D. sessiliflora, D. zeyheri, 
and D. saligna). 

Other sections, such as Steirocoma DC. (D. ca- 
pensis and D. niccolifera, but not D. elegans) and 
Rhigiothamnus (Less.) DC. (D. picta and D. fruti- 
cosa), may be monophyletic; however, and apart 
from the monotypic sections Hochstetteria (DC.) O. 
Hoffm. (D. schimperi) and Eudicoma DC. (D. to- 
mentosa), the species of my D. tomentosa group do 
not appear to form coherent monophyletic groups 
assignable to any of the sections previously de- 
scribed. 

Broadly speaking, the results of the present anal- 
ysis (Figs. 10—12) suggest that taxa with primitive 
characters, like those of the D. carbonaria group, 
may be precursors of the genus Dicoma that have 
survived as relict populations in Madagascar. It 
seems reasonable to hypothesize that these precur- 
sors gave rise to a lineage of xeromorphic taxa with 
small, spiny, coriaceous leaves (such as D. spinosa, 
D. relhanioides, 1). grandidieri, and D. cana). The 
remaining species of the D. sessiliflora group, with 
scarcely coriaceous, scarcely spiny, and more or 
less broad leaves (D. sessiliflora, D. saligna, and D. 
zeyheri) probably originated from that lineage in ad- 
aptation to moister, shadier conditions. The origin 
of this group, and of the genus as a whole, would 
thus appear to have been in Madagascar and south- 
ern África, as found by ЕЈдепаз and Andenberg 
(1996) for the genus Anisopappus (which has a sim- 
ilar distribution and ecology to Dicoma). On this 


hypothesis, the remaining species of the genus 


arose, probably later, generally in open dry, includ- 
ing sub-desert and desert, environments. 


Literature Cited 


Anderson, L. E. 1954. Hoyer's solution as a rapid mount- 
ing medium a bryophytes. КА 57: 242-241. 
Baagge Microc Ћагас te 

Compositae. Po. 119-139 
borne & B. L. Turner Are The Biology and (hem: 
зе of 5 Compositae ademic Press, London. 
—— Каш ide ipa of ligule micro- 
charac ‘ters in Compositae, П: Arctotideae, Astereae, 
Galen leas: Eremothamneae, Inuleae, Libeae, Mutis- 
ieae, and Senecioneae. Bot. Tidsskr. 72: 125-174. 
EUR K. 1987. Tribal о of the Astera- 
eae. Cladistics 3: 210— 
993a. ente are 
tation of the center of origin concept. Syst. 


436—445. 


аз— А cladistic reinterpre- 


Biol. 41: 


3b. Intercontinental relationships of African 
and South American Asteraceae—A cladistic biogeo- 
graphic analysis. Pp. 104—135 in P. Goldblatt (editor), 
Biological Relationships between Africa and Sout 
America. Yale Univ. Press, New Haven. 
1994. Asteraceae. Cladistics & Classification. 
Timber ce е Огероп. 
Cabrera, А. L. 9. Revision of the genus died 
Vi oie d Mus. La Plata, Secc. Bot. 9: 21— 
00. 


1977. (roni н review. Рр. 1039— 


1066 їп 


1 V. Н. Heywood, J-B. Harborne & В. L. Turner 
ru The Biology and Е hems of the Compositae. 
mic Press, Lo 


Candolle, 3 P de. 1838. [e ON о панаа 
& ‚ Ра 


Anatomy of т Guyana Mutisieae. 
Mem. New York Bot. Gard. 9: 4 

Dormer, 5 J. 1962. The fibrous ey in ihe anthers of 
Compositae. New Phytol. 61: 150- 

Eldeniis, P. & A. А. Anderberg. 1996. А eladisti pr 
of Anisopappus (Asteraceae: Inuleae). РІ. vol 
199: 167-192. 

Farris, J. S. 1969. A successive approximation aproach to 
character weighting. Syst. Zool. 18: 374—385. 

/. A. Albert, M. Killersjé, D. Lipscomb 
Kluge. 1996. Parsimony jac -a outperforms neigh- 
bor-joining. Mog 12( 124. 

Felsenstein, J. ви dimit its on phylogenies: 
An i using и bootstrap. а 39: 783- 
791. 


Grau, J. e Testa der Mutisieae und ihre syste- 
пиће Bedeutung Mitt. Bot. Staatssaml. Miinchen 
9-332 


Hansen, H. V. 199 1. Phylogenetics eod in Compositae 
tribe Mutisieae. Opera Bot. 109: 1— 

Harvey, W. H. 1865. Compositae. Pp. wen in W. H. 
Tao & O. Sonder, Flora Capensis 3. Hodges, Smith, 
Dublin. 

Hoffmann, " е Tubiflorae—Mutisieae. Рр. 333-350 
in Н. С. A. Engler & К. A. Е. Prantl, Die а 
нЕ 4(5). №. на, Leipzig 

1893b. Compoitas africanae. I. Tribus Mutis- 
ieae. Bot. Jahrb. Syst. 15: 536 6 547. 

Humbert, Н. 1963. Flora de Madagascar et des Comores. 

Ш. 189° famille—Composées. Muséum National 


Volume 87, Number 4 
2000 


Ortiz 
Dicoma and Related Genera 


a Naturelle, Laboratoire de Phanérogamie, Par- 


Jeffrey, C. 1967. Notes on Compositae. П. The Mutisieae 
in East cog Africa. Kew Bull. 21: 177-224. 

Karis, P. O. 1993. Morphological phylogenetics of the As- 
Herode Анн Шил e Po on character evolu- 
tion. Pl. se Evol. 186: 6 

M. Kallersjé & К. бан тег. 1992. ir раке 

analysis of the Cichorioideae (Asteraceae), with empha 
sis on the Mutisieae. Ann. Missouri Bot. Gard. 79: - 416 
27. 


Гапе, М. А. 1996. dris biology P Compositae. Pp. 
61-71 in Р. D. S. Caliga . Hind (editors), 
Compositae: Biology & “Utilization. Pob de of the 
International Compositae Conference, Kew, Vol. 2. The 
Royal Botanic Gardens, Kew. 

830. The idi iie i arm — Be- 
rolinensis, ои secunda. Linna 

. 1832. Synopsis Generum аю заны Dunak- 
er & Humblot, Berlin. 

Lisowski, S. 1991. Les Asteraceae dans la € а roe 
я Fragm. Flor. Geobot. 36(1), Suppl. 1: 251– 


PAR J. Donoghue & D. R. Maddison. 
1984. и adus and parsimony. Syst. Zool. 33: 
83-103. 


Moore, S. 1906. Alabastra diversa. Part XIII. J. Bot. 44: 
145-150. 


Ortiz, S. & Rodríguez- -Оиђта. 1994. Dicoma = 
and Dicoma cinerea (Asteraceae), two new species from 
Somalia. Canad. J. Bot. 72: 1478-1481. 

————— & ————. 1996. Two new species of Dicoma 


Cass. иет from Somalia and Ethiopia. Nordic 
J. Bot. 16: 2 
& 


«& . À new taxonomic кзы ти 
of the Dicoma psc complex (Asteraceae, Mutis 
ieae). Nordic J. Bot. 16: 583-584. 

M. Tadesse. 1998. A taxonomic revi- 
sion al Dicoma (Asteraceae: Cichorioideae: Mutisieae) 
n the Horn of Africa. Ann. Missouri Bot. Gard. 85 

440-459. 


Pope, G. V. 1991. Notes on Dicoma Cass. (Compositae). 
Kew gro at 699-709. 
· Flora Zambesiaca, vol. 6, р 1. Flora 


new species 
of the genus Dicoma (Asteraceae) from Somalia. Bot. J. 
Linn. Soc. 119: 59-64, 


1997. A new species of Pleiotaxis 
Steetz (Asteraceae) from Angola. Nordic J. Bot. 16: 
477—479. 


dien P. F. 1980. pop к) of character 
es. Annual Rev. Ecol. Syst. 11: 333-358 
Swofford, D. L. 1998. PAUP*. аке Analys is Us- 
ing Parsimony (*and other methods). Version 4. Sinauer 
Associates, Sunderland. 
Watrous, L. E. & O Wheeler. 1981. The out-group 
comparison method of character analysis. Syst. Zool. 30: 


Whitehead, D. R. 1969. Wind pollination in the angio- 
sperms: Evolutionary and environmental considerations. 
Evolution 23: 28-35. 

Wilson, F. C. 1923. Revision of the genus Dicoma. Bull. 
Misc. Inform. Kew 1923: 377—388. 


THE EVOLUTION OF 
NON-CODING CHLOROPLAST 
DNA AND ITS APPLICATION 
IN PLANT SYSTEMATICS! 


Scot A. Kelchner? 


ABSTRACT 


icle reviews several proposed mec hanisms of molec ular evolution operating in non-coding regions of the 
I 


This 
chloroplast genome and argues 
реза analysis of 
a Ban anii d inserti 


on a collective understanding of genic DN 
suggest an approach to the 


essential К 


ructured, nonrandom, and non- independent events. Established methodologies are based in large part 
NA evolution and may need modification when applied to non-coding sequence 
phylogenetic study of non-coding cpDNA that inc orporates identification of 


‹ 

ШШК mechanisms in alignment and homology assessment of indels. I к discuss repercussions of non-coding 

sequence evolution for such aspects of phylogeny estimation as maximum likelihood, distance, and parsimony analysis, 

the i inclusion of indels as phylogenetic characters, and bootstrapping, jackknifing, and “decay” analysis as measures 
1. 


of к suppor 
wo 


ords: 
phyloge netic analysis, secondary structure. 


alignment, intergenic spacers, introns, molecular evolution, mutational biases, mutational mechanisms, 


There is growing interest in comparative analysis 
of non-coding chloroplast (non-coding cpDNA) se- 
quences for plant systematic studies at low taxo- 
nomic levels. Recognition of the limitations of cod- 
ing (genic) DNA for resolving relationships at these 
levels inspired the probing of chloroplast introns 
and intergenic spacers for phylogenetic utility. Un- 
derlying this effort was the reasonable premise that 
non-coding regions experience limited or no selec- 
tive pressure and are likely to evolve at rates far 
surpassing those of genic ш (e.g., Curtis & 
d 1984; Wolfe et al., 1987; Palmer, 1987, 

991; Olmstead & Palmer, end óhle et а|., 
Ted There was also an expectation that non-cod- 
ing regions should experience random and inde- 
pendent mutations, both in mode and distribution. 

For these reasons, a remarkable number of plant 
systematics studies currently in progress include a 
molecular component of comparative analysis of 
non-coding cpDNA sequences. A considerable 
amount of work already published has demonstrat- 
ed the potential phylogenetic utility of discrete non- 


coding regions in the chloroplast: the trnL-trnF 
spacer (e.g., Gielly & Taberlet, 1994; Mes & t'Hart, 


spacer (Воће et al., 1994, 1997; Small et al., 
1998), the rpoA-petD and rpsll-rpoA spacers (Pe- 
terson & Seberg, 1997), the atpB-rbcL spacer (Go- 
lenberg et al., 1993; Hodges & Arnold, 1994; Na- 
tali et al., 1995; Samuel et al., 1997; Savolainen et 
al., 1997; Setoguchi et al., 1997; Hoot & Douglas, 
998), the rbcL-psal spacer (Morton & Clegg, 
1993), the psbA-trnH spacer (Aldrich et al., 1988; 
Sang et al., 1997), the accD-psal spacer (Small et 
al., 1998), the rpl16-rpl14 and rps8-rpl14 spacers 
(Wolfson et al., 1991), the intron surrounding matK 
(Johnson & Soltis, 1994), the rpoC1 intron (Downie 
et al., 1996a, 1996b; Asmussen & Liston, 1998; 
Downie et al., 1998), the rp/16 intron (Jordan et al., 
1996; Kelchner, 1996; Kelchner & Clark, 1997; 
Schnabel & Wendel, 1998; m et al. 
Small et al., 1998), the trnL intron (Sang et a 
1997; Bayer & Starr, 1998; Kajita et al., 1998; Bay- 


! | thank Conny Asmussen, Lyn Cook, Steve 


for review 
whose insightful comments contributed to the clarity ar 

Centre for | "lant Biodiversity Research, 
Canberra, A.C ; 
sity, Canberra, “Australia, scot.kelchner@pi.csiro.au. 


ANN. Missour! Bor. 


M. D. Crisp, J. G. Wes 
w of the manuscript; Victoria Hollowell for e careful editing and commentary; E the anonymous reviewers, 
id presentation of ideas in this pa 

ош Nation 


Dickie, Jonathan Wendel, Rich Cronn, and Randy Small for many 
interesting and insightful discussions on ihe topic P ake -coding seque 
comments on an earlier version of this paper; R. J. Bayer, 


nce evolution; R. G. bes: for his valuable 
Kelchner, and B. Oxelman 


al Herbarium, CSIRO Plant dad G.P.O. Box 1600, 


Australia. Second affiliation: Division of Botany and Zoology, The Australian National Univer- 


GARD. 87: 482—498. 2000. 


Volume 87, Number 4 
2000 


Kel 483 
pite Me Chloroplast 
DNA Evolution 


er et al., 2000), the rps16 intron (Liden et al., 1997; 
Oxelman et al., 1997), and the ndhA intron (Small 
et al., 

The literature above not only reveals profound 
differences between the evolution of non-genic and 
genic cpDNA, but critically contradicts initial as- 
sumptions of constraint-free evolution in non-cod- 
ing regions. Recurring difficulties associated with 
non-coding sequence data inclu 
alignment possibilities of insertions and deletions 
(indels), regions of length mutation in which ho- 
mology assessment is questionable or impossible, 
and the occurrence of localized *hot spots" of in- 
ferred excessive mutation, frequently to the point 
of saturation and loss of phylogenetic signal. How 
best to proceed with the phylogenetic analysis of 
such regions should be a € of considerable con- 
cern (see Golenberg et al., 1993; Downie et al., 
1996a; Kelchner & Clark, 1997; Sang et al., 1997; 
Downie et al., 1998). 

It is now evident that sequence evolution in non- 
coding regions of the chloroplast is far more com- 
plex than previously supposed. Both introns and 
intergenic spacers are thought to embody a consid- 


e alternative 


erable degree of sequence structure, sometimes in 
a manner similar to that of ribosomal DNA (rDNA). 
This structure may generate either regionalized se- 
quence conservation or mutational hot spots of both 
nucleotide substitutions and insertion/deletion 
events. Sequence-directed initiators of mutational 
events may persist as “mutational triggers" (Kel- 
chner, 1996; Kelchner & Clark, 1 

ly increasing the possibility of reversal or parallel 
gain of mutations, particularly length mutations or 
minute inversions. 


997), dramatical- 


ence, there exist essential vi- 
olations of the assumptions of randomized and in- 
dependent character evolution embedded in much 
of the current phylogenetic methodology for com- 
parative sequence analysis—methodology that is 
based largely on observational comparative study 
of coding sequence data. Considering that these are 
today's commonly employed tools for phylogeny es- 
timation based on DNA sequences, there has been 
as yet remarkably little controversy in the literature 
about their application to non-genic sequence data. 
here are ways to account for mutational patterns 
observed in non-coding DNA. Comparative studies 
of non-coding cpDNA sequences during the past 
decade in particular (e.g., Palmer, 1985; Blasko et 
al., 1988; vom Stein & Hatchel, 1988; Wolfson et 
al., 1991; Golenberg et al., 1993; Gielly & Taberlet, 
1994; Morton, 1995a; Downie et al., 1996a; Kel- 
chner & Wendel, 1996; Kelchner & Clark, 1997; 
Sang et al., 1997) have allowed inference of spe- 
cific underlying mutational mechanisms responsi- 


ble for generating sequence diversity in non-coding 
regions of the chloroplast genome. Unfortunately, 
these mechanisms are often invoked, but rarely in- 
corporated, into the analysis. 

Recognition of the potential of structured molec- 
ular evolution in non-coding cpDNA regions to im- 
prove alignment and assessment of phylogenetic re- 
lationships is, critical for 
development of functional molecular systematic re- 
search based on non-coding sequence data. Toward 
this end, I endeavor here to illustrate the following: 
(1) non-coding regions are highly structured and 
their elements evolve non-randomly and non-in- 
dependently; (2) this structure may be used to align 
the sequence matrix and better assess homology; 
3) the resulting gaps in the aligned matrix may 
contain phylogenetically important information and 
should be used in a phylogenetic analysis; and (4) 
the mode of non-coding sequence evolution de- 
scribed here may have potentially serious reper- 
cussions for the accuracy of genetic-distance, max- 
imum likelihood, and parsimony analyses, and for 
bootstrapping and jackknifing techniques. A de- 
scription of proposed mechanisms of non-coding 


e leve, 


— 


sequence evolution is followed by a discussion of 
the appropriateness of current alignment and anal- 
ysis procedures, with the expectation that it may 
provide a more informed approach to the applica- 
tion of non-coding sequence data in plant system- 
atics researc 

is uds is not intended to be a complete re- 
view of literature pertaining to the evolution of in- 
trons and intergenic spacers in all genomes of an 
organism. Instead, it serves as a brief review of 
current literature on non-coding cpDNA regions, 
and summarizes mutational mechanisms suggested 
to occur in these regions. Discussed are some of 
the serious implications this manner of molecular 
evolution has for the assumptions underlying mod- 
els employed today by plant molecular systematists. 


MECHANISMS OF NON-CODING SEQUENCE 
EVOLUTION 


The strength of any phylogenetic estimation rests 
on the accuracy of character homology assessment. 
Thus, the molecular systematist strives to maximize 
character homology by the careful alignment of 
DNA sequences in a data matrix. F мр сечи to 
any alignment procedure of non-coding cpDN 
quence data should be a familiarity with Pisis 
mechanisms directing molecular evolution in non- 
coding regions. Recognition of these mechanisms 
as generators of specific mutations can be a pow- 
erful tool for the placement of gaps and for the 


484 


Annals of the 
Missouri Botanical Garden 


assessment of probable homology of insertions and 
deletions (Kelchner, 1996; Kelchner & Clark, 
1997) 


SLIPPED-STRAND MISPAIRING (SSM) 


A widely reported mechanism of length mutation 
in non-coding regions of the chloroplast is slipped- 
strand mispairing (SSM). SSM is thought to be a 
major, even principal, factor in length mutations 
within non-coding regions of the chloroplast, mi- 
tochondrial, and nuclear genomes (e.g., Levinson & 
Gutman, 1987; Hancock, 1995; Wolfson et al., 
1991; Kelchner & Clark, 1997; Sang et al., 1997). 
Length mutations are important components of non- 
coding sequence evolution and have been suggest- 
ed to occur at least as frequently as base substi- 
tutions in some chloroplast non-coding regions 
(Curtis & Clegg, 1984; Wolfe et al., 1987; Zurawski 
& Clegg, 1987; Clegg & Zurawski, 1992; Golen- 
berg et al., 1993; Gielly & Taberlet, 1994; Clegg 
et al., 1994). 

Slipped-strand mispairing is thought to proceed 
by a localized mispairing of single-stranded DNA 
in regions of sequence repeats, as either a string of 
mononucleotide repeats or tandemly arranged mul- 
tibase repeat units (Palmer, 1991; Wolfson et al., 

; Cummings et al., 1994; Hancock, 1995; re- 
viewed by Levinson & Gutman, 1987). Diagrams of 
proposed SSM mechanics can be found in Levinson 
and Gutman (1987) and Wolfson et al. (1991). Be- 
cause A/T-rich regions of bacterial genomes аге 
particularly susceptible to slipped-strand mispair- 
ing (Levinson & Gutman, 1987), one could expect 
a similar effect in the A/T-rich non-coding regions 
of the chloroplast genome (Wolfson et al., 1991). 
This is not to imply that SSM acts uniquely on A 
and T nucleotides; aligned non-coding sequence 
matrices often infer inserted repeats containing G 
and C nucleotides, sometimes as pure strings of G 
or C mononucleotide repeats. 

Strings of mononucleotide repeats, particularly of 
А or T, appear frequently in non-coding cpDNA, 
and slipped-strand mispairing may potentially gen- 
erate length mutations within these strings. The dif- 

iculty in assessing homology of length variation in 
long strings of repeats, whether mononucleotide or 
multinucleotide repeats, derives from the increas- 
ing potential for further length mutation relative to 
string length (Streisinger & Owen, 1985; Golenberg 
et al., 1993; Kelchner & Clark, 1997; Sang et al., 
1997). Subsequent SSM activity may either gener- 
ate additional repeats of the initial sequence or de- 
lete sequence susceptible to slipped-strand mis- 
pairing. Perhaps an equilibrium might exist 


between the probability of inserting subsequent 
length mutations and the probability of removing 
sequence from the repeat string. Whether such an 
equilibrium is present or not, there may be a com- 
petitive phenomenon that keeps the length of tan- 
dem repeated sequence units continually in flux. 
Representation of long repeat strings in non-coding 
sequence alignments would therefore be а “snap- 
shot” of sequences experiencing continual inser- 
tions and deletions at that locality. 

It follows that a point substitution within a long 
string of mononucleotide repeat units could act as 
a stabilizing factor, disrupting its previous unifor- 
mity and lowering the probability of further SSM 
events. Such a substitution would directly influence 
ensuing mutations in the region and is one example 
of a non-independent character mutation in non- 
coding DNA. If the situation were reversed, with a 
non-homogeneous sequence becoming a string of 
repeat units, the likelihood of an SSM event would 
increase and could induce further non-independent 
mutations by the addition or removal of repeated 
sequence by slipped-strand mispairing. 

As an aid to alignment, SSM-generated inser- 
tions and deletions can be used to position and 
determine number of gaps. A quick study of a re- 
peat unit or the flanking sequence of a gap may be 
enough to determine if slipped-strand mispairing is 
the likely progenitor of an observed length muta- 
tion. Occasionally, evidence of an SSM event may 
not be apparent, particularly if a deleted sequence 
is not a direct repeat of its flanking sequence, or if 
a subsequent length mutation due to another mech- 
anism obscures an earlier SSM event (Kelchner, 
1996). 


STEM-LOOP SECONDARY STRUCTURE 


Striking to both intergenic spacers and introns 
in the chloroplast genome is the presence and num- 
ber of probable secondary structures referred to as 
“stem-loops.” Stem-loops are believed to occur dur- 
ing single-stranding events when inverted repeats 
meet to form a region of pairing (the stem) sur- 
mounted by their interceding sequence (the loop). 
Such structures have been widely discussed for ri- 
bosomal DNA, with ITS and 185 rDNA regions be- 
ing of particular interest to the plant systematist 
(see Baldwin et al. (1995), Soltis et al. (1997), and 
Soltis & Soltis (1998) for discussion of secondary 
structures in these regions and their phylogenetic 
implications). 

robable stem-loop secondary structure is com- 
monly reported in non-coding regions of organellar 
genomes (e.g., Michel et al., 1989; Buroker et al., 


Volume 87, Number 4 
2000 


Kelchner 
Non-Coding Chloroplast 
DNA Evolution 


485 


1990; Golenberg et al., 1993; van Ham et al., 1994; 
Gielly & Taberlet, 1994; Natali et al., 1995; Rigaa 
et al., 1995; Downie et al., 1996b; Kelchner & 
Wendel, 1996; Kelchner & Clark, 1997; Sang et 
al., 1997; Downie et al., 1998). Gielly and Taberlet 
(1994) reported several probable stem-loops in the 
trnL-trnF region of the chloroplast genome, includ- 
ing nine highly probable structures within the trnL 
intron itself. All other introns in the chloroplast ge- 
nomes of land plants are classified as Group II in- 
trons and share a diagnostic secondary structure of 
six well-defined stem-loop domains (Kohchi et al., 
1988; Michel et al., 1989; Downie et al., 1996b; 
Downie et al., 1998). Diagrams of putative single- 
stranded secondary structure of introns may be 
found in Michel and Dujon (1983), Michel et al. 
(1989), and Downie et al. (1998). 

Loop regions of stem-loop secondary structures 
are often associated with hot spots for mutation in 
non-coding regions, both of nucleotide substitutions 
and indel events (vom Stein & Hatchel, 1988; Al- 
drich et al., 1988; Golenberg et al., 1993; Gielly & 
Taberlet, 1994; van Ham et al., 1994; Clegg et al., 

4; Ferris et al., 1995; Downie et al., 1996b; 
Kelchner & Clark, 1997). Indels located in prob- 
able loop sequence are frequently inserted or de- 
leted repeat units likely the result of SSM. How- 
length mutations attributable to 
slipped-strand mispairing often occur within loop 


ever, not 
sequences as well and may be remnants of recom- 
bination events. 

Although indels are most common in the termi- 
nal loop, they may occur anywhere along a second- 
ary structure. For example, Kelchner and Clark 
(1997) detected what appeared to be an entire de- 
letion of a small sub-loop positioned partway up the 
stem of an rp/16 intron stem-loop in Oryza sativa. 
Such side loops, when present, may be removed in 
some taxa without compromising the favorability of 
a stem formation. Occasionally, small segments of 
the stem itself will be deleted, decreasing the stem 
length, though perhaps not to an extent that would 
annihilate possible secondary structure formation. 

Very large loops are often associated with regions 
of chaotic or “labile” length variation characteristic 
of many non-coding cpDNA sequence matrices 
(e.g., Golenberg et al., 1993; Downie et al., 1996a; 
Soltis et al., 1996; Kelchner & Clark, 1997; Baum 
et al., 1998). Homology assessment here can be 
difficult or impossible, and the conservative ap- 
proach of removing these regions from the data ma- 
trix before phylogenetic analysis is frequently 
adopted. 

In contrast to the loop of stem-loop secondary 
structures being highly susceptible to nucleotide 


substitutions and length mutation, the inverted re- 
peated sequence composing the stem is frequently 
conserved in character (Learn et al., 1992; Gielly 
& Taberlet, 1994; Downie et al., 1996a, 1996b; 
Kelchner & Clark, 1997), particularly when stems 
are long and possess highly favorable energy of for- 
mation values (AG values; see Kelchner & Wendel, 
1996; Dumolin-Lapégue et al., 1998). A sequence 
involved in stem formation is bs available for sub- 
stitution and length mutation because it is paired 
with its sister repeat; this can engender non-ran- 
domly and non-independently evolving sequence 
units. 

Similar to ribosomal RNA and rDNA secondary 
structure (e.g., Curtiss & Vournakis, 1984; Wheeler 
& Honeycutt, 1988; Dixon & Hillis, 1993; Soltis & 
Soltis, 1998), a nucleotide substitution occurring in 
a stem sequence of a non-coding cpDNA region 
could compromise secondary structure formation. 
Compensatory mutation may then occur to preserve 
the potential for structure formation (Kelchner, 
1996; Kelchner & Clark, 1997). Although se- 
quence conservation may be present merely as a 
function of sequence pattern (perhaps the case in 
intergenic spacers), the degree of secondary struc- 
ture conservation in a chloroplast Group II intron 
suggests secondary structures are integral to proper 
functioning of the intron (Clegg et al., 1986; Learn 
et al., 1992; Downie et al., 1996a). Experimental 
evidence has shown some of this structure is es- 
sential for auto-splicing mechanisms in Group I 
and II introns (Bonnard et al., 1984; Kohchi et al., 
1988; Dujon, 1989; Cech, 1990; Michel & Westhof, 
1990; Hibbett, 1996). 

Identification of probable secondary structure 
can be valuable when aligning and analyzing non- 
coding sequences by improving gap positioning and 
the appraisal of character homology. Gaps flanked 
by inverted repeats and regions relatively rich in G 
and C content are suspect as possible stems of sec- 
ondary structures. Аз noted, regions of chaotic 
length mutations are correlated with loops, so the 
boundaries of a chaotic region will frequently cor- 
respond with inverted repeats that can form a stem, 
even if they do not directly neighbor the chaotic 
region. Computer programs such as OLIGO (Ry- 
chlik & Rhoads, 1989), MULFOLD (Jaeger et al., 
1989; Zuker, 1989), and GCG's Stemloop (Genetics 
Computer Group, Madison, Wisconsin) can assist 
in the detection of secondary structure in non-cod- 
ing sequences. А search can be conducted by hand, 
particularly if a published data set exists for the 
region. Free energy of formation values (AG) can 
be calculated with some of the prior software as an 
appraisal of the likelihood of formation of a partic- 


486 


Annals of the 
Missouri Botanical Garden 


ular secondary structure (see Kelchner & Wendel 
) for an example where AG values were ap- 
plied to parallel inversion events in their data). 


MINUTE INVERSIONS 


Minute inversions of four to six base pairs have 
been linked to small stem-loop secondary struc- 
tures commonly referred to as hairpins (Kelchner 
& Wendel, 1996). Hairpins consist of a stem com- 
posed of nearly adjacent inverted repeats producing 
a stem-loop structure with a particularly small loop. 
This loop may become inverted by recombination, 
and the inversion may be so small that it either 
escapes notice during alignment (Kelchner & Wen- 
del, 1996; Kelchner & Clark, 1997), or the inverted 
sequence matches particular bases of the uninvert- 
ed sequence, resulting in a confusing array of mi- 
nute gaps (see Golenberg et al., 

Identifying minute inversions can require careful 
attention when aligning sequence data, particularly 
if alternative gap weighting schemes of an align- 
ment program have not been rigorously explored. 
Candidates for a hidden inversion are several ad- 
jacent nucleotide substitutions, a series of tiny 
gaps, or a gap that demonstrates no repeat aspect 
to its sequence structure. Alternatively, one could 
investigate these probable secondary structures by 
hand or with a secondary structure computer pro- 
gram. Failure to recognize minute inversions in a 
sequence data set has several repercussions for 
phylogenetic analysis, discussed fully in Kelchner 
and Wendel (1996) and summarized here in Anal- 
ysis of Non-Coding Sequence Data. 

Finally, small inversions associated with hairpins 
may be highly susceptible to reversal and parallel- 
ism within a study group, even at the interspecific 
level (Kelchner & Wendel, 1996; Kelchner & 
, 1997; Dumolin-Lapégue 
998). This Maes to reversal or par- 
allelism is due to the persistence of the mutational 
trigger (Kelchner & Clark, 1997)—the nearly ad- 
jacent inverted repeats—after the initial inversion 
event. 


NUCLEOTIDE SUBSTITUTIONS 


Nucleotide substitutions are generally reported 
as being more common in non-coding than in cod- 
ing regions (Wolfe et al., 1987; Zurawski & Clegg, 
1987; Olmstead & Palmer, 1994; Hoot & Douglas, 
1998; however, see Sang et al., 1997, for an excep- 
tion). Surprisingly, a number of studies report nu- 
cleotide substitutions as being just equal to or less 
frequent than length mutations in closely related 
taxonomic groups (Curtis & Clegg, 1984; Wolfe at 


al., 1987; Zurawski & Clegg, 1987; Clegg & Zu- 
rawski, 1992; Golenberg et al., 1993; Gielly & Ta- 
berlet, 1994; however, see Small et al., 1998). 

Percent AT content is quite variable in non-cod- 
ing cpDNA regions, though it is generally higher 
than the average value for the chloroplast genome 
(Shimada & Sugiura, 1991; Downie at al., 1996a; 
Small et al., 1998). Because of their high AT con- 
tent, non-genic regions must make a significant 
contribution to the high overall frequency of A and 
T in the chloroplast genome. Kajita et al. (1998) 
reported an AT content of 67% in the trnL-trnF 
spacer and trnL intron, Kelchner and Clark (1997) 
reported 70.5% AT composition in the intron of 
chloroplast gene тр 6 in bamboos, and Small et al. 
(1998) found an incredible 77.1% AT content in 
the intergenic spacer trnT-trnL in Gossypium. Un- 
doubtedly, this unequal tendency toward AT rich- 
ness in non-genic chloroplast DNA has several as 
yet undetermined implications for phylogenetic 
analysis of non-coding sequence data. At a mini- 
mum, it introduces a strong base composition bias 
into the analysis. 

Substitutions may demonstrate rather high levels 
of homoplasy in non-coding cpDNA regions due to 
the frequency of inferred multiple-hit sites (nucle- 
otide sites experiencing multiple substitution 
events). Multiple-hit sites occur even at very low 
estimates of percent sequence divergence (Kel- 
chner, 1996; Kelchner & Clark, 1997), suggesting 
that the accepted coding region estimates of 
"around 10-15%” sequence divergence for optimal 
phylogenetic signal may be inadequate measures 
for phylogenetic utility of a non-coding region. 

Precise understanding of mechanisms underlying 
multiple-hit substitutions in non-coding DNA is 
lacking. However, attributes of the molecular evo- 
lution of non-coding regions influence the manner 
of nucleotide mutation or the distribution of nucle- 
otide substitution events in an intron or intergenic 
spacer. Stem sequence and loop regions may dif- 
ferentially permit mutations, resulting in non-ran- 
domly distributed and non-independent nucleotide 
substitutions. Statistical significance of differential 
mutation rates in loops relative to stems may be 
tested for an adequate distribution model (see Olm- 
stead et al.’s (1998) test for stochastic mutation in 
the chloroplast genes ndhF and rbcL), yet has гаге- 
ly, if ever, been performed on non-coding cpDNA 
data sets. 

In addition to secondary structure affecting the 
random distribution of nucleotide substitutions, 
there may be constraints on the type of mutation 
an individual site experiences. For example, there 
is a correlation between transition/transversion ra- 


Volume 87, Number 4 
2000 


Kelchner 487 
Non-Coding Chloroplast 


DNA Evolution 


tios and neighboring base composition in non-cod- 
ing regions (Morton, 1995a, b; Morton et al., 1997; 
Savolainen et al., 1997). The correlation suggests 
that nucleotides flanked by A and/or T will dem- 
onstrate a significant tendency toward transversion 
mutations. Such a tendency limits possible nucle- 
otide replacements at these sites, increasing the 
chance of parallelism and reversals, particularly if 
the site experiences multiple hits. One would also 
expect transversion substitutions to be more com- 
mon in data sets of high AT content. 


INTRAMOLECULAR RECOMBINATION 


Intramolecular recombination on an extra-re- 
gional or genomic scale has been suggested be- 
tween adjacent or nearby repeats in the chloroplast 
genome (Howe, 1985; Palmer et al., 1985; Palmer 
et al., 1987; Blasko et al., 1988; Ogihara et al., 
1988; Milligan et al., 1989; Kanno & Hirai, 1992; 
Kanno et al., 1993; Morton & Clegg, 1993; Hoot & 
Palmer, 1994). In the context of non-coding se- 
quence comparison, such a large-scale recombi- 
nation involving the particular region of study could 
result in indels of surprising size that contain se- 
quence content not readily identifiable in origin. 

Recombination events may operate on a finer 
scale within a discrete non-coding region. Occa- 
sionally one infers extensive deleted sequence in 
an alignment with no apparent mechanistic expla- 
nation, presence of a small or moderately sized in- 
version, or a large insertion showing little congru- 
ence with surrounding sequence pattern. Such 
mutations suggest intramolecular recombination, 
and they frequently occur in the loop regions of 
probable secondary structures. Sequences involved 
in stem-loops may be particularly susceptible to re- 
combination events due to the conserved inverted 
repeats and mutationally flexible loop. Therefore, 
such structures could experience interactive recom- 
bination with other stem-loops, particularly with 
those existing in complementary sequence position. 

Recombination involving the entire loop of a sec- 
ondary structure may occur, particularly in struc- 
tures with long stems, resulting in minute or mod- 
erate-sized inversions in both intron and intergenic 
spacer regions (Natali et al., 1995; Kelchner & 
Wendel, 1996; Kelchner & Clark, 1997; Sang et 
al., 1997). Such incidents are often homoplasious 
(Kelchner & Wendel, 1996; Kelchner & Clark, 
1997; Sang et al., 1997; Dumolin-Lapégue et al., 
1998) due to the persistence of the mutational trig- 
ger; in this case, the hairpin stem. 

Intramolecular recombination is a notable alter- 
native to slipped-strand mispairing as a source for 


certain inserted or deleted tandem-repeat length 
mutations (Palmer, 1985; Blasko et al. 1988). 
However, Wolfson et al. (1991), Sang et al. (1997), 
and Kelchner and Clark (1997) suggested SSM is 
a more likely mechanism for length mutation in 
their studies of chloroplast introns and intergenic 
spacers. 


ALIGNMENT 


There are many philosophies for sequence align- 
ment, and much of the literature centers on the 
proper application of computer software for this 
purpose. The structure present in a non-coding 
cpDNA sequence makes it an excellent example for 
discussing what I believe to be the fundamental 
problem of most computer alignment programs: de- 
fining the nucleotide as a discrete and independent 
character. The identification of secondary structure 
and mutational mechanisms in the data may greatly 
improve on current algorithmic alignments of gaps, 
and thus on assessment of character homology. 

Many have found software, particularly versions 
of CLUSTAL (Higgins et al., 1992; Thompson et 
al., 1994), to be of help at least initially with the 
alignment of non-coding sequences. The alignment 
is then subjected to an “improvement by hand” to 
w gaps (e.g., Samuel et al., ; Downie et 

1998; Bayer & Starr, 1998; Kajita et al., 1998). 
This procedure saves time if the sequences are sim- 
ilar in length, but when indels become numerous 
in the data matrix the difficulties of alignment dra- 
matically increase. This is because most alignment 
software initially regards each character in the ma- 
trix as an independent unit, unless otherwise spec- 
ified by particular position or gap weighting 
schemes defined by the user. The software is in- 
capable of determining when mutations other than 
substitutions have arisen, such as non-independent 
insertions, deletions, or inversions correlated with 
SSM and secondary structure. Appropriate weight- 
ing for these mutations that could be incorporated 
into an alignment algorithm is, at present, unde- 
veloped. 

The Elision method of Wheeler et al. (1995) at- 
tempts to improve gap placement and indel homol- 
ogy by alignment software. The Elision method uses 
standard alignment algorithms to produce a series 
of competing alignments based on varying gap 
weighting schemes. These competing alignments 
are then combined in a single matrix and an anal- 
ysis is performed, with the effect that support is 
increased for aligned regions that most frequently 
appear among the various gap-weighting schemes. 
This method aims at objectivity, but makes no im- 


Annals of the 
Missouri Botanical Garden 


provement on the alignment algorithm’s inability to 
assess mutation types other than independent point 
substitutions. Mutations in non-coding regions are 
influenced by surrounding sequence structure and 
frequently occur not as independent base mutations 
but as linked multinucleotide mutation events, like 
the insertion of a repeat unit (Kelchner, 1996; Kel- 
chner & Clark, 1997). The likelihood that many 
non-coding mutations are derived from sequence 
fragments that are inserted, deleted, inverted, or 
otherwise rearranged, negates the assumption of 
discrete, independent nucleotide characters under- 
lying all alignment algorithms, as well as any ex- 
tension of those algorithms like the Elision method. 

At a minimum, those using sequence alignment 
programs to establish putative homology of char- 
acters in their data matrix should experiment with 
a wide variety of gap-weighting options. These op- 
tions, however, may not reveal the underlying mu- 
tational mechanisms occasioning sequence rear- 
rangements in chloroplast non-coding regions. They 
may, however, facilitate the rapid alignment of seg- 
ments of the matrix that share consistent sequence 
integrity and thus pinpoint regions of variable 
length that require special consideration. 

Alternatively, some have avoided alignment pro- 
grams entirely and describe aligning sequences by 
hand (e.g., Golenberg et al., 1993; Hodges & Ar- 
nold, 1994; Kelchner & Clark, 1997). This ap- 
proach facilitates a careful study of the matrix as 
it forms and increases the researcher's familiarity 
with mutations in the sequences. However, align- 
ment by hand, especially when dealing with con- 
siderably divergent taxa or with the presence of a 
great number of nn mutations, can be tedious 
and time consumin 

Kelchner and Clark (1997) suggested that aware- 
ness of the proposed mutational mechanisms active 
in non-coding regions can be useful for inferring 
and positioning gaps and ultimately in assessing 
homology. Golenberg et al. (1993) were the first to 
detail a criterion for aligning gaps in non-coding 
cpDNA matrices. Based on their example, Kel- 
chner (1996) and Kelchner and Clark (1997) mod- 
ified the alignment criterion for chloroplast rp/16 
intron sequences. Hoot and Douglas (1998) also re- 
vised Golenberg et al.'s (1993) method of gap align- 
ment, framing the beginnings of a nomenclatural 
procedure for defining gap categories. Although a 
nomenclatural system is not requisite for gap treat- 
ment in a phylogenetic analysis, it may be useful 
in collating information of inferred mutational 
mechanisms if universally applied in non-coding 

studies. 


ALIGNMENT ISSUES: EXAMPLES FROM NON-CODING 
CpDNA DATA 


Here I present examples (Kelchner & Wendel, 
1996; Kelchner & Clark, 1997; Kelchner, unpub- 
lished data) to illustrate the inference of mutational 
mechanisms in non-coding cpDNA sequences and 
demonstrate the practice of applying mechanistic 
explanations to alignment and homology assess- 
ment. Nucleotides in lower-case bold print are in- 
ferred insertions; underlined nucleotides indicate 
the probable progenitor sequence of an insertion or, 
in Examples 4 and 5, call attention to a particular 
sequence of interest. 

А common type of insertion in non-coding 
cpDNA is a direct repeat of a neighboring sequence 
("Type la” gap; Golenberg et al., 1993; Hoot & 
Douglas, 1998). These often take the form of vari- 
able-length strings of a mononucleotide repeat unit 


(Example 1). 


EXAMPLE 1. 


l. TTAAAAAAAAA---TTGA 


2. ТТАААААААААА--ТТСА 
3. TTAAAAAAAA----TTGA 
4. TTAAAAAAAAAAAATTGA 


Homology can be highly uncertain for these re- 
peated nucleotides. Therefore, such regions are ei- 
ther removed from consideration as potential phy- 
logenetic characters (a conservative approach) or 
included as coded gap characters corresponding to 
length of the repeat string (often becoming highly 
homoplasious in the context of a resulting topology). 
Uncertainty of homology is exacerbated by potential 
inaccuracies of enzymatic processes during PCR 
amplification and sequencing, which can also gen- 
erate variable-length repeat strings independent of 
the template's sequence constitution. When strings 
of adjacent mononucleotide repeats are highly var- 
iable in length in a matrix and reach or exceed the 
range demonstrated above, they become more likely 
to experience further SSM mutation. For this rea- 
son, it 1s perhaps most reasonable to remove such 
areas from consideration in a phylogenetic analysis 

Insertions can also be multinucleotide repeat 
units of a neighboring sequence, as demonstrated 
in Example 2 by the inserted repeat unit ataaa 
("Type 1b” gap; Golenberg et al., 1993; Hoot & 
Douglas, 1998) 


Моште 87, Митбег 4 
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Kelchner 489 
Non-Coding Chloroplast 


DNA Evolution 


EXAMPLE 2. 


An inserted repeat of this nature could be exten- 
sive in length and may be difficult to recognize as 
a repeat unit during alignment (for example, I have 
identified a 73 bp inserted repeat [unpublished 
data] in the trnT-trnL intergenic spacer in Myopor- 
aceae). A repeat unit by its very nature shares nu- 
cleotide content and order with flanking sequence; 
therefore, multiple gaps may be inferred by pairing 
segments of an inserted repeat with its progenitor 
sequence. This is particularly problematic if the in- 
sertion or its progenitor has experienced subse- 
quent nucleotide substitutions. 

ven when a single gap is inferred, positioning 
of the gap may hide evidence that the insertion is 
a repeat unit. Example 3 is reproduced from Kel- 
chner and Clark (1997) and demonstrates how a 
repeat unit may be obscured in a sequence matrix. 


EXAMPLE 3. 


A. 
1. GGTTATGA ----- ATTAACA 
2. GGTTATAA ----- ATTAACA 
3. ССТТАТАА tataa ATTAACA 
4. GGTTATAA tataa ATTAACA 
B. 
l. GGTTAT-- --- GA ATTAACA 
2. GGTTAT-- --- AA ATTAACA 
3. GGTTATAA tataa ATTAACA 
4. GGTTATAA tataa ATTAACA 
С. 
l. GGTTA--- -- TGA ATTAACA 
2. GGTTA--- -- TAA ATTAACA 


3. GGTTATAA tataa ATTAACA 
4. GGTTATAA tataa ATTAACA 


Alignment possibilities A, B, and C were equally 
probable using CLUSTAL W (Thompson et al., 
1994). Only alignment A reveals the insertion is a 


repeat unit—a common mutation type in non-cod- 
ing regions. If alignment options B or C were used 
for phylogenetic analysis, the content of the inser- 
tion would be of unexplainable origin (though still 
possible) and the potential of incorrectly assessing 
nucleotide homology in the region may be consid- 
erable. 

Any of the gap positions in this particular ex- 
ample would not affect a topology generated from 
these four taxa, but gap positioning may have a 
significant effect in a larger matrix of more distantly 
related taxa. The position of the gap in alignment 
3A and detection of the repeat unit may also be 
relevant in determining a weighting scheme for 
these non-independent characters. 

Length mutations may overlap with one another 
to create a progressive-step indel. In the more ex- 
treme cases, appraisal of homology in these regions 
can be very difficult, or impossible (Palmer et al., 

85; Downie et al., 6b; Kelchner & Clark, 
1997). Example 4 demonstrates a probable pro- 
gressive-step indel in which two possible place- 
ments exist for the repeat TTGA. Note that the un- 
derlined sequence is a direct repeat of the 
preceding sequence TCGTAATTGA in the matrix. 


EXAMPLE 4. 


1. AATCGTAATTGA ---------- ---- AACAGA 
2. AATCGTAATTGA ---------- ---- AACAGA 
3. AATCGTAATTGA TCGTAATTGA ----AACAGA 
4. AATCGTAATTGA TCGTAATTGA ----AACAGA 


5. AATCGTAATTGA TCGTAATTGA ttgaAACAGA 


If part of the underlined ТТСА in sequences 3 
and 4 is moved from its current position to align 
with ttga in sequence 5, the possibility that the 
ttga sequence is a direct repeat of the preceding 
sequence may be obscured; however, this alignment 
choice would not be impossible. As the preceding 
sequence to the underlined 10 bp repeat does not 
contain this additional ttga repeat, we can infer 
that two separate events have given rise to an initial 
10 bp insertion in sequences 3 and 4, followed by 
an additional 4 bp insertion in sequence 5. Wheth- 
er ttga itself or the preceding ТТСА is the sub- 
sequent inserted mutation is impossible to deter- 
mine. In this case, either alternative alignment of 
the TTGA unit would cause no effect in a phylo- 
genetic analysis; it is most important here to dis- 
cern the two length mutation events. If any poten- 
tially informative nucleotide substitutions were 
present in either of the repeat units in Example 4, 
these substitutions should be excluded from a phy- 


490 


Annals of the 
Missouri Botanical Garden 


logenetic analysis on the basis that nucleotide ho- 
mology of the repeats is not discernable. 

The example above suggests that homology may 
be indicated by the length of insertions or deletions 
in a gap, although such an assumption is not with- 
out risk. Example 5 below demonstrates multiple 
possible alignments of the gatt repeat unit (repre- 
sented individually by sequences 2, 3, and 4) with 
the insertion in sequence 


EXAMPLE 5. 


1. CAGALIGAIIGALIIALIIAIACIGALIIAIGC 


2. CAGATT gattATGC 
3. CAGATTgatt ATGC 
4. CAGATT gatt ATGC 
9. CAGATT ATGC 


Again, actual homology is impossible to assess 
with confidence, for there exist three GATT repeat 
units in the insertion in sequence 1. In cases like 
this, homology is often inferred on the basis of 
length of indel and minimum number of gaps re- 
quired to position the repeat. Hence, the gatt re- 
peats in sequences 2, 3, and 4 would be aligned 
one above the other and on one side of the gap to 
reduce the number of inferred indel events. When 
coding indels as characters, this would be a rea- 
sonable solution in lieu of other evidence for indel 
origin, and the repeat gatt would be treated as ho- 
mologous for those sequences that contain it. 

Equal length of insertions may not be strong ev- 
idence of their homology (Kelchner, 1996; Kel- 
chner & Clark, 1997; Hoot & Douglas, 1998). Con- 
sider the insertions in Example 6A. 


EXAMPLE 6A. 
. GGTTAAT tctat TCTATCT 


— 


N 


. GGTTAAT ttaat TCTATCT 


> 


GGTTAAT ttaat TCTATCT 


TCTATCT 


~ 
Q 
Q 
= 
= 
> 
= 


ел 


ССТТААТ ТСТАТСТ 


Alignment of the insertions in Example бА ге- 
sults in the probably mistaken homology of indels 
in sequences 2 and 3 with that of sequence 1. The 
insertion in sequence 1 likely arose from ап ш- 
serted repeat of the sequence to the right of the 
gap, TCTAT. This would be a more parsimonious 
explanation, in terms of total number of mutation 
events, than to infer a single inserted repeat fol- 
lowed by two adjacent nucleotide substitutions in 


sequence 1. Sequences 2 and 3 probably share a 
similar origin as a repeat of the preceding sequence 

AT. The events, aligned as they are in Example 
6A, are probably non-homologous. A re-alignment 
could be performed to accommodate the two sepa- 
rate indel events (Example 6B), even though the 
insertions are of the same length and the alignment 
infers an additional gap (see Hoot & Douglas, 

98) 


EXAMPLE 6B. 
1. GGTTAAT ----- tctat TCTATCT 
2. GGTTAAT ttaat ----- TCTATCT 
3. GGTTAAT ttaat ----- TCTATCT 
4. GGTTAAT ----- ----- TOTATCT 
5. ССТТААТ ----- ----- ТСТАТСТ 


There is а hazard that minute inversions (Kel- 
chner & Wendel, 1996) can be completely ob- 
scured in a matrix if they introduce no gaps during 
alignment, particularly if alternative gap-weighting 
schemes have not been rigorously pursued. If pre- 
sent and unrecognized in a data matrix, minute in- 
versions may overweigh a particular mutation by 
interpreting the single mutation event (an inversion) 
as multiple apomorphies of adjacent nucleotide 
substitutions. Example 7 below illustrates a situa- 
tion in which sequences 2 and 3 share the inversion 


TTGG to CCAA (from Kelchner & Wendel, 1996) 
EXAMPLE 7. 

TAATATT TTGG AATATTA 

TAATATT CCAA AATATTA 


TAATATT CCAA AATATTA 


eS cp = 


TAATATT TTGG AATATTA 
5. TAATATT TTGG ААТАТТА 


If the inversion is of sufficient length to introduce 
multiple gaps in the matrix (see Golenberg et al., 
; Sang et al., 1997), two possibilities can oc- 
cur: the gaps will be misaligned to parts of the in- 
verted sequence sharing spurious sequence simi- 
larity with the uninverted sequences; or, there will 
be inference of an inserted sequence of unknown 
origin (in reality, the inverted nucleotides), which 
corresponds with a deletion in the homologous un- 
inverted sequences. Each possibility will lead to 
inaccurate assessment of homology and may poten- 
tially have a considerable effect on phylogeny es- 
timation. 
Regions in the matrix demonstrating many in- 
dependent variable-length insertion and deletion 


Volume 87, Number 4 
2000 


Kelch 
Non- pute Chloroplast 
DNA Evolution 


491 


events will likely be associated with secondary 
structures, specifically with loop regions of stem- 
loops (Kelchner, 1996; Downie et al., 1996b; Kel- 
chner & Clark, 1997). Identification of flanking se- 
quences involved in possible stem formation could 
locate the boundaries for the region and aid in 
aligning the indels. Discerning probable SSM-sus- 
ceptible sites can also be informative for the infer- 
ence of parallel and reversed insertions or dele- 
tions. 

Perhaps methods of gap or character weighting 
and alignment based on mechanisms of mutation 
can be incorporated into software designed for non- 
coding sequence alignment, particularly by includ- 
ing an evaluation of AG values for probable sec- 
ondary structures. However, the diversity of rates 
and types of molecular evolution in non-coding re- 
gions may be profound. As with coding DNA, we 
are far from understanding all forces directing non- 
coding molecular evolution to a degree that we can, 
with any certainty, assign probabilities to individual 
mutations. 

Considering that alignment of sequence data is 
fundamental to the entire phylogeny estimation pro- 
cess, authors should more fully describe the steps 
taken to align their sequence data in order to pro- 


vide necessary information for the assessment of 


their proposed reconstructions of phylogenies. 


ANALYSIS OF NON-CODING SEQUENCE DATA 


The mechanisms of evolution described above 
have a number of significant implications for the 
phylogenetic analysis of non- о sequence data. 
Among these are the follow 

(1) Slipped-strand mispairing can be the result 
of persistent mutational triggers (especially when 
the trigger sequence is located in the stem of a 
stem-loop secondary structure). This can introduce 
homoplasy from parallelisms and reversals into any 
phylogenetic estimations that include gap-coded 
characters in the matrix. Multiple indel events in a 
localized region may obscure homology of length 
mutations. Non-independence of these mutations 
introduces the issue of relative weighting of nucle- 
otide characters linked in a repeat unit, if each 


issue if the unit is included in the analysis as a 
coded gap character 

(2) Secondary structure shows nonrandom mu- 
tation in the form of compensatory mutation and 
possible homogenization of sequence necessary for 
stem formation. Loop sequence is available for mul- 
tiple mutations in the form of inversions, length 


mutations, and multiple-hit point substitutions, any 
of which may obscure evolutionary history. 

(3) Inversions may show high levels of parallel- 
ism and reversal, and their phylogenetic utility may 
not be particularly robust. Undetected minute in- 
versions may be buried within a data matrix and 
consequently treated as multiple base substitution 
synapomorphies instead of a single mutational 


(4) Nucleotide substitutions may be under pe- 
culiar constraints not fully understood. There is ev- 
idence of a bias in non-coding regions involving 
transition/transversion substitution ratios due to the 
influence of neighboring bases. A particular base 
may experience substitution events multiple times 
in closely related lineages, reaching saturation long 
before the expected saturation level for the remain- 
ing sequence. А base-composition bias toward A/T 
content is clearly present in non-coding cpDNA. 

Selective pressures exerted on non-coding re- 
gions may be largely a function of the physical 
structure of the sequence and possible functionality 
of introns and intergenic spacers. Reliance on 
methodology developed for coding sequence, which 
includes estimates of constraints on coding se- 
quence evolution, transition/transversion ratios, and 
mutation probabilities, is inappropriate for the 
analysis of non-coding regions. 

Phylogenetic estimations based on genetic dis- 
tance measures of non-coding cpDNA sequences 
must be approached with care. Superficial appli- 
cation of models for maximum likelihood (ML; Fel- 
senstein, 1981) or neighbor-joining (NJ; Saitou & 

. 1987) could easily produce erroneous phylo- 
ien estimations if several key assumptions un- 
derlying the methodology are violated. 

For example, most models consider a nucleotide 
site as the unit of evolution (Ritland & Eckenwald- 
er, 1992), a consideration that is contradicted by 
the mode of non-coding sequence evolution. Sim- 
plistic models based on the commonly calculated 
Kimura estimates (Kimura, 1980) and Jukes-Cantor 
estimates (Jukes & Cantor, 1969) assume an equal 
25% frequency for each nucleotide type throughout 
the sequence and generate base mutation proba- 
bilities from this assumption. Because non-coding 
cpDNA regions can demonstrate much higher A/T 
content, this assumption is clearly contradicted. 
Furthermore, transition/transversion ratios in non- 
coding regions can differ considerably from coding 
ones (see Hoot & Douglas, 1998), and may even 
vary between discrete non-coding regions of the 
chloroplast genome. Among-site mutation rate het- 
erogeneity is highly probable, especially if regions 
of conservation and hot spots for mutation exist in 


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Annals of the 
Missouri Botanical Garden 


the data. The presence of multiple gaps in an 
aligned matrix presents an additional hurdle for 
distance analysis, and indels themselves are diffi- 
cult to incorporate as additional characters. 

Countering such complications can be involved 
and computationally demanding. Modification of 
the initial Jukes-Cantor estimates to allow for vary- 
ing base frequencies (e.g., Tajima & Nei, 1984 
should be employed. Transition/transversion ratios 
can be estimated directly from the non-coding se- 
quence matrix by pairwise sequence comparisons 
(e.g., Yang & Yoder, 1999), eliminating the circu- 
larity occasioned by measures derived from a to- 
pology. More refined distance models that incor- 
porate these problems stand a better chance of 
reflecting the underlying manner of molecular evo- 
lution in non-coding sequence data. Such refined 
models may therefore estimate a more accurate 
phylogeny that better recovers the evolutionary his- 
tory of the characters. 

With ML, transition/transversion estimates are 
dependent on whether among-site rate variation has 
been incorporated in the model and can be sensi- 
tive to the accuracy of the topology used for their 
estimation (Sullivan et al., 1996). Among-site rate 
heterogeneity in the data is often assumed to fi 
either a negative binomial or gamma distribution 
function, and confirmation can be assessed statis- 
tically. Such rate heterogeneity is likely present in 
non-coding sequence data due to the effects of sec- 
ondary structure on mutation likelihoods. Rates of 
variation at sites are usually expected to fit a gam- 
ma distribution model (Yang, 1996), and a param- 
eter (a) can be determined to define the shape of 
that underlying function in an ML analysis (see 
Yang (1994) and Yang (1996) for thorough expla- 
nation). However, Sullivan et al. (1996) suggested 
a estimates are strongly affected by the topology 
used for their estimation. Therefore, to improve the 
ability of a model incorporating gamma distribution 
to recover the “correct” phylogeny, а must be cal- 
culated directly from the data matrix; this should 


м 


Eb 


be done by pairwise comparison, which can be a 
computationally intensive or even impossible pro- 
cedure as the number of taxa increases in the ma- 
trix (Yang, 1996; Sullivan et al., 1996). Poor esti- 
mation of « can easily result in a misleading 
phylogenetic hypothesis (Yang, 1996; Sullivan et 


Other problems associated with non-coding 
cpDNA sequence data may be very difficult to ad- 
dress. If at least some of the mutation in non-coding 
sequences occurs in linked units, then the non- 
independence of these nucleotide characters di- 
rectly affects the subsequent analysis. At present, 


there is no reliable parameter estimate to incorpo- 
rate such non-independent characters in a distance 
model. Most work on parameter estimates for mod- 
els has been based on coding sequence observa- 
tions, and thus may not reflect the unique aspects 
of molecular evolution in non-coding regions. 

Determining probabilistic estimates for non-cod- 
ing cpDNA mutations is, at this time, difficult; 
Де е: the accurate assessment of the underly- 
ing mode of evolution for maximum likelihood anal- 
ysis may be impossible. As Yang et al. (1995) dis- 
cussed in detail, the accuracy of ML in recovering 
an evolutionary history is strongly dependent on the 
evolutionary model applied. Thus, for non-coding 
cpDNA sequence data (as well as genic sequence 
data), deeper understanding of the manner of evo- 
lution in these regions is required before an accu- 
rate model for ML phylogenetic analysis can be ap- 
plied. 

The frequent alternative to distance measures 
and maximum likelihood is parsimony analysis. 
Heuristic parsimony searches can be considerably 
faster and less computationally intensive than a 
maximum likelihood analysis with the parameter 
adjustments described above; however, they are of- 
ten much slower than a distance analysis. Parsi- 
mony analyses that contain no weighting schemes 
for transition/transversion bias and non-indepen- 
dent mutation of matrix characters may be as vul- 
nerable to recovery of an inaccurate phylogeny as 
similarly simplistic distance models. 
suggested that parsimony’s potential in some cases 
to recover a correct topology decreases significantly 


t has been 


when among-site rate heterogeneity exists in the 
data (Tateno et al., 1994; Kuhner & Felsenstein, 
1994; Huelsenbeck, 1995). Such rate heterogeneity 
could arise from the structured sequence patterns 
described here in non-coding c A. And though 
it has been proposed that the reliability of parsi- 
mony estimates increases with increasing number 
of taxa included in an analysis (e.g., Wakeley, 1993; 
Sullivan et al., 1995; Yang, 1996), it is unclear if 
this effect is independent of possible among-site 
rate variation. 

Parsimony specifies no particular probabilistic 
evolutionary model, but like all phylogenetic esti- 
mation methods it is influenced by non-indepen- 
dence of characters. This problem can be alleviated 
to a degree if mutations such as inversions and in- 
serted or deleted repeats are recognized as поп- 
independent events and are either excluded from 
the analysis or coded separately as described be- 
ow. Апу non-independent evolution of neighboring 
nucleotides in a sequence would create an artificial 
weighting effect for these positions in a parsimony 


Volume 87, Number 4 
2000 


Kelchner 493 
Non-Coding Chloroplast 


DNA Evolution 


analysis that considers each nucleotide an inde- 
pendently evolving character. 

Various weighting schemes have been proposed 
to counter this effect. Weighting has been applied, 
for example, to compensatory mutations associated 
with secondary structure in rDNA (e.g., Wheeler & 
Honeycutt, 1988; Dixon & Hillis, 1993; Baldwin et 
al., 1995; Soltis et al., 1997; Soltis & Soltis, 1998). 
Trial weighting schemes have also been applied to 
non-coding sequence data from the chloroplast 
(e.g., Downie et al., 1996a; Liden et al., 1997). 
However, Olmstead et al. (1998) reasoned that an 
erroneous weighting model increases the chance 
that the correct topology is excluded from the most 
parsimonious topologies recovered. In their opin- 
ion, a more general model such as equal weighting 
of characters may limit resolution, but would in- 
crease the chance that the “true” tree is recovered 
by the analysis. Development of defensible weight- 
ing schemes for non-coding sequence data would 
necessarily come from evidence provided by com- 
parative analysis of non-coding regions throughout 
the chloroplast genome, and may be specific to in- 
dividual data sets. The likelihood of misdiagnosing 
an appropriate weighting scheme for subsets of the 
data may still be high. Therefore, it is perhaps sen- 
sible for now to apply equal weighting to non-cod- 
ing sequence characters until we have further evi- 
dence to support a particular weighting scheme. 

Insertions and deletions have been shown to be 
of considerable phylogenetic value (e.g., Golenberg 
et al., 1993; Mes & Hart, 1994; Natali et al., 1995; 
Downie et al., 1996a; Kelchner & Clark, 1997; Ox- 
elman et al., 1997; Sang et al., 1997; Liden et al., 
1997; Downie et al., 1998; Bayer & Starr, 1998), 
and one should consider including gaps as coded 
(present/absent) characters appended to the se- 
quence matrix (e.g., Hodges & Arnold, 1994; Kel- 
chner & Clark, 1997; Sang et al., 1997; Downie et 
а!., 1998; Hoot & Douglas, 1998; Bayer & Starr, 
1998). Selection of gaps to be included in the anal- 
ysis, however, is somewhat subjective in that opti- 
mally only those length mutations arguably homol- 
ogous based on size, composition, and related 
mechanistic origin should be included. 

e exclusion of gaps and removal of coded gap 
characters from a non-coding sequence matrix can 
e an interesting and informative approach to 
studying the degree of resolution provided by point 
substitution information alone (e.g., Kelchner, 
1996; Kelchner & Clark, 1997). A similar analysis 
can be conducted by including coded gap charac- 
ters only and excluding all other characters in the 
matrix. Coupled with mapping characters onto a to- 
pology produced from a complete matrix, these par- 


titioned analyses may prove useful in locating and 
determining the degree of problematic homoplasy 
affecting resolution in competing topologies. 

Minute inversions should be identified and re- 
moved from the analysis, to be added as present/ 
absent characters at the end of the matrix (Kelchner 
& Wendel, 1996; Kelchner & Clark, 1997). This 
eliminates potential scoring of multiple non-homol- 
ogous synapomorphies that are artifacts of an in- 
version mutation. 

Of some concern is the tendency to treat nucle- 
otide gap characters of taxa that do not share an 
insertion (i.e., have only spaces present at the in- 
sertion position in the matrix) as missing characters 
when conducting parsimony analysis. This results 
in inferred nucleotide homology for characters in 
the inserted sequences, which leads to cladistic as- 
sessment of their base substitutions. Such an ap- 
proach should be applied only when evidence of 
the homology of inserted sequences is convincing. 
Chaotic regions or other areas where homology as- 
sessment is deemed impossible should be excluded 
from the data matrix before analysis (see Liden et 
al., 1997) to avoid this mistaken claim of nucleotide 
homology. 

Bootstrap (Felsenstein, 1985) and jackknife 
(Farris et al., 1997) analyses, frequently misunder- 
stood to be direct measures of phylogenetic accu- 
racy, are only as sound as their underlying analysis 
procedure. As with coding sequences (see Trueman, 
1993; Hillis & Bull, 1993; Bremer, 1994; Mishler, 
1994; Brown, 1994), both support measures can be 
affected by the non-independent structure present 
in non-coding sequences. The structure invalidates 
a requirement of the statistic that each nucleotide 
be a discrete and independent character. 

Bootstrap and jackknife analyses are a re-sam- 
pling of the data matrix in an effort to statistically 
measure how robustly the data in the matrix sup- 
port a particular topology. The concept is sound, 
but the statistical integrity of both measures relies 
on the assumption that each nucleotide is an in- 
dividual character, that each character evolves ran- 
domly and independently, and that the matrix rep- 
resents a sample of a much larger population of 
characters evolving in identical fashion (Felsen- 
stein, 1985). Due to the non-independent structure 
existing in non-coding regions, and the probably 
unique series of evolutionary constraints acting not 
only on individual non-coding regions but also on 
partitions of a region, each of these assumptions 
may be violated. Sampling from within such a data 
set equates to sampling a nonrandom and non-in- 
dependent subset of a non-existing larger popula- 
tion. А large number of bootstrap replicates should, 


494 


Annals of the 
Missouri Botanical Garden 


in theory, cover all possible error due to reduced 
character sampling in each replicate, but the 
strength of the bootstrap test is weakened if the 
characters are not accurately defined. If a character 
in some cases is not an individual nucleotide but 
a suite of nucleotides, the conditions that would 
make bootstrapping and jackknifing accurate as 
measures of data support for a topology are not sat- 
isfied. An analysis would produce an unequal 
weighting effect on subsets of the data in each re- 
sampling due to the frequent localized violation of 
character definition. 

À non-resampling technique that allows assess- 
ment of data support for individual clades is the 
Bremer Support measure (BS, or *decay" analysis; 
Bremer, 1988, 1994; Donoghue et al., 1992; for 
application to large data sets, see Baum et al., 
1994; Morgan, 1997). The measure is a function 
only of the recoverability of clades in topologies 
progressively one step longer. Bremer support has 
the possibility of sidestepping the effects of char- 
acter definition issues discussed above for boot- 
strapping if the model underlying the phylogeny es- 
timation considers the variable nature of character 
definition in a nucleotide set 

Oxelman et al. (1999) demonstrated that boot- 
strapping and BS evaluate different parameters of 
the data matrix, and are thus not directly compa- 
rable measures (though BS values, when high, may 
be imperfectly correlated with bootstrap and jack- 
knife values). BS values cannot be viewed as prob- 
abilistic estimates themselves (Oxelman et al., 
1999), and an inability to adapt the measures to a 
standard scale that is universally applicable ren- 
ders the technique of dubious worth to some sys- 
tematists. However, the innovation by Oxelman et 
al. (1999) that includes minimal branch length val- 
ues with each BS value does, in a non-standard 
way, improve the comparative information capacity 
of the measure. This procedure may be more mean- 
ingful and informative than bootstrap and jackknife 
values for non-coding cpDNA data. 


CONCLUSIONS 


In summary, great care should be given to the 
alignment and assessment of non-coding sequence 
data. There is considerable evidence now that non- 
coding regions are highly structured, non-randomly 
evolving DNA; thus, alignment by current random- 
ized algorithmic software is rarely adequate. An un- 
derstanding of the proposed mechanisms of muta- 
tion acting on non-coding sequences is critical for 
the positioning of gaps and the better assessment 
of homology of indels and point substitutions. Prob- 


able secondary structure should be routinely iden- 
tified and used as an important source of informa- 
tion to aid in aligning chaotic or labile regions of 
the data matrix. Prior to phylogenetic analysis, all 
matrices should be carefully reviewed for obscured 
mutational events, such as minute inversions or 
misaligned repeat units. 

Important for understanding molecular evolution 
in non-coding DNA is the concept of the mutational 
trigger (Kelchner, 1996; Kelchner & Clark, 1997), 
a specific sequence pattern that creates the foun- 
dation for a mutational event. Such triggers often 
remain intact after generating a mutation, and their 
presence can easily occasion a repeated, paralleled, 
or reversed mutation event. Triggers may likely be 
responsible for much of the homoplasy of gap char- 
acters inferred in studies at any taxonomic level; 
those applying non-coding sequence data to molec- 
ular ideo d should be aware of their occur- 
rence and eff 

соң of the kind presented here can in- 
crease the predictive value of mutational events in 
non-coding DNA. For example, Kelchner and Wen- 
del (1996) suggested that minute inversions asso- 
ciated with hairpin secondary structures described 
in non-coding cpDNA could occur in similar situ- 
ations in other genomes. Dumolin-Lapègue et al. 
(1998) recently reported just such an event in the 
mitochondria of oak populations of southern 
France. Hence, recommendations proposed in this 
paper for the phylogenetic analysis of non-coding 
cpDNA sequences may likely apply to data from 
non-coding regions of nuclear, and particularly mi- 
tochondrial, genomes. 

Choosing an appropriate non-coding region for a 
particular taxonomic level is essential for maximiz- 
ing its utility as a phylogenetic tool, but there is no 
infallible method for determining what that “prop- 
er” degree of mutation is for a particular study. A 
region’s utility may vary between plant groups that 
are assumed to occupy the same evolutionary level, 
and data from multiple non-coding regions, when 
applied to one taxonomic group, can vary remark- 
ably in phylogenetic utility (see Small et al., 1998). 
In light of the mutational mechanisms outlined in 
this article, at least one concern seems justified: if 
the taxonomic level is too high, one would expect 
saturation of multiple hit sites and concealment of 
multiple hit indels in any non-coding region, de- 
creasing its utility as a phylogenetic tool. 

The perceived intricacies of molecular evolution 
and their bearing on phylogenetic analysis, both in 
non-coding and coding regions (for genes have 
well-known mechanistic biases as well—the codon 
position being just one example) can be discour- 


Моште 87, Митбег 4 
2000 


Kelchner 495 
Non-Coding Chloroplast 


DNA Evolution 


aging. However, the phenomena outlined in this ar- 
ticle have solutions in most cases, and attention to 
alignment and analysis should enhance the phylo- 
genetic utility and accuracy of non-coding cpDNA 
data. It should be noted that in almost all system- 
atic studies based on non-coding cpDNA sequenc- 
es, the authors profess to have found sufficient phy- 
logenetic information in their data to warrant its use 
in lower-level phylogenetic analyses. 

Clearly there is a need to develop an understand- 
ing of molecular evolution in non-coding cpDNA 
regions similar to that which exists for chloroplast 
genic DNA. Continued research into non-coding se- 
quence evolution may eventually produce a more 
balanced process for the alignment and phyloge- 
netic analysis of non-coding sequence data. Future 
software may be able to measure and assess prob- 
abilities associated with particular mutational 
mechanisms and incorporate this information into 
the alignment process. This would be an immense 
aid to those systematists who wish to apply non- 
coding molecular tools to the field of plant system- 
atics. 


Literature Cited 


s J., B. W. Cherney, E. Merlin & Г. Christopherson. 
. The role of insertion/deletions i in the evolution of 
d trnH in the 


ere Liston. 1998. Chloroplast DNA 

aracters, ке њено classification of Lathyrus 
bFabsoeac) Amer. J. Bot. mage ae 

Baldwin, B. G., M. J. ae eh J. M. Po 
jeiechowski, С. S. Campbell & M. J. Dono: 1995. 
The ITS region of nuclear ribosomal DN valuable 
source of evidence on angiosperm "уар: n Mis- 
souri ы Gard. 82: –277. 

Baum, D. A., К. L. Small & J. F. Wendel. 1998. Bioge- 
ography jt floral evolution of baobabs (Adansonia, 
Bombacac TU as inferred from multiple data sets. Syst. 

бе 


Biol. 47: 


та & Р. С. Hoch. 1994. A phylogenetic 

analysis of paco ра зида и based on nuclear 
ribosomal DNA sequences. Syst. Bot. 19: 363—388. 

Bayer & J. R. Starr. 1998. Tribal phylogeny of the Aster- 
aceae based on two non-coding chloroplast sequences, 
the trnL intron and tr po intergenic spacer. Ann. 
Missouri Bot. Gard. 85: 242-256. 

, C. F. Puttock & S. з Kelchner. 2000. Phylogeny 
of South African Gnaphalieae (Asteraceae) based on two 
non-coding chloroplast sequences. Amer. J. Bot. 
259-272. 

Blasko, K., S. A. Kaplan, K. G. se R. Te B. 
B. Sears. 1988. Variation in copy number of a 24-base 
pair ие repeat in the кемен DNA of Oenothera 
hookeri strain Johansen. Curr. ias 14: 287-292 

. Е. Martin. 1994. 

Non- vacating chloroplast DNA for hd molecular sys- 

tematics at the infrageneric level. Pp. 391—403 in B. 

Schierwater, B. Streit, G. P. Wagner & R. DeSalle (ed- 


нога), Molecular Ecology and Evolution: Approaches 
and Applications. Birkhauser Verlag, Basel. 
— & W. F. 


artin. 1997. Island colonizati 
and pvolutiog of the insular woody habit in Кымын) L. 
канашы, Proc. Natl. Acad. За. U.S.A. ӨЗ: 
11740-11 


Bonnard, G., i Michel, J. H. Weil & A. Steinmetz. 1984. 
Nucleotide sequence of the split tRNA(Leu/UAA) gene 
from Vicia faba chloroplasts: Evidence for structural ho- 
mologies of the ela tRNA(Leu) intron with the 
intron from the autosplicable Tetrahymena ribosomal 

A precursor. Molec. Gen. aie 194: 330-336. 

Bremer, K. 1988. The limits of amino acid sequence data 
in angiosperm phylogenetic Se Evolution 
42: 795-803. 

. 1994. Branch support and tree stability. Cladis- 
tics 10: 295-304. 

Brown, J. К. М. 1994. Bootstrap еи tests for m 
lutionary trees and other dendrograms. Proc. Natl. 
Acad. Sci. U.S.A. 91: 12293-122 

Жк М. E., J. В. Brown, T. A. Gilbert, P. J. O’Hara, 

Bee копља W. K. Thomas & M. J. Smith. 1990. 
ga heteroplasmy of sturgeon mitochondrial DNA: 
An illegitimate elongation model. Genetics 124: 157— 
163. 


Cech, T. R. 1990. кү of Group I introns. Annual 
Rev. Biochem. 59: 543-568. 

Clegg, M. T. & G. Zurawski. 1992. Chloroplast DNA and 
the study of нея phylogeny: Present ee i future 
prospects. Pp. 1-13 in P. S. Soltis, D. E. Soltis & J. J. 
Doyle (editors), Molecular баа ог s Chap- 
man x Hall, New York. 

Ritland & G. Zurawski. 1986. Processes of 

d DNA evolution. Pp. 275-294 in S. Karlin 

& E. Nevo (editors), Evolutionary Processes and Theory. 

Academic Press, we w York. 

. B. S 


Gaut, G. H. Learn, Jr. . Morton. 
1994, Seah ae eet of c xia DN + нетен 
Proc. Natl. Sci. U.S.A. 91: 6795-68( 


Cros, Ј., М. С. Сите Р. Trouslot, x dran S. Hamon, 
A. Charrier & P. Lashermes. 199 С anal- 
ysis of о DNA e а in Coffea L. Molec. 
Phylogenet. -— 117. 

Cummings, ee Ка & Е. A. Kellogg. 1994. 

Slipped- же Жер, А та с сЕ гроС2 in 
grasses (Poaceae). Molec. Biol. Evol. 1-8. 

Curtis, S. E. & M. T. Clegg. 1984. nc a бүле of 

_ DNA sequences. Molec. Biol. Evol. 


ue W. € . N. Vournakis. 1984. Quantification of 
base зба? in eukaryotic 55 rRNA: Selection for 
the е of RNA secondary structure. J. Molec. 


Evol. 
Dixon, i T r D. M. Hillis. 1993. M RNA sec- 
ondary structure: Compensatory mutations and impli- 


mpen 
cations for phylogenetic дите Molec. Biol. Evol. 10: 
256-207. 

Donoghue, M. Ј., . Olmstead, J. F. Smith & J. D. 
Palmer. 1992. Piles и of ya 
based on rbcL sequences. Ann. Missouri Bot. Gard. 7 
333-345. 

Downie, S. R., D. S. Katz-Downie & K.-J. Cho. 1996a. 
Phylogenetic analysis of Apiaceae subfamily Apioideae 
using nucleotide sequences from га и rpoC1 


intron. f Phylogenet. Evol. 6 


Llanas & D. S. Katz- iden ЛЕ Multiple 


Annals of the 
Missouri Botanical Garden 


independent losses of the гроС1 uum in angiosperm 

chloroplast DNA's. Syst. Bot. 21: 135-151. 

и. Ramanath, D. S. Katz- P & E. Llanas. 
1998. Molecular systematics of Apiaceae subfamily 
Apioideae: Phylogenetic analyses of nuclear ribosomal 
DNA internal transcribed spacer and plastid rpoCl in- 
tron sequences. Amer. J. Bot. 85: у 

Dujon, B. 1989. Group I introns as mobile genetic sed 
ments: vi and mechanistic speculations 
Gene 8 а 

нас Tem 


А revie 


, M.-H. Pemonge & R. J. Petit. 1998. 
ла! ion between chloroplast and mitoc hondrial lin- 
Molec. ney Evol. 15: 1321 , 
ie + Albert, М. Kallersjé, D. Lipscomb & 
997. ies jackknifing outperforms 
vinione ed 12: 99-124. 
Felsenstei 1981. Evolutionary trees from DNA se- 
quences: А maximum likelihood approach. J. Molec. 
Evol. 17: 368—376. 
985. Confidence limits on phylogenies: 2 ap- 
proach using the bootstrap. о 39: 783-7 
Ferris, С., В. Р. Oliver, A. J. Davy & С. М. Hewitt 
Using chloroplast DNA to nae Pen ЕСЕ 
routes of oaks into Britain. Molec. Ecol. 4: –738. 
Gielly, L. & P. Taberlet. 1994. The use a DNA 
to resolve plant РА. Yu -COC a versus rbcL 
sequences. Molec. Biol. Evol. 11: 769- 
Golenberg, E. Ма М. | Clegg, М Ya Durbin, J. Doebley 
). P. Ма. 1993. Evolution a a non-coding region of 
the ШОС genome. Molec. Phylogenet. Evol. 2 
52-04 


Ham, R. van, Н. t'Hart, T. H. M. Mes & J. M. 
ЗА ИЕ 1994. Mole cular evolution е non- coding те- 
gions of the chloroplast genome in the 
related species. a ;enet. 25: TETA 

Hancock, J. M. 1995. The contribution of slippage- p 
processes to genome evolution. J. Mole 
1038-1047. 

Hibbett, D. S. 1996. Phylogenetic evidence for horizontal 
transmission of Group I introns in the nuclear ribosomal 
о of mushroom-forming fungi. Molec. Biol. Evol. 13 

903—917 

в D. € 

TAL 


ec. Evol. 


A. J. Bleasby & R. Fuchs. 1992. CLUS 

V: die software for multiple sequence dis 

ment. A ura Applic. 9—19]. 

Hillis, D. M. & J. J. Bul 93. An empirical test of 
bu as a method Е ен к ш 'e in 
di ari analysis. Syst. B 82-19 

Hodges, S. A. & M. L. Arnold. 1994. Combines: A geo- 
een widespread species flock. Proc. Natl. Acad. 
Sci. U.S.A. € 2. 

Hoot, S . Douglas. 1998. Phylogeny of the 
Proteaceae en on atpB and atpB-rbcL intergenic 
spacer region sequences. Austral. Syst. Вог. 11: 301– 
320. 


Biosci. 8 
19 


—5l: 


). Palmer. 1994. Structural rearrangements, 
inc ludo. parallel i inversions, within the chloroplast ge- 
nome of Anemone and related genera. J. Molec. Evol. 
38: 274—281. 
we, C. J. 1985. The endpoints pm an inversion in wheat 
chloroplast DNA are associatec short repeated se- 
е es containing homology to att- b Curr. Gen- 

. 10: 139-145. 
muerta J. P. 1995. The performance of phylogenetic 
methods in simulation. Syst. Bio : 17-48. 


Jaeger, J. A., D. H. Turner & M. Zukef 1989. Improved 


predictions of secondary structures for RNA. Proc. Natl. 
Acad. Sci. U.S.A. 86: 7706-7 
Johnson, L. А. & D. E. Soltis. 1994. так DNA sequenc- 
es and phylogenetic reconstruction in Saxifragaceae 5. 
43-156. 


. M. W. Courtney & J. E. Neigel. 1996. Low 
levels it И genetic variation : a rapidly 
evolving chloroplast DNA locus in Nor 
duckweeds (Lemnaceae). Amer. J. Bot. 83. 430—439. 

Jukes, T. H. & C. R. Cantor. 1969. Evolution of protein 
molecules. Pp. 21-132 in H. N. Munro (editor), Mam- 
malian Protein Metabolism. Academic Press, New York. 


American 


Kajita, T. туа, К. Nakamura, Н. Tachida, R. Wick- 
neswari, Y. Tsumura, Н. Yc aru & T. Yamazaki 
1998. Molecular phylogeny of Dipterocarpaceae in 


rnL 
pls DNA. Molec. Phylogenet. Evol. 10: 202- 
bon A. & A. Hirai. 1992. Comparative studies of the 
structure of chloroplast DNA from four species of Oryza: 
Cloning and physical maps. Theor. Appl. € 83: 
791-798 


;enet. 

————,. N tanabe, 1. Nakamura & A. Hirai. 1993. 
Ма рола in chloroplast DNA from rice (Oryza sativa): 
Differences between deletions mediated by short direct- 
repeat sequences within a single species. Theor. Appl. 
Genet. 86: 579-584. 

Kelchner, S. A. 1996. Molecular evolution pir phyloge- 


netic utility of the chloroplast rp/16 intro Chusquea 
and the Bambusoideae (Poaceae). PE nesis ba 
lowa State University, Ames 


partment of Botany, 
& Clark. 1997. Molecular panne an a 

йалан utility of the chloroplast Lise intron in 

a ai is the Bambuse ideae (Poaceae). Molec 

Phylog Evol. 8 5-397. 

— R) Wendel. 1996. Hairp ins create minute in- 

versions in non- -coding regions of chloroplast DNA. 

262. 


A simple method for estimating evolu- 
tionary rates of base substitutions through comparative 
studies of nucleotide sequences. J. Molec. Evol. 1 
111-120. 

Kohchi, T., Y. Ogura, K. Umesono, Y. Yamada, T. Komano, 
H. Ozeki & K. Ohyama. 1988. Ordered processing and 
splicing in a polycistronic transcript in liv 
roplasts. odd . Genet. 14: 147-154 

Kuhne . Felsenstein. 1994. A simulation of 
пи algorithm under equal and unequal evolu- 
tionary rates. Molec. Biol. Evol. 9-468. 

Learn, С. Н,, Jr.. J. S. Shore, С. "urnier, С. Zurawski 
& M. T. Clegg. 1992. Constraints on the ies of 
ied introns: The group и intron т Шер gene encoding 
tRNA-Val(UAC). Molec. E 56—871. 

к. С. & С. А. Сштап. 1987. Slipped- -strand mis- 

jor mechanism for DNA sequence evolu- 

- Biol. Evol. 4: 203-221. 

Liden, M., T. Fukuhara. J. Rylander & B. Oxelman. 1997. 
Phylogeny and classification of Fumariaceae, with em- 
phasis on Dicentra s.l., based on the plastid gene rps16 

intron. PI. Syst. Evol. 206: 411—420. 

Mes, T. H. M. РНам. 1994. Sedum ictu and 
5. jaccardianum (Coals eae) share a unique 70bp 


erwort chlo- 


=< 
5 
= 
= © 


deletion in the chloroplast DNA LUAM trn F(GAA) 
intergenic spacer. Pl. Syst. Evol. 193: 2 
Michel B. Dujon. 1983. Co inen dnd А sec- 


ondary structures in two intron families including mi- 


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Kelchner 497 
Non-Coding Chloroplast 


DNA Evolution 


tochondrial-, chloroplast-, and nuclear-encoded mem- 
bers. EMBO J. 2: 33-38. 

. Westhof. 1990. Modelling of the three-di- 
mensional architecture of кш Baci introns 
based on comparative seq s. J. Molec. Biol. 


`. Um dnd H. Ozeki. 1989. Comparative E 
MINH anatomy of group II catalytic introns—A r 
view. — 82: 5-30. 

Milligan, B. G., J. N. Hampton & J. D. Palmer. 1989. 
Dispersed hadas and structural reorganization in sub- 
clover chloroplast DNA. Molec. Biol. Evol. 6: 355-368. 

Mishler, B. D. 1994. Cladistic anam. of molecular and 
morphological data. Amer. J. Phys. Anthropol. 94: 143— 
156. 


gan, D. R. 1997. Decay analysis of large sets of phy- 

ры. data. Taxon 46: 509-517. 

Morton, B. R. 1995a. _ Neighboring base а and 

of rice and 

oroplast non- one regions. чн Natl. Acad. 

Sci. U. = 92: 9717-9 

995b. эе bese composition is strongly 

Pei with base substitution bias in a r а n = the 

chloroplast genome. J. Molec. Evol. 41: 597—6 

. Clegg. 1993. A chloroplast e muta- 

tional hotspot and gene conversion in a non-coding re- 

gion near rbcL in the grass family (Poaceae). Curr. Gen- 

et. 24: 357-365. 

. V. M. Oberholzer & M. T. Clegg. 1997. The 
"ДЕА of Mir neighboring bases on ааш еп 
bias in сна к of the plant chloroplast ge- 
nome. J. Molec. Evol. 4 7-231. 

Natali, А., J.-F. ae Кге 1995. Phylogeny 
of the Rubiaceae—Rubioideae, in particular the tribe 
pP Evidence from a non-coding aper DNA 

quence. Ann. Missouri Bot. Gard. 82: 

Ogihara, Y.. T. Terachi & T. Sasakuma. теч Intramolec- 
ular rec а ог chloroplast genome mediated by 
short direct-repeat sequences in wheat sp Proc. 
Natl. Acad. Sci. U.S.A. 85: aped 

Olmstead, R. G. & * D. Palmer. 1994. Chloroplast DNA 

i view of one gud data analysis. 

81: 1205-12 

Reeves & | Yen. 1998. Patterns of 


dequenee pi поп an a Pe 


ecies. 


xad "ái 


Hall. New York. 

. M. Backlund & B. Bremer. 1999. Rela- 
но e the Buddlejaceae s.l. investigated using 
parsimony jackknife and branch support analysis of 
chloroplast ndhF and rbcL sequence data. Syst. Bot. 
24: 164-182. 


. Liden & D. Berglund. 1997. Chloroplast rps16 
intron hea of the vee с (Caryophylla- 
ceae). Pl. Me Evol. 206: 393 
Palmer, J. D. 1985. E nization of chloro- 
plast ximo Annual Rev. Gen 9: 325-354. 
987. CU plat DNA Dac and biosyste- 
бане uses of ве DNA variation. Amer. Natu- 
ralist "m (cup): 56–52 
—. Teng а Structure and еуо- 
lution. pee n L. Bogorad & 1. K. Vasil (editors), 
Cell Culture and Бари Cell Genetics of Plants, Vol. 
7A: The Molecular Biology of Plastids. Academic Press, 
Orlando, Florida 


—— R. A. Jorgensen & W. E Thompson. 1985. Chlo- 
roplast DNA variation and evolution in Pisum: Patterns 
of change and ae ig analysis. б а 109: 
195—213. 


‚ 1. М gent & L. А. Неђоп. 1987. Unusual 
бше of geranium chloroplast DNA: А triple-sized 
inverted repeat, extensive gene duplications, multiple 
inversions, peg epeat families. Proc. Natl. Acad. 
Sci. U.S.A. 84: 169-71 73. 
Peterson, G. & O. Seberg. 1997. Phylogenetic analysis of 
the Triticeae Poata ‘) based on DUM sequence data. 
ае pos Evol. 7: 217-2 
aa, 


mitochondrial DNA: 2 replication origin features. 
J. Molec. Evol. 41: 189-195. 

Ritland, K. & J. E. срна 1992. EON 
hybridization, and apos evolutionary rate in molec- 
ular phylogenies. 404—428 in P. S. Soltis, D. 

& Doyle (editors), Molec ular Svatentiatin 5 of 


computer program 
for choosing optimal oligonucleotides for filter hybrid- 
ization, sequencing and in vitro E ше ation of DNA 
Nucleic Acids Res. 17: 8543-8 

Saitou, N. & M. Nei. 1987. The ie method: 
A new method for reconstructing phylogenetic trees. 
Molec. Biol. Evol. 4: 406—425. 

Samuel, R., W. Pinsker & M. Kiehn. 1997. Phylogeny of 
some species of Cyrtandra (Gesneriaceae) inferred from 
the "e cpDNA intergene region. Bot. Acta. 110 
503-5 

d T., Crawford & T. F. Stuessy. 1997. Chloroplast 

NA г reticulate evolution, and biogeography 
a Paeonia (Paeoniaceae). Amer. J. Вог. 84: 1120-1136. 

Savolainen, V., R. Spichiger & J.-F. Manen. 1997. Poly- 
phyletism of Celastrales deduced from a chloroplast 
noncoding DNA region. Molec. Phylogenet. Evol. 7: 
145-157. 

Schnabel, A. & J. F. Wendel. 1998. Cladistic biogeogra- 
phy of Gleditsia (Leguminosae) based on о. па 
rpl V6 chloroplast gene sequences. Amer. J. 

753-176 "x 

uenis hi, . Doi, H. Koyama & M. Tsuda. 

gs ular I of ЗРЯ ее 
се ie based on har atpB-rbcL intergenic spacer of the 
"Морач DNA. J. Pl. Res. 110: 469—484. 

Shimada, Н. & М. Sugiura. 1991. Fine structural features 
of the nd genome: Comparison of the se- 
quenced chloroplast genomes. Nucleic Acids Res. 19: 

3-995. 

Small, R. L., J. A. Ryburn, R. C. Cronn, T. Seelanan & 
J. F. Wendel. 1998. The tortoise and the = Choosing 

ar ADH 


Doyle (editors), Molec те 

о ~ ся П: DNA Sequencing. Chapman 

and Hall, pe 

^ jj Це п & С. Looney. 1996. Discordance 
between ITS and vinis topologies in = Boykinia 
group (Saxifragaceae). Syst. Bot. 21: 169-18 

—— ———, P. S. Soltis, D. L. Nickrent, L. А. Drm W. J. 


498 Annals of the 
Missouri Botanical Garden 
Hahn, S. B. Hoot, J. A. Sweere, » K. Kuzoff, К. A. — Wakeley, J. 1993. Substitution rate variation among $ 


Kron, M. W. Chase, S. M. Swe „А. Zi immer, S.- 

M. Chaw, L. J. Gillespie, W. " Kre <. J. Sytsma. 

1997. Angiosperm phylogeny inferred from 185 ribo- 

somal DNA sequences. Ann. Missouri Bot. Gard. 84: 
€ 


1-49. 

Stein, J. vom & Ш о 1988. Deletions/insertions, 
short inverted г sequences resembling att-lamb- 
da, and frame- Mic mutated open reading ne are 
involved in chloro A differences in the genus 
Oenothera ien Munzia. Molec. Gen. Genet. 213: 

3—5 

né J. Owen. 1985. Mechanisms of sponta- 
luced frameshift mutation in bacterio- 

T4. ms 109: 633—659. 

. E. Holsinger & C. Simon. 1995. erp 

site rate variation and phylogenetic analysis of 12 

rRNA in Sigmodontine rodents. Molec. Biol. Evol. p 

988-1001. 


5 
Sireisinger, ( 


. 1996. The effect of topology 
on m E oe би гаје variation. J. Mole 
Evol. 42: 308- 

Tajima, ч & M. "Ne ei. 1984. Estimation of evolutionary 
distance enl nuc de sequences. Molec. Biol. 

Evol. 1: 269— 

Tateno, Y., N. т Nei. 1994. Relative ећ- 
ciencies of the maximum likelihood, neighbor joining, 
and maximum parsimony methods о и 
rate varies w ы Biol. Evol. 61-277. 

ве Ј. у. Higgins & T. J. € ro in 1994. 
CLUSTAL t us ће sensitivity of progressive 
multiple sequence alignment through mei weight- 
ing, position-spec "fic gap penalties and weight matrix 
choice. np leic 313—468t 

Trueman, J. 3. Randomization confounded A 
response to i asd Cladistics 9: 101— 


in Jac ag region 1 of human ise 'hondrial DNA. 

. Molec. Evo |. 37: 613-623. 

Wheeler, W.C.& = L. Honeycutt. 1988. Paired sequence 
difference in ribosomal RNAs: Evolutionary and phy- 
logenetic ips: Molec. Biol. Evol. 5: 96. 

. J. €. Gatesy & R. Desalle. 1995. Elision: A 
commodating multiple molecular se- 
акына sites. 


— 


method 

(ends levies wit 

. Phylogenet. Evol. 4 

Wolfe, K. H., W.-H. Li & РМ M 1987. Rates of 
nucleotide substitution vary ке among plant mito- 
chondrial, ран, апа лыс саг DNAs. Proc. Natl. 
Acad. Sci. U.S.A. 84: 9058. 

Wolfson, R., K. G. Higgins & B. B. Sears. 1991. Evidence 
for replication slippage in the evolution A Oenothera 
Mp den st DNA. Molec. Biol. Evol. 8: 709—720. 

Yang, 7 904., Maium likelihood ia esti- 
mation from DNA sequences with variable Pos over 

s: Approximate nds J. Molec. Evol. 39: 306— 


1996. Among-site rate variation and its impact 
on ühylogimede s Tree 11(9): 367-372. 

— а г. 1999. Estimation of the | 
transversion rate bias and species sampling. J. Mol 


Evol. 4 


Goldman & A. Friday. 1995. Maximum like- 
lihood trees from DNA sequences: A Зе iar statistical 
estimation problem. Syst. Biol. 44: : 9. 

Zuker, М. 1989. On finding all вена foldings of an 
RNA molecule. Science 48-52. 

Zurawski, G. . T. Clegg. 1987. Evolution of higher- 
plant ies DNA-encoded genes: Implications for 
pU e function and phylogenetic studies. Annual 

‚. Pl. Physiol. 38: 398—418. 


ALLPAHUAYO: FLORISTICS, Rodolfo Vásquez Martínez? and 
STRUCTURE, AND Oliver L. Phillips? 
DYNAMICS OF A 

HIGH-DIVERSITY FOREST 

IN AMAZONIAN PERU! 


ABSTRACT 


This paper describes the results of a floristic inventory at the Allpahuayo Reserve, near Iquitos in Amazonian Peru. 
Two long-term one-hectare plots were established using a pre-determined sampling grid, with each individual tree and 
liana over 10 cm diameter collected at least once, except for palms. The plots were re-censused after 5 years to quantify 
forest ум Floristic analysis shows that the Allpahuayo ruunt: is among the most diverse site yet inventoried, with 
281 t п species per hec "are, and at least 466 species and 61 families in the 1277-stem two-hectare "ais. 


family in Ds plots is Fabaceae sensu lato, with 231 stems and 89 species; no other family re epresents more than 7% 

of the species or 10% of the stems. In contrast to the exceptional floristic dinde both the structure and = dynamics 
of the Allpahuayo forest are similar to those recorded from other old-growth neotropical forests. Many tree and liana 
canopy species were previously unknown to both the Iquitos area and to Amazonian Peru, which г тибин ће 
significance of Amazon ecological studies to systematic botany. 


RESUMEN 


ste documento describe el estado actual de los resultados de un inventario florístico a largo plazo, en la Reserva 
T aja de Allpahuayo, cerca de Iquitos, en la Amazonía Peruana. Allí se establecieron dos parcelas de una hect. 
cada una, usando un muestreo pre-determinado, cada árbol, palmera o liana, mayores o iguales que as cm de diám. 
fué marcado y colectado, excepto las palmeras que fueron colectadas solo una vez por especie. Las parcelas fueron 
re-censadas despues de 5 años para cuantificar el proceso de la dinámic `а del bosque. El análisis de Tu resultados 


ee 
En contraste a la excepcional diversidad florística, tanto la: estructura del bosque como la tasa de mortalidad en 
Allpahuayo, están dentro los е de otros sitios neotropicales. El valor de los estudios ecológicos para la botánica 


sistemática, se demuestra en ta о que, antes de que empezáramos a tomar datos ecológicos e instalar parcelas 
permanentes, muchas especies ph árboles y lianas eran desconocidas tanto en el área de Iquitos, como en la 
eruana. 


Key words: Amazonia, floristics, inventory, Neotropics, species richness, species turnover. 


! We gratefully acknowledge the help of institutional support from Instituto de Investigaciones de la Amazonía Peruana 
(ПАР) for permission to work in Allpahuayo Reserve and for logistical support for our work; the John D. and Catherine 
T. MacArthur Foundation for long-term support of the Missouri Botanical Garden's floristic and ecological research at 
Allpahuayo and elsewhere in Peruvian Amazonia; the National Geographic Society for support for forest dynamics 
research in Amazonian Peru (grant #5472-95), and the U.K. Natural Environment Research Council for a Research 
Fellowship (OP). Melchor Agua aes Grandez, Rosa Ortiz de Gentry, Nestor Jaramillo, Dennis Milanowski, John J. 
Pipoly III, Peggy Stern, Henk van der Werff, and Arturo Vasquez Martinez assisted in establishing, inventorying, and 
re-censusing the Allpahuayo ке Herbarium determinations were provided by numerous systematists at MO and other 
herbaria, and we are especially grateful to: C. C. Berg (BG); T. Pennington, Sir G. Prance (K); A. C Tyr tiir т. 
Gereau, R. Liesner, J. J. Pipoly HI, G. Schatz, C. Taylor, M. Timaná, H. van der Werff, and the late A. H. Gen dm 
D. Daly, S. Mori, and M. Sen (NY). The maps were drawn by A. Mansoni at the School of Geography rei Unit, 
University of Leeds. We thank V. Hollowell, Sir G. Prance, and H. van der Werff for their constructive suggestions to 
inipro ve our manuscript. 

? Missouri Botanical Garden, Apartado 020, Jaén, Cajamarca, Peru 
* Centre for Biodiversity and Conservation, School of Geography, University of Leeds, Leeds LS2 9JT, U.K. 


ANN. Missouni Вот. GARD. 87: 499—527. 2000. 


Annals of the 
Missouri Botanical Garden 


1 
73°15'W 


~ ито 


| А 
уде 
ћ 
Ù ) 
2 
A 
% » 
> y 
73°24'36"W 
10Km 
Figure 1. Map showing location of study region in relation to Iquitos and Peru. —A. Peru. —B. Iquitos region, 
showing location of Allpahuayo. Modified from Kalliola et al. (1998). 


Biodiversity is at risk in much of the tropics. Yet 
biological research and conservation initiatives are 
hampered by inadequate baseline information. 
Thus, sufficient data on the numbers, kinds, and 
abundance of most major plant and animal taxa are 
not available. Moreover, knowledge of species as- 
sociations and biogeographic distributions of taxa 
remain meager (McNeely et al., 1990; National Sci- 
ence Board, 1990; National Research Council, 
1992; Phillips & Raven, 1996). 

In the Neotropics, the extraordinary species rich- 
ness, our limited taxonomic understanding, and the 
physical inaccessibility of most areas are formida- 
ble barriers to phytogeographic investigation. Tra- 
ditional biological inventory efforts that rely mostly 
on ad hoc collection efforts and lengthy monograph 
studies cannot address the knowledge gap alone. 
Alternative methods are critical to improve our un- 
derstanding of the factors determining species com- 
position and the ecological dynamics of tropical for- 
est ecosystems. In recent years there has been 
growing scientific interest in more ecological, plot- 
based work as a means of understanding tropical 
forests (e.g., Gentry, 1988a, b; Phillips & Gentry, 
1994; Phillips et al., 1994; Dallmeier & Comiskey, 
1998a, b). This approach can contribute greatly to 


floristic understanding by generating large numbers 
of new collections that are associated with site-spe- 
cific ecological information. Botanical institutions 
play a vital role in initiating and supporting this 
work, and contribute essential expertise for ensur- 
ing accurate voucher determination. Equally, eco- 
logical inventories generate much biogeographical 
data of relevance to monographic and phylogenetic 
studies. Such eco-floristic research has recently 
been initiated in several tropical countries, includ- 
ing Madagascar (Lowry et al., 1997; Rakotomalaza 
& Messmer, 1999), Colombia (Rudas, 1996), and 
Peru (Gentry, 1988a, b). This paper reports the re- 
sults of a similar research program at the Allpa- 
huayo Reserve, near Iquitos in northern Peru. 


STUDY SITE 


The Allpahuayo Reserve (3°57'S, 73*26' W) lies 
southwest of Iquitos in Amazonian Peru, between 
the blackwater Rio Nanay on the northwest and the 
Iquitos-Nauta road to the southeast (Fig. 1). This 
2750-ha reserve is administered by the Peruvian 
Institute for Amazonian Research (IIAP). The cli- 
mate is humid and hot (with the mean annual pre- 
cipitation about 3000 mm and an average temper- 


501 


Vasquez Martinez & Phillips 


Volume 87, Number 4 


2000 


High-Diversity Peruvian Forest 


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Annals of the 
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ature of 26°C; Marengo, 1998). Allpahuayo Reserve 
lies between 110 m and 180 m above sea level. 
Edaphic conditions are variable, representing a mo- 
saic of patches ranging in texture from clayey to 
almost pure sand, and in drainage from water- 
logged swamps to well-drained hill tops. Vegetation 
is mature, old-growth forest, although some palm 
trees (Оепосатрих bataua Mart.) have been cut to 
harvest their fruits. Allpahuayo and neighboring 
Mishana comprise one of three locations within 100 
km of Iquitos where the Missouri Botanical Garden 
has been conducting a long-term floristic inventory 
effort with the institutional collaboration of IIAP 
and Explorama Tours S.A. (Vásquez, 1997). The 
other two sites, Reserva Ecológica Explorama 
Lodge (Yanamono) and Reserva Ecológica Sucusari 
(ACEER), have rather different soil conditions and 
floristic composition. The 3750 ha of Allpahuayo- 

ishana is extremely species-rich, with more than 
10% of the plant species reported from the whole 
of Peru (Brako & Zarucchi, 1993) recorded here 
(Vásquez, 1997). 


METHODS 


Sampling took place within a 500 X 500 m grid 
in the Allpahuayo Reserve, sufficiently far from the 
Iquitos-Nauta road (> 2 km at its nearest point) to 
minimize human disturbance, yet close enough to 
the research station (3 km) to allow us to commute 
daily. Within the sampling grid, two long-term plots 
were established in a pre-determined location with- 
in the grid (Fig. 2) to eliminate subjective bias. 
Each plot is 20 X 500 m, with each plant = 10 
cm DBH (diameter at breast height, 1.3 m) marked 
with sequentially numbered aluminum tags. Addi- 
tional narrower samples of 2 X 500 m with all 
plants = 2.5 cm DBH collected but not tagged were 
made at pre-determined points in the grid: the re- 
sults of these inventories will be reported in a fu- 
ture publication. The linear nature of all our plots 
means that they each traverse a mixture of edaphic 
formations. Soil conditions within the grid ranged 
from poorly to adequately drained clay soils (“Ѕһа- 
paja") to well-drained white sand soils at topo- 
graphic high points (*Varillal"). 

In the long-term plots, every tagged plant's 
height was visually estimated and its diameter mea- 
sured at 1.3 m with diameter tape. Herbarium col- 
lections (Vásquez 14592-15829) were made in 
1990 and 1991 from every individual plant (except 
for palm tree species, which were only collected 
once each) using extendable aluminum collecting 
poles and, where necessary, spiked climbing irons 
for climbing trees. A full set of duplicates is de- 


posited in two Peruvian herbaria (AMAZ, USM) 
and at MO, with partial collections existing at IIAP 
and the Universidad Agraria La Molina (MOL), as 
well as distributions to family specialists world- 
wide. 

Fertile collections were made on occasion when- 
ever flowers or fruits of plot species were seen, but 
in common with other tropical plot inventories most 
species (9096) are represented only by sterile ma- 
terial. At every collection number was first 
identified to family by the first author and Ron 
Liesner, with each family set then distributed to 
family specialists (Anacardiaceae, Apocynaceae, 
Bignoniaceae, Burseraceae, Chrysobalanaceae, Dil- 
leniaceae, Fabaceae, Lauraceae, Lecythidaceae, 
Malpighiaceae, Melastomataceae, Meliaceae, Mor- 
aceae, Myrtaceae, Rubiaceae, Sabiaceae, Sapinda- 
ceae, Sapotaceae, Simaroubaceae, Violaceae). For 
most species at least three fertile Peruvian collec- 
tions were noted for verification (MO), most deriv- 
ing from the Iquitos Florula project (Vásquez, 
1997 


Re-census of both plots was done in February 
1996, with the re-measurement of all trees and li- 
anas. Plants that had died in the intervening period 
of 5.25 years were noted as such, with evident 
cause of death. Each new recruit into the 10 cm 
DBH class was measured, tagged, and given a 
unique number. New recruits were collected and 
vouchers distributed to herbaria as for the original 
collections. 

In order to maximize potential comparability 
with other forests, diversity values for each plot 
were expressed in terms of both species richness 
and Fishers Alpha, using various subsets of our 
data. We estimated "species richness"—the sum of 
the number of tree species—as this measure is eas- 
ily understood and widely reported from other for- 
ests. We also estimated Fishers Alpha, where: 


Alpha * In(1 + stems/Alpha) 


species richness — 


Fishers Alpha values from small tropical forest 
tree samples provide good estimates of the overall 
diversity of each forest (Condit et al., 1998). Both 
species richness and Fishers Alpha values were 
calculated on a per-area basis (i.e., for each one- 
hectare plot), and on an area-independent basis 
(i.e., for the first 500 stems encountered in each 
plot) in order to remove the complicating effect of 
varying stem density. Finally, each diversity index 
was also estimated separately for all woody stems, 
and for trees alone. 

In order to compare stem densities of individual 
species and families, average per-hectare values 
were calculated. The most speciose families across 


Volume 87, Number 4 
2000 


Vasquez Martinez & Phillips 
High-Diversity Peruvian Forest 


Table 1. 


Allpahuayo forest diversity in 1990, for stems = 10 cm diameter in 1-ha plots 


. The values given for 


species richness and Fisher Alpha are for the minimum and the most likely (underlined) estimates, respectively. The 
diversity values are calculated for (A) all woody stems (i.e., trees and lianas), and (B) for trees only. See text for further 


etails. 


Species per Fisher's 
Fisher's first 500 Alpha, first 
Stems Species Alpha stems 00 stems 
(A) All woody stems 
Plot 1 643 281/293 190/208 233/242 170/185 
lot 2 634 306/311 233/242 250/255 199/208 
Plots 1 and 2 combined 1277 466/480 264/280 N/A N/A 
(B) Trees only 
Plot 1 616 204/275 175/191 228/237 162/176 
Plot 2 608 290/295 218/226 243/248 186/195 
Plots 1 and 2 combined 1224 433/444 239/250 N/A N/A 


our whole sample were tabulated on the basis of 
the total species recorded in both hectare plots. The 
degree of departure from randomness in species’ 
distribution with respect to soil type was explored 
using a binomial test based on the relative fre- 
quency with which individuals were recorded in 
clay soil and sandy soil forests. 

Standard measures of forest structure (density of 
stems and total basal area of stems = 10 cm DBH 
were computed for each hectare plot. Annual mor- 
tality and recruitment rates were estimated using 


~ 


standard procedures that use logarithmic models 
that assume a constant probability of mortality and 
recruitment through each inventory period (Phillips 
et al., 1994; Swaine & Lieberman, 1987), and com- 
puted separately both for woody stems and for 
woody basal area. Turnover rates for each period 
were represented by the mean of recruitment and 


mortality rates (Phillips, 1996). 


RESULTS AND DISCUSSION 
(A) TAXON IDENTIFICATION 


Of the 1277 plants in the plots in 1991, 100% 
were identified to family, 1249 (97.8%) to genus, 
and 1168 (91.5%) to species (Appendix 1). We 
made 1160 collections from the two plots, of which 
1053 (90%) were sterile, 22 (2%) were in bud, 33 
(3%) were in flower, and 52 (5%) were in fruit. The 
high proportion of sterile material complicated the 
identification process. Near-complete identification 
was possible only because of the large number of 
fertile collections from Allpahuayo and nearby lo- 
calities available for comparison at MO. For most 
of the unidentified species, it was possible to allo- 
cate collections to morphospecies (i.e., to allocate 
collections to a morphological species concept, al- 


beit unnamed), but for a few even this was not pos- 
sible. 


(B) SPECIES RICHNESS AND DIVERSITY 


Because of the difficulties with identification 
there is some uncertainty about the exact number 
of species in our samples. We estimated a “most 
likely value” for species richness (5) by multiplying 
the number of unidentified plants per sample that 
could not be allocated to morphospecies (U) by the 
ratio of species:individuals for the plants in that 
sample that were identified to species or morphos- 
pecies (R), and adding this value to the number of 
morphospecies and species concepts actually dis- 


tinguished (D). 
Thus, (S = (U * R) + D. 


This estimate probably errs on the conservative 
side because the most difficult plants to identify 
tend to be the rarest, and therefore it is likely that 
the ratio of species to individuals will be greater 
for non-identified plants than it is for identified 
plants. 

Diversity values for each plot and for both plots 
combined are presented in Table 1, with results 
given separately for all stems and for trees alone. 
These reveal that the Allpahuayo plots are some of 
the richest 1-ћа samples ever reported. The best 
estimate of Fisher’s Alpha for trees in our plot 2 
(226) is the greatest value ever recorded for trees 
= 10 cm DBH in 1-Ва plots, comparing with 221 
at Yanamono in Peru (Condit et al., 1998), 211 at 
Cuyabeno in Ecuador (Condit et al., 1998), and 191 
in Allpahuayo plot 1 (this study), all of which are 
upper Amazonian sites. In terms of published re- 
sults of numbers of species per 500 stems, the All- 


504 


Annals of the 
Missouri Botanical Garden 


pahuayo plots are surpassed only by Yanamono 
(Phillips et al., 1994). There are also some pub- 
lished inventory results of, variously, lianas, hemi- 
epiphytes, trees, and saplings from this part of 
Amazonia (e.g., Gentry, 1988a, b, 1992; Valencia 
et al., 1994; Clinebell et al., 1995; Duivenvoorden 
& Lipps, 1995). These confirm that the region’s un- 
flooded forests generally have globally exceptional 
levels of diversity for woody plants. 
Within-community (*alpha-") diversity in neo- 
tropical forests is closely correlated with climatic 
conditions, with the richest forests found in the hot 
and aseasonal equatorial lowlands (Clinebell et al., 
1995). High levels of local diversity may also be a 
consequence of close juxtaposition of с to- 
pographical and soil formations in western Ama- 
zonia (Tuomisto et al., 1995) with pe effect" 
processes causing species to spill over from adja- 
cent edaphically defined communities. Each All- 
pahuayo plot traverses contrasting substrates, of 
sandy and clayey soils, which contributes to the 
high diversity values, as some species are appar- 
ently specialized on each particular soil type (cf. 
(c) below). However, the Allpahuayo plots were 
placed without regard to forest type, so these di- 
versity values are presumably typical for most 1-ha 
patches within the Allpahuayo forest. Data are also 
available from one 0.1-ћа sample of plants = 2.5 
cm DBH, purposively laid out at Allpahuayo by A. 
Gentry and the first author so as to only sample 
clay soil forest (Clinebell et al. 1995; http.// 


www.modbot [ research/ 


gentry.html). Here 275 species of (сени. lianas, and 
hemiepiphytes were found in 401 individuals, giv- 
ing a Fisher’s Alpha value of 386, the highest spe- 
cies richness and Fisher’s Alpha value of all Gen- 
try’s 227 0.1-ћа samples worldwide (Phillips & 
Miller, in prep.). 


(C) FLORISTIC COMPOSITION 


The Allpahuayo forest is not only very species- 
rich, but also has a very low degree of dominance 
by any one species. The ten most common species 
together represent less than 20% of all stems (Table 
2A). Most of the families that physically dominate 
the forest (Table 2B), such as Fabaceae, Myristi- 
caceae, Euphorbiaceae, Sapotaceae, and Moraceae, 
do so by virtue of having many species almost all 
of which are present at a very low density. The palm 
family is atypical in that its status as the second- 
most stem-dense family is due mostly to the high 
density of one species (Oenocarpus bataua Mart.). 

most speciose families in our entire Allpa- 
huayo sample (Table 2C) are the same families that 


dominate other Amazonian forests (Gentry, 1988a). 
Fabaceae are by far the most species-rich, and even 
when treated individually the three legume subfam- 
ilies each have about as many species as the other 
top-five largest families (Moraceae, Lauraceae, An- 
nonaceae, Burseraceae). 

The plots each traverse two contrasting soil 
types, richer clay soils (the “Shapaja” series) and 
poor white sand soils (the “Varillal” series), and the 
strong local edaphic contrasts might be expected to 
result in some floristic differentiation within the 
plots. Because of the high diversity, few species 
were sampled frequently enough in the two plots to 
allow us to test the extent to which they are habitat 
specialists or generalists. However, among the 56 
species that were relatively common, with five or 
more individual plants in the two hectares, a bi- 
nomial test reveals that a greater number are con- 
fined to one soil type or another than would be 
expected under a null model of random distribution 
(Table 3). This analysis reveals a large number of 
species that are apparent specialists on the poor 
white sand soils (for example, Macrolobium micro- 
calyx, Ocotea aciphylla, Tachigali ptycophysca, Ta- 
ble 3A), and a smaller group mostly confined to the 
relatively rich clay soils (for example, Leonia gly- 
cycarpa, Lindackeria paludosa, Senefeldera skutch- 
iana, Table 3B). Note that these results are not 
proof of specialization, since apparent specialists 
could arguably be constrained to one patch of forest 
by chance alone, especially if they have poor dis- 
persal mechanisms. However, a parallel study at 
Allpahuayo (Vormisto et al., 2000) demonstrated 
that the local-scale distributional pattern of soil 
types is closely related to plant distributional pat- 
terns, not only for tree species but for other plant 
groups as well, and these spatial patterns are con- 
gruent among different groups. This research con- 
firms that the local distributional patterns of many 
plants within the Allpahuayo plots are influenced 
by edaphic conditions. The high diversity of the 
Allpahuayo plots is therefore not only due to high 
intra-community diversity, but also derives from the 
contrasting edaphic conditions evident within each 
plot. 


(0) STRUCTURE AND DYNAMICS 


Although the Allpahuayo plots are exceptionally 
species-rich, they do not appear to be particularly 
remarkable in terms of their physical structure. 
This is indicated by the fact that both their stem 
density and tree basal area appear to be well within 
the typical ranges for most lowland Amazonian for- 


ests (Table 4). However, we could find no published 


Volume 87, Number 4 Vasquez Martinez & Phillips 
2000 High-Diversity Peruvian Forest 


505 


Table 2. Dominant woody plant taxa at Allpahuayo. 


A. The 10 species with the highest density of stems = 10 cm diameter per hectare, on average and by soil type. 


The two plots included 1.16 ha of clay soil forest, and 0.84 ha of sandy soil forest 


Stem density, 


clay soi sandy soil 
Family Genus Stem density forest 
Palmae Oenocarpus bataua 39.0 26.3 55.2 
subsp. bataua 

Myristicaceae E на ulei 14.5 8.8 20.8 
Myristicaceae Virola pavonis 11.5 11.3 14.6 
Euphorbiaceae Hevea nm 10.0 1.3 18.8 
Fabaceae Macrolobium microcalyx 9.5 0 19.8 
Аппопасеае Diclinanona tessmannii 7.5 0 15.6 
Euphorbiaceae Micranda elat 7.9 0 15.6 
Euphorbiaceae Senefeldera P 7.5 17.5 1.0 
Apocynaceae Aspidosperma excelsum 7.0 1.3 12.5 
Palmae Astrocaryum macrocalyx 7.0 16.3 1.0 
Тор 10 species 121.0 82.8 175.9 
All species 638.5 637 640 
B. The 10 families with the highest density of stems = 10 cm diameter per hectare. 
Family Stem density 
Fabaceae: 115.5 

Caesalpiniaceae 53.0 

Mimosaceae 23.5 

Papilionaceae 36.5 

Indet. Fabaceae 2.5 
Palmae 61.5 
Myristicaceae 59.0 
Euphorbiaceae 56.5 
Sapotaceae 33.0 
Moraceae 30.5 
Аппопасеае 27.0 
Lauraceae 26.5 
Burseraceae 21.5 
Lecythidaceae 16.0 
Top 10 families 420.0 
АП families 638.5 
C. The 10 families with the кез number of species = 10 cm diameter sampled throughout our 2-ha sampled area. 

The values given are for the minimum and most likely handled inet number of species per family. 
Total species 

Family inventoried 
Fabaceae: 82/89 

Caesalpiniaceae 27/29 

Mimosaceae 25/28 

Papilionaceae 28/29 

Indet. Fabaceae 3 
Могасег 31/31 
Lauraceae 29/29 
Annonaceae 27/27 
Burseraceae 23/24 
Euphorbiaceae 23/23 
Sapotaceae 22/23 
Myristicaceae 21/21 
Lecythidaceae 14/14 
Myrtaceae 10/12 


Top 10 families 262/273 
All families 433/444 


506 Annals of the 
Missouri Botanical Garden 
Table 3. Edaphic in 'ialists in the Allpahuayo plots. For all species with five or more individuals, a binomial test 


hypothesis of random distribution with respect to soil type. The и was rejected 
* (P < 0.01); *** (P < 0.001)). See text for further deta 


was conducted to test the 
for half the 56 species teste ted (* (Р < 0.05); * 


A. Specialists on white sand (“Varillal” soils). 


Probability 
Total Shapaja Varillal of random 
Family Taxon stems stems distribution 
Annonaceae Diclinanona tessmannii 15 0 15 one 
Apocynaceae Aspidosperma excelsum 14 1 12 ** 
Bignoniaceae Jacaranda macrocarpa 5 0 5 * 
Euphorbiaceae еа guianensis 20 1 18 ioa 
Euphorbiaceae Micranda elata 15 0 15 v» 
Fabaceae (Caes.) Macrolobium bifoli 8 0 8 ** 
Fabaceae (Caes.) Macrolobium microcalyx 19 0 19 же» 
Fabaceae (Caes.) Sclerolobium bracteosum 5 0 5 * 
Fabaceae (Caes.) Tachigali ptychophysca 10 0 10 жеж 
Fabaceae (Caes.) Tachigali tesmannii cf. 13 0 13 mee 
Fabaceae (Pap.) Swartzia racemosa var. Мири 11 1 10 я» 
Icacinaceae Emmotum floribundum 5 0 5 * 
Lauraceae Ocotea aciphylla 8 0 8 жж 
Malpighiaceae Byrsonima stipulina 8 0 8 ыы 
Myristicaceae Iryanthera ulei 29 7 20 * 
Nyctaginaceae ea floribunda 8 1 7 * 
Palmae Oenocarpus bataua subsp. bataua 78 21 53 “чы 
Ңозасеае Prunus detrita vel sp. aff. 5 0 5 " 
Rubiaceae Ferdinandusa chlorantha 5 0 5 " 
Sapotaceae Chrysophyllum bombycinum 9 0 9 ** 
В. Specialists on clay (“Shapaja” soils). 
Euphorbiaceae Conceveiba rhytidocarpa 7 6 1 * 
Euphorbiaceae Nealchornea yapurensis 9 8 1 Бы 
Euphorbiaceae Senefeldera skutchiana 15 14 1 v 
Flacourtiaceae Lindackeria paludosa 6 5 0 * 
Monimiaceae Siparuna decipiens 5 5 0 * 
Imae Astrocaryum macrocalyx 14 13 | ane 
Palmae lriartea deltoidea 13 12 1 ке. 
Violaceae Leonia glycycarpa var. glycycarpa 8 6 0 * 
Table 4. Stem density and basal area of some lowland Amazonian forests, for stems =10 ст diameter es ha. Data 


hana, Tambopata, Yanamono: Gentry, 1988b; 
1994; Cusco Amazónico: Phillips Fe va un- 


are from this study (Allpahuayo, 1990 census) or other sources (Mis 
B 


fiangu, Jatun Sacha, Belém, San Carlos de Rio Negro: Phillips et al., 


published data). Stem density values are per ha. Basal area values аге m? per ha. 


Liana and Liana and 
Tree stem strangler Tree basal strangler 
Site density stem density area basal area 
Belém, Brazil 572 27.7 
Јашп Sacha, Ecuador 724 30.5 
Afiangu, Ecuador 734 23.1 
Allpahuayo Plot 1, Peru 616 21 26.71 0.42 
Allpahuayo Plot 2, Peru 608 26 27.33 0.27 
Cusco t o Plot 1, Peru 89 45 25.9 
Mishana, Peru 842 16 28.7 
Tonho Plot 1, Peru 585 17 26.9 
Yanamono, Peru 580 26 32.7 


San Carlos de Rio Negro, Venezuela 744 23.0 


Volume 87, Number 4 
2000 


Vasquez Martinez & Phillips 
High-Diversity Peruvian Forest 


507 


Table 5. 


Annual rates of natural stem and basal area mortality, recruitment, and increment, for stems = 10 cm 


diameter, between 1990 and 1996, All results are based on 0.84 ha (plot 1) and 0.96 ha (plot 2). 


A. Stem dynamics. 


Annual turnover 


Annual nnual (mean of loss and 
mortality recruitment ain) 

Plot 1: Tree stems 1.89% 1.57% 1.73% 

Plot 2: Tree stems 1.84% 2.04% 1.94% 

Plot 1: Liana and strangler stems 4.41% 1.89% 3.15% 

Plot 2: Liana and strangler stems 4.87% 8.02% 6.44% 


B. Basal area dynamics. Note that the annual gain in basal area is the sum of new recruitment into the 10 cm diameter 
size-class, plus the growth of pre-existing stems = 10 cm diameter, and annual turnover is the mean of mortality 


and gain rates. 


Annual nnual 
mortality recruitment Annual gain Annual turnover 
Plot 1: Tree basal area 1.76% 0.48% 2.08% 1.92% 
Plot 2: Tree basal area 2.21% 0.57% 2.32% 2.26% 
Plot 1: Liana and strangler basal area 5.69% 1.03% 5.72% 5.71% 
Plot 2: Liana and strangler basal area 8.29% 7.24% 8.57% 8.43% 


data for liana basal area (lianas are rarely system- 
atically censused in ecological plots, and liana bas- 
al area is even more rarely reported), so it is not 
possible to compare Allpahuayo with other forests 
in this respect. Elsewhere (Phillips et al., 1998) we 
reported that tree basal area values have been in- 
creasing in the majority of Amazonian plots cen- 
sused since the mid-1970s, which we interpreted 
as being a possible effect of long-term fertilization 
by rising atmospheric concentrations of carbon di- 
oxide. In this context it is interesting to note that 
by 1996, both plots at Allpahuayo had experienced 
small net increases in basal area (by 0.1% and 
1.9%) over the 1990 values shown in Table 4, in 
spite of some illegal felling of palm trees within a 
few of the sub-plots. Clearly a longer census inter- 
val will be needed to confirm whether the small 
change in forest structure is part of a long-term 
trend at Allpahuayo, or simply part of a pattern of 
random fluctuation around a long-term stable state. 

To estimate annual natural mortality and growth 
rates we excluded the sub-plots where palm trees 
were cut (Table 5). Stem turnover and basal area 
turnover functions measure slightly different attri- 
butes of the stand dynamics—stem turnover is con- 
cerned with population dynamics (i.e., the mean of 
population mortality and recruitment rates), while 
basal area turnover is concerned with basal area 
dynamics (i.e., the mean of basal area mortality and 
recruitment plus growth of existing trees). Over the 
long-term an old-growth forest would be expected 
to have similar values of each, and at Allpahuayo 
the turnover rates of tree stems and basal area both 


averaged close to 2% per year in the first half of 

Os. This rate is not unusual by the standards 
of western Amazonian forests, but it is higher than 
the average turnover rate for other tropical moist 
forests (~ 1.5% per year based on studies at 40 
different sites; Phillips, 1996). Based on our small 
sample of lianas and stranglers, large climbing 
plants appear to turn over notably faster than trees, 
about 5% a year, which indicates that these organ- 
isms may have shorter canopy residence times than 
most trees. Whether this pattern is repeated else- 
where remains to be seen, but if faster turnover of 
lianas and stranglers is a general property of Am- 
azon forests it would have implications for plot 
studies, most of which still ignore climbers. We may 
be overlooking a component of the forest that is 
more significant ecologically than has been appre- 
ciated. 


CONCLUSIONS 


Our floristic and ecological results at Allpahuayo 
well demonstrate the ecological and systematic 
benefits that can result when we concentrate our 
joint efforts on inventories in fixed plots. For the 
ecologist the benefits are clear—without the collab- 
oration of botanists in the field and the herbarium 
it is impossible to characterize patterns of diversity 
and floristic composition in most tropical forests, 
let alone explore the factors that determine these. 
As forests become more vulnerable to widespread 
environmental stresses such as fragmentation and 


climate change (Laurance et al., 1997; Phillips, 


Annals of the 
мо. Botanical Garden 


1997), plots are also needed to monitor these im- 
pacts on biodiversity. Systematists at botanical gar- 
dens therefore offer essential expertise for under- 
standing the biological effects of global change. 

For the systematist or floristic monographer, es- 
tablishing permanent sample plots can help in un- 
derstanding the local flora. The precise rigor of eco- 
logical sampling forces researchers to look equally 
at every plant that meets pre-determined criteria. 
In contrast, peripatetic botanizing may catalog the 
weedy, common, obvious, and accessible species 
while missing rarer or larger plants, especially can- 
opy trees, lianas, and epiphytes. Rigorous plot in- 
ventories can therefore reduce the spatial, taxonom- 
ic, life-form, and even seasonal biases prevalent in 
herbaria (e.g., Nelson et al., 1990), especially when 
integrated into a larger intensive collecting effort. 
Thus, a comparison of taxa in Vasquez (1997) with 
those listed in Brako and Zarucchi (1993) shows 
that the intensive collecting effort in permanent 
plots and surrounding forest at several Iquitos sites 
has yielded nearly 250 taxa new to Peru 
pahuayo, new tree taxa have been recognized in 
Annonaceae (2 species; Chatrou, 1998) and in 
Lauraceae (3 species including a new genus; van 
der Werff, 1993, 1997), while the repetition of plot 
visits has also allowed us to collect fertile material 
confirming new herb and shrub taxa (e.g., Kallunki, 
1994; Vasquez, 1997) 


> 


Literature Cited 


Brako, L. & J. L. Zarucchi. 1993. Catalogue of the Flow- 
ering Plants and Gymnosperms of Peru. Monogr. Syst. 
ot. ee же Gard. 45. 
үөү. genera: Syst 
Afric: `ап panes Herbar 


1: 


I, О. L. Phillips, A. H. Gentry, N. Stark 
mue 1995. Prediction of neotropical tree and 
liana m ies richness from | and climatic data. Bio- 
diversity and о 
Condit, R., . Foster, 5. Р. ‘Hubbell, R. Sukumar, Е. 
G. Le d N. visi S. Loo de Lao, J. V. LaFrankie 
Ashton. 1998. Assessing forest diversity on 
small lots: Calibration E. а ies-individual curves 
from 50-ha plots. Pp. –267 in F. Dallmeier & J. 
Comiskey (editors), b. Пе erm Monitoring of Biologic val 
“SCO, P. 


d Modeling: Concep- 
tual Background and Old World Case Studies. MAB 
Series Vol. 20. pà VES CO, Paris. 

& ditors). 1998b. Forest Biodiversity in 
North, Central de South America, and the Caribbean: 
Research and Monitoring. MAB Series Vol. 21. 
UNESCO, Paris. 

Duivenvoorden, J. F. & H. Lipps. 1995. A land- ecological 
study of soils, vegetation, and plant diversity in Col- 
ombian Amazonia. Tropenbos Series 12: 1—438. 

Gentry, A. H. 1988a. Changes in plant community diver- 


sity and floristic composition on environmental and geo- 
graphical gradients. Ann. Missouri Bot. Gard. 75: 1—34. 
1988b. Tree species richness of upper Amazo- 
nian deed Proc. Natl. Acad. Sci. U.S.A. 85: 156-159. 
992. Diversity and floristic composition of An- 
dean forests of Peru and adjacent countries: Implica- 
tions for their conservation. Mem. Mus. Hist. Nat. 
(Lima) 21: 11-19. 
eris R., K. Ruokolainen, H. Tuomisto, A. Linna & S. 


Flores Paitán (editors), онај у Desarrollo Ama- 
zónico. Turku, Finland. 
Kallunki, J. A. 1994. Revision of age Aubl. (Cuspa- 
riinae, Rutaceae). iinet 46: 2 
Laurance, W. F., S. N. bo M. Ran- 
ki Lovejoy. 1997. Bio 


Amazonian forest fragments. Senga 


Lowry, Р.Р, IL, G. E. Schatz & P. B. Phillipson. 1997. 


The classification of natural and anthropogenic vegeta- 
: n Ana о Solis 


Marengo, J. 1998. : Climatologí de la zona de Iquitos, 
Perú. Pp. 35-57 т В. Kalliola & S. Flores Paitán (ed- 
itors), Geoecología y Desarrollo Amazónico. Turku, Fin- 
land. 


McNeely, J. A, K. R. Miller, X V. Reid, R. A. Mittermeier 
& 


Werner serving the World's Biodi- 
versity. IUCN, Gland, и. and Island Press, 


bos es Cantor nia. 


sity: А Global Crisis Roadie international Solutions. 

National Science Board, оа D.C. 
Я ета, М. Е da Silva & М. L. 

nc а centres, refugia and botan- 
ical collecting denaity in Brazilian Amazonia. Nature 
345: 714—716. 

Phillips, O. L 1996, Long-term environmental change in 
tropical forests: Increasing tree turnover. кыша 
= 23: 235-248. 

7. The changing ecology of ра al forests. 

cuba and m 11. 
& А. Н. Gentry. 994. b s is 
through time in tropical hd Science 263: 954—958. 
& J. 5. Miller. In prep. Global Patterns i Forest 
Diversity: The ge of Alwyn Gentry. Monogr. Syst. 
Bot. Missouri Bot. Gard. 
P. en: 1996. A strategy 2 sampling 

ОГ forests. Pp. 141-165 in A. С. Gibson (edi- 
or), Neotropical Biodiversity and Conservation. Occas. 

Publ Mildred E. Mathias Botanical Garden 1. Mildred 

E. Mathias Botanical Garden, Р Angeles. 

‚ P. Hall, A. Н. Gentry, S. A. Sawyer & В. Vás- 
quez. 1994. Dynamics and uds richness of tropical 


rain forests. Proc. Natl. Acad. Sci. U.S.A. 91: 2805- 


Y. Malhi, N. Higuchi, W. F. Laurance, P. Nufiez 

SR. Vásquez M., 5. С. Laurance, L. V. Ferriera, M. 
A ern, 5. Brown & J. Grace. 1998. Changes in the car- 
bon balance of tropical forest: Evidence from long-term 
plots. Science 282: 439—442. 


Моште 87, Митбег 4 
2000 


Vasquez Martinez & Phillips 
High-Diversity Peruvian Forest 


509 


Rakotomalaza, P. J. = N. Messmer. 1999. Structure and 
floristic composition of the vegetation in the Réserve 
Naturelle Integrale d Andohahela, Madagascar. Fieldi- 
ana: Zoology, n.s., 94: 51–96 

Rudas L., A. 1996. Estudio Floríatioo y de la Vegetación del 
Parque Nacional Natural Amacayacu Amazonas-Colombia. 
M.Sc. Thesis, Universidad Nacional de Colombia, Bogotá. 

Swaine, M.D & D. Lieberman. 1987. Note on the cal- 

Бле H., K. Ru cler. R. Kalliola, A. p jen К? 
Danjo 7. Rodrigues 1995. Dissecting Amazonian 
bodie CEA. 269: 63—66. 

Valencia, R., H. Balslev & С. Paz у Miño. 1994. High 


tree alpha-diversity in Amazonian Ecuador. Biodiversity 
and Conservation 3: 21— 
Vásquez M., R. 1997. Flórula de las Reservas Biológicas 
de Iquitos. I Syst. Bot. Missouri Bot. Gard. 63. 
Vormisto, y ide . Phillips, K. Dm H. Tuomisto 
R. ez M. 2000. A comparison of small-scale 
distribution patterns of four ыл groups in an Ата- 
zonian — Ecography 23: 349-359. 
Werff, H., 3. A revision of the genus Pleu- 
enter ганні Ann. Missouri Bot. Gard. 80: 
18. 


extonia, a new genus of Lauraceae from 
South Але. Novon 7: 436—439. 


Missouri Botanical Garden 


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Annals of the 


512 


Missouri Botanical Garden 


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High-Diversity Peruvian Forest 


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Volume 87, Number 4 


2000 


"рэпицио) ‘| xipueddy 


Missouri Botanical Garden 


Annals of the 


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2000 


High-Diversity Peruvian Forest 


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Missouri Botanical Garden 


Annals of the 


516 


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Volume 87, Number 4 


2000 


Vasquez Martinez & Phillips 


High-Diversity Peruvian Forest 


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Annals of the 


518 


Missouri Botanical Garden 


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High-Diversity Peruvian Forest 


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Volume 87, Number 4 


2000 


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Volume 87, Number 4 


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Volume 87, Number 4 


2000 


Vasquez Martinez & Phillips 


High-Diversity Peruvian Forest 


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А SYSTEMATIC TREATMENT 
OF ACACIA COULTERI 
(FABACEAE, MIMOSOIDEAE) 
AND SIMILAR SPECIES IN 
THE NEW WORLD! 


n T. Jawad,? David S. Seigler,’ 
John E. Ebinger* 


ABSTRACT 


Detailed descriptions, habitat preferences, geographic ranges, and representative specimens are given for the 13 taxa 


of the Acacia coulteri group from Mexico, 


of the 


Central America 
distinct group within Acacia series Vulgares, lacking prickles and usually having persistent stipules. A principal 
ponents analysis (PCA) of vegetative and floral features shows tha i 
first three principal components. The groups established by PCA mostly coincide with previously described 
species. The taxa within this group are phenetically similar, sharing many morphological features. These da 


, and the 


southwestern United States. These species form a 
com- 
t the specimens examined form discrete units in plots 


ta also 


suggest that there is occasional gene flow between species, but that hybrids are not common. About half the species 


have restricted ranges (А. compacta, А. dolichostachya, A. durangensis, A. millefolia, А 
willardiana), but the remainder are wide-ranging, either from Oaxaca and Puebla 


. russelliana, А. sericea, and А 
north into central and northern Mexico 


(A. acatlensis, А. coulteri, А. mammifera, and А. salazari), or south into Central America (А. centralis and A. изита- 


cintens A 
Key w 


species delineation, hybridization. 


Acacia coulteri group, Acacia series Vulgares, Fabaceae, phytogeography, principal components analysis, 


Acacia Miller series Vulgares Benth. (Fabaceae: 
Mimosoideae), consists of about 200 species dis- 
tributed in tropical and subtropical wet to dry hab- 
itats, in both the Old and New Worlds. Series Vul- 
gares broadly corresponds to subgenus Aculeiferum 
Vassal (1972), according to Pedley (1978). Later, 
Pedley (1986) suggested that this subgenus should 
be elevated to generic status under the name Se- 
negalia Raf. More recent systematic studies suggest 
that Acacia series Vulgares may not represent a 
a е group (Chappill & Maslin, 1995; 
Clarke, 5). 

Most species of Acacia series Vulgares are of di- 
verse habitat, habit, morphology, and chemistry. 
Most are shrubs or small trees, though some are 
tree-lianas, with branches scrambling over other 
vegetation. Most species are armed with prickles 
on the stems, leaf petioles, and rachises. Leaves are 
bipinnately compound with deciduous stipules, and 


numerous pairs of pinnae and leaflets. Peduncles 
are solitary, or in racemose clusters, from the leaf 
axils, and flowers form globose heads or cylindrical 
spikes. Legumes of series Vulgares are relatively 
large, strongly flattened, usually dehiscent, and 
with a single row of relatively large seeds. This se- 
ries is often separated from two other Acacia series, 
Gummiferae and Filicinae, by the presence of 
prickles that are scattered along the stems and 
sometimes the leaf rachises (Bentham, 1875). 
Acacia series Vulgares can be divided into sev- 
eral informal “species groups” based on overall 
similarities of habit, stipule persistence, the pres- 
ence or absence of prickles, petiolar gland shape 
and structure, as well as inflorescence shape. At 
present, however, these informal groups within the 
series have not been established to be monophy- 
letic units. The taxa of one of these groups of phe- 


netically similar species, “the Acacia coulteri 


' We acknowledge support by the National Science Foundation (NSF BSR 82-15274 and NSF-PCM-82-17114), the 


United States Departmen 


t of Agriculture (OICD 58-319R-0-011 
University of Illinois Research Board (1994), and the Rupert Aii Award by the 


), the American ге Society (DSS 1992), the 
w York Botanical oli for 


DSS in 1997. We also thank the curators of the many herbaria that sent specimens, and Lo a ico an o Sousa 
for making manuscripts of unpublished work available. ат Jera gratefully acknowledges support as ај гуш 
dis inten by the Howard Hughes Foundation. istance of David Clarke, Kurt Potgieter, Karin "Re adel, 


Catherine Glass, and Claudio Gratton with field portions of pp res 


earch is greatly appreciated. The comments and 
r are much rae iate 


cester Point, Virginia 23062. Us 
rtment of Plant Bios iden of Illinois, Urbana, Illinois т U.S.A.; author for correspondence, 


* Emeritus Professor of Botany, Eastern Illinois University, Charleston, Illinois 61920, U.S.A. 


ANN. Missour! Bor. 


GARD. 87: 528—548. 2000. 


Volume 87, Number 4 
2000 


Jawad et a 529 


Acacia coulteri Group 


group" (Maslin & Stirton, 1997), are erect shrubs 
or small trees with persistent stipules and flowers 
in cylindrical spikes. Except for minor differences 
in flower size and occasionally pubescence, flowers 
are quite similar, with a tubular calyx and corolla, 
each 5-lobed, numerous separate stamens, and a 
short-stalked pistil. The 13 species of the A. coul- 
teri group range from Arizona, south through Mex- 
ico into Costa Rica. They are morphologically dis- 
tinct from other species of Acacia series Vulgares 
in that they always lack prickles, are never lianas, 
and by a number of other features. Nonetheless, 
taxonomic treatments have not dealt with this group 
as a separate unit within the Acacia series Vulgares 
(Bentham, 1875; Standley, 1920; Britton & Rose, 
1928) 


Many sterile and fruiting specimens of species 
of the A. coulteri group have been misidentified, 
adding to the taxonomic confusion. Flowering spec- 
imens of Albizia, Lysiloma, and Piptadenia can eas- 
ily be distinguished from spicate species discussed 
in this work. In contrast to the A. coulteri group; 
Albizia and Lysiloma have capitate inflorescences. 
Although inflorescences of Piptadenia species are 
spicate, these flowers have 10 stamens with strap- 
like filaments and oblong anthers that are larger 
than those of Acacia. Stamen filaments of flowering 
specimens of Albizia and Lysiloma have stamens of 
about the same number and size as those in Acacia, 
but the filaments are united into a distinct tube. 
Capitate species are normally evident, even in fruit- 
ing condition. Most fruiting specimens of Piptad- 
enia can be recognized by stamen remnants at the 
base of the fruits; its strongly venose fruits appear 
distinct from those of Acacia species, especially if 
comparative material is available. Careful search of 
most Piptadenia specimens reveals the presence of 
prickles, which are completely absent from all 
members of the A. coulteri group, as well as from 
members of the genera Albizia and Lysiloma. Two 
spicate species of the latter genus, L. acapulcense 
and L. auritum, are sometimes confused with mem- 
bers of the A. coulteri group. For these two species, 
the two parallel sutural ribs are fused around the 
periphery of the pod; these ribs often separate from 
the valves (Barneby & Grimes, 1996). Vegetative 
specimens of several Lysiloma species may be con- 
fused with Acacia species of this group. Some Lys- 
iloma species have cordate-auriculate stipules (at 
least on young branches) that are never found on 
the species treated in this work. Finally, vegetative 
specimens of Albizia usually can be differentiated 
by the palmate/pinnate venation present in the leaf- 
lets (Barneby & Grimes, 1996). 


The only other member of Acacia series Vulgares 


with which species of the A. coulteri group are eas- 
ily confused is A. macilenta Rose. Often, herbarium 
specimens of this last species lack prickles, and 
those specimens that lack prickles are superficially 
similar to some members of the A. coulteri group. 
Their separation is straightforward, however, as A. 
macilenta has petioles that are consistently less 
than 25 mm long, and the single petiolar gland is 
dark brown and shallowly volcano-shaped. In large- 
leaved species of the A. coulteri group with which 
A. macilenta might be confused, the petioles exceed 
25 mm in length and have petiolar glands that are 
light green or yellow and not volcano-shaped. 

Concepts and identification of taxa of the A. coul- 
teri group have been unclear in the past. No func- 
tional keys that permit accurate identification of 
these taxa have been published. Surprisingly, in 
most herbaria, up to half the specimens of this 
group of species were misidentified. Further, a cla- 
distic analysis based on molecular data from chlo- 
roplast restriction site analyses (Clarke, 1995), and 
cladistic analyses by Catherine Glass and David S. 
Seigler based on morphological data (unpublished) 
suggest a more ancient origin of these taxa than 
other species of Acacia series Vulgares. Otherwise, 
there are no comprehensive studies on the phylo- 
genetic relationships of the Acacia coulteri group. 
The present study was undertaken to clarify the 
taxonomic status and discreteness of the taxa in- 
volved, as well as to provide accurate descriptions 
of and functional keys to these species. 


MATERIALS AND METHODS 


About 500 Acacia specimens, obtained from 27 
herbaria (A, ARIZ, ASU, BM, BR, CM, DS 
F, G, GH, ILL, ISC, LL, MEXU, MIN, MO, MU, 
NY, POM, RSA, SD, TEX, UC, US, VT, WIS), were 
studied to determine the geographic ranges and 
morphological variation of the species in the A. 
coulteri group. These specimens were initially sep- 
arated into groups based on the similarity of mor- 
phological characteristics. After removal of dupli- 
cate and incomplete specimens, only 15 to 
specimens of most groups remained available for 
subsequent analysis. Only unambiguous specimens 
were scored and are indicated in the exsiccatae by 
an asterisk. Of the original specimen pool, 237 
were scored for 1 floral and 14 vegetative charac- 
ters (Table 1). All characters were measured for 
each specimen (three or more measurements) and 
plotted to confirm that gaps existed in order to per- 
mit the use of scored characters. The data were 
then analyzed by principal components analysis 


(PCA) using NTSYS-pc version 1.70 (Rohlf, 1993). 


530 Annals of the 
Missouri Botanical Garden 
Table 1. Characters scored for the principal compo- Table 2. Principal components for the data set for the 


nent analysis of the 13 species of the Acacia coulteri 


1. Bark of trunk (1 = not exfoliating, 2 = exfoliating). 
2. Petiole pubescence (1 — glabrous or nearly so, 2 — 
puberulent with appressed hairs, 3 — woolly pubescent). 


3. cun length (1 = 0-10 mm, 2 = 710-99 mm, З 
= mm). 


4. pe purple glands on petiole, rachis, and pinnae 
b se 


ipn es (1 — absent, 2 — 


5. Petiole gland structure ü — sessile, 2 — stalked, 3 


absent). 
6. Petiole gland apex (1 = flattened, 2 = doughnut- 


shaped, 3 — globose, 4 — absent). 
7. Rachis length (1 = absent, 2 = 1-20 mm, 3 = >20 
mm). 


8. Number of pinna pairs (1 = 1 to 6, 2 = 7 or more). 
9. Petiolule length (1 = 2.4 mm, 2 = 72.4 mm). 

10. Distance between leaflets (1 = $1.5 mm, 2 = >1.5 

mm). 

11. Number of leaflet pairs per pinna (1 = <36, 2 = 
36) 


12. Leaflet s (1 = 4-36 mm, 2 = >36–65 mm). 

13. Leaflet apex (1 = obtuse to broadly acute, 2 = nar- 
rowly acute M ac Иа 
14. Leaflet pubescence ( 
pubescent with appressed hairs beneath, 3 = 


= glabrous or nearly so, 2 = 
pubescent 
with appressed hairs on both surfaces, 4 = long hairs 
clustered at the base of midrib beneath). 

15. Perianth pubescence (1 = glabrous or nearly so, 2 = 


pubescent with appressed hairs, 3 = woolly pubescent). 


Data were transformed by a square root transfor- 
mation (Sokal & Rohlf, 1969). Groups were ana- 
lyzed os in various combinations, and si- 
multaneou 

ecause cyanogenesis has previously been found 
in many Acacia species, and is sometimes ше 
taxonomically (Seigler & Ebinger, 19 
Clarke et al. 1989, 1 , all specimens were test- 
ed for the presence of cyanogenic compounds by 
the Feigl-Anger method (Feigl & Anger, 1966; Tan- 
tisewie et al., 1969). Seigler et al. (1978) reported 
a few herbarium specimens of A. acatlensis that re- 
leased cyanide, and Conn et al. (1989) reported 
weakly positive cyanide tests for both A. acatlensis 
and A. coulteri using fresh, air-dried leaf material. 
During the present study, no material examined 
gave a positive-test for cyanide, even after the ad- 
dition of emulsin. 


RESULTS AND DISCUSSION 


When the entire set of 237 specimens was ana- 
lyzed using PCA, the first three principal compo- 
nents accounted for 28%, 18%, and 17% respec- 


Acacia coulteri group. 


Component number 


aracter 
number 1 2 3 
1 0.704 0.113 0.203 
2 —0.638 – 0.033 0.678 
3 0.698 0.244. 008 
4 0.651 0.040 0.526 
5 0.396 0.782 0.250 
6 0.352 0.810 0.435 
7 —0.446 0.428 —0.272 
8 —0.139 0.564. — 0.409 
9 0.744 0.144 0.024 
10 0.427 0.444 0.589 
11 —0.280 —0.189 —0.761 
12 0.356 —0.368 0.353 
13 0.469 0.545 —0.188 
14 – 0.404. 0.394. 0.441 
15 – 0.773 0.006 0.189 
Percent of variance 28.2 17.9 17.2 


tively, or 63% of total variance (Tables 1, 2). The 
amount of variance contributed by the remaining 
components diminished slowly from the third prin- 
cipal component onward. The best perspective for 
display of the phenetic groupings was seen in the 
ordination based on the second and third principal 
components (Fig. 1). This figure presents all taxa, 
clearly resolved in one view. In this plot, 13 distinct 
roups, corresponding to the 13 previously defined 
species can be observed, although the data clusters 
of some groups are proximal. Perianth pubescence, 
petiolule length, and exfoliating bark (characters 
, 9, 1) were the most important characters for 
determining the component score for the first prin- 
cipal component axis (Table 1). Petiolar gland apex, 
petiolar gland structure, and the number of pinna 
pairs (characters 6, 5, 8) were most important for 
the second principal component axis. The most im- 
portant characters for the third principal compo- 
nent axis were leaflet pairs per pinna, petiole pu- 
bescence, and the distance between leaflets 
(characters 11, 2, 

e clusters on this plot do not contain recog- 
nizable subgroups, and, except for A. usumacinten- 
sis and A. dolichostachya, which nearly overlap on 
this plot, each of the clusters is clearly separated 
from the others (Fig. 1). In addition, the OTUs of 
each group are closely spaced, indicating that the 
species are homogeneous. 

In addition to those herbarium specimens bor- 
rowed, the present treatment is also based on ex- 
tensive collections and observations of these taxa 


Моште 87, Митбег 4 Jawad et al. 531 
2000 Acacia coulteri Group 


-1.0 -0.5 0.0 0.5 1.0 
. I < ‚ • 1 А . а K Ж A I 
1.0- = 
. А. mammifera 
* A. durangensis Q 
0.5- = 


A. sericea 


О А. compacta 


A. willardiana А 
R А. coulteri Я 
А А. salazari 1 
0.0- x 
А А. Оу Г 
Р russelliana 
. А. dolichostachya 
5 Жы centralis 
a. Бе 1; 
acatlensis 


A. usumacintensis 


A 

X • 

т. 

5. 

3. 

40- 
. I е е е е . . . I . . . . . . . I . LJ • • . . . I . . . . . . . LJ I . 
-1.0 AXIS 2 -0.5 0.0 0.5 1.0 
Figure 1. Plot of axis 2 versus З of a р components analysis, using one floral and 14 vegetative variables 


from 237 specimens of the Acacia coulteri grou 


in Mexico and Central America by one of the au- the PCAs are included in the list of representative 
thors (DSS) on 15 trips during 1975-1998. Only specimens (designated by an asterisk after the her- 
the most reliable, diagnostic characteristics were barium abbreviation). Synonyms and types are in- 
used to distinguish the taxa. Although floral char- cluded, along with maps showing the geographic 
acteristics were measured carefully, and are in- distribution of each species (Figs. 2, 3) 

cluded in the descriptions, flower variation and Occasionally, a few flowering specimens lack 
character overlap precludes using them for distin- leaves, or have extremely small leaves, making it 
guishing species. Extensive lists of representative difficult to determine the number of leaflets per 
specimens are included, as specimens are com- pinna and the number of pinnae per leaf. Even with 
monly misidentified. Also, all specimens used in these specimens, the number of pinnae per leaf of- 


532 


Annals 


of the 
Missouri Botanical Garden 


$ а" Y X ii |“ 
gm dá је ~ s M^ * Gs 
• yv T о зс а өгө до v Н 
ә 
: че LA 
3 e X e one 
| "ux ХХ» 
А А ~ Дар < 
• Acacia acatlensis 00 x | 
о Acacia centralis 
* Acacia dolichostachya 
x Acacia sericea 
+ Acacia usumacintensis 


Distribution of Acacia acatlensis, А. centralis, А. dolichostachya, А. sericea, and А. usumacintensis in 


Figure 2. 


g 
Mexico, Belize, and Guatemala. The distribution of A. vient in the rest of Central America is not shown 


are only lightly appressed pubescent, whereas, in 
the latter, the calyx, corolla, and floral bracts are 
densely sericeous pubescent. Two 
have these small purple glands and occur within 
the same geographic range. Acacia compacta is a 
small shrub with short shoots, and A. mammifera 
is a small tree with flowers that have cream-colored 


ten can be determined from small emerging leaves, 
or by counting the number of pinna scars on an old 
rachis that may be present. In excess of 500 spec- other species 
imens were examined in this study and few lacked 
leaves altogether. Most of these were either Acacia 
acatlensis or A. sericea. Minute purple glands on 
the young stems and peduncles of both taxa distin- 
guish them from other members of this group of corollas with а purplish tinge. However, these are 
species. Acacia acatlensis is then separated from А. only uncommonly found in flowering condition 


sericea because the former’s calyx and floral bracts without leaves. 


KEY TO SPECIES 
la. Pinnae mostly with more than 36 pairs of leaflets, especially those near the middle of the rachis 
Petiolar gland(s) flattened, usually located on the lower third of the petiole; leaflet apex Pee to broadly 
acute. 
Ja. Leaflets 1.2-1.9 mm wide; leaves with more than 6 pairs of pinnae 1. А. usumacintensis 
3b. Leaflets mostly less than 1.2 mm wide; most leaves with 6 or deat pairs of su е 
anes а, КН 
2Ь. Petiolar lando a sauc ет [о cup-shaped, m dnd: on the upper half of the itd rarely 
absent; ini apex narrowly acute to acumina 
nute purple glands common at the base <a leaflets and usually along the rac кн leaflet 
је king long hairs on the lower side at the Базе . A. ман үз 
4Ь. Minute a glands absent; leaflets E with long hairs on the lower side at the ia 
ect - 4. A. же 
lb Pinnae mostly with lewe than 36 | pairs of leaflets, or pinnae pres 


Leaves less than 30 mm long, some clustered on short shoots 5. А. compacta 
eaves more than 30 mm long; short shoots absent; pinnae sometimes s absent in Acacia йал. 
Leaflets appressed to erect pubese ent on both surfaces, usually densely so; minute purple glands 
common on the rachis and pinna rachises. 

'etiole and rachis densely pubescent with erect hairs about 0.3 mm long: p less 
than 2.1 mm long; fruit pubescent s. . sericea 
Petiole and rachis iib or with short, appressed hairs; petiolules more than 20 mm 
ong; fruit glabrous. 


5b. 


ao 


7b. 


Volume 87, Number 4 Jawad 533 
2000 


et al. 
Acacia coulteri Group 


TIE NIRE. 


Фех + Ho ж: 


Acacia willardiana 


! e • бебу у Ra 
А E 2 ~ # 
s aren dere, et ee Ч 
РА Li 
oc te lee 
• f * 
A 
~ 
i 
~ 


e 3. Distribution of Acacia compacta, A. coulteri, A. durangensis, А. mammifera, A. millefolia, A. russelliana, 
A. cni and A. williardiana in the U.S.A., Mexico, Belize, and Guatemala. 


8a. р glands raised, the apex bulbous; most leaves with fewer than 7 pairs о 

Steet 7. А. mammifera 

8b. Petiolar glands se ом and with an irre gularly raised apex; ‘most leaves with ame than 

0 pairs of pinna Я 

Ob. Leaflets Pru or lightly о pubese ent bene ath; minute purple glands absent. 
9a. 


s mostly with a single pair of pinnae (rarely 2 or 3); many petioles more me 100 п 


A. durangensis 


OTN a с. at P . A. uin M 
E mostly with 4 or more pairs of pinnae; petioles less than 70 mm lo 


ong. 
ae gland between the upper pinna pair stalked; shrub or small tree less a 4n 
10. A. milla 
10b. Rachis gland between the upper pinna pair sessile, usually saucer-shaped, cup-shaped 
or absent; large shrub or tree; more than 4 m tall. 


9b. 


= 


of trunk and larger branches exfoliating and papery; petiolar pans absent 
many petioles; leaflet apex acuminate 11. A. salazari 
lib. Bark of trunk and larger branches smooth to furrowed, not exfoliating; petiolar 
glands present; leaflet apex broadly acute to obtuse. 
12a. Leaflets appressed pubescent beneath; rachis and pinna rachises puberulent 
above; perianth pubescent 12. A 


. A. coulteri 
12b. Minna glabrous beneath rachis and pinna rachises as well as perianth 
labrous or nearly s 18. 


A. russelliana 


Annals of the 
Missouri Botanical Garden 


Figure 4. 


mifera. —Е. A. coulteri.—F. A. centralis. —G. А. 


1. Acacia usumacintensis Lundell, Contr. Univ. 
Michigan Herb. 4: 8. 1940. TYPE: Mexico. 
Tabasco: Boca Cerro on the Rfo Usumacinta 
above Tenosique, 1-5 July 1939, Е. Matuda 
3550 (holotype, MICH!; K!, LL, 
MEXU!, MICH!, NY!). 


isotypes, 


Tree to 30 m tall with bark dark gray, shallowly 
furrowed; twigs brown to greenish brown, not flex- 
uous, glabrous to lightly puberulent; short shoots 
absent. Leaves alternate, 90-200 mm long; stipules 
herbaceous, light brown, narrowly triangular, to 2.2 

5 mm near the base, glabrous to lightly pu- 
berulent, d petiole adaxially shallowly 
m long, glabrous to lightly pu- 
berulent; pide svi solitary, usually located on 
the lower third of the petiole, sessile, commonly 
elliptical, 1-6 mm long, flattened, glabrous (Fig. 
4B); rachis adaxially grooved, 50-140 mm long, 
glabrous to lightly puberulent, a sessile, saucer- 
shaped gland, 1.0—1.9 mm across, between the up- 
per pinna pair; pinnae 7 to 15 pairs E leaf, 55— 
90 mm long, 5-12 mm between na pairs; 
petiolules 1.7-3.0 mm long; leaflets (3336 to 55 
pairs per pinna, opposite, 1.0—1.6 mm between leaf- 


grooved, 3 


Petiolar glands. —A. Acacia ране — 


williardiana. —H. 


А. usumacintensis. —C. А. dolichostachya. —D. А. mam- 
A. compacta. 


lets, oblong, 4-7 X 1.2-1.9 mm, glabrous, lateral 
veins obvious, with a midvein and 1 to 3 smaller 
veins from the base, base oblique, margins ciliate, 
apex acute to obtuse. Inflorescence a loosely flow- 
ered cylindrical spike 50—110 mm long, 1 to 3 from 
the leaf axil, or sometimes in terminal racemose 
clusters; peduncle 7-15 X 0.7-1.1 mm, puberu- 
lent; involucre absent; floral bracts linear, to 1 mm 
ong, puberulent, early deciduous. Flowers sessile, 
creamy-white; calyx 5-lobed, 1.1-1.5 mm long, 
densely appressed pubescent; corolla 5-lobed, 1.8— 
2.5 mm long, densely appressed pubescent; stamen 
filaments 4—6 mm long; ovary glabrous to lightly 
pubescent, on a stipe to 0.3 mm long. Legumes 
light yellowish brown to dark brown, straight, flat- 
tened, oblong, 90-250 x 20-33 mm, cartilaginous, 
transversely striate, glabrous, eglandular, dehis- 
cent; stipe to 22 mm long; apex acute. Seeds un- 
iseriate, no pulp, dark reddish brown, oblong to 
oval, strongly flattened, 8.8—10.0 х 7.0-9.0 mm, 
smooth; pleurogram U-shaped, 3-4 mm across. 
Flowers: April-June. 


Distribution. Moist tropical forests, along 
streams, and moist disturbed sites below 500 m 


Моште 87, Митбег 4 
2000 


Jawad et al. 
Acacia coulteri Group 


535 


elevations in Belize, Guatemala, and the states of 
Campeche, Chiapas, Oaxaca, Tabasco, and Vera- 
cruz, Mexico (Fig. 2). 

Acacia usumacintensis is a tall tree that is a com- 
mon component of lowland tropical rainforests in 
Guatemala and southern Mexico. No specimens of 
this species have been observed by the present au- 
thors farther south than Guatemala and Belize. 
Specimens labeled A. usumacintensis from Nicara- 
gua are the result of confusing this taxon with var- 
ious species of Lysiloma. Acacia usumacintensis 
specimens are commonly misidentified as L. aca- 
pulcensis (Kunth) Benth. Thompson (1980), in a re- 
vision of Lysiloma, annotated the type of А. изи- 

acintensis as L. acapulcensis. This specimen (E. 
Matuda 3550), however, is A. usumacintensis, hav- 
ing numerous separate filaments remaining at the 
base of some of the fruits. 


Representative specimens. (*used in PCA analysis) 
BELIZE. El Cayo Dist., Benque Viejo, Río Mopán, Con- 
ин 7089 (CAS, F, LL, МО“, NY); vicinity Georgeville, 

estern hwy., Dwyer 12662 (MO, US*); Valentin, El Cayo. 
ae 6356 (F, GH, iy и GUATEMALA. Izabal: 
Puerto Barrios, ca. 20 km from town on the road to Ma- 
chacas, Marshall, Castillo & Marshall 423 (MO. NY*). 
Petén: Lacandón, about 2 km S, Contreras 3467 (CAS, 
F, LL*, MO); е uU 5479 (F, MICH*, US); 
Dos Lagunas, 19 Apr. 1969, Contreras 8356 (F*, LL, MO); 
Tikal, in se on Main Plaza, Lundell 15348 (CAS*, 
Е LL, c Tikal, around Aguada Tikal, Lundell 
15973 bd X. NY); Sayaxche, in corozal about 5 km 5 
of village, Lundell 18071 (F, LL, MO, NY*); Cadenas, on 
river bank bordering Río Gracios, Lundell & Contreras 
19033 (F*, LL, NY). MEXICO. Campeche: El tormento 
Escárcega, Sousa, Cabrera, Davidse & Chater 12369 (IL E 
MO*). Chiapas: W side of Laguna Miramar E of S 
Quintín, Breedlove 33387 (DS*, dup MO); 20 km al SE 
de la desviación San Javier Frontera Echeverría hacia Río 
ЦЫ santum, Chimal, Durán & Quintanilla 755 (F, ILL, 

MO*); a 5 km al E de Crucero Corozal, camino a Frontera 
Corozal, Ocosingo, Martínez 7291 (MO, NY*); a 15 km al 
E de Nvo. San Juan Chamula camino a Ixcan, Martínez & 
Girón 6288 (MO*, MU). Oaxaca: Hincón Bamba, al O 
de Salina Cruz, Martínez 629 (MO*). Tabasco: Mun. Bal- 

ncán, por la carretera O, por el Poblado Arroyo del Triun- 
% и 2385 (F*, NY); La Palma, Balancán, Matuda 

4 (Е, MICH*, WIS). руке Mpio. Minatitlan, Lo- 
mas SW de La Garganta, 5.8 km al W de шашар; 
Wendt, Villalobos & raid 4127 (F, MICH, MO, TEX). 


— 


2. Acacia dolichostachya S. F. Blake, Proc. Biol. 
Soc. Washington 34: 43. 1921. Senegalia dol- 
ichostachya (S. F. Blake) Britton & Rose, N. 


umer & Sons 23329 (holotype, F!; isotypes, 
F!, G!, K!, NY photo!) 


Small tree to 15 m tall with bark dark brown to 
dark gray, scaly in rectangular plates; twigs light 


brown to greenish brown, not flexuous, glabrous; 
short shoots absent. Leaves alternate, 40-120 mm 
long, stipules herbaceous, light brown, narrowly tri- 
angular, to 1.3 X 0.3 mm near the base, glabrous, 
persistent; petiole adaxially shallowly grooved, 30— 
80 mm long, glabrous; petiolar gland solitary, lo- 
cated on the lower half of the petiole, sessile, cir- 
cular to more commonly elliptic, 1.0–2.5 mm long, 
flattened or with slightly raised margins, glabrous 
Fig. 4C); rachis adaxially grooved, 20-62 mm long, 
lightly puberulent, a sessile, saucer-shaped gland, 
0.6-1.5 mm across, between the pinnae of the up- 
per 1 to 2 pinna pairs; pinnae 2 to 6(8) pairs per 
leaf, 40—75(90) mm long, 4—9 mm between pinna 
pairs; petiolules 2.5—4.0 mm long; leaflets 36 to 65 
pairs per pinna, opposite, 0.7—1.1 mm between leaf- 
lets, oblong, 3.5—5.5(7.5) X 0.8-1.3 mm, glabrous, 


lateral veins usually not obvious, only one vein 


— 


from base, base oblique, margins ciliate, apex acute 
to obtuse. Inflorescence a loosely flowered cylin- 
drical spike 20—90 mm long, 1 to 3 from the leaf 
axil, or rarely in terminal racemose clusters; pe- 
duncle 3-10 x 0 
berulent; involucre absent; floral bracts linear, to 1 
mm long, glabrous to lightly pubescent, usually not 
deciduous. Flowers sessile, creamy-white; calyx 5- 
lobed, 0.5-1.0 mm long, lightly appressed pubes- 
cent; corolla 5-lobed, 1.2-2.2 mm long, lightly ap- 
pressed pubescent; stamen filaments 3—5 mm long; 
ovary glabrous, on a stipe to 0.3 mm lon 


.5—0.9 mm, glabrous to lightly pu- 


g. Legumes 
light yellowish green to light brown, straight, flat- 
tened, oblong, 80-130 х 13-20 mm, cartilaginous, 
transversely striate, glabrous, eglandular, dehis- 
cent; stipe to 12 mm long, apex acute to acuminate. 
Seeds not seen. Flowers: April—July. 


Distribution. Common in thorn-scrub thickets, 
and disturbed dry forests in the lowlands of the 
states of Campeche, Quintana Roo, and Yucatán 
Fig. 2). 

Restricted to the Yucatán Peninsula, Acacia dol- 
ichostachya appears to be a relatively common 
component of thorn-scrub thickets and disturbed 
sites. The fact that most leaves have fewer than 
seven pinna pairs separates А. dolichostachya from 
most other species of this group. Other species with 
six or fewer pinna pairs differ by having pinnae 
with fewer than 35 pairs of leaflets, along with 
smaller leaflets (А. compacta), much longer petioles 
(A. willardiana), or pubescent leaves (А. mammi- 
fera and A. sericea). 

Considering the restricted distribution of this 
species, the only other taxon that A. dolichostachya 
might be confused with is А. usumacintensis. Both 
taxa have more than 35 pairs of leaflets on each 


~ 


536 


Annals of the 
Missouri Botanical Garden 


pinna, flattened petiolar glands on the lower half of 
the petiole, and petiolules that are more than 2.5 
mm long. They are easily separated as A. dolichos- 
tachya has fewer pinna pairs per leaf (mostly six or 
fewer), smaller stipules (less than 1.3 mm long), 
less distance between the leaflets on the pinna ra- 
chis (less than 1.2 mm), and smaller sepals (mostly 
less than 1.0 mm long). 


пирето specimens. MEXICO. Campeche: 4 
km despues de Sho laguna Xmaben, Mpio. Hopelchen 
= & pe 1023 (UC*); a 12 km al E de Constitución, 
o sea a 15 km al W de Conhuas, Sousa, Ramamoorthy, 
nano Rico & Basurto 12229 (MO*). Quintana Roo: En 
Rancho San Felipe, a ne / de la desviación a Puerto 
Morelos, por la carretera Ca 
Me )*); a 15 km al N de la ейде 'ión a к 

зоһге 1а ЕЕ ПА мп, Са 
3158 (MO*); а 
Vigia Chico, Leia Cabrera 3561 (MO*); a 35 km al 
NW de Carrillo Puerto, i 


е rdín Botánico Е Коо, 
Duran, Olmsted & бн 85 (МОФ); 3 i. 5 of Filipe 
Carril o Puerto, Dwyer, Spellman, | p Wunderlin 

(MO ;rether, 


E de Filipe Carrillo Puerto, Sousa, Cabrera, Dav- 
idse & Chater 12441 (MO*); al NW del entronque Che- 
tumal-F. Carrillo Puerto, sobre carretera Mérida via Corta, 
Téllez & Cabrera 2012 (MO*); a 10 km al S de Tulum, 
Téllez & Cabrera 2032 (MO, NY*); a 15 km al N de Ba- 
éllez & Cabrera 2366 (NY*); 10 km antes de la 
entrada a Buenavista, de Chetumal rumbo a Felipe Car- 
по Puerto, Ucan & Flores 985 (UC*). Yucatán: 11.2 
km N of Dzibilchaltum turn on road from Mérida to Pro- 
greso, Bradburn & Darwin 1294 (MO, NY*); 4.6 km al E 
de Sucila, sobre la carretera Tizimin-Mérida, Cabrera & 
Cabrera 13203 (MO*); near Piste, Lundell & Lundell 7344 

„ MICH*, MO, US); 7 km al $ de Yaxcaba, Vara, 
Arias & [es 574 (C AS, F*, MO) 


3. Acacia acatlensis Bentham, London J. Bot. 1: 
513. 1842. Senegalia acatlensis (Bentham) 
Britton & Rose, N. Amer. Fl. 23: 112. 1928. 
TYPE: Mexico. Puebla: Acatlán, May 1830, G. 
Andrieux 396 (holotype, К!, F photo!, MICH 
photo!, NY photo!, TEX photo!; isotypes, G!, 
US!) 


TM venale Britton & Rose, N. Amer. Fl. 23: 

13. TYPE: Mexico. Oaxaca: Cerro San An- 

с, a 1800 m, 12 Oct. 1907, С. Conzatti 2046 

ypes, F!, MEXU!, NY!). [Britton 

(1928) ud the o as Conzatti 25,346. 

Jn the 1 NY isotype, Bri “the number of this 
specimen was жна uaa 25,346."] 


Shrub or small tree to 15 m tall with bark dark 
gray, shallowly furrowed; twigs light brown to 
greenish brown, not flexuous, glabrous to lightly pu- 


berulent; short shoots absent. Leaves alternate, 50— 
150 mm long; stipules herbaceous, light brown, 
narrowly linear, to 3 X 
ally glabrous, persistent; petiole adaxially shallowly 
grooved, glabrous, but com- 
monly with minute purple glands; petiolar gland 
solitary, located between the lower pinna pairs or 
along the upper half of the petiole, sessile, nearly 
circular to slightly elongated, 0.7-2.3 mm long, 
saucer- to cup-shaped, glabrous, rarely absent; ra- 
chis adaxially grooved, 20-110 mm long, glabrous 
to lightly puberulent and usually with minute pur- 
ple glands, a sessile, cup-shaped gland, 0.4—0.9 
mm across, between the upper pinna pair; pinnae 
7 to 30 pairs per leaf, 25-60 mm long, 3-7 mm 
between pinna pairs; petiolules 1.0-2.4 mm long; 
leaflets 36 to 60 pairs per pinna, opposite, 0.5—1.2 
mm between leaflets, linear, 2. x 0.7-1.1 
mm, glabrous and commonly light greenish purple 
l to 2 


smaller veins from the base, base oblique, margins 


mm near the base, usu- 


mm long, + 


above, lateral veins obvious, a midvein and 


ciliate, apex narrowly acute to acuminate. Inflores- 
cence a loosely flowered cylindrical spike 40—100 
mm long, 1 to 4 from the leaf axil; peduncle 5-10 
X 0.5-1.0 mm, glabrous to lightly puberulent and 
commonly with scattered minute purple glands; in- 
volucre absent; floral bracts linear, to 1 mm long, 
early deciduous. Flowers sessile, creamy-white; ca- 
lyx 5-lobed, 1.0—1.6 mm long, lightly appressed pu- 
bescent; corolla 5-lobed, 2.1-2.8 mm long, lightly 
appressed pubescent; stamen filaments 4.5—6.5 mm 
long; ovary glabrous, on a stipe to 0.4 mm long. 
egumes light to dark brown, straight, flattened, ob- 
long, 80-170 Х 13-25 mm, cartilaginous, trans- 
versely striate, glabrous, eglandular, dehiscent; 
stipe to 20 mm long; apex broadly acute. Seeds 
uniseriate, no pulp, dark reddish brown, oval, 
strongly flattened, 7-10 X 4.5-7.2 mm, smooth; 
pleurogram U-shaped, 1.2-2.3 mm across. Flowers: 
arch-June 

Distribution. Dry, deciduous, tropical forests 
and thorn-scrub forests between 500 an m 
elevations in the states of Chiapas, Guerrero, Jal- 
isco, México, Michoacán, Oaxaca, and Puebla, 
Mexico (Fig. 2). 

Acacia acatlensis is very similar morphologically 
to A. centralis. Both are widely distributed in south- 
ern Mexico, although A. centralis has been collect- 
ed more frequently in the very south of Mexico and, 
unlike A. acatlensis, it occurs across the border in 
Guatemala and countries to the south (see Fig. 2). 
These two species are difficult to separate, and it 
is possible that they should be considered as sub- 
species of a single species. The two can be sepa- 


Моште 87, Митбег 4 
2000 


Jawad et al. 537 


Acacia coulteri Group 


rated based on the presence or absence of minute 
purple glands that are common at the base of the 
leaflets, in the grooves of the rachis and petiole, 
and not uncommon along the axis of the inflores- 
cence in A. acatlensis. Rarely these glands are 
clear, lacking the purple color, but are easily ob- 
served under magnification. None of these minute 
glands were found on specimens of A. centralis ex- 
amined. No other characteristics could be found 
that consistently separate these two species, though 
most specimens of A. centralis have leaflets with 
long hairs at the base on the ventral surface. These 
hairs are particularly evident on young leaflets, 
sometimes falling off as the leaves mature. 

Acacia acatlensis is also similar to A. usumacin- 
tensis, differing by having narrower leaflets (less 
than 1.1 mm across) and lacking the large flattened 
petiolar gland of that taxon. Acacia acatlensis is 
sympatric with A. sericea and A. mammifera in Oa- 
xaca; these two taxa also have minute purple glands 
on the leaves. Acacia acatlensis is easily separated 
from A. mammifera, which has fewer than 33 pairs 
of leaflets per pinna, leaflets that are commonl 
more than 1.5 mm wide, and a stalked petiolar 
gland. The dense, erect pubescence of A. sericea 
petioles and fruits usually separates this species 
from A. acatlensis. 

MEXICO. Chiapas: a 4 km 
Ramamoorthy, Cortés & Her- 
TEX). : Tario, Coyuca 
Dist, Hinton 7755 (DS, GH, MICH, NY, US); en el Puerto 
El Salado, a 7 km al N de Пара camino a m 
Martínez 1070 (MO, TEX*). Jalisco: 2 km 
Mirador, 10 km al NE de El Corcobado, carretera Autlán- 
оаа Magallanes 2169 (MO*); а 20 km al NW de 

San Patricio, Magallanes 2420 (MO*); 20 km NE of Au- 
Пап, Mc газ 23271 (MICH*, MO, NY); Barranca de Los 
anques, desviación al camino San Martín de las Cafias, 
Villarreal 6121 (MICH*). México: Pungarancho, Temas- 
caltepec Dist., Hinton 3747 (F, GH*, МО, NY, US). Mi- 
choacán: 3 km al S de Jungapeo, Soto, домы. Silva 
& Pizarras 8573 (MEXU*). Oaxaca: 5 km sobre la des- 
viación а Barranca de Los Calabazos, piscis 220 (NY*); 
3 km al NE de Tonala, carretera a Hua ajuapán, Calzada 
& Campos 18320 (CAS*, MICH, MSC); 
Mitla, near pass to Díaz icu со 10 (08%). 2.5 i 
al N de Yosocuta en la c uapán-Juxtlahuaca, 
Magallanes, Ramos & шла li (CAS*, LL. МО); 3 
m al NW de Santo Domingo Tonala, carretera a Ина 
рап, Orttz-Bermudez 291 А Төшеп Сапуоп, 
Pringle 5885 (GH, MICH, US*, VT); сегса de Tonala, 
Rzedowski 34920 (CAS". MO); Subida a Pa bien del 
Cerro Guiengola, Torres, Cortéz & Martínez, 928 
(MO*); La ue иде ин ee n Guiengola, Torres Torres, 
Téllez & Martínez 400 (MO*). Puebla: Tepeji de Rodrí- 
guez, Felger 85-38 (MO*); 29 mi. Ap ~ Acatlán on hwy. 
190, Seigler & Fdo 12692 (EIU*, ILL). 


Representative specimens. 


= 

— 
У 

r 

= 

= 


c 
— 


4. Acacia centralis (Britton & Rose) Lundell, 
Contr. Univ. Michigan Herb. 4: 7. 1940. Se- 


negalia centralis Britton & Rose, N. Amer. Fl. 
23: 113. 1928. TYPE: El Salvador. Near Зап 
Salvador, 1923, S. Calderón 1774 (holotype, 
NY!; isotypes, BM!, Е!). 


Tree to 25 m tall with bark dark grayish brown, 
vertically fissured, rough and scaling; twigs light 
brown to greenish brown, not flexuous, mostly gla- 
brous; short shoots absent. Leaves alternate, 70— 
180 mm long; stipules herbaceous, light brown, 
narrowly linear, to 4.5 X 0.6 
glabrous, persistent; petiole adaxially shallowly 
grooved, mm long, glabrous to lightly pu- 
berulent; petiolar gland solitary, located on the 
middle part of the petiole, sessile, usually circular, 


mm near the base, 


1.0-2.6 mm across, saucer- to cup-shaped, gla- 
brous; rachis adaxially grooved, 40—150 mm long, 
glabrous to puberulent, a sessile, saucer- to dough- 
nut-shaped gland, 0.6—1.3 mm across, between the 
pinnae of the upper 1 to 2 pinna ig (Fig. 4F); 
pinnae (7)11 to 24 pairs per leaf, 30-70 mm long, 
—9 mm between pinna pairs; de 0.8-1.5 
mm long; leaflets 40 to 60 pairs per pinna, oppo- 
site, 0.5—1.2 mm distance between leaflets, linear, 
3.0-5.5 X 0.6-1.1 mm, glabrous except for usually 
some long hairs at the base beneath, lateral veins 
obvious, with a midvein and 1 to 2 smaller veins 
from the | base oblique, margins ciliate, apex 
narrowly acute to acuminate. Inflorescence a loose- 
ly flowered аа spike 60—140 mm long, 1 to 
3 from the leaf axis, or in terminal racemose clus- 
ters; peduncle 4-10 x 0.5-1.0 mm, glabrous to 
lightly puberulent; involucre absent; floral bracts 
linear, to 1 mm long, pubescent, early deciduous. 
Flowers sessile, creamy-white; calyx 5-lobed, 0.7— 
1.3 mm long, lightly appressed pubescent; corolla 
5-lobed, 1.8-2.5 mm long, lightly appressed pu- 
bescent; stamen "p 4.5—6.5 mm long; ovary 
glabrous, on a stipe m long. Legumes light 
7 straight, DEP. Sons: 100-160 x 16- 
mm, cartilaginous, transversely striate, glabrous, 
eglandular, dehiscent; stipe to 20 mm long; apex 
roadly acute to obtuse and usually apiculate. 
Seeds uniseriate, no pulp, dark reddish brown, 
nearly circular, strongly flattened, 6-9 mm across, 
smooth; pleurogram U-shaped, 1.5—3.0 mm across. 
Flowers: April-August, and sporadically through- 
out the year when moisture is avai 


Distribution. Lowland forests, and moist dis- 
turbed sites below 1300 m in Guatemala, El Sal- 
vador, Honduras, Nicaragua, Costa Rica, and the 
states of Chiapas, Jalisco, Oaxaca, and Sinaloa, 
Mexico (Fig. 2). 

tall tree, sometimes entering the canopy of 
moist lowland forests, Acacia centralis is also a 


Annals of the 
Missouri Botanical Garden 


common component of disturbed habitats at lower 
elevation through most of Central America. Most 
collections are from roadsides, disturbed pastures, 
and gallery forests. In addition to its close morpho- 
logical similarity to Acacia acatlensis discussed 
above, A. centralis is also sympatric with five other 
acacia species in southern Mexico. Of these, it is 
easily separated from A. compacta and A. mammi- 
fera, as these species have fewer pinna pairs per 
leaf (fewer than 7) and fewer leaflets per pinna 
(fewer than 36). The erect hairs of A. sericea, the 
larger leaflets of A. usumacintensis, and the exfoli- 
ating bark of A. salazari can be used to distinguish 
these species from A. centralis. 

The authors have frequently encountered speci- 
mens of A. centralis that were annotated Albizia nio- 
poides (Benth.) Burkart. Flowering material of these 
two taxa is easily separated by the filament-tube 
and fewer stamens in Albizia. Fruiting material and 
sterile specimens also can be separated, as in A. 
centralis, the leaflets are less than 5.5 mm long and 

ave an acuminate apex, whereas in Albizia то- 
poides, the leaflets are longer, mostly more than 5.5 
mm long, and the apex is broadly acute to obtuse. 


Representative a COSTA RICA. Guana- 
caste: vicinity of Саћаз, Daubenmire 775 (F*); Santa 
Rosa National Park, Janzen 10355 (MO*). Puntarenas: 
San Luis village, Haber & Bello 1628 (MO*). EL SAL- 
VADOR. vic € of Comasagua, Allen & Armour 7257 (Е, 
LL, NY, US*). GUATEMALA. Petén: Laguna Yaxja, Har- 
mon & Dwyer 2763 d zu Zacapa: Gualán, Deam 
6281 (А, Е, GH*, MO, NY, UC, US, VT). HONDURAS. 
Moresin: Alrededores i | Ciudad Universitaria, Torres 
183 (NY*). Paraiso: 15 kms S of El Paraiso, Molina 
18420 (NY, US*). MEXIC O. Chiapas: Siltepec, ере и 
1584 (A, MICH*, NY, US). Jaliseo: 5 km al SE de 
Manzanilla, carretera Puerto Vallarta—Barra de Navidad, 
Masculina 906 (Е, MEXU). Oaxaca: Pto. San Bartolo, 
Yautepec, MacDougall H230 (MICH, NY*); between Li- 
món and Zapote, MacDougall s.n. (NY*, US). Sinaloa: 
Машо, Los Tepemesquites, Опера 5717 (GH, US). NIC- 
ARAGUA. L(on: Mares ен near Punta El Dia- 
blo, Neill 480 (MO, NY). Ma 8 El Cru- 
cero, en ta 96 oe vicinity of Managua, Garnier 957 
аи * US); Managua, Punta Chiltepec, Grijalva, Hn 

gas Ps Sánchez 3105 (CM, MO*); Esquipulas, Hall & 
Bockus 7981 (MO, NY*, UC); Peninsula de Chiltepec, 
Punta Chiltepe, Moreno 16917 (СМ, МО“); along hwy. 8, 
km 28, Stevens 3922 (MO*); 1.9 km from hwy. 2 on road 


ubes, 


along ridge of Sierra de Managua, Stevens 4753 (MO*); 1 
k X Laguna Apoyeque, Vincelli 779 (MO*). Ma- 
Laguna de Ap 21308 (MO). Rivas: 


Quebrada Las Cafias, near Río Escalante, Stevens 9681 
(MO). 


5. Acacia pe Rose, Contr. U.S. Natl. 
Herb. 8: 31. 1903. ade: compacta (Rose) 
Britton _ olt N. Am . 29: 111. 1928. 
TYPE: Mexico. Мерк ‘Tomellin Canyon, 24 


June 1899, J. N. Rose & W. Hough 4680 (ho- 
lotype, US!; isotypes, GH!, K!, NY!). 


Lysiloma standleyana Britton & Rose, 2 Amer. Fl. 23: 
81. 1928. 5 Mexico. Oaxaca: Tomellin, Sept. 
1905, J. N. Rose 10082 Oe NY *!, F photo!, 
MO photo!). 


Shrub or small tree to 3(4) m tall with bark dark 
gray, flaking off in thin strips; twigs light brown to 
dark reddish brown, slightly flexuous, pubescent to 
glabrous, when young with minute purple glands; 
short shoots commonly present above the nodes, to 
3 mm long, covered with acuminate stipules and 
old leaf bases. Leaves alternate, also commonly 
clustered on the short shoots, 5—30 mm long; stip- 
ules herbaceous, light brown, narrowly linear, to 3 
X 0.4 mm near the base, usually glabrous, persis- 
tent; petiole adaxially grooved, 2.5-14.0 mm long, 
usually pubescent and with minute purple glands; 
petiolar gland solitary, located at or just below the 
lower pinna pair, sessile, circular, 0.4—1.] mm 
across, doughnut-shaped, glabrous (Fig. 4H); rachis 
adaxially grooved, 0—20 mm long, usually pubes- 
cent and with minute purple glands, a sessile, sau- 
cer-shaped gland, 0.2—0.6 mm across, occasionally 
present between the upper pinna pair; pinnae 1 to 
6 pairs per leaf, 6-18 mm long, 2-5 mm between 
pinna pairs; petiolules 0.5—1.1 mm long; leaflets 9 
to 22 pairs per pinna, opposite, 0.5—0.9 mm dis- 
tance between leaflets, oblong, 1.4—3.0 х 0.5-0.9 
mm, glabrous above, usually lightly pubescent be- 
neath with long 
only one vein from the base, base oblique, margins 
usually ciliate, apex acute to obtuse. Inflorescence 
a loosely flowered cylindrical spike 30—70 mm 
long, solitary (rarely 2 to 3) from the leaf axil; pe- 

uncle X 0.4—0.7 mm, usually pubescent and 
with minute purple glands; involucre absent; floral 
bracts linear, to 1.5 mm long, pubescent, early de- 
ciduous. Flowers sessile, creamy-white; calyx 5- 


airs, lateral veins not obvious, 


lobed, 1.1-1.7 mm long, densely appressed pubes- 
cent; corolla 5-lobed, 2-3 mm long, densely 
appressed pubescent; stamen filaments 5.5-7.5 mm 
long; ovary glabrous, on a stipe to 0.4 mm long. 
Legumes light yellowish brown, straight, flattened, 
oblong, 50-120 X 10–16 mm, cartilaginous, trans- 
versely striate, glabrous, eglandular, dehiscent; 
stipe to 8 mm long; apex acuminate and usually 
beaked. Seeds uniseriate, no pulp, purplish brown, 
near circular, strongly flattened, 5-8 mm across, 
smooth; pleurogram U-shaped, 1.3-2.2 mm across. 


Flowers: April-Jul 


Distribution. Thorn-scrub forests, thickets, 
rocky slopes and washes between 500 and 1600 m 


Volume 87, Number 4 
2000 


Jawad et al. 539 


Acacia coulteri Group 


elevation in the states of Puebla and Oaxaca (Fig. 


Acacia compacta, a much-branched shrub that 
rarely exceeds 3 m, has a very restricted distribu- 
tion, occurring in southeastern Puebla and adjacent 
Oaxaca. Even there it does not appear to be a com- 
mon species, relatively little material being avail- 
able for study. All material examined is from xeric 
habitats, usually on rocky slopes and in washes, 
where A. compacta forms small thickets (Rico Arce 
& Rodriguez, 1998). The short shoots at many of 
the nodes, small leaflets (mostly less than 2.5 mm 
long), leaves that are less than 30 mm (mostly less 
than 20 mm) long, and fruits with the apex acu- 
minate and beaked, separate A. compacta from oth- 
er members of this group. The presence of short 
shoots is probably an adaptation to xeric conditions. 

Acacia compacta shows variation with regard to 
several important characteristics. Leaf size is highly 
variable: the leaves that develop on the short shoots 
are usually smaller and have shorter petioles and 
fewer pinna pairs; whereas the leaves that form on 
fast-growing shoots are larger, the petioles some- 
times reach a length of 14 mm, and 5 or 6 pinna 
pairs develop along the rachis. The shape and 
structure of the petiolar gland is also somewhat var- 
iable. The majority of the specimens have a sessile 
petiolar gland that is doughnut- or torus-shaped, 
although a few specimens have a stalked petiolar 
gland with a bulbous apex. Generally, these glands 
are found on young leaves forming on the short 
shoots; other leaves have sessile, doughnut-shaped 
glands. It is possible that Acacia compacta may 
rarely hybridize with A. mammifera, the only other 
species of this group with stalked petiolar glands. 


Representative specimens. MEXICO. Oaxaca: 52 km 
al S de Tecomavaca por la carretera rumbo a la Ciudad 


km al W de San Gabriel, < 
(ARIZ*, Е, MEXU, RSA); 14 km al N de Cuicatlán, vis 
Martínez, Téllez & Magallanes 5394 (CAS*, US, WIS); a 

3 km al NW de Guadalupe Los Obos, o sea a 10 km al 
NW de Cuicatlán, Sousa, Téllez, Magallanes & Delgado 
6912 (MICH*). Puebla: Alrededores del lado W del 
pueblo de Axusco, Chiang, Salinas & Dorado F-2466 
(ARIZ, MO, RSA*); en la desviación a San Luis Atoloti- 
tlán y Los Reyes Metzontla, Medrano, Chiang, Davila & 
Villasefior и (ARIZ*, МО»); 18 km al NW de Теон Ап 
del Camino, а Tehuacán, Medrano, Jaramillo, Villaseñor, 
Ruiz & Singer F-1172 (MO*); po з San Luis Tulti- 

O, 


tlanapa, Purpus 3193 (F, GH*, M Y, UC, US); 3 km 
al SE de San Rafael, Rico, Ramos E Hernández 246 
(CAS*, a aide e La Escalera, Ejido de San José 


os & танак 248 (CAS*, МО); 1 km 
al NE-E del limite estatal Oaxaca-Puebla por la carretera 
ehua cán, Salinas & Dorada F-3199 
Rafael, cerca de los limites 


Puebla-Oaxaca, Sousa & Sousa 10405 (ARIZ*, TEX); а 2 


km‘al SE de San Rafael, Sousa, Sousa & Basurto 10430 
(CAS*, TEX); Cerro Tepetroja al SW de Axusco, Tenorio 
& Romero 9043 (CAS*); Jardín Botánico de Cactáceas y 
Suculentas de Zapotitlán de las Salinas, Valiente-Banuet 
& Maeda 674 (RSA, TEX*); Jardín Botánico de Cactáceas 
y Suculentas de Zapotitlán de las Salinas, Valiente-Banuet 
& Maeda 706 (RSA* 


6. Acacia sericea Martens & Galeotti, Bull. Acad. 
Roy. Sci. Bruxelles 10(2): 311. 1843. Sene- 
galia sericea (Martens & Galeotti) Britton & 
Rose, N. Amer. Fl. 23: 111. 1928. TYPE: 
Mexico. Puebla: Tehuacán, alt. 6000 ft., May 

, H. Galeotti 3345 (holotype, ВЕ!; iso- 
types, K!, Р!, MICH photo!, NY photo!). 

Acacia кн Brandegee, Univ. оа cng Bot. 
4: 85. . TY AR Pues Puebla: o de So- 
lunte, xb ‚ June 1909 es pu Purpus 
3863 (holotype, om m photo!; арек, ВМ! 
МО!, NY!, US!). 


Shrub or small tree 3 to 4(6) m tall with bark 
dark gray, cracked and fissured, breaking away and 
leaving dark purplish brown, smooth areas; twigs 
dark brown to purplish brown, not flexuous, gla- 
brous to lightly pubescent; short shoots absent. 
Leaves alternate, 30— mm long; stipules her- 
baceous, light brown, narrowly linear, to 4 X 0.4 
mm near the base, glabrous to pubescent, some- 
times tardily deciduous; petiole adaxially grooved, 
10—30 mm long, densely pubescent with erect hairs 
and scattered minute purple glands; petiolar gland 
solitary, located along the upper half of the petiole, 
sessile, circular to elliptic, 0.6-1.2 mm long, sau- 
cer-shaped to cup-shaped, glabrous, sometimes ab- 
sent; rachis adaxially grooved, 1 mm long, 
pubescent with erect hairs and scattered minute 
purple glands, a sessile, saucer-shaped gland, 0.5— 
1.1 mm across, between the pinnae of the upper 1 
or 2 pinna pairs; pinnae 5 to 13 pairs per leaf, 25— 
49 mm long, 4—8 mm between pinna pairs; petio- 
lules 1.5-2.1 mm long; leaflets 14 to 35(44) pairs 
per pinna, opposite, 0.7—1.3 mm interval between 
leaflets, linear, 2.5-5.0 х 0.8-1.5 mm, pubescent 
on both surfaces with appressed to erect hairs, lat- 
eral veins obvious with only one vein from the base, 
base oblique, margins ciliate, apex acute to obtuse. 
Inflorescence a densely flowered cylindrical spike 

0 mm long, solitary from the leaf axil; pedun- 
cle 3-20 X 0.7-1.2 mm, densely pubescent with 
erect hairs; involucre absent; floral bracts linear, to 
3.5 mm long, densely pubescent, deciduous. Flow- 
ers sessile, creamy white; calyx 5-lobed, 1.5-2.2 
mm long, densely pubescent with erect hairs; co- 
rolla 5-lobed, 2.0-3.0 mm long, densely pubescent 
with erect hairs; stamen filaments 6-8 mm long; 
ovary glabrous, on a stipe to 0.3 mm long. Legumes 


540 


Annals of the 
Missouri Botanical Garden 


light yellowish brown, straight, flattened, oblong, 

170 x 4 mm, cartilaginous, transversely 
striate, pubescent, eglandular but usually with mi- 
nute purple glands when young, dehiscent; stipe to 
14 mm long; apex acuminate and apiculate to 5 
mm long. Seeds uniseriate, no pulp, dark purplish 
brown, nearly circular, strongly flattened, 5.0—8.5 
mm across, smooth; pleurogram U-shaped, 1.5-2.5 
mm across. Flowers: February-June. 


Distribution. Rocky desert and dry thorn-scrub 
forests from 1100 to 2000 m elevation in Puebla 
and Oaxaca, Mexico (Fig. 2). 

A small tree, mostly 3 or 4 m tall, Acacia sericea 
is known from southeastern Puebla and adjacent 
Oaxaca. It occurs at higher elevation, usually above 
1100 m elevation, in desert and thorn-scrub forests. 
Most collections are from roadsides, usually in dry, 
disturbed habitats, and many are from the Tehu- 
acán valley (Rico Arce & Rodríguez, 1998). Based 
on the number of specimens available for study, 
this taxon is not a common component of the veg- 
etation. 

Acacia sericea is distinct from most other mem- 
bers of this group. The dense, erect pubescence on 
most parts of the plant makes it easy to distinguish 
this taxon. The leaf rachis, the pinna rachis, and 
usually the petiole are densely pubescent with erect 
hairs that exceed 0.3 mm in length. The leaflets are 
mostly pubescent with erect to slightly appressed 
pubescence, whereas the mature fruits are short pu- 
bescent. Also, the calyx and corolla are pubescent 
with erect hairs, as are the floral bracts, which are 
commonly longer than those found in other mem- 
bers of this group. 

Acacia sericea possibly hybridizes with A. aca- 
tlensis in areas where they are sympatric. Occa- 
sional specimens were encountered with reduced 
pubescence and many leaflet pairs per pinna, char- 
acteristics usually associated with A. acatlensis. 


За еа € MEXICO. Oaxaca: Cuest 
inferior de Sa , Districto de Cuicatlán, Conzatti 5321 
(NY*); Lien de pila Vieja, Smith 430 (GH*); a 8 km al 
NE de Teotitlán del Camino, carretera a Huautla, Sousa 
€: (CAS, MEXU, MO, UC*); а 12 km al N de Tonal- 

tepec, Sousa & Magallanes 8917 е а 8 km al МЕ 
de Теош Ап del Camino, en el camino a Huautla, Sousa, 
Martínez, Téllez & Magallanes 5396 (CAS, MEXU*, US, 
WIS); a 9 km al NE de Teotitlán del Camino, Sousa, Mar- 
tínez, Téllez & Magallanes n (CAS*, US); a 9 km al 
i . Sousa, Ramos & Téllez 
6131 (CAS, MO, SD*, WIS); a 8 km al E-NE де Тео ап 
del Camino, camino а Huautla, Sousa, Téllez, Germán & 
Rico 8083 (CAS*); Cafiada de Carrizalillo, Cerro Verde, 
MEXU*); 
retera a они а de Jiménez, 
Torres & Martinez 6499 (MEXU*. MO). Puebla: Cerros 
calizos al NE de Tehuacán, Chiang, ИЦазейог & Durán 


F-2024 (MO, NY*); Tehuacán, Conzatti 2176 (F*, US); 3 
mi. N of the city limits of Tehuacán, Hansen, Hansen & 
Nee 1728 (LL, MICH, US*, WIS); about 10 mi. N of Te- 
huacán on main road toward Orizaba, Hughes, Lewis & 
Contreras 1320 (MEXU, NY*); Cerro de Solumta, Purpus 
3863 (MO*); Tehuacán, Purpus 5845 (F, MO, NY*, UC, 
US); Tehuacán, Purpus 10682 (NY, US*); Ac aie pec, SW 
Zapotitlán, Sousa 2668 (CAS*, US); Ladera W de Cerro 
Grande Mpio. Caltepec, Tenorio & Romero 5424 (CAS*); 
Barranca de Los Membrillos, al SW de Caltepec, Tenorio, 
Torres & Romero 3784 (CAS*) 


T 


4. Acacia mammifera Schlechtendal, Linnaea 
. 1838. Senegalia mammifera (Schle- 
chtendal) Britton & Rose, N. Amer. Fl. 23: 
112. 1928. TYPE: Mexico. Hidalgo: Barranca 
de Acholoya, C. Ehrenberg s.n. (holotype, 
HAL!; isotype, UC!). 


Shrub or small tree to 5 m tall with bark dark 
gray, shallowly fissured; twigs light brown to pur- 
plish brown, not flexuous, usually puberulent and, 
when young, commonly with minute purple glands; 
short shoots absent. Leaves alternate, 30-130 mm 
long; stipules herbaceous, light brown, narrowly tri- 
angular, 2.5 X 0.6 mm near the base, glabrous to 
puberulent, persistent; petiole adaxially grooved, 
8–50(70) mm long, usually lightly puberulent and 
commonly with minute purple glands; petiolar 
gland solitary, located between the lower pinna pair 
or rarely along the upper half of the petiole, 
stalked, circular, 0 8 mm across, apex globose, 
glabrous (Fig. 4D); rachis adaxially grooved, 10-70 
mm long, puberulent and commonly with minute 
purple glands, with a stalked gland with a globose 
apex, 0.4—0.6 mm across, between most pinna 
pairs; pinnae 1 to 6(9) pairs per leaf, 30-85 mm 
long, 6-15(25) mm between pinna pairs; petiolules 
2.0-2.8(4.5) mm long; leaflets 10 to 26(33) pairs 
per pinna, opposite, 1-5 mm between leaflets, ob- 
long, 4—12 X 1.5-3.5(4.5) mm, loosely pubescent 
on both surfaces with appressed hairs, commonly 
purplish above, light green to purplish green be- 
neath, lateral veins obvious with a midvein and 1 
to 3 smaller veins from the base, base oblique, mar- 
gins ciliate, apex obtuse to broadly acute. Inflores- 
cence a loosely flowered cylindrical spike 90 
mm long, solitary (rarely 2) from the leaf axil; pe- 
duncle 6-15 X 0.7-1.1 mm, puberulent and with 
minute purple glands; involucre absent; floral 
bracts linear, to 1.4 mm long, pubescent, early de- 
ciduous. Flowers sessile, creamy-white; calyx 5- 
lobed, 1.3-2.0 mm long, lightly appressed pubes- 
cent; corolla 5-lobed, 2.2-3.5 mm long, P 
appressed pubescent; stamen filaments 6.5-8. 
long; ovary glabrous, on a stipe to 0.3 mm lor 
Legumes light yellowish brown to dark greenish 


Моште 87, Митбег 4 


Jawad et al. 
Acacia coulteri Group 


541 


brown, straight, flattened, oblong, 80-240 x 18- 
35 mm, cartilaginous, transversely striate, glabrous, 
eglandular, dehiscent; stipe to 12 mm long; apex 
acuminate and usually beaked. Seeds uniseriate, no 
pulp, dark brown, nearly circular, strongly flat- 
tened, 8.0—10.5 mm across, smooth; pleurogram U- 
shaped, 3—4 mm across. Flowers: April-June. 


Thorn-scrub forests and from the 
pinyon-juniper zone in dry thickets, and rocky 
slopes from 1300 to 2700 m elevation in eastern 
Mexico from the states of Tamaulipas and Nuevo 
León south to Oaxaca (Fig. 3). 

Acacia mammifera is widely distributed through- 


Distribution. 


out the central part of Mexico from the state of 


Oaxaca, north to Tamaulipas and Nuevo León. It 
does not appear to be a common species, however, 
many of the collections being from near the same 
localities in the various states. All collections ex- 
amined are from above 1300 m, many from near 
the pinyon-juniper zone, or from rocky slopes and 
dry thickets, mostly associated with thorn-scrub 
vegetation. The few pinna pairs, the large leaflets 
with appressed hairs on both surfaces, the dull pur- 
ple color on the upper surface of the leaflets, the 
narrowly triangular stipules, and the stalked peti- 
olar glands can be used to separate Acacia mam- 
mifera from other members of this group. 

This wide-ranging species is not extremely var- 
iable morphologically, but one unusual specimen 
from Puebla, Mexico, was encountered [E. M. Mar- 
tínez S. 21665 (MEXU)]. This was a small, compact 
plant with the small leaves and leaflets of A. com- 
pacta, but was similar to A. mammifera in lacking 
short shoots, having narrowly triangular stipules, a 
stalked petiolar gland with a bulbous apex, and leaf- 
lets that were purplish and pubescent on both sur- 
faces. Occurring at an elevation of 2300 m, this 
specimen was probably outside the natural range of 
A. compacta. The status of this possible hybrid must 
await additional field studies. 

Representative specimens. MEXICO. Guanajuato: 
Rancho Beltrán, 10 km al 5 de Xichá, Ventura & López 
6778 (CAS, MEXU*). Hidalgo: Barranca de Tuzanapa, 
Zacualtipán, González 824 (MEXU*); Barrancas de Tolan- 
tongo, Cardonal, Hernández 3775 (MO*); 6 km al N de 
Zoquital, Hernández, Cortés & Hernández 6036 (CAS, 
MO*); a 3 km al N de Molanguito, Tolantongo, Cardonal, 
Medrano, Hiriart & Ortíz 10083 (MEXU*, МО); Sierra de 
la Mesa, Rose, Painter & Rose 9126 (GH, NY, US*). Nue- 
vo León: а 24 km al S de Са. Linares, Castillón 687 


Chilapa, Mendoza 930 (CAS, MO, MU*, WIS); a 4 km al 
SW de home ть sobre la carretera Tamazulapán-Chi- 
lapa de Díaz, Rico, Torres & Cedillo 333 (CAS*, MO). 
Puebla: Barranca Tends al NW de Caltepec, Tenorio & 


Romero 5777 (ARIZ*, CAS, CM, TEX, WIS); Cerro Te- 
pearco, al E de el Rancho | Tlacuiloltepec, Tenorio & 
Romero 8814 (MO*, RSA, TEX). Querétaro: about 80 
km NE of Querétare on uM to Pinal de Amoles, McVaugh 
10364 (GH, LL, MICH*, MO, TEX, US); 3-4 km al Pon- 
iente de La Parada, Servin 117 (CAS*, MEXU); Cuesta 
Colorada, km 21 ae la carretera Vizarrón-Jalpán, Tenorio 
& Hernández 272 (CAS, ILL*, MO). San Luis Potosí. 5 
km W of jct. of hwy. 86 with roads to Rayón and Cárdenas, 
82 km W of Valles, Roe & Roe 2220 (NY*, WIS). Ta- 
maulipas: ca. 5 km N of La Joya de Salas, trail to Car- 
abanchel, Paid 129 (MICH*); 8 mi. E of Dulces Nom- 
bres, Meyer & Rogers 2640 (GH, MO*); 15 km al N de 
Tula, Puig 4741 (MEXU*). 


8. Acacia durangensis (Britton & Rose) Jawad, 
Seigler & Ebinger, comb. nov. Basionym: Se- 
negado durangensis Britton & Rose, N. Amer. 

1. 23: 112. 1928. TYPE: Mexico. Durango: 
San Ramón, 21 Apr.-18 May 1906, E. Palmer 
107 (holotype, NY*!, MEXU photo!; isotypes, 
F!, MO!, UC!, 05!). 


Shrub or small tree to 5 m tall with bark of main 
trunk dark gray, shallowly fissured; twigs light 
brown, not flexuous, puberulent; short shoots ab- 
sent. Leaves alternate, 65—160 mm long; stipules 
herbaceous, light brown, narrowly triangular, to 2.5 

.7 mm near the base, puberulent, persistent; 
petiole adaxially grooved, 30-50 mm long, puber- 
ulent and with erect hairs to 0.2 mm long; petiolar 
gland solitary, located near the middle of the peti- 
ole, sessile, elliptical, 1.1-2.2 mm across, apex ir- 
regularly raised, glabrous (Fig. 4A); rachis adaxi- 
ally grooved, 5 0 mm long, puberulent and 
usually with minute purple glands, a sessile, flat- 
tened gland, 0.4–0.8 mm across, between the pin- 
nae of the upper 1 to 2 pinna pairs; pinnae 6 to 13 
pairs per leaf, 60-85 mm long, 8-14 mm between 
pinna pairs; petiolules 2.5—4.0 mm long; leaflets 28 
to 36 pairs per pinna, opposite, 1.3-2.1 mm be- 
tween leaflets, linear, 5.0-7.5 X 1.3-2.1 mm, 
loosely pubescent on both surfaces with appressed 
hairs, commonly purplish above, light green to pur- 
plish green beneath, lateral veins obvious with a 
midvein and 1 to 2 smaller veins from the base, 
base oblique, margins ciliate, apex obtuse to acute. 
Inflorescence a loosely flowered cylindrical spike 
60-120 mm long, solitary (rarely 2) from the leaf 
axil, or rarely in short racemose clusters; peduncle 
5-15 x 1.0-1.8 mm, puberulent; involucre absent; 
floral bracts linear, to 1 mm long, pubescent, early 
deciduous. Flowers sessile, creamy-white; calyx 5- 
lobed, 1.0-1.4 mm long, densely appressed pubes- 
cent; corolla 5-lobed, 2.0-2.5 mm long, densely p 
pressed pubescent; stamen filaments 5.5-7.5 m 
long; ovary glabrous, on a stipe to 0.4 mm ia 
Legumes dark reddish brown, hah flattened, 


Annals of the 
Missouri Botanical Garden 


oblong, 80-120 X 16-22 mm, cartilaginous, trans- 
versely striate, glabrous to lightly puberulent, 
eglandular, dehiscent; stipe to 10 mm long; apex 
acuminate and usually beaked. Seeds not seen. 
Flowers: April-June. 


Distribution. Thorn-scrub forests and dry thick- 
ets, 1600 to 2200 m elevation in the states of Du- 
rango and Chihuahua, Mexico (Fig. 3). 

Few specimens of Acacia durangensis are avail- 
able for study; we saw the type and two others. This 
is the only species of this group known from Du- 
rango, and it occurs in a region that has been poorly 
collected. Superficially, A. durangensis is similar to 

. mammifera, the leaflets being purplish above, 
light green beneath, with obvious veins and loosely 
pubescent on both surfaces with appressed hairs. 
Leaves with 6 to 13 pinna pairs, pinnae with 28 to 
36 leaflets, and the sessile petiolar gland that is 
elliptical in outline and with a raised apex separate 
this taxon from A. mammifera. The petiolar gland 
is the most distinctive feature of A. durangensis. 
Other species of this group have a flat or doughnut- 
shaped gland, or the gland is stalked. In A. dur- 
angensis, in contrast, the sessile gland appears as 
an elliptical mound, with a few indentations. On 
herbarium specimens, this gland has a purple color, 
and rarely a few long hairs on its surface. 

Though none were observed on the few speci- 
mens available, it is possible that plants of Acacia 
durangensis occasionally may have prickles. If pre- 
sent, this would suggest that A. durangensis is more 
closely related to A. macilenta and other members 
of the Acacia series Vulgares that commonly have 
prickles. Also, the petiolar gland of A. durangensis 
is similar to those found in many members of Aca- 
cia series Vulgares. 


Representative specimens. MEXICO. Chihuahua: be- 
tween El Tejebán and Río Unique, Bye, Davis, Randolph 
& Gerson 12774 (MEXU*) еа El Pino 20 km E 
el entronque a Sapioris, con la Brecha Coyotes-San 

uel de Cruces, Mpio. Tayoltita, 2 105?49' W, Ten- 
orio, Romero & Ramamoorthy 6323 (TEX*). 


9. Acacia willardiana Vue in ir sey & Rose, 
Contr. U.S. Natl. Herb. 1: 88. 1890. TYPE: 
Mexico. Sonora: rocky is and ledges on 
the coast of Guaymas, 1-2 Apr. 1890, 
Palmer 164 (holotype, US!). 


Prosopis о Bentham, London J. Вог. 5: 82. 
1846. "ed А (Bentham) Britton & 
Rose a Amer. Fl. 23: . 1928. : Mexico. 
Sonora: Alta, 1830. T. us г s.n. фур, TCD) 
[not Acacia heterophylla Willdenow, 1806]. 


Tree to 10 m tall with bark smooth, white to red- 


dish yellow, exfoliating and papery; twigs light gray, 
becoming dark reddish purple, not flexuous, gla- 
brous; short shoots absent. Leaves alternate, 30— 
400 mm long; stipules herbaceous, light brown, 
narrowly linear, 1.1 х 0.2 
brous, tardily deciduous; petiole flattened, not 
grooved, 20—400 mm long, usually glabrous; peti- 
olar gland solitary, located between to just below 
the lower pinna pair, sessile, circular, 0.3—0.7 mm 
across, doughnut-shaped, glabrous (Fig. 4G); rachis 
flattened, not grooved, 0— 


mm near the base, gla- 


mm long, glabrous, 
glands absent; pinnae 1 (rarely 3) pair per leaf, 16— 
80 mm long; petiolules 2.5-8 mm long; leaflets 4 
to 20 pairs per pinna, opposite, 1-5 mm distance 
between leaflets, oblong to elliptic, 3.0-7.5(12.0) x 
1.0-2.3 mm, glabrous to rarely lightly pubescent 
with appressed hairs on both surfaces, lateral veins 
not obvious with only one vein from the base, base 
oblique, margins lightly ciliate, apex narrowly acute 
to acuminate. Inflorescence a loosely flowered cy- 
lindrical spike 30-90 mm long, solitary from the 
leaf axil, or in short racemose clusters. Peduncle 
5-25 X 0.4—0.8 mm, glabrous or nearly so; invo- 
lucre absent; floral bracts linear, to 1 mm long, gla- 
brous to lightly pubescent, early deciduous. Flow- 
ers sessile, creamy-white; calyx 5-lobed, 1.3-2.2 
mm long, glabrous; corolla 5-lobed, 2.4—3.6 mm 
long, glabrous; stamen filaments 6—8 mm long; ova- 
ry glabrous, on a stipe to 1 mm long. Legumes light 
yellowish brown, straight, flattened, oblong, 70-180 
mm, chartaceous, irregularly striate, gla- 
brous, eglandular, dehiscent; stipe to 14 mm long; 
apex obtuse. Seeds uniseriate, no pulp, dark brown, 
nearly circular, strongly flattened, 6-11 mm across, 
smooth; pleurogram usually absent, when present, 
U-shaped, about 2 mm across. Flowers: February— 
une. 


Distribution. Arid hills, rocky slopes and wash- 
es in desert scrub vegetation between sea level and 
500 m elevation in Sonora, Mexico (Fig. 3). 

A common species at lower elevations in the 
state of Sonora, Acacia willardiana is a very obvi- 
ous component of the desert scrub of this region 
because of it nearly white, to yellowish, to almost 
reddish, papery, exfoliating bark. Being so obvious, 
this species is commonly collected; numerous spec- 
imens are from the vicinity of Guaymas. The ma- 
jority of the specimens lack pinnae, which are early 
deciduous; only the elongated, flattened petioles 
persist. Although flowering is common from Feb- 
ruary through May, it may occur at other times if 
moisture is available. 

Acacia willardiana is easily separated from other 
members of this group. The most obvious differ- 


Volume 87, Number 4 
2000 


Jawad et al. 543 


Acacia coulteri Group 


ences are the elongated, flattened petioles that may 
reach 400 mm in length, and the leaves with usu- 
ally only one pair of pinnae, though rarely two or 
three may be present. This species has very small, 
tardily deciduous stipules (1.1 mm long or less), 
and the fruit valves are papery with more irregular 
striations than those found in other members of this 
group. The characteristic papery, exfoliating bark is 
shared only with A. salazari, a species of this group 
restricted to central Mexico. 

Bentham (1846) tentatively assigned this taxon 
to Prosopis heterophylla based on a single fruiting 
specimen. He suggested that the general habit of 
the plant was more like that of Prosopis than any 
other genus, and mentioned the almost phyllodi- 
nous vertical expansion of the petiole. Based on 
flowering material, Vasey and Rose (1890) realized 
that this taxon was an Acacia and used the name 
A. willardiana, as Bentham’s name was preoccu- 

ied. 

Vassal (1972) noted that A. willardiana is a spe- 
cies with authentic phyllodes and suggested that, if 
he had seen flowering material, Bentham would 
have placed this species in the “Juliflorae.” Vassal 
considered this species to be a member of his sec- 
tion Heterophyllum, subsection Spiciferae, and to 
have affinities to this predominantly Australian 
group. In our view, homology between the phyllodes 
of that group and the apparent petioles of A. wil- 
lardiana should be examined more thoroughly. Fur- 
ther, in most other characteristics, y species dif- 
fers little from other members of the A. coulteri 
group discussed in the present study 

Representative specimens. MEXICO. Sonora: rocky 
hill N of Guaymas, Blakley B-820 (ASU*); % mi. from top 
of microwave tower hill at Guaymas, Brunner 57 (ARIZ* 
Bahfa de San Carlos, ca. = km NW of Guaymas, Carter 
& Kellogg 3246 (SD*); ca. % mi. SW of Hotel Playa de 
Cortés, Miramar, Felger 5513 (ARIZ*); Isla Tiburón, Fel- 
ger 9151B (ARIZ*, SD); Sierra Seri, Hast Eemla, Felger, 
Drees & Moser 74-8 (ARIZ*); Bahía Colorado, arroyo at E 
base of Morro Colorado, Felger & Hamilton 15638 
(RSA*); Ensenada Grande, San Pedro Bay, Felger, Russell 
& Kleine 11588 (ARIZ, SD*); Sierra Bojihuacame SE of 
Cd. Obregón, Gentry 14483 (ARIZ*, US); 16 mi. S of Her- 
mosillo, uri : ^c e 551A (ASU*); 5 mi. N of Guay- 
mas near hwy. 15, Henrickson 1565 (RSA*); Bahía San 
Carlos, Fouad т 5рһоп s.n. (RSA*); Hermosillo, Jones 
Pm (MO, POM*); microwave tower, 60 mi. N of Navi- 
joa, Joseph s. n. (ARIZ*); hillside N of Guaymas, Knobloc h 


, MO, RSA); 
ry SE of Hermosillo, Moris s.n. (ARIZ*); 
Ardilla Island, ure Harbor, Moran 4017 (SD*, 
WIS); rte. 15, са. 25 mi. S of Hermosillo, Pinkava P12788 
(ARIZ, ASU*); dry hillside slopes near Torres, Whitehead 
M250 (ARIZ*). 


10. Acacia millefolia S. Watson, Proc. Amer. 


Acad. Arts 21: 427. 1886. Senegalia millefolia 
(S. Watson) Britton & Rose, N. Amer. Fl. 23: 
111. 1928. TYPE: Mexico. Chihuahua: Haci- 
enda San lags near Batopilas, Aug. 1885, Е. 
Palmer 45 (lectotype, designated es Isely 
(1969), an. жч m MEXU!, NY!). 


Shrub or small tree to 3 m tall with bark gray, 
smooth when young, becoming fissured into square 
plates 1-2 cm across; twigs light brown to greenish 
brown, not flexuous, usually lightly puberulent; 
short shoots absent. Leaves alternate, 60-230 mm 
long; stipules herbaceous, light brown, narrowly 
linear, 6.5 X 0.5 mm near the base, usually gla- 
brous, persistent; petiole adaxially grooved, 30—75 
mm long, usually glabrous; petiolar gland absent; 
rachis adaxially grooved, 50-190 mm long, gla- 
brous to lightly pubescent; a stalked gland with a 
globose apex, 0.3—0.9 mm across, between the pin- 
nae of the upper 1 to 2 pinna pairs; pinnae (2)6 to 
14 pairs per leaf, 30-55 mm long, 10-28 mm be- 
tween pinna pairs; petiolules 2.0—4.0 mm long; leaf- 
lets 20 to 35(37) pairs per pinna, pe 0.8-1.6 
mm between leaflets, oblong, 2.0—6.5 X 0.7-1.4 
mm, glabrous above, lightly pubescent beneath 
with appressed hairs, lateral veins not obvious with 
only one vein from the base, base oblique, margins 
sometimes ciliate, apex acuminate. Inflorescence a 
loosely flowered cylindrical spike 30-75 mm long, 
solitary (rarely 2 to 3), from the leaf axil; peduncle 
5-15 X 0.3-0.8 mm, glabrous to lightly puberu- 
lent; involucre absent; floral bracts linear, 10 1.3 
mm long, glabrous to lightly pubescent, early de- 
ciduous. Flowers sessile, creamy-white; calyx 5- 
lobed, 1.1-1.6 mm long, glabrous; corolla 5-lobed, 
2.0-2.7 mm long, glabrous; stamen filaments 4.5— 
6.5 mm long; ovary glabrous, on a stipe to 0.4 mm 
long. Legumes light yellowish brown, straight, flat- 
tened, oblong, 70-170 х 12-21 mm, chartaceous, 
irregularly striate, glabrous, eglandular, dehiscent; 
stipe to 12 mm, apex acute to obtuse. Seeds un- 
iseriate, no pulp, dark brown, nearly circular, 
strongly flattened, 6.2-9.5 mm across, smooth; 
pleurogram U-shaped, 2-3 mm across. Flowers: 
June—August. 


Distribution. Desert grasslands, rocky slopes, 
subtropical scrub, and open oak woodlands from 
700 to 1700 m elevation in southern Arizona, south 


through Sonora to western Chihuahua, Mexico (Fig. 


A shrub or small tree, mostly less than 3 m tall, 
Acacia millefolia is relatively common in desert 
grassland and desert scrub vegetation in extreme 
southern Arizona and adjacent Sonora, Mexico. 
Commonly collected along the steep sides and 


544 


Annals of the 
Missouri Botanical Garden 


floors of canyons, A. millefolia is rarely a dominant 
member of the vegetation, most collections indicat- 
ing scattered individuals. 
ough numerous specimens are available from 

throughout most of the geographic range of this tax- 
on, no specimens, other than the type collection, 
are known from Chihuahua. This taxon may be ex- 
tremely rare in southwestern Chihuahua, or it is 
possible that the collecting data on the type spec- 
imens are incorrect. This collection is more than 
100 km east of any specimens of A. millefolia seen 
by the present authors 

Acacia millefolia is distinct from other members 
of this group, as it lacks a petiolar gland and pos- 
sesses a stalked rachis gland between the upper 
pairs of pinnae. The only other taxa of this group 
within the range of A. millefolia are A. willardiana 
and A. russelliana. Acacia millefolia is easily dis- 
tinguished from A. willardiana, which has papery, 
exfoliating bark and leaves with only 1 to 3 pinna 
pairs. Leaflets with appressed hairs, the stalked ra- 
chis glands, and the persistent stipules of A. mil- 
lefolia separate it from A. russelliana. In addition, 
A. millefolia is normally a shrub found above 700 
m elevation, whereas the other species are usually 
trees and commonly occur at lower elevations. 


ey oan a MEXICO. Sonora: 3 km by 


road 5 of Nac 1, Felger 3596 (ARIZ*, MICH, SD); 4 


mi. N of Coloris ye, Hastings & Turner 65-25 (ARIZ, 
SD*); 5 mi. E of Mina Verde, Shreve 6753 w | MICI, 
MO); Río Bavispe, Colonia Oaxaca, White 6 9 (ARIZ 


GH, MICH); Саћоп ps Bavispe, White 3019 (ARIZ. G H, 
MICH*); 9 mi. W of La Angostura, White 4034 (ARIZ, 
GH, MICH*); б де! Molino preg Е of Colonia 
Morelos, White 4511 (ARIZ*, GH, MIC 
isaderos, Wiggins 7457 (A, ARIZ, CH, Е, 

.S.A. Arizona: Cochise Co.: foothills of the Ei illo 
Mts., T2285 КЗ2Е 532, Kluever s.n. (ARIZ*). Pima Co.: 
Box Canyon, Santa Rita Mts., Goodding 89-53 (ARIZ*); 
Chimney Creek, Rincón Mts., Kearney & Peebles 10461 
(ARIZ*, MICH, UC, US); Box Canyon, N end of Santa 
Rita Mts., McKeighen s.n. (ARIZ, ASU*, UC); 7 mi. W of 
hwy. 83 on Greaterville-Madera Canyon Road, Mc- 
Laughlin 64 (ARIZ*); vic. of Helvetia, Santa Rita Mts., 
rapida ы (ARIZ, ASU*); Greaterville Road, 5.4 mi. 

| 3, Pinkava, Keil & Lehto 14487 (ASU*); foot- 
hills of the pi Rita Mts., Pringle s.n. (А, F, GH, MICH, 

Total Wreck Mines, о 107 (МО, РОМ, 
UC*, U 3) Ophir Gulch, 4 mi. N and 2 mi. W of Sonoita, 
Tramontano T-25 (ARIZ*); din National Monument, 
urner & Gunzel 78-111 (ARIZ*, SD). Santa Cruz Co.: 
about % mi. N of Sonoita Creek above Rio Rico, Kaiser 
1394 (ARIZ*). New Mexico: Hidalgo Co.: about 0.5 mi. 
E of Arizona, about 50 m N of Mexico border, wedge Sec. 
24 1245, R22W, Spellenberg & Repass 5371 (ММС). 


11. Acacia salazari (Britton & ien SFA 
Contr. Univ. Michigan Herb. 4: 8. 1940 
negalia salazari Britton & Rose, " ке г 


23: 113. 1928. TYPE: Mexico. Michoacán: 
Xochiapa, 13 Apr. 1912, Е Salazar s.n. (ho- 
lotype, US!; isotypes, MEXU!, МУ!). 


Tree to 15 m tall with bark yellow to red or light 
gray, exfoliating and papery; twigs greenish brown 
to light reddish brown, not flexuous, usually gla- 
brous; short shoots absent. Leaves alternate, 50— 
180 mm long; stipules herbaceous, light brown, 
narrowly linear, 4.5 X 0.6 mm near the base, gla- 
brous, persistent; petiole adaxially grooved, 20— 
50(60) mm long, mostly glabrous; petiolar gland 
solitary, located between the lower pinna pair, ses- 
sile, nearly circular, 0.5—1.3 mm across, globose to 
doughnut-shaped, absent on most leaves; rachis 
adaxially grooved, 35-140 mm long, glabrous to 
lightly puberulent, a sessile, doughnut- to saucer- 
shaped gland, 0.5—1.0 mm across, between the pin- 
nae of the upper 1 or 2(3) pinna pairs; pinnae 7 to 
18 pairs per leaf, 35—63 mm long, 4–12 mm ђе- 
tween pinna pairs; petiolules 2.4—4.0 mm long; leaf- 
lets 20 to 38 per pinna, opposite, 0.8—1.5 mm be- 
tween leaflets, 0.9-1.3 mm, 
usually lightly pubescent with appressed hairs be- 
neath, commonly light greenish purple and gla- 
brous above, lateral veins usually not obvious with 
only 1(2) vein(s) from the base, base oblique, mar- 
gins usually ciliate, apex narrowly acute to acu- 
minate. Inflorescence a loosely flowered, cylindrical 
spike 45-110 mm long, 1 to 3 from the leaf axil, 
or rarely in terminal racemose clusters; peduncle 
5-10 X 0.5-1.0 mm, lightly puberulent; involucre 
absent; floral bracts linear, to 1 mm long, puberu- 
lent, early deciduous. Flowers sessile to rarely short 
stipitate, creamy-white; calyx 5-lobed, 1.5-2.3 mm 
long, lightly appressed pubescent; corolla 5-lobed, 

5-3.5 mm long, lightly appressed pubescent; sta- 
men filaments 4.5—7.0 mm long; ovary glabrous, on 
a stipe to 0.3 mm long. Legumes yellowish brown, 
straight, flattened, oblong, 115-180 x 21-35 mm, 


cartilaginous, transversely striate, glabrous, eglan- 


linear, 2. 


dular, dehiscent; stipe to 13 mm long; apex obtuse. 
Seeds uniseriate, no pulp, dark reddish brown, cir- 
cular to oblong, strongly flattened, 10.0-14.5 x 7– 
12 mm, smooth; pleurogram U-shaped, 1.2-3.0 mm 
across. Flowers: April-June. 


Distribution. Thorn-scrub thickets, and dis- 
turbed dry forests from near sea level to 1800 m 
(but mostly above 1000 m) elevation in the states 
of Guererro, México, Michoacan, Morelos, Oaxaca, 
and Puebla, Mexico (Fig. 3 

A small tree, not exceeding 15 m in height, Aca- 
cia salazari is restricted to dry habitats, particularly 
thorn-scrub forests of southern Mexico. Many col- 
lections are from above 1000 m elevation, but a 


Моште 87, Митбег 4 
2000 


Jawad et al. 
Acacia coulteri Group 


few individuals are reported from near sea level. 
Acacia salazari is similar to A. acatlensis with 
which it is sympatric nearly throughout its entire 
range. Both have leaves with many pinna pairs, and 
leaflets of similar size that are lightly pubescent 
with appressed hairs beneath and commonly light 
greenish purple and glabrous above. Acacia aca- 
tlensis, however, has minute purple glands along the 
petiole, rachis, and at the base of most leaflets; pin- 
nae with more than 36 pairs of leaflets; petiolules 
that are mostly less than 2.0 mm long; and dark 
gray, fissured bark. Acacia salazari, in contrast, 
lacks the small purple gland and usually has fewer 
than 38 pairs of leaflets per pinna, petiolules that 
usually exceed 2.5 mm in length, 
reddish bark that is papery and exfoliating. Some 
specimens with a mixture of characteristics of these 
two species suggest that they probably hybridize in 
Guerrero. Determination must await field studies. 
Although A. salazari is similar to other species 
of this group from southern Mexico, particularly A. 
centralis and A. usumacintensis, the papery, exfoli- 


and yellow to 


ating bark allows for easy separation. Also, both of 


these species have pinnae with more than (36)40 
leaflet pairs, whereas A. salazari has 38 or fewer 
leaflet pairs per pinna. Acacia usumacintensis has 
large, flattened petiolar glands that are located on 
the lower half of the petiole, while Acacia salazari 
commonly lacks petiolar glands. Occasionally, pet- 
iolar glands are found on a few leaves of some spec- 
imens; these are globose to doughnut-shaped, and 
located between the lower pair of pinnae. These 
glands are not common; most specimens lack pet- 
iolar glands altogether. On the remaining speci- 
mens, only one or two of the leaves have petiolar 
glands, and many are globose, not doughnut-shaped 
as is typical in most taxa of this group. Acacia sal- 
azari and A. millefolia are the only taxa in this 
group that usually lack petiolar glands. 


Representative specimens. MEXICO. Guerrero: creek 
banks N of Chilpancingo, Clark 7235 (MO*); 21 fen al 
NE de Chilpancingo, Delgado & Garcta 1085 (C } 
MEXU, MO, NY, WIS); a 20 km al N de eal питам 
camino a Iguala, M 557 (F*); N de Zumpango del 
Río, Rico & Funk 204 (CAS*); 24 km al N de Zumpango 
del Río hacia Iguala, Torres, Tenorio & Romero 1243 

ic u 


teaga, McVaugh 22520 (CAS, MICH*, MO, NY 
Cañas, cerca de la desviación al Infiernillo, carretera Nue- 
va Italia—Playa Azul, Soto 1081 (CAS, МОЖ); carretera 
Nueva Italia—Arteaga, Soto & Aureolea 7745 (MEXU*). 
Morelos: yard, Apuyeca, Clausen 6036 (CU*). Oaxaca: 
Rancho El Mezquite, a 7 km al S de Teotitlán, Sousa 6934 
(CAS, MO, SD, UC*); Teotitlán del Camino, en la orilla 
del Pueblo, Sousa, Téllez, Germán & Rico 8060 (CAS, 


МО“); a 3 km al E-NE де Teotitlán del Camino, carretera 
a Huautla, Sousa, Téllez, Germán & Rico 8079 (CAS*, 
TEX). Puebla: 20 mi. SE of Tehuacán on road to Teotitlán 
del Camino, Anderson & Anderson 5332 (MICH*); 3.5 km 
al N de Teotitlán, por el camino rumbo a Vigastepec, Cor- 
nelo 25 (MO*); Piaxtla, Huerta s.n. (NY*); just N of Cox- 
catlán along hwy. 131, Lavin, Sundberg, Hardison & Whit- 
temore 4610 (TEX*); El Papayo, a 16 km al NW de 
Amatitlán, Sousa, Pérez & Ruiz 4339 (NY*); Ahuehuetilla, 
a 20 km al NW de Acatlán, Sousa, Téllez & Magallanes 
5418 (MICH*, MO); hwy. 190, 17 mi. from the Oaxaca 
border in Puebla, Trott, Case, Thurm, Dunn, Hess & Dzie- 
kanowski 140 (MO, NY*). 


12. Acacia coulteri Bentham, in A. Gray, Pl. 

. 1852. pu coulteri (Ben- 

tham) Britton & Rose, N. Am : 112 

1928. TYPE: Mexico. Hidalgo: Пой Т. 

Coulter s.n. (holotype, К!, Е photo!, СН photo!, 

MEXU photo!, MICH photo!, MO photo!, NY 
photo!, US photo and fragment!). 


Shrub or small tree to 15 m tall with bark dark 
gray, shallowly furrowed; 
greenish brown, not flexuous, glabrous to lightly ap- 
pressed puberulent; short shoots absent. Leaves al- 
ternate, 50— mm long; stipules herbaceous, 
light brown, narrowly linear, 2.1 X 0.4 mm near 
the base, glabrous, tardily deciduous; petioles 
adaxially shallowly grooved, 25—55 mm long, usu- 
ally lightly appressed puberulent; petiolar gland 
solitary, located on the upper third of the petiole 


twigs light brown to 


and commonly just below the first pinna pair, ses- 
sile, nearly circular, 0.5—1.6 mm across, doughnut- 
shaped, glabrous, rarely absent (Fig. 4E); rachis 
shallowly grooved adaxially, 20-1 
lightly puberulent, a sessile, cup-shaped gland, 
0.4–0.9 mm across, between upper pinna pair; pin- 
nae 5 to 11 pairs per leaf, 40-90 mm long, 6–12 
mm distance between pinna pairs; petiolules 3—5 
mm long; leaflets 18 to 35 pairs per pinna, oppo- 
site, 1.5-2.3 mm interval between leaflets, oblong, 
4.5-7.5 X 1.4-2.1 mm, glabrous above, lightly ap- 
pressed pubescent beneath, lateral veins obvious 
with a midvein and occasionally one other vein 


mm long, 


from the base, base oblique, margins ciliate, apex 
broadly acute to obtuse. Inflorescence a loosely 
flowered cylindrical spike 50-90 mm long, 1 to 4 
from the leaf axil, or rarely in terminal racemose 
clusters; peduncle 7-13 X 0.5-1.0 mm, usually 
puberulent; involucre absent; floral bracts linear, to 
1 mm long, puberulent, early deciduous. Flowers 
sessile, creamy-white; calyx 5-lobed, 1.2-1.6 mm 
D lightly appressed pubescent; corolla 5-lobed, 

1.9-2.6 mm long, pop appressed pubescent; sta- 
men filaments 5.0—6.5 mm long; ovary glabrous, on 

a stipe to 0.4 mm b Legumes light yellowish 
brown to dark brown, straight, flattened, oblong, 


546 


Annals of the 
Missouri Botanical Garden 


100-185 X 16-25 mm, cartilaginous, transversely 
striate, glabrous, eglandular, dehiscent; stipe to 15 
mm long; apex acute to acuminate. Seeds uniser- 
iate, no pulp, dark reddish brown, circular to nearly 
oblong, strongly flattened, 7.3-10.5 X 5.5-8.5 mm, 
smooth; pleurogram U-shaped, 2.2-3.5 mm across. 
Flowers: April—August. 

Distribution. Open dry forest, dense thorn- 
scrub thickets, and dry rocky slopes below 1800 m 
elevation in the foothills and mountains of north- 
eastern Mexico in the states of Coahuila, Guana- 
juato, Hidalgo, Nuevo Гебп, Querétaro, San Luis 
Potosí, and Tamaulipas (Fig. 

Abundant in northeastern Mexico, Acacia coul- 
teri is a common component of relatively dry forests 
and thorn-scrub thickets. Many of the collections 
are from roadsides and rocky pastures. This taxon 
is abundant in the states of Tamaulipas and San 
Luis Potosí, becoming less common to the south. 
Acacia coulteri is similar morphologically to A. rus- 


selliana, which is common in Sonora and parts of 


Sinaloa. Acacia russelliana is usually a small shrub 
or understory tree that rarely exceeds 4 m in height. 
Acacia russelliana has usually been considered as 
conspecific with A. coulteri, Britton and Rose 
(1928) being the only authors that recognized the 
two as distinct. Both of these taxa have leaves with 
fewer than 12 pinna pairs, pinnae with fewer than 
35 leaflet pairs, petiolules that exceed 2.4 mm in 
length, and identical petiolar glands. Though sim- 
ilar in many traits, these taxa are easy to separate, 
A. coulteri having appressed pubescence on the 
lower leaflet surface, perianth, and the rachis and 
pinna rachises; A. russelliana, in contrast, being 
glabrous throughout. 

The stipules of Acacia coulteri seedlings are de- 
cidedly spinescent, but are only weakly rigid after 
the first leaf stage, and become progressively small- 
er and less rigid on older plants (Vassal, 1972). 


Because this species lacks prickles and because of 


correlations of the cotyledonary petiole and shape, 
as well as certain other features, Vassal placed A. 
coulteri in his subgenus Acacia, a group roughly 
equivalent to Bentham's (1842) Acacia series Gum- 
miferae. However, А. coulteri differs in some sig- 


nificant features from members of that series. Chief 


among them, the pollen grains of A. coulteri are 
porate, whereas those of other members of Ben- 
tham's series Gummiferae are colporate (Vassal, 


1972). 


Representative specimens. MEXICO. Coahuila: Саћоп 
: Puerto, Sierra de Santa бон, Villarreal, Vásquez, Gu- 
terrez & Urbina 5958 . Guanajuato: between Au- 
rora and Xichu, Mc Vaugh 14866 (MICH*); Mina de La 


=: 
tz 


Liga, Ventura. = oe 8935 (MEXU*). Hidalgo: 10 km 
al NW de Zimapán, González 2358 (DS, LL, MICH*). 
Nuevo León: кын N of Linares, Clark 6819 (MO*); 
El Cercado, Santiago, Hinton 24102 (NY*, TEX); Sierra 
de la Silla, Pringle 2549 (A*, CM, F, G, GH, MO, NY, 
PH, UC, US, VT. WIS); N slope of La Silla, White & 
Chatters 8 (GH, MICH*, TEX). Querétaro: 15 km al SE 
de Agua Zarca, Rubio e реј MO, TEX, WIS). 
San Luis Potosí: 10 mi. W of Tamuin on the Tampico- 
Valles hwy., Crutchfield & Johnston 5401 (MICH*, dr 
Minas de San Rafael, Purpus 5 ‚ С, GH, MO 
UC, US); 16 km W of Tamuin, Seigler, "Clarke & Puget 
13010 (EIU*, ILL). Tamaulipas: Ejido Los Angeles, 
Diaz, Cedilla & Jiménez 484 (ME XU): Camino a bá mina 
El Berrinche, Victoria, Jiménez 67 (MEXU*); 60 mi. N of 
Victoria, Hitchcock & Stanford 6867 (CU, DS, F, GH, 
ILL*, ISC, MO, NY, UC, US); along route 70 а 3 mi. 
S of Victoria, King 4498 (F, MICH, NY, TEX*,UC, US); 
2 km WNW of Gómez Farfas, Martin 71C MICH ү Г*); 
Puerto de la Angostura, km 658—60 between ај апа 
Mante, Moore, Jr. & ger | eid in BH, MICH, UC); 
Victoria, Nilson 4421 (GH : 33 mi. | of Mante, 
Seigler, Richardson & aoa 11627 (EIU*, ILL). 


13. Acacia russelliana (Britton & pu Lundell, 
Contr. Univ. Michigan Herb. 4: 7. 1940. Se- 
negalia russelliana Britton & Ro N. Amer. 

23: 112. 1928. TYPE: Mexico. Sinaloa: vi- 
cinity of San Blas, 22 Mar. 1910, J. N. Rose, 
P. C. Standley & P. G. Russell 13204 (ве 
US!; isotype, GH!, NY!). 


Shrub or small tree to 8 m tall with bark dark 
gray, shallowly furrowed; twigs light brown to 
greenish brown, not flexuous, glabrous; short shoots 
absent. Leaves alternate, 60—140 mm long; stipules 
herbaceous, light brown, narrowly linear, to 2.5 X 
0.4 mm near the base, glabrous, tardily deciduous; 
petiole adaxially shallowly grooved, 20-50 mm 
long, glabrous; petiolar gland solitary, located near 
the middle of the petiole to just below the lowest 
pinna pair, sessile, usually circular, 0.4—1.5 mm 
across, doughnut- to urn-shaped, 
times absent; rachis shallowly grooved adaxially, 

mm long, glabrous, rarely a sessile, dough- 
nut-shaped gland, mm across, between the 
upper pinna pair; pinnae (2)4 to 11 pairs per leaf, 
35-70 mm long, 5-12 mm between pinna pairs; 
petiolules 2.0—3.5 mm long; leaflets 18 to 34 pairs 
per pinna, opposite, 0.9-1.8 mm distance between 
leaflets, oblong, 4.0—7.5 X 1.3-1.8 mm, glabrous, 
lateral veins obvious with a midvein and 1 to 3 
smaller veins from the base, base oblique, margins 


glabrous, some- 


sometimes lightly ciliate, apex obtuse to broadly 
acute. Inflorescence a loosely flowered cylindrical 
spike 25—60 mm long, solitary (rarely 2 to 3) from 
the leaf axil, or rarely in terminal racemose clus- 
ters; peduncle 1-10 X 0.4—0.7 mm, glabrous; in- 
volucre absent; floral bracts linear, to 1 mm long, 


Моште 87, Митбег 4 
2000 


Jawad et al. 547 
Acacia coulteri Group 


glabrous, early deciduous. Flowers sessile, creamy- 
white; calyx 5-lobed, 1.0-1.4 mm long, glabrous; 
corolla 5-lobed, 1.7–2.5 mm long, glabrous; stamen 
filaments 5—7 mm long; ovary glabrous, on a stipe 
2-3 mm long. Legumes greenish brown to dark 
brown, straight, flattened, oblong, 55-170 X 16- 
27 mm, cartilaginous, transversely striate, glabrous, 
eglandular, dehiscent; stipe to mm long, apex 
acute to obtuse, sometimes beaked. Seeds uniser- 
iate, no pulp, reddish brown, circular to oval, 
strongly flattened, 6.8—9.6 х 5.1–8.0 mm, smooth; 
pleurogram U-shaped, 2-3 mm across. Flowers: 
March—August. 


Distribution. Dry, deciduous, tropical forests to 
thorn-scrub and desert-scrub vegetation, mostly on 
rocky slopes, from near sea level to about 700 m 
elevation in southwestern Sonora, and extreme 
northern Sinaloa, Mexico (Fig. 3). 

A small shrub or understory tree not exceeding 
8 m in height, Acacia russelliana is a common spe- 
cies of Sonora and parts of Sinaloa. Occurring at 
elevations below 700 m, it appears to be a common 
component of desert and thorn-scrub vegetation, 
and is also found as an understory tree in tropical 
deciduous forests. 

Britton and Rose (1928) were the first to recog- 
nize this taxon, all previous and subsequent authors 
considering it to be conspecific with Acacia coulteri, 
a species of northeastern Mexico. It is easily dis- 
tinguished from this taxon by being essentially gla- 
brous; A. coulteri, in contrast, has leaves that are 
lightly pubescent beneath with appressed hairs, 
and the petiole and rachis are lightly puberulent, 
as are many other vegetative parts of the plant. 
Also, the perianth of most flowers of A. coulteri is 
puberulent, but it is glabrous in A. russelliana. 

The only acacia species of this group found with- 
in the range of Acacia russelliana are A. willardiana 
and A. millefolia. Both taxa are easily separated 
from A. russelliana; A. willardiana by its extremely 
long, flattened petioles topped with 1 to 3 pairs of 
pinnae, and A. millefolia by the stalked gland with 
a bulbous apex between the upper pinna pair, the 
long, usually persistent stipules, and the missing 
petiolar gland. 

It is quite probable that Acacia russelliana oc- 
casionally hybridizes with A. willardiana in thorn- 
scrub forests on the arid, rocky slopes at lower el- 
evation in extreme northern Sinaloa, Mexico. The 
few specimens available suggest that the probable 
hybrid is similar to A. willardiana in being a small 
tree with exfoliating, papery bark, petioles that 
commonly exceed 100 mm in length, and pinnae 
with fewer than 26 pairs of leaflets that are rela- 


tively widely spaced. The proposed hybrid is sim- 
ilar to A. russelliana in having stipules to 2.5 mm 
long, petioles that are round in cross section and 
slightly adaxially grooved, a doughnut-shaped pet- 
iolar gland, and pei "i 2 to 11 pairs of pinnae. 
Few specimens are available for study [Gentry 

14337 (MICH, US), ne 14420 (LL, MICH, US), 
Gibson & Gibson 2101 (ARIZ, ASU), Rose, Standley 
& Russell 13317 (05)]; field studies will be nec- 
essary to determine the status of these specimens. 


Representative specimens. зет Sinaloa: vicinity 
of Culiacán, Brandegee s.n. (GH*, UC); Cerros de Na- 
vachiste about Bahía тра Gentry 14309 (US*); 
Cerros del Fuerte, 18-24 of Los Mochis, Gentry 
17635 (LL*); near г Yao ht Hotel Topolobampo, Hastings & 
Turner 64-103 (SD*); vicinity of Culiacán, Rose, Standley 
& Russell 14994 (US*). Sonora: 34 km E of Hermosillo 
on road to Sahuaripa, Felger, Aronson & Shmida 84-204 
(SD, TEX*); foothills at S end of Sierra Libre, 12.3 mi. S 
of La Palma on hwy. 15, Felger & Reichenbacher 85-1084 
(TEX*); SW me Ures, Felger 3000 cree LL*); Río So- 
nora, 22.2 mi. by road E of Mex. 15 on road to Ures, 
m 3628 (ARIZ*): Lake Mocuzari, үн оп 191-77 
(CAS*); Cerro Prieto, 8.6 пи. E of Navajoa, Sanders, Guzy, 
Way, Charlton & Moos 4618 (NY*, TEX); summit of 
Cerro Prieto, E of Navajoa, А Ballmer, 
Charlton, Clarke & pee 9286 (SD*), Rfo 


mi. of Alamos, Y^ ce Ballmer, а Clarke & 


(CAS*, MO, TEX, WIS); Alamos, Cerro La Luna, Sanders, 
Friedman, Spenger & Kossack 13260 (CAS*, MO, TEX); 
at crossing of Río Sonora, 23 mi. МЕ of El Sacatón, 19 
Sep 1934, Shreve 6703 (GH*, MICH, MO); Cerro Prieto, 
15 km al E de Navajoa, Tenorio, Romero, Ignacio & Dav- 
ila 10202 (F, CAS, MO, NY*); near the Mirador, Alamos, 
Devender & Lindquist 94- 828 (NY*); crossing the Rfo So- 
nora, 13 mi. S : 
UC, US*); 
Guaymas, Wiggins & Rollins 228 (A, LI 
МО, NY, UC). 


DS*, MICH, 


Literature Cited 


Вагпеђу, R. С. & J. W. Grimes. 1996. Silk Tree, Guana- 
caste, Monkey’s Earring. Mem. New York Bot. Gard. 
ol. 74, Part 1. 

Bentham, G. 1842. Notes on Mimoseae, with a synopsis 
of species. о Bot. 1: 318-392, 194—528. 

Notes оп a with a synopsis of 

TN London T Bot. 5: 75-108. 

————. 1875. Revision of the order Mimoseae. Trans. 

Linn. don: УЕ 5—664. 

Britton, N. L. & J. М. Rose. 1928. Mimosaceae. 
American T 23(2 ): 1-194. 

Chappill, J. A . R. Maslin. К т phylogenetic 
assessment of tr ibe  Асасіе еае. Рр. 7 99 in М. Crisp & 
J. Doyle (editors), Advances in Жыны enin ан 

“ла Royal Botanic Gardens, Kew. 

Clarke, H. . Systematics and evolutionary studies 
of eae series билион (Fabaceae: Mimosoideae). 
Ph.D. Dissertation, Department of os Biology, Uni- 
a of Illinois, Urbana—Champaig 

— ———, D. S. Seigler & J. Е. Ebinger "1980. Acacia far- 


North 


Annals of the 
Missouri Botanical Garden 


nesiana (Fabaceae: Mimosoideae) and related species 
from Mexico, the southwestern U. S., and the Caribbe- 
ап. Du Bot. 14: 549—564. 

& Acacia constricta (Fa- 
baceae: Mimosoideae) and related species from the 
southwestern U.S. and Mexico. Amer. J. Bot. 77: 305- 
315 


Conn, E. E., D. S. Seigler, B. R. Maslin & J. Dunn. 1989. 
Cyanogenesis in Acacia subgenus Aculeiferum. Phyto- 
chemistry 28: 817—820. 

Feigl, F. & V. A. Anger. 1966. Replacement of benzidine 
by copper ethylacetoacetate and tetra base as spot-test 
reagent for hydrogen cyanide and cyanogen. Analyst 91: 

82-284 

Isely, D. 1969. Legumes of the United States: I. Native 
Acacia. SIDA 3: 365-386. 

Maslin, B. R. & C. H. Stirton. 1997. Generic and infra- 
generic classification in Acacia (Leguminosae: Mimo- 
soideae): A list of critical species on which to build a 
comparative data set. Bull. Groupe Int. Étude Mimo- 
soideae. 20: 22-44. 

Pedley, L. 1978. A revision Lunas Mill. in Queensland. 
оа 75- 

ee ae dispersal of Acacia (Legu- 
Беу with particular reference to Australia, and the 
recognition of Senegalia and Racosperma. Bot. J. Linn. 
Soc. hie 219-254. 

Rico . & A. Rodrfguez. 1998. Mimosaceae R. Br. 
tim је 'acieae Benth. Fascículo 20. Flora del Valle de 
Tehuacán-Cuicatlán. Instituto de Biología, UNAM 


Rohlf, Е L 


1993. Numerical Taxonomy Systems-pc 


(NTSYS- Po) Version 1.70, Applied Biostatistics, Setau- 
ket, New 

Seigler, D. S vu E. Ebinger. 1988. Acacia macracantha, 
A. pennatula, and A. cochliacantha (Fabaceae: Mimo- 
soideae) species complexes in Mexico. Syst. Bot. 13: 7- 
15. 


995. Taxonomic revision of the ant- 
acacias (Fabaceae, Mimosoideae, Acacia, series Gum- 
miferae) of the New World. Ann. Missouri Bot. Gard. 

. Dunn, E. E. Conn & C. L. Holstein. 1978 
Acac peralin Бош six сы of Acacia of Mexic о and 
Texas. Рћугос TL 17: 445-446. 
ies В. В. & Е J. Rohlf. a Biometry. W. H. Free- 
n, San Francisco, Califor 
Standley, P. C. 1920. b ies and А of Mexico. Contr. 
U.S. Natl. бы: 23: 


Тапйвезпе, B., H. W. Г. 
Di 


ocn & R. Hegnauer. 1969 


ле п Sippen. Pharmaceu- 
Weekblad MA Nederland 104: 1341-1354. 
‚е, | L. 1 Revision of the Genus Lysiloma 
(Le ae). Ph D. Dissertation, Department of Bot- 
any, ео с University at Carbondale, Illinois. 
Vasey, G . 1890. List of plants collected by 
Dr. Edward Palmer in Lower rade and western 
Mexico in 1890. Contr. U.S. Natl. Herb. 1: 63-90 
Vassal, J. 1972. Apport des rec 'herches ontogéniques et 
seminologiques 
hylo 


. 1806. Species Plantarum. Berlin. (Aca- 
cia) 4 on 1049—1093. 


А REVIEW OF GAMOSEPALY 
IN THE BRASSICACEAE AND 
A REVISION OF DESIDERIA, 
WITH A CRITICAL 
EVALUATION OF RELATED 
GENERA! 


Ihsan А. Al-Shehbaz? 


ABSTRACT 


Gamosepaly i is reported in 12 genera of the Brassicaceae and is considered to have evolved jn зиду as many 


times. It is concluded that gamosepaly is not a useful character for the circumscription of gene 


pinnatifi 
As herein delimited, Christolea consists only two 


Desideria, Christolea, Ermania, Eurycarpus, Leiospora, 
Key won 


8 
BS 
=: 2 
= 
* 


da. с. scaposa, Desider ria pamirica, dein bifaria, E. kachoori, E. килиш, апа Ё. 
species; Erma 
Ermaniopsis and Oreoblastus are и to synonymy of Desideria 

Melanidion, жий Solmslaubachia are discussed. 


eight new synonyms (Christolea karakorumensis, C. 


ania 18 reduced to synonymy of Mela 
The MI RON and distinguishing characters of 


Brassicaceae, Christolea, Desideria, Ermania, Eurycarpus, gamosepaly, Leiospora, Melanidion, Solmslaubachia. 


During work on the Brassicaceae (Cruciferae) for 
the Flora of China, Flora of Nepal, and Flora of Ka- 
zakstan, it became evident that the limits of several 
Himalayan and Central Asian genera needed critical 
evaluation, and the nomenclature of many species and 
infraspecific taxa needed adjustments. The genera ad- 
dressed in the present paper are Christolea Cambess., 
Desideria Pamp., Ermania Cham. ex Botsch., Erman- 
iopsis H. Hara, Eurycarpus Botsch., Leiospora (C. A. 
Mey.) Dvořák, Melanidion Greene, Oreoblastus Sus- 
lova, and Solmslaubachia Muschl. They exhibit over- 
lapping similarities in several characters, and their 
limits have often been confused. 

Because Desideria was based on a species with 
a gamosepalous calyx, a review of gamosepaly in 
the Brassicaceae is presented to determine whether 
or not this character alone is sufficient to establish 
genera. The study led to the revision of Desideria 
and also critically evaluated the limits of several 
presumably related genera. 


GAMOSEPALY IN THE BRASSICACEAE 


Gamosepaly has been reported in at least 12 
genera of the Brassicaceae from Asia and South 


America. It was first reported by Oliver (1893) in 
Braya uniflora Hook. f. & Thomson. Hooker and 
Thomson (1861) and Hooker and Anderson (1872) 
did not report gamosepaly in the species even 
though the type collection has all flowers and fruits 
with persistent, united sepals. Schulz (1924) trans- 
ferred the species to the monotypic Pycnoplinthus 
O. E. Schulz, a genus restricted to China and Kash- 
mir (Jafri, 1973; Kuan, 1987; Hajra et al., 1993). 
Desideria mirabilis Pamp. (China, Kashmir, Ta- 
jikistan) is the second species reported to have a 
nd it too was placed in a 
monotypic genus (Pampanini, 1926, 1930). Hedge 
9 described | Sisymbrium | gamosepalum 
Hedge and Arabidopsis gamosepala Hedge, both of 
which are endemic to Afghanistan, but the latter 
species was transferred by Al-Shehbaz and O'Kane 
1997) to Neotorularia Hedge & J. Léonard. Sis- 
ymbrium L. (ca. 50 species; Al-Shehbaz, unpub- 
lished) is represented by indigenous species on all 
continents except Australia and Antarctica (Al- 
Shehbaz, 1988), whereas Neotorularia includes 
about 15 species distributed primarily in Central 


Asia and the Middle East (Al-Shehbaz, unpub- 


gamosepalous calyx, a 


=~ 
ке 


—. 


! | am most grateful to Zhu Guanghua and Song Hong for their help in the translation of Chinese text and herbarium 


labels, to Tatyana Shulkina for help with the Russian literature, and to Henk van 
nomenclatural problems. I am profoundly thankful to A. R. 


Turland, and Michael Gilbert for their advice on some 


Naqshi for sending duplicates of type material. Suzanne I. 
the manuscript. I also thank the curators and directors of the herbaria cited in this 


thanked for their critical review of 


der Werff, 


Gerrit Davidse, Nicholas 


Warwick, Peter Heenan, and an anonymous reviewer are 


paper. I am grateful to Oliver Appel and Juan Martínez-Laborde for bringing to my attention gamosepaly in Catadysia 


rosulans and Eudema friesii, respectively. 


? Missouri Botanical Garden, P.O. Box 299, St. Louis, Missouri 63166-0299, U.S.A. 
ANN. Missouni Bor. GARD. 87: 549—563. 2000. 


550 


Annals of the 
Missouri Botanical Garden 


lished). Sisymbrium and Neotorularia each includes 
only a single species with a gamosepalous calyx. 

Two additional species of Desideria, D. pamirica 
from Tajikistan (Suslova, 1973) and D. nepalensis 
from Nepal (Hara, 1975), were described with a 
gamosepalous calyx. The reports of gamosepaly in 
Christolea scaposa by Jafri (1973), C. karakorumen- 
sis by Wu and An (1994), and D. pamirica are 
shown in the present study to be erroneously based 
on plants of D. mirabilis. 

Gamosepaly was first reported from South Amer- 
ica by Al-Shehbaz (1990b) in Brayopsis Gilg & 
Muschl., a genus of six species of which only B. 
gamosepala Al-Shehbaz (Bolivia) has united sepals. 
An examination of other South American species 
revealed gamosepaly in Catadysia rosulans 0. Е. 
Schulz (Appel, pers. comm.) and Eudema friesii O. 
E. Schulz (Martínez-Laborde, pers. comm.). Eude- 
ma Humb. & Bonpl. includes six species distrib- 
uted from Ecuador into Argentina and Chile (AI- 
Shehbaz, 1990a), of which only Е. friesii has a 
gamosepalous calyx, whereas Catadysia O. E. 
Schulz is a monotypic genus endemic to Peru 
(Schulz, 1929, 1936). 

Gamosepaly has recently been discovered in one 
of six species of the Himalayan Pegaeophyton Hay- 
ek & Hand.-Mazz., P. watsonii Al-Shehbaz of Sik- 
kim (Al-Shehbaz, 2000а), and in one of six species 
of the Himalayan and Central Asian Phaeonychium 
О. E. Schulz, P. дари Al-Shehbaz (Al-Shehbaz, 
2000b), although the type collection of the latter 
has plants with free and united sepals. Solmslau- 
bachia xerophyta (W. W. Sm.) Comber (China) also 
has calyces with either free or completely united 
sepals, whereas S. gamosepala Al-Shehbaz & С. 
Yang (China), which is known only from the type 
collection, has united sepals (Al-Shehbaz & Yang, 
2000b). 


At least one of the approximately 150 species of 
Erysimum L., E. siliculosum (M. Bieb.) DC., has a 
gamosepalous calyx. The species was previously 
recognized in Syrenia Andrz., а genus that I place 
in the synonymy of Erysimum. It is likely that some 
of the species related to E. siliculosum also have 
gamosepalous calyces, but I have not examined ad- 
equate material of those. 

In all four species of Pugionium Gaertn. (north- 
ern China, Mongolia, and adjacent Siberian Russia) 
the sepals are connate. As the fruit develops, the 
calyx ruptures basally along the lines of sepal con- 
nation. 

Finally, the genus Gamosepalum Hausskn. was 
initially thought to have a gamosepalous calyx 
(Schulz, 1927b, 1936). However, careful examina- 


tion of its component species revealed that the se- 


pals are free, but they appear connate because of 
interlocking stellate trichomes (Dudley, 1964). 

In conclusion, two generalizations can be made 
regarding gamosepaly. First, the union of sepals 
evolved independently several times in the Bras- 
sicaceae. It is not known whether it evolved one or 
more times within Desideria, but a phylogenetic 
study based on molecular data should reveal that. 
With the critical examination of more genera of 
Brassicaceae, it is likely that more species with 
gamosepalous calyces will be found. Second, ga- 
mosepaly alone cannot be used to define the bound- 
aries of genera because it occurs in several genera 
in which the majority of species have free sepals. 
Therefore, in the present delimitation of Desideria 
gamosepaly is ignored as a generic character, and 
the overall similarities and relationships of species 
are emphasized. 

Nothing is known about the inheritance of ga- 
mosepaly in the family, but the occurrence of plants 
with free and united sepals in the same population 
of Phaeonychium jafrii is a good lead for conduct- 
ing a simple experiment to test the genetic basis of 
this character. 


GENERIC RELATIONSHIPS AND CIRCUMSCRIPTIONS 
DESIDERIA 


Pampanini (1926) established the monotypic De- 
sideria solely on the basis of having a gamosepalous 
calyx. Although he indicated that D. mirabilis re- 
sembles what was then known as Cheiranthus him- 
alayensis Cambess., Schulz (1927a, 1936), Bot- 
schantsev (1955, 1956), and Jafri (1955) regarded 
gamosepaly as an anomaly and reduced D. mira- 
bilis to synonymy of C. himalayensis, a species that 
Schulz and Botschantsev assigned to Ermania and 
Jafri to Christolea. However, these authors over- 
looked the significant features (see below) that dis- 
tinguish these two species. With the description of 
two additional species in Desideria (Suslova, 1973; 
Hara, 1975), the genus was recognized as distinct 
in subsequent floristic works (e.g., Czerepanov, 
1995; Hara, 1979; Pachomova, 1974; Yunussov, 
1978), and it remained to be delimited primarily 
on the basis of having a gamosepalous calyx. 

A critical evaluation of all genera related to De- 
sideria in this paper leads to the conclusion that 
the genus should include 6 of the 10 species treat- 
ed in Ermania by Schulz (1936), 8 of the 10 spe- 
cies recognized in Ermania by Botschantsev 
(1955), and 5 of the 13 species assigned to Chris- 
tolea by Jafri (1955). The species recognized by 
these authors in Christolea or Ermania and exclud- 
ed from Desideria in the present account are: Par- 


Моште 87, Митбег 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


rya villosa Maxim. and Cheiranthus albiflorus T. 
Anderson, which now belong to Phaeonychium (А1- 
Shehbaz, 2000b); Draba parryoides Cham. and Me- 
lanidion boreale E. L. Greene, which are assigned 
to Melanidion (see below); Christolea crassifolia 
Cambess., which is retained in Christolea; and Par- 
rya lanuginosa Hook. f. & Thomson, which is 
placed in Eurycarpus Botsch. (Al-Shehbaz & Yang, 
2000). 

As herein delimited, Desideria consists of 11 Hi- 
malayan, Chinese, and Central Asian species char- 
acterized by having well-defined basal rosettes, 
slender and rhizome-like caudices, orbicular or fla- 
bellate to broadly ovate or obovate, often dentate 
and palmately veined basal leaves, simple and/or 
forked trichomes, linear to linear-lanceolate lati- 
septate fruits rectangular in cross section, nonto- 
rulose and strongly veined valves with distinct mar- 
ginal veins, valve apices united with the replum, 
often obsolete styles, 2-lobed stigmas, and accum- 
bent cotyledons. A combination of fruits rectangu- 
lar in cross section, valves with prominent marginal 
veins, valve apices united with the replum, often 
obsolete styles, and dentate leaves often palmately 
veined readily distinguish Desideria from the other 
genera discussed in this paper. 


CHRISTOLEA, ERMANIA, AND MELANIDION 


In his description of Draba parryoides, Chamisso 
(1831: 533) stated, *DRABA? parryoides n. sp. vel 
potius novum genus e solo fructu, deficiente flore, 
haud rite definiendum. Drabis dolichocarpis sub- 
jungimus pro tempore plantam aliquando fors jure 
meritoque nomine inventoris ERMANIAM parryoi- 
dem slutandam." Several workers (e.g., Schulz, 
1936; Botschantsev, 1955; Hedge, 1968a; Suslova, 
1972; Ovezinnikov & Yunussov, 1978; Greuter et 
al., 1993) considered the above statement as a valid 
publication of the genus Ermania, while others 
(e.g., Jafri, 1955, 1973; Jurtsev, 1975; Berkutenko, 
1988; Czerepanov, 1995) did not. According to Ar- 
ticle 34 of the Code (Greuter et al., 2000), Cham- 
isso's statement does not constitute valid publica- 
tion of the genus. Despite Schulz's (1936) detailed 
description of Ermania, it was in German, and the 
genus remained invalidly published until Botschan- 
tsev (1956) provided the Latin diagnosis. Therefore, 
all transfers to Ermania proposed by Schulz 
(1927a, 1933a, b, c) and Botschantsev (1955) re- 
mained invalid. As it is presently delimited, Er- 
mania includes only E. parryoides (Cham.) Botsch., 
the generic type, and all other species assigned to 
it belong to other genera. Еттата does not occur 
in the Himalayas and Central Asia and, therefore, 


regardless of the interpretation of its effective date 
of valid publication, it does not affect the nomen- 
clature of the unrelated taxa herein placed in De- 
sideria. 

Although superficially resembling some species 
of Desideria and Christolea, Ermania parryoides is 
most closely related to Melanidion boreale E. L. 
Greene. Both species have Arctic and subarctic 
distribution (the Russian Far East for the former 
and Alaska, Yukon, and Northwest Territories for 
the latter) and are similar in habit, foliage, pubes- 
cence, flowers, and fruit morphology. Hultén (1945) 
was the first to point out this close relationship, and 
he transferred M. boreale to Ermania, but his trans- 
fer was illegitimate because Ermania was invalidly 
published. The principal difference between these 
species is that E. parryoides has latiseptate fruits 
(flattened parallel to the septum) and M. boreale has 
angustiseptate fruits (flattened at a right angle to 
the septum), but this difference is not as significant 
as once thought because there are many genera of 
the Brassicaceae with both fruit types. Drury and 
Rollins (1952) and Rollins (1993) reduced Melan- 
idion to synonymy of Smelowskia C. A. Mey., but 
their circumscription of the North American Sme- 
lowskia was so broad that some of the species rec- 
ognized are doubtfully congeneric. If M. boreale 
and Е. parryoides were kept in a genus distinct 
from Smelowskia, as I presently support, then Er- 
mania. would have to be abandoned and the earlier 
published Melanidion recognized. These two spe- 
cies will be dealt with in a subsequent publication. 

In his original description of Cheiranthus hima- 
layensis and Christolea crassifolia, Cambessédes 
(1844) did not indicate anything about their rela- 
tionship or similarities to each other. However, Jafri 


+“ 


1955) placed them and several other species in 
Christolea and adopted a broad generic concept 
that included species presently assigned to the gen- 
era Christolea, Desideria, Eurycarpus, Melanidion, 
Parrya В. Вг., and Phaeonychium. With such а 
broad delimitation, several additional genera, es- 
pecially Pegaeophyton and Pycnoplinthus, could 
have easily been included in Christolea without ex- 
panding the generic limits any further. Unfortu- 
nately, Jafri’s delimitation of Christolea was closely 
followed in some of the more recent floras (e.g., An, 
1987, 1995; Hajra et al., 1993; Huang, 1997b; 
Kuan, 1985). 

Ovezinnikov and Yunussov (1978) also adopted 
a rather broad concept of Ermania by including 
Christolea and Oreoblastus as sections. These au- 
thors differed from Jafri (1955) primarily in their 
decision about the effective date of valid publica- 
tion of Ermania. In my opinion, their vastly het- 


552 


Annals of the 
Missouri Botanical Garden 


erogeneous generic circumscriptions of Christolea 
or Ermania are unacceptable. Christolea consists of 
two species, the Himalayan C. crassifolia and the 
Chinese endemic С. niyaensis 2. Х. An, and it dif- 
fers from Melanidion (including Ermania) by hav- 
ing many-leaved stems, nonrosulate lower leaves, 
exclusively simple trichomes, incumbent cotyle- 
dons, apiculate anthers, and transversely oriented 
seeds. By contrast, Melanidion has leafless stems, 
well-developed basal rosettes, dendritic trichomes 
mixed with simple ones, accumbent or obliquely 
accumbent cotyledons, obtuse anthers, and longi- 
tudinally oriented seeds. 

Desideria differs from both Melanidion and 
Christolea by having fruits rectangular in cross sec- 
tion, valves with prominent marginal veins, and 

ve apices united with the replum. From Chris- 
tolea, Desideria differs by having a well-developed 
basal rosette, usually leafless stems, slender and 
rhizome-like caudices, often palmately veined 
leaves, nontorulose fruits, longitudinally oriented 
biseriate seeds, and accumbent cotyledons. By con- 
trast, Christolea has nonrosulate lower leaves, leafy 
stems, compact and woody caudices, pinnately 
veined leaves, strongly torulose fruits, transversely 
oriented uniseriate seeds, and incumbent cotyle- 
dons. The Himalayan and Central Asian Desideria 
also differs from the Arctic and subarctic Melani- 
dion by lacking the dendritic trichomes and having 
sessile 2-lobed stigmas, smooth fruits, biseriate 
seeds, and toothed nectaries lacking the median 
glands. Melanidion has dendritic trichomes, entire 
and capitate stigmas on distinct styles, torulose 
fruits, uniseriate or rarely subbiseriate seeds, and 
annular nectaries with well-developed median 
glands. A comparison of Desideria with the presum- 
ably related genera is summarized in Table 


EURYCARPUS 


In establishing the genus Eurycarpus, Botschan- 
tsev (1955) separated it from Ermania by having 
biseriate instead of uniseriate seeds, broadly lan- 
ceolate instead of linear fruits, entire instead of 
dentate leaves, and leafless instead of leafy scapes. 
However, he probably compared only the type spe- 
cies of both genera because most of the differences 
above do not hold if one compares Eurycarpus with 
the ten species Botschantsev recognized in Erman- 
ia. As indicated above, eight of Botschantsev's ten 
species of Ermania are presently assigned to De- 
sideria. A comparison of Desideria with Eurycarpus 
(two species) sensu Al-Shehbaz and Yang (2000a 
shows that the latter differs by having entire and 
pinnately veined leaves, obscurely veined valves 


Мм 


without marginal veins, broadly lanceolate to ob- 
long fruits narrowly elliptic in cross section, well- 
defined subconical styles, and minute, entire stig- 
mas much narrower than the style. By contrast, 
Desideria almost always has dentate, palmately 
veined leaves, prominently veined valves with well- 
developed marginal veins, linear to linear-lanceo- 
late fruits rectangular in cross section, obscurely 
differentiated or cylindric styles, and distinct, often 
2-lobed stigmas as broad as the style 


OREOBLASTUS 


Although Jafri (1973) admitted the artificiality of 
his delimitation of Christolea, he (p. 155) correctly 
stated that, “Even if, Christolea Camb. (s. str.) and 

rmania Cham. ex [Botschantsev] Schulz (s. str.) 
are considered as separate genera, there can be no 
doubt that Oreoblastus Suslova is congeneric with 
Desideria Pamp., where most of our species would 


go. 

Suslova (1972) separated Oreoblastus from De- 
sideria by having free instead of united sepals, a 
deciduous instead of persistent calyx, and septate 
instead of eseptate fruits. However, she must have 
overlooked the persistent calyx in several speci- 
mens that she annotated as Oreoblastus, and the 
holotype of her D. pamirica (Suslova, 1973) has 
septate instead of eseptate fruits, though the septa 
are perforated but never lacking. Except for having 
free instead of united sepals, Oreoblastus is indis- 
tinguishable from Desideria. As indicated above, 
sepal connation alone is insufficient for the estab- 
lishment of genera and, therefore, Oreoblastus is 
reduced herein to synonymy of the earlier pub- 
lished Desideria. 


ERMANIOPSIS 


The presence vs. absence of a tooth on the me- 
dian stamens was considered by some (e.g., Schulz, 
1936; Hara, 1974; Golubkova, 1976) as an impor- 
tant generic character. In my opinion, this feature 
alone does not justify the segregation of genera. 
Toothed and toothless filaments are found in Don- 
tostemon Andrz. ex C Mey. (Al-Shehbaz & 
Ohba, 2000), whereas winged or wingless, toothed 
or toothless, and appendaged or unappendaged fil- 
aments are found in Alyssum L. (Al-Shehbaz, 1987; 
Dudley, 1964) 

Although Hara (1974) provided a detailed dis- 
cussion to distinguish Ermaniopsis from Ermania 
and related genera, the single character that sets 
Ermaniopsis apart is the presence of a lateral tooth 
on the filaments of median stamens. On the basis 
of all other characters, Ermaniopsis pumila H. Hara 


553 


Al-Shehbaz 


Volume 87, Number 4 


2000 


Gamosepaly in Brassicaceae 


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554 


Annals of the 
Missouri Botanical Garden 


is perfectly at home in Desideria. In fact, Hara in- 
dicated that E. pumila resembles Desideria (as Par- 
rya) pumila in vegetative characters. A close ex- 
amination of flower and fruit characters clearly 
shows that the two species are congeneric, and Er- 
maniopsis is reduced herein to synonymy of Desi- 
deria. Unfortunately, both species have the same 
epithet, and E. pumila is named hereafter as D. 
haranensis. The median filaments of both D. pumila 
and D. haranensis are dilated, and only the latter 
species shows a minute to prominent tooth on the 
median staminal filaments. 


SOLMSLAUBACHIA AND LEIOSPORA 


On the basis of fruit morphology, Desideria is 
most closely related to Solmslaubachia (9 spp.: 
endemic to China and 1 extending also into Bhutan 
and Sikkim) and the Central Asian and Himalayan 
Leiospora (6 spp.). All three genera have fruits 
readily detached from the pedicel, and their valves 
are adnate apically to the replum. Upon maturity, 
the fruit falls off the plant and its apex remains 
tardily dehiscent. The three genera also have ob- 
solete or no styles, and their fruit valves are strong- 
ly angled at the margins and completely conceal 
the replum. These combinations of characters are 
not found in any Himalayan or Central Asian gen- 
era of the Brassicaceae. 

Desideria is easily separated from Solmslauba- 
chia by having palmately veined leaves apically 3- 
to 9(to 11)-toothed, ovate to oblong anthers often 
less than 1 mm long, and forked trichomes some- 
times mixed with simple ones. By contrast, Solms- 
laubachia has entire, pinnately veined leaves, lin- 
ear-oblong anthers more than 1 mm long, and 
exclusively simple trichomes. 

Desideria is readily distinguished from Leiospora 
by having wingless seeds, equal sepals with the lat- 
eral pair nonsaccate, palmately veined leaves api- 
cally 3- to 9(to 11)-toothed, oblong-linear anthers 
0.4—1(-1.6) mm long, and capitate, slightly 2-lobed 
stigmas with neither decurrent nor connivent lobes. 
Leiospora often has winged or margined seeds, un- 
equal sepals with the lateral pair strongly saccate, 
entire or marginally dentate leaves, linear anthers 

3 mm long, and conical, prominently 2-lobed 
stigmas with connivent, decurrent lobes. 


TAXONOMIC TREATMENT 


KEY TO THE SPECIES OF DESIDERIA 
la. 


© 


Petals 11-13 X 5-6 mm; calyx 5-6 n 


Sepals united, persistent till or after fruit е ‘ence; septum absent ог reduced to а rim 
2a. n long; flowers 2-4, appearing solitary; Мера al 
2b. Petals and calyx smaller; flowers more ibas 4. in distinct racemes; China, Kashmir, о 


Desideria Pamp., Bull. Soc. Bot. Ital. 1926: 111. 
1926. TYPE: Desideria mirabilis Pamp. 


Ermanioosis H. Hara, J. Jap. um 49: 198. 1974. TYPE: 
"rm niopsis punila H. 

Oreoblastus bye ova, Bot. Zhur n (Ме w & Leningrad) 
57: 648. 1972. “TYP E: Oe fan. PIE (Regel) 
Suslov 


Herbs perennial, with a slender, often many- 
branched, rhizome-like caudex often covered with 
remains of basal rosettes. Trichomes simple and/or 
mixed with short-stalked forked ones. Stems sim- 
Basal 
3- to 9(to 11)- 
toothed, often palmately veined, persisting whole or 
only petioles persistent. Cauline leaves similar to 
basal ones, entire or toothed, subsessile or petio- 
late, or absent. Racemes 3- to 30-flowered, dense 
or lax, bracteate throughout or ebracteate, corym- 
bose, elongated or not elongated in fruit, sometimes 
flowers solitary on pedicels originating from basal 


ple, leafy or leafless, sometimes absent. 
leaves petiolate, rosulate, simple, 
P Р 


rosette. Sepals ovate to oblong, free or united, de- 
ciduous or persistent, erect, equal, base of inner 
pair not saccate, margins membranous. Petals pur- 
ple, purple-green, or rarely white, sometimes yel- 
koh at base of blade; blade obovate to spatulate, 
apex obtuse to subemarginate; claw strongly differ- 
entiated from blade, subequaling or longer than se- 
pals. Stamens 6, erect, tetradynamous; filaments 
wingless or rarely winged, toothless or rarely 
toothed, free, dilated at base; anthers ovate to ob- 
long, not apiculate at apex. Nectar glands 2 and 
lateral, or 1 and confluent outside bases of all sta- 
mens; median nectaries present or absent. Ovules 
10 to 70 per ovary. Fruit dehiscent siliques, linear 
to lanceolate, latiseptate, rectangular in cross sec- 
tion, not inflated, sessile; valves papery, with a 
prominent midvein and distinct marginal veins, gla- 
brous or pilose, smooth, adnate with replum at fruit 
apex; replum rounded, often concealed by valve 
margin; septum complete, perforated, or reduced to 
a rim, membranous, translucent, veinless, rarely 
absent; style obsolete; stigma capitate, slightly 2- 
lobed. Seeds uniseriate or biseriate, wingless, ob- 
long to ovate, often flattened; seed coat obscurely 
reticulate, not mucilaginous when wetted; cotyle- 
dons accumbent. 

Eleven species: Himalayas, western China, and 
adjacent central Asia. 


11. D. nepalensis 


10. D. mirabilis 


Volume 87, Number 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


555 


lb. e free, caducous or rarely persisting till about fruit maturity; septum complete or rarely perforated 


apica 
F ш solitary from a basal ro 


se 
4a. Fruit ovate to broadly lanceolate, 6-9 mm wide, soset reticulate veined . 


4b. Fruit linear to linear-lanceolate, 2-5 m 


. 9. D. baiogionensis 
wide, obscurely 


5a. Leaf trichomes forked не ien replum ны M yat valves glabrous; sepals 3—4 mm 


ong; petals 6-8 mm lon 


. pumila 


5 
5b. Leaf trichomes exclusively simple; replum and valves pilose to villous; sepals 6-7 mm a lend 
long 


etals 11—14 mm lon 


w 
> 


Flowers (3 to)6 to 30 in a raceme. 
Racemes bracteate throughou 
7a. Stem and pedicel trichomes forked 


7. D. prolifera 


2. D. stewartii 


7b. Stem and pedicel trichomes exclusively simple or absent 


Fruit lanceolate to linear-lanceolate, а 
.8-2(-2.3) х | 1. D. 
.8-)1-1. es mm im NUM 4—5(-5.5) X 1.5-2.5 mm; ow uni- 


seriate, 0.81 ] x 0.5-0.8 m 


6b. Racemes ebract 
a 


mm wide; petals (6— 


)6.5-8 х 3-4 


himaloyenst 


3. D. linearis 


eate 
Fila ments p nd о, petals 6.5-8 mm long; leaf trichomes minutely 


forked, mixed with short 


imple one 


aranensis 


9b. cob. terete, esa petals i 18 mm long; leaf trichomes either кейий simple 


= oe rked. 


. Plants canescent; leaf trichomes almost exclusively branched; leaves 3(to 5)-toothed 


D. incana 


9. 
10b. Plants greenish; leaf trichomes exclusively simple, to 1.5 mm long; leaves e ny 


9(to 11)-toothed 


D. flabellata 


1. Desideria himalayensis (Cambess.) Al-Sheh- 
baz, comb. nov. Basionym: Cheiranthus him- 
alayensis Cambess., in Jacquemont, Voy. Inde 

: 14. 1844. Ermania himalayensis (Cambess.) 
0. Е. irons Notizbl. Bot. Gart. Berlin-Dah- 

. Oreoblastus himalayensis 

(Cambess.) a. Bot. Zhurn. (Moscow & 

Leningrad) 57: 652. 1972. TYPE: [У Tibet.] 

“In declivitate orientali jugi vulgd Kioubrung- 

ghauti in Tartariá sinensi,” Victor Jacquemont 


1782 (holotype, P!; isotypes, K!, P!). 


Plants 4–20 cm tall, densely pilose throughout 
to subglabrous. Trichomes simple, to 1.5 mm long. 
Stems simple, pilose or glabrous. Basal leaves not 

eshy, pilose or glabrous, persistent; petiole 0.4— 
1.6(-3) cm long, not ciliate; leaf blade broadly ob- 
ovate to spatulate, 4-14 X 3-9 mm, base cuneate 
to attenuate, margins (3 to)5-toothed, apex acute. 
Stem leaves similar to basal or linear to lanceolate, 
5-17 X 14 mn, often entire, short petiolate to 
subsessile. Racemes 6- to 25-flowered, bracteate 
throughout; bracts similar to stem leaves but small- 
er, sometimes adnate to pedicel. Fruiting pedicels 
ascending, straight or curved, 3-10 mm long, pilose 
or glabrous. Sepals free, oblong, 3—4 X 1.2-1.5 
mm, caducous, pilose or with a terminal tuft of 
hairs, base not saccate, margins membranous. Pet- 
als purple or lilac with yellowish center, broadly 
spatulate, (6—)6.5—8 X 3—4 mm, apex subemargin- 
ate; claw 3—4 mm long. Filaments white, slightly 
dilated at base, median pairs 3—4 mm long, lateral 


pair 24 mm long; anthers ovate, ca. 0.6 mm long. 
Ovules 7 to 12 per locule. Fruit lanceolate to lin- 
ear-lanceolate, (1.7-)2-3.5(-4) ст X (3-)4—6 mm, 
strongly flattened; valves pilose or glabrous, dis- 
tinctly veined; septum complete, membranous; 
style obsolete; stigma 2-lobed. Seeds brown, ovate, 
(1.5—)1.8–2(–2.3) 


reticulate. 


X 1-1.4 mn, biseriate, minutely 


Phenology. Flowering June through August. 
Fruiting July to mid October. 
Habitat and distribution. 
hills, sandstone scree; 4300— 


Alpine tundra, open 
00 m. China (Qing- 
hai, Xizang), India, Kashmir, Nepal. 

Selected specimens examined. CHINA. Qinghai: Lei- 


xie-wu-den, Wu, po & Yang K-890 (HNWP). Xizang: 
Baingoin Xian, Whale Lake, Wu, Ohba, Wu & Fei 4075 


(KUN, MO, ТІ); E of Moincer, 31°14’N, 80°56'Е, С. & S. 
Michie 9643/17 (СОЕТ, MO); NW Tibet, 34^55'N, 
82°24’E, Pike 842 (К); Aksu, Deasy 92 (BM); Shuang Ho 


Xian, Qinghai- Xizang ен 12009 (РЕ); Ritu Xian, Qing- 
hai-Xizang Team 76-9061 (KUN, PE), Li Bosheng & 
ral Du 10980 (PE). INDIA. Punjab: Lahul, Kangra, 
Lach La, Koelz 6738 (GH). KASHMIR. Harnag, Up- 
per ade: valley, Stewart 9349 (B, G, K, MO). NEPAL. 
Dhaulagiri Himal, hidden valley, between Dhampus pass 
and French pass, Wald 65 (BM); Thorong La, Marsyandi 
Valley, Me Beath 1486 (E); hia ai Marsyandi, McBeath 
1 406 п N Annapurna Gla- 
nnapurna Glacier icefall, 
170-175 km N of Pokhara, Komarkova 18 (GH) 


Desideria himalayensis is most frequently con- 
fused with D. linearis, and some authors (e.g., Jafri, 
1955) considered them to be conspecific. Three 


556 


Annals of the 
Missouri Botanical Garden 


Е of Desideria linearis (Lyon 44, Stainton 

3055, Stainton 3241) were cited by Jafri (1973) as 
Christolea himalayensis, and the first two were list- 
ed by Hedge (1968a) as Ermania himalayensis. The 
two species can be readily separated by petal size, 
fruit width, and seed arrangement and size (see 
key). Mixed collections of the two species (e.g., 
Koelz 6738) are not uncommon, but no intermedi- 
ates have been found. Both species can be distin- 


guished from the related D. stewartii by having leaf 
and stem trichomes exclusively simple instead of 


forked. 

Desideria himalayensis was reported (as Chris- 
tolea) from Xinjiang by An (1995), but I have not 
seen any material from that part of China. One col- 
lection (Polunin, Sykes & Williams 37) was cited 
by Hara (1979) as this species, but this collection 
clearly belongs to Desideria linearis. 

Desideria himalayensis was erroneously illustrat- 
ed in Jafri (1973) with ebracteate inflorescences. It 
is likely that the plant illustrated belongs to D. fla- 
bellata, a species that occurs in bordering Afghan- 
istan, China, Kyrgyzstan, and Tajikistan but is not 
yet reported from Kashmir. Desideria himalayensis, 
D. stewartii, and D. linearis are the only three spe- 
cies of Desideria that consistently have racemes 
bracteate throughout. 


2. Desideria stewartii (T. Anderson) Al-Shehbaz, 
comb. nov. Basionym: Cheiranthus stewartii T. 
Anderson, in J. D. Hooker, Fl. Brit. India 1: 
132. 1872. Ermania stewartii о О. 
E. Schulz, Bot. Jahrb. Syst. 66: 1933 
Christolea stewartii (T. она Notes 
Roy. Bot. Gard. Edinburgh 22: 53. 1955. Or- 
eoblastus stewartii (T. Anderson) Suslova, Bot. 
Zhurn. (Moscow & Leningrad) 57: 653. 1972. 
TYPE: Kashmir. Ladak, 15,000—16,500 ft., J. 
L. Stewart s.n. (holotype, K!; isotype, E!). 


Plants 8-20 cm tall, densely pilose. Trichomes 
stalked forked, rarely some simple near the stem 
base. Stems simple, pilose or glabrous. Basal leaves 
subfleshy, pilose, persistent; petiole 2-10 mm long, 
not ciliate; leaf blade broadly obovate to spatulate, 
2-15 х 2-10 mm, base cuneate to attenuate, mar- 
gins 3- to 5-toothed or subentire, apex acute. Stem 
leaves similar to basal or linear to lanceolate, often 
entire. Racemes 8- to 15-flowered, bracteate 
throughout; bracts similar to stem leaves but small- 
er, often adnate to pedicel. Fruiting pedicels as- 
cending, straight or slightly curved, 4-12 mm long, 
pilose. Flowers not seen. Ovules 7 to 12 per locule. 
Fruit lanceolate to lanceolate-linear, 1.7-3.5 cm X 
3—5 mm, strongly flattened; valves pilose or gla- 


brous, distinctly veined; septum complete, mem- 
branous; style obsolete; stigma 2-lobed. Seeds 
brown, ovate, 1.4—2.2 х 0.8-1.1 mm, biseriate, mi- 
nutely reticulate. 


Phenology. Flowering unknown. Fruiting in 
August. 

Habitat and distribution. Scree slopes; 4100- 
5000 m. China (Xizang), India, Kashmir. 

Selected specimens examined. CHINA. Xizang: Ali, 
Geji, Qinghai- е $e 76-8652 (PE). INDIA. Punjab: 

Lacha Pa . Cooper 5490 (E). Himachal 

Pradesh: Zingzingbar, MeBeath 2105 (Е). 


Desideria stewartii is a very rare species known 
thus far from the few collections cited above. Re- 
ports of the species from China (Kuan, 1985; An, 
1987) are most iid based on misidentified plants 

of D. himalay 

Jafri (1973) пи the distinction of Desideria 
stewartii from D. himalayensis (both as Christolea), 
and he confused the limits of the two species by 
citing one collection, Stewart 9349, under the for- 
mer instead of the latter species. I have not seen 
any flowering material of the species, and the de- 
scription of the flowers by Jafri (1973), which was 
followed by Hajra et al. (1993) and An (1987), was 
almost certainly based on a small flowering branch 
of D. linearis mounted on the holotype sheet of D. 
мешати. 

In overall aspects of foliage and fruit, Desideria 
мешати most closely resembles D. himalayensis. 
However, D. stewartii is readily separated by the 
presence of forked, stalked, intermingled trichomes 
instead of exclusively simple straight ones. 


3. Desideria linearis (N. Busch) Al-Shehbaz, 
comb. nov. Basionym: Christolea linearis N. 
Busch, in Komarov, Fl. URSS. 8: 636. 
Ermania linearis (N. Busch) Botsch., Bot. Ma- 
ter. Gerb. Bot. Inst. Komarova Akad. Nauk 
S.S.S.R. 17: 166. 1955. Oreoblastus linearis 
(N. Busch) Suslova, Bot. Zhurn. (Moscow & 
Leningrad) 57: 652. 1972. TYPE: Tajikistan. 
Pamir: Schugnan, Abchary, 2 Aug. 1904, B. 
Fedtschenko s.n. (holotype, LE!). 

Еттата parkeri О. E. Schulz, Керем. Sp. Nov. Regni 
Veg. 31: 333. 1933. poss parkeri (O. E. Schulz) 
Jafri, Notes Roy. Bot. n Edinburgh 22: 52. 1955. 
Oreoblastus parkeri (О. Schulz) Suslova, = 
Zhurn. (Moscow & Leningrad) 57: 653. 1972. 
nov. TYPE: Kashmir. Sonamarg, Luderw 
m 11 Aug. 1928, R. R. ema 98744 шл к 


M kashmiriana Dar & Naqshi, J. Bombay Nat. Hist. 
Soc. 87: 274. 1990. Syn. nov. TYPE: Kashmir. Shal- 
imar, Sonamarg (Sind Valley), 3900 m, 
1983, G. H. Dar 7786 (holotype, KASH). 


20 Aug. 


— 


Volume 87, Number 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


557 


т“ kachrooi Паг & Naqshi, J. Bombay Nat. Hist. 
7: 277. 1990. Syn. nov. TYPE: Kashmir. Bal- 

tal, Sonamarg (Sind Valley), 3200 m, 2 Sep. 1982, 
3934 (holotype, KASH; isotypes, KASH, 


Plants 4—15 ст tall, densely pilose throughout 
to subglabrous. Trichomes simple, to 1.5 mm long. 
Stems simple, pilose or glabrous. Basal leaves not 
fleshy, pilose or glabrous, persistent; petiole 2-7 
(-12) mm long, not ciliate; leaf blade broadly ob- 
ovate to spatulate, 4-15 X 2-12 mm, base cuneate 
to attenuate, margins 3- to 5-toothed or rarely sub- 
entire, apex acute. Stem leaves similar to basal or 
linear to lanceolate, 5-10 X 1-3 mm, often entire, 
short petiolate to subsessile. Racemes 8- to 20- 
flowered, bracteate throughout; bracts similar to 
stem leaves but smaller, often adnate to pedicel. 
Fruiting pedicels ascending, straight, 2–8(–12) mm 
long, pilose or glabrous. Sepals free, oblong to 
ovate, 2—3 X mm, caducous, pilose or with 
a terminal tuft of hairs, base not saccate, margins 
membranous. Petals purple or lavender with paler 
base, narrowly spatulate, 4—5(-5.5) X 1.5-2.5 mm, 
apex rounded; claw 2-2.5 mm long. Filaments 
white, slightly dilated at base, median pairs 2.5— 
3.5 mm long, lateral pair 1.8-2.5 mm long; anthers 
ovate, 0.4—0.5 mm long. Ovules 8 to 13 per locule. 
Fruit linear, (1.5-)2-3.5(-4.2) cm X (0.8—)1-1.7 
(-2) mm, flattened; valves pilose or glabrous, dis- 
tinctly veined; septum complete, membranous; 
style obsolete; stigma 2-lobed. Seeds brown, ovate, 

.8 mm, uniseriate, minutely retic- 
ulate. 


Phenology. Flowering June through August. 
Fruiting July through September. 

Habitat and distribution. Gravelly or sandy 
slopes, scree, gravelly moraine below glacier; 
3200-5200 m. China (Xinjiang, Xizang), Kashmir, 
Nepal, Tajikistan. 


Selected specimens examined. CHINA. Xinjiang: 
Yecheng Xian, Li Bosheng et al. 11278 (PE); Tagdum- 
basch-Pamir, Pistan near Saryokol, Alexeenko 2729 (LE). 
Xizang: without locality, Коргоп s.n. (ВМ, С, GH, К 

P); Ali, Qinghai-Xizang Team 76-7948 (HNWP). INDIA. 
Punjab: Lahul, Kangra, Bara Lach La, Koelz 6738 (GH). 
KASHMIR. Ishkuman Aghost, Schmid 2449 (G). Chitral: 
Laspur (Hachin), 36°2’N, 72°27'Е, Lyon 44 (A, E); Siro- 
gol, S of Shah Jinali Pass, Stainton 3055 (E); Dorah Pass, 
Lutko valley, Stainton 3241 (E); Amarnath, io s 8709 
(E); Sonamarg, Luderwas, Stewart 9874 (B, G, MO). Ka- 
rakorum: оен Glacier base camp, 13 ті. E of oni 
Polunin 6133 (BM); Karakorum, Oct. 1877, Clarke 
(K). Ladak: above Stok, Maxwell 92 g Morgens. lacie 
Sentik, 34°N, 76°, Delouche 27 (P). T 
200 km NW. of Gilgit, near watershed edi. Cilgit 
from Chitral, Broadhead 39 (E); Taklung La, Koelz 6500 
(GH). NEPAL. Naur Pass, Lowndes 1159 (BM); 5 mi. S of 


» 


Saldanggaon, Polunin, Sykes & Williams 37 (BM). TAJI- 
KISTAN. Pamir: N slope, river Zor-Chechekty, 12 Aug. 
1948, Stanjukovich & Kishkovsky s.n. a) Chechekty, riv- 
er Zor-Chechekty, Raikova 228 


Although I have not seen the holotype of Er- 
mania kashmiriana, the original description and il- 
lustration, as well as the examination of a paratype 
(Dar 8301), clearly support the placement of the 
species in synonymy of Desideria linearis. Dar and 
Naqshi (1990) compared E. kashmiriana and E. 
kachrooi with D. stewartii and D. himalayensis (all 
as Ermania), but they failed to relate their novelties 
to D. linearis. In my opinion, E. kachrooi is only a 
glabrous form of D. linearis, a species within a giv- 
en population of which one can find glabrous and 
pubescent plants. In general, plants of Desideria, 
including D. linearis, that grow in partly shaded 
areas, especially under large boulders, often have 
the apex of the caudex elongated so that the leaf 
rosette appears much less congested. 

Several authors (e.g., An, 1987, 1995; Kuan, 
1985) followed Jafri (1955) in listing Desideria li- 
nearis as a synonym of Christolea himalayensis, but 
these authors erroneously recognized C. parkeri as 
a distinct species. In my opinion, the last species 
is only a glabrescent form of D. linearis. In fact, C. 
parkeri is based on Ermania parkeri, an invalidly 
published species assigned to the invalid Ermania 
(see Greuter et al. (2000) under Article 43.1). 

Although Jafri (1973) maintained Christolea par- 
keri, he correctly indicated that it is not different 
from the earlier-published Desideria (as Christolea) 
linearis, a species that he did not recognize for Pak- 
istan and Kashmir. However, Suslova (1972) main- 
tained both species (as Oreoblastus) and separated 
them mainly by the presence in the former of a 
subapical tuft of hairs on the sepals instead of its 
absence in D. linearis and D. himalayensis. Obvi- 
ously, this distinction is artificial, and all taxa have 
pubescent sepals that often are more densely hairy 
below the apex. The restriction of trichomes to the 
sepals and leaf apices is quite frequent in glabres- 
cent forms of D. linearis and D. himalayensis. 

Schulz (1931) considered Desideria linearis (as 
Ermania parkeri) to be closely related to E. albiflo- 
ra A т О. E. Schulz, but the nearest rel- 

e of the first is D. himalayensis. As shown by 
AL ‘Shehbaz mode E. albiflora belongs to the ge- 
nus Phaeonychium O. E. Schulz 

Desideria linearis is extremely variable in the oc- 
currence and density of the indumentum, fruit 
width, length, and indumentum, and number of leaf 
teeth. However, an examination of material from the 
various parts of the species range clearly negates 


558 


Annals of the 
Missouri Botanical Garden 


the need to subdivide the species into infraspecific 
taxa. 


4. Desideria flabellata (Regel) Al-Shehbaz, 
comb. nov. Basionym: Parrya flabellata Regel, 
Bull. Soc. Imp. Naturalistes Moscou 43: 261. 
1870. Christolea deep eem N. Busch, 
in Komarov, 8: 330. 1 
ар (Regel) a chulz, Bot. Jahrb. 

: 98. 1933. Каш н flabellatus (Re- 
D Bulova, Bot. Zhurn. (Moscow & Lenin- 
grad) 57: 651. 1972. TYPE: Southern Tian 
Shan, Dschaman-Daban, Sewerzow s.n. (holo- 


type, LE!). 


39. Ermania 


е ен iege К. F. Huang, е Phytotax. Sin. 

35: 5: ›. 1997. Syn. nov. о hina. Qinghai: Ма- 

"n, "d oT Mt., m, m June 1981, R. F 
Bone CG-81-154 кшн HNWP)). 


Plants greenish, 4—15 cm tall. Trichomes simple, 
straight, to 1.5 mm long. Stems distinct, 
densely pilose. Basal leaves subfleshy; petiole 2—7 
mm long, pilose; leaf blade flabellate to broadly 
obovate, rarely spatulate, 0.6–2.5 X 0.3-2.5 cm, 


simple 


» 


pilose, base cuneate to attenuate, margins (3 to)5- 
to 9(to 11)-toothed, rarely lowermost entire, apex 
acute; teeth to 10 X 3 mm. Stem leaves similar to 
basal. Racemes 7- to 12-flowered, 
Fruiting pedicels ascending, straight to curved, 
(0.5—)0.7—1.5(-2.5) cm long, spreading pilose. Se- 
pals free, narrowly oblong, 5-8 X 1.5-2.5 mm, of- 
ten persistent, pilose, base not saccate, margins 


ebracteate. 


membranous. Petals purple, broadly spatulate, 1.1— 
1.5 ст X 3.5—6 mm, apex subemarginate; claw 7— 
9 mm long. Filaments white to mauve, slightly di- 
lated at base, median pairs 4.5—6 mm long, lateral 
pair 3—4 mm long; anthers oblong, 0.9-1.3 mm 
long. Ovules 7 to 12 per locule. Fruit lanceolate to 
lanceolate-linear, (1.7-)2.5-3.5(-4.5) em X 2.5-5 
mm, strongly flattened; valves pilose, distinctly 
veined; septum complete, membranous; style ob- 
solete; stigma 2-lobed. Seeds brown, ovate, 1.3—2 
X 0.9-1.2 mm, uniseriate, minutely reticulate. 


Phenology. Flowering early July and August. 
Fruiting late July through early September. 

Habitat Alpine gravelly 
slopes, moraine slopes; 33 


and distribution. 
О m. Afghanistan, 
China (Xinjiang), Kyrgyzstan, Tajikistan. 


ected specimens | examined. AFGHANISTAN. 
Tien above s haie Gibbons 623 (MO); Hindu 
Kush, Gilbert 88 (E). Parvan: Panjshir, Hedge & Wen- 
ads 5451 (E). Kapisa: Mir S Samir area, Gibson 211 (Е). 
: Khost-o-Fereng, valley Echani-Tai, Е of Chun- 

duk, Podlech 11832 (G). CHINA. Xinjiang: Kashgaria, 
ian Shan, 1889, Roborowski s.n. (LE), Merzbacher 354 
LE): Kashani. Billuli Pass, 13 June 1909, Divnogor- 


skaya s.n. (LE, MO); Akto Xian, e ng, 5. б. Wu, Y. 
H. Wu & Y. Fei 4641 (HNWP, КОХ). KYRG YZSTÀN. 

echie: Przewalsk, river Kayche, 30 July 1913, 
Shishkin s.n. (LE), 2 Aug. 1913, Capoznnikov s.n. (MO). 
Tianshan: Glacier Каїпаї, кені 338 (С). ТАЈІКІЅ- 

. Shugnan: Е Bukhara, near Pass Garm-Chashmy, 
Пион 150 (A, LE). Pamir: bc Glacier, Gorbanov 
191 (LE). 


In every aspect of trichome morphology, flower 
size and color, and habit, Christolea pinnatifida is 
indistinguishable from plants of Desideria flabella- 
ta. The type of the former has no fruits and is rather 
immature. It differs only slightly from typical plants 
of D. flabellata by having slightly elongated, spat- 
ulate leaves instead of typically flabellate ones. 
Huang (19972) considered C. pinnatifida to be re- 
lated to C. karakorumensis, but the latter is a syn- 
onym of D. mirabilis and has sepals typically united 
instead of free. He indicated that the ovaries are 
glandular mamillate, but this observation was based 
on developing trichomes, and neither Desideria nor 
Christolea has any glandular trichomes or papillae. 


5. Desideria incana (Ovcz.) Al-Shehbaz, comb. 
nov. Basionym: Christolea incana Ovcz., Sov- 
etsk. Bot. 1941(1 & 2): 151. 1941. Ermania 
incana (Ovcz.) Botsch., Bot. Mater. ges) Bot. 
Inst. Komarova Akad. Nauk S.S.S.R. 17: 164. 

Oreoblastus incanus (Ovez.) Sova 

. (Moscow & rina ia 57: 

197 2. TYP E: Tajikistan. Darvaz: Mt. en 

glacier Abdul Gassan, 11.000—12.000 ft., 23 

July 1899, V. I. Lipsky 1936 (holotype, LE!). 


Plants 4—15 cm tall, densely tomentose through- 
out. Trichomes short-stalked forked and simple, to 
1 mm long. Stems simple, tomentose. Basal leaves 
subfleshy, canescent, densely tomentose, persistent; 
petiole 0.5—2 mm long, not ciliate, unexpanded and 
not papery at base; leaf blade broadly obovate to 
spatulate, 4-13 X 2-8 mm, base cuneate to atten- 
uate, margins 3(to 5)-toothed, sometimes subentire 
on sterile branches, apex acute. Stem leaves similar 
to basal. Racemes 6- to 20-flowered, only basally 
bracteate. Fruiting pedicels ascending, straight, 2— 
7 mm long, tomentose. Sepals free, narrowly ob- 
long, 5-7 X 
tose, base not saccate, margins membranous. Petals 
purple with paler or yellowish base, spatulate, 12— 
18 X 4—6 mm, apex rounded; claw 7-10 mm long. 
Filaments white, slightly dilated at base, median 
pairs 5—6 mm long, lateral pair 3—4 mm long; an- 
thers narrowly oblong, 1.2-1.5 mm long. Ovules 25 
to 35 per locule. Fruit linear, 3—6.5 cm X 2.5-3.5 
mm, strongly flattened; valves tomentose, distinctly 
veined; septum complete, membranous; style ob- 


5—2 mm, caducous, densely tomen- 


Моште 87, Митбег 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


559 


solete; stigma 2-lobed. Seeds brown, oblong, 1.2- 
1.5 X 0.7-1 mm, minutely reticulate. 


Phenology. Flowering July. Fruiting July and 
August. 

Habitat and distribution. Alpine gravelly areas, 
3300—4600 m. Endemic to Tajikistan. 

Selected specimens examined. TAJIKISTAN. Pamir- 
Alay: Sauk-Dara valley, [konnikov 17878 (LE). Bukhara: 
Darvaz, range of Peter-the-Great, glacier Vereshkay, 29 
July 1899, Lipsky s.n. (G, LE). 


6. Desideria haranensis Al-Shehbaz, nom. nov. 
Replaced name: Ermaniopsis pumila H. Hara, 
J. Jap. Bot. 49: 200. 1974, not Desideria pum- 
ila (Kurz) Al-Shehbaz. TYPE: Nepal. Ca. 5 mi. 
SW of Saldanggaon, 26 June 1952, very loose 
scree, 19,500 ft., N. Polunin, W. R. Sykes & 
L. H. J. Williams 24 (holotype, ВМ!; isotypes, 
A!, BM!, Е!) 


Plants 2-6 cm tall. Trichomes simple, straight, 
to 0.5 mm long, mixed on leaves with short-stalked, 
unequally branched forked ones. Stems erect, sim- 
ple, pilose to hirsute. Basal leaves fleshy, persis- 
tent; petiole 2-12 mm long, sparsely to densely pi- 
lose with simple trichomes, ciliate at base, not 
expanded or papery at base; leaf blade broadly 
3-11 mm 
sparsely to densely pubescent, base cuneate or ob- 
tuse, margins l- to 5-toothed, apex obtuse. Stem 
leaves absent. Racemes 3- to 8-flowered, ebracte- 
ate. Pedicel divaricate, straight, 4—12 mm long, pi- 
lose. Sepals free, oblong, 3.5—4.5 X 1.7-2 mm, ca- 
ducous, pilose, base not зассаје, 
membranous. Petals white tinged with greenish 
blue, obovate, 6.5—8 X 3—4 mm, apex obtuse; claw 

—4 mm long. Filaments white, flattened, subapi- 
cally toothed, median pairs 3—4 mm long, lateral 
pair 2-3 mm long; anthers oblong, 0.9-1.1 mm 
long. Ovules 5 to 7 per locule. Immature fruit lin- 


ovate, suborbicular, to obovate, 3-13 X 


margins 


ear, flattened, sessile, straight, retrorsely pilose; 
septum complete; style-like apex glabrous, to 1.5 
mm long; stigma capitate, subentire. Seeds not 
seen. 

Phenology. Flowering in June. 

Habitat and distribution. Scree slopes; 5000— 
5900 m. Endemic to Nepal. 

Additional specimen examined. NEPAL. me Sya 
Gompa, 29°10'N, 82°59'E, Stainton 4332 (BM, E). 


Desideria haranensis is named in honor of Hi- 


roshi Hara (5 January 1911-24 September 1986), 


an eminent Japanese botanist and the discoverer of 


this species and D. nepalensis. The new name is 
proposed because the transfer of Ermaniopsis pum- 


ila to Desideria would create a homonym of D. pum- 
ila (Kurz) Al-Shehbaz, which is based on the earlier 
published Parrya pumila (Kurz, 1872). 

Desideria haranensis is a very rare species 
known thus far only from the two collections cited 
above. It is most closely related to D. pumila, from 
which it is distinguished by having papery instead 
of thickish petiolar bases, toothed instead of tooth- 
less filaments, subentire instead of 2-lobed stigmas, 
and several-flowered racemes instead of solitary 
flowers. 


Desideria prolifera (Maxim.) Al-Shehbaz, 
comb. nov. Basionym: Parrya prolifera Max- 
im., Fl. Tangutica 56. 1889. Ermania prolifera 
(Maxim.) O. E. Schulz, Bot. Jahrb. Sys 

98. 1933. Christolea prolifera (Maxim.) Ovcz., 
Sovetsk. Bot. 1941(1 & 2): 151. 1941. Oreob- 
lastus proliferus (Maxim.) Suslova, Bot. Zhurn. 
(Moscow & Leningrad) 57: 652. 1972. Chris- 
tolea prolifera (Maxim.) Jafri, Notes Roy. Bot. 
Gard. Edinburgh 22: 53. 1955. TYPE: China. 
Tibet: Kon-chun-ua, 14,500 ft., 3 July 1984, 
N. M. Przewalski s.n. (holotype, LE!; isotypes, 
K!, P!, РЕ!). 


Plants scapose, villous to pilose. Trichomes sim- 
ple. straight, to 1.5 mm long. Stems absent. Basal 
leaves subfleshy; petiole (0.2—)0.8—2(-3) cm long, 
persistent, sparsely to densely pilose or villous, cil- 
iate, somewhat papery at base; leaf blade broadly 
ovate, suborbicular, obovate, to spatulate, 2—10 
(-15) x 2-9(-12) mm, villous or pilose, base ob- 
tuse to cuneate, margins (3 to)5- to 9-toothed, rarely 
subentire, apex subacute. Stem leaves absent. 
Flowers solitary from basal rosette. Pedicel ascend- 
ing-divaricate, straight, (0.2—)0.5—1.5(-2.5) ст 
long, villous. Sepals free, oblong, 6–7 X 2-2.5 mm, 
usually persistent, pilose, base not saccate, margins 
membranous. Petals purplish green, broadly ob- 
ovate, 1.1–1.4 cm X 4—5 mm, apex subemarginate; 
claw 6-7 mm long. Filaments white, dilated at 
base, toothless, median pairs 4—6 mm long, lateral 
pair 3—4 mm long; anthers 1.2-1.6 mm long. Fruit 
linear to linear-lanceolate, (2.5—)4–6.5(–7.2) cm X 
(3—)4—5 mm, flattened, sessile, straight; valves ob- 
scurely veined; replum and valves pilose to villous; 
septum complete; style obsolete; stigma capitate, 2- 
lobed. Seeds oblong, 2.5-3.5 X 1.4—1.7 mm 


Phenology. Flowering July and August. Fruit- 
ing July through September. 

Habitat and distribution. Scree slopes, sili- 
ceous shist; 4700—5900 m. Endemic to China 
(Qinghai, Xizang). 


Annals of the 
Missouri Botanical Garden 


Selected specimens examined. CHINA. Qinghai: Bay- 
adoi Xian and Chindu Xian, on 


road between Madoi and Yushu, 34°7'N, 97°39'Е, Hi 
Bartholomew, Watson & Gilbert 1684 (BM, CAS, E 
HNWP, MO); Tangula Shan, Tangula s, 32°53'N 

Xue Shan 


91°54'Е, С. ve S. s Miele 9436/08 (GOET, МО); 
HN 


Community, An 


tangula Shan, 

Miehe, 9495/14 (G ОЕТ, 
pass E of Zongang/Wangda, 29942" N. 

8866 (GOET, MO); Demula Shan, Basu Xian, varied 
Tibet Team 73-1253 (KUN, PE); Bi у 5 
Qinghai-Xizang Team 11172 (КИМ); Zhongba, Qinghai. 
Tibet Team 6537 (KUN); peak of Sengge, near Shingkyer 
Yubrong, 24 July 1951, Aufschnaiter s.n. (BM) 


Maximowicz (1889) compared Desideria prolifera 
(as Parrya) with D. himalayensis and D. flabellata 
and discussed their distinguishing characters. Both 
D. prolifera and D. flabellata have similar flower 
size and their calyces tend to persist. The principal 
feature separating them is that the flowers in D. 
flabellata are arranged in distinct racemes, whereas 
in D. prolifera they are solitary from the basal ro- 
sette. 


8. Desideria pumila (Kurz) Al-Shehbaz, comb. 
nov. Basionym: Parrya pumila Kurz, Flora 55: 
285. 1872. Christolea pumila (Kurz) Jafri, Fl. 
West Pakistan 55: 157. 1973. Vvedenskeyella 
pumila (Kurz) Botsch., Bot. Mater. Gerb. Bot. 
Inst. Komarova Akad. Nauk S.S.S.R. 17: 176. 
1955. Solmslaubachia pumila (Kurz) Dvořák, 
Folia Рпгодоуса. Fak. Univ. Purkyne Brne, 
Biol. 13(4): 24. 1972. TYPE: Kashmir (as Ti- 
bet). Rupschu, 15,000-18,000 ft., F. Stoliczka 
s.n. (holotype, CAL?; isotype, K!). 


Ermania koelzii O. E. Schulz, Repert. Sp. Nov. e 94 
31: 332. 1933. TYPE: Kashmir. Rupshu, Куеп 
19,000 ft., 9 July 1931, Walter Koelz 2231 үс 
B!). 


Ermania уала Botsch., Bot. Zhurn. (Moscow & Lenin- 

: 730. 19 56. Based on the invalidly риђ- 
‚. bifaria acne Bot. Mater. 
Nauk S.S.S.R. 17: 
1955. Oreoblastus bifarius (Botsch.) Suslova, 
Bot. Zhurn. (Moscow & Leningrad) 57: 652. 1972. 
Syn. nov. о. China. Xinjiang: Kuen-Lun, Нит- 
boldt Range, Ulan-Bulak, 4200 m, 30 June 1894, W. 
Roles. s.n. (holotype, 


Plants scapose, pilose to tomentose. Trichomes 
simple, straight, to 0.5 mm long, mixed on leaves 
with short-stalked forked ones. Stems absent. Basal 
leaves fleshy; petiole 2-10 mm long, persistent, 
densely pilose with simple trichomes, ciliate, ex- 
panded and papery at base; leaf blade broadly 
ovate, suborbicular, obovate, to spatulate, 2-14 X 
1-11 mm, densely tomentose or pilose, base ob- 
tuse, margins 3- to 7-toothed to repand, apex ob- 


98°0'E, Dicko ré 


tuse. Stem leaves absent. Flowers solitary from bas- 
al rosette. Pedicel ascending-divaricate, straight, 
3-10 mm long, pilose. Sepals free, oblong, 3—4 х 
1.5-2 mm, caducous, pilose, base not saccate, mar- 
gins membranous. Petals creamy white to purplish 
green, broadly obovate, 6-8 X 3-4.5 mm, apex 
subemarginate; claw 3—4 mm long. Filaments 
white, dilated at base, toothless, median pairs 3—4 
mm long, lateral pair 2–2.5 mm long; anthers паг- 
rowly oblong, 0.9-1.2 mm long. Ovules ca. 7 per 
locule. Immature fruit oblong-linear to linear-lan- 
ceolate, 1-2 cm X 2-3 mm, flattened, sessile, 
straight, retrorsely pilose along replum; valves gla- 
brous; septum complete; style obsolete; stigma cap- 
itate, 2-lobed. Seeds not seen. 


Phenology. Flowering June and July. 

Habitat and distribution. Limestone, mica 
shist; 4200-5800 m. China (Xinjiang,? Xizang), 

ashmir. 

Botschantsev's (1955) description of Ermania bi- 
faria was invalid because he placed the species in 
what was then an invalidly published genus. When 
he (Botschantsev, 1956) validated Ermania, he list- 
ed E. bifaria with full reference to his earlier work. 
Therefore, the correct date of the valid publication 
of E. bifaria should be Botschantsev's 1956 instead 
of 1955 work. 

Botschantsev (1955) recognized two species in 
Vvedenskeyella Botsch., of which the generic type, 
И kashgarica Botsch., has been transferred to 
Phaeonychium (Al-Shehbaz, 2000b). The second 
species, which is based on Parrya pumila, is as- 
signed here to Desideria. Apparently, Botschantsev 
did not examine the type material of P. pumila, as 
evidenced from his description of the same species 
as Ermania bifaria. Phaeonychium differs from De- 
sideria by having a thick and compact instead of 
slender and rhizome-like caudex, pinnately veined 
instead of often palmately veined leaves, incumbent 
instead of accumbent cotyledons, and fruit valves 
without instead of with prominent marginal veins. 

Desideria pumila was said to occur in Xizang 
(Kuan, 1985; An, 1987), but I have not seen any 
material other than the types cited above, which 
were collected from Rupshu, Kashmir. Jafri (1973) 
considered the species (as Christolea) to be very 
closely related if indeed different from what he 
called C. lanuginosa (Hook. f. & Thomson) Ovez. 
However, the last species is clearly unrelated to D. 
pumila and has been treated in Eurycarpus by Al- 
Shehbaz and Yang (20004). 


Volume 87, Number 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


9. Desideria baiogoinensis (K. C. Kuan & Z. X. 

An) Al-Shehbaz, comb. nov. Basionym: Chris- 
tolea baiogoinensis K. C. Kuan & Z. X. An, 
C. Y. Wu, Fl. Xizang. 2: 388. 1985. TYPE: 
China. Xizang: Baiogoin, 5100 m, 18 June 
1976, K. ¥ Lang 9460 (holotype, PE!; isotype, 
РЕ!). 

Plants scapose, villous. Trichomes simple and 
short-stalked forked, straight, to 1 mm long. Stems 
absent. Basal leaves subfleshy; petiole 0.4—1.6 cm 
long, persistent, villous, ciliate, somewhat papery 
at base; leaf blade broadly ovate, suborbicular, or 
obovate, 4-8 X 3-6 mm, villous, base obtuse to 
cuneate, margins 3- to 7-toothed, apex acute. Stem 
leaves absent. Flowers solitary from basal rosette. 
Pedicel ascending-divaricate, straight, 0.5-2 cm 
long, villous. Sepals free, oblong, 4—6 X 1.5-2.5 
mm, usually persistent, pilose, base not saccate, 
margins membranous. Petals purplish, broadly ob- 
ovate, 7-1.2 cm X 3 
ate; claw 4—6 mm long. Filaments white, dilated at 
base, toothless, median pairs 3.5—5 mm long, lat- 
eral pair 2-2.5 mm long; anthers 1-1.2 mm long. 
Ovules 15 to 20 per locule. Fruit ovate to lanceo- 
late, 1-2.5 cm X 6-9 mm, flattened, sessile, 
straight; valves prominently reticulate veined; re- 


mm, apex subemargin- 


plum and valves villous; septum complete; style 
0.5-1 mm long; stigma capitate, 2-lobed. Seeds ob- 
long, 1.5-2 X 0.8-1.1 m 


Phenology. Flowering June and July. Fruiting 
July and August. 

Habitat and distribution. Open sand and grav- 
el; 4700—5600 m. Endemic to China (Qinghai, Xiz- 
ang). 

Selected specimens examined. ere Xizang: Amdo 
Xian, Tao Deding 10819 (HNWP, KUN); Baingoin Xian, 
Lang Kaiyong 9469 (PE), 9487 d Tumain, Yang 
inxi (KUN), Qinghai- SEMAINE Team 177 


g-Tanggulashanqu, 34°35'N, 92?44' E, С. & S. Miehe 


oli 
9416/00 (GOET, MO). 


The species was included in the Flora of Qing- 
hai (Huang, 1997b), but I have not seen any ma- 
terial from that province. 


10. Desideria mirabilis Pamp., Bull. Soc. Bot. 
Ital. 1926: 111. 1926. Christolea mirabilis 
(Pamp.) Jafri, Fl. West Pakistan 55: 160. 1973. 
TYPE: [Kashmir.] Karakorum; above Caracash 
Valley, Chisil Gilgha Pass, 5360 m, 28 June 
1914, G. Dainelli & O. Marinelli 2 (lectotype, 
here designated, FI, photo!). 


pu ee Jafri, Notes Roy. Bot. Gard. Edinburgh 
: 58. 1955. Syn. nov. TYPE: Kashmir. Shaksgam 


Valley, di m, 3 July 1926, R. C. Clifford 7 (ho- 
lotype, 
picos hoan umensis Y. H. Wu & Z. X. An, Acta 
ax. Sin. 32: 577. 1994. Syn. nov. TYPE: Chi- 
na. JXinjiang: Pishan (Guma), less onis 5250 m 
y 19 n Expedition 5100 


uslova, Novosti Sist. Vyssh. Rast. 10: 
163. 3. Syn. nov. TYPE: Tajikistan. Pamir: above 
nd near Zor 4900 m, 10 Aug. 1970, T. Sus- 


Acad. Im er. ч 4: 89 
1896. TYPE: same as that of Desideria pamirica. 


Plants 2-10 cm tall. Trichomes simple and to 1.5 
mm long, rarely mixed with forked ones. Stems dis- 
tinct, simple, densely pilose. Basal leaves subfle- 
shy; petiole 3-15 mm long, densely pilose, not ex- 
panded or papery at base; leaf blade flabellate to 
spatulate-orbicular, (2-)5-15 X 3-9(-15) mm, pi- 
lose, base cuneate, margins 8- to 10-toothed, apex 
acute; teeth to 8 mm long. Stem leaves similar to 
basal. Racemes 8- to 20-flowered, 
Fruiting pedicels ascending, straight to curved, 5- 
10 mm long, spreading pilose. Sepals united, (2.5—) 
3.5-5.5 X 1.5-2.5(-3) mm, persistent, densely pi- 
lose, base not saccate; calyx lobes ovate, unequal, 

2 mm long, margins membranous. Petals pur- 
ple to purplish green with yellowish base, obovate, 
5-8 X (1.5-)2.5-3 mm, apex obtuse; claw 2.5-4 
mm long. Filaments white, slightly dilated at base, 
median pairs (3—)4—5 mm long, lateral pair (2- 
)2.5-3.5 mm long; anthers oblong, 0.5—0.8 mm 
long. Ovules 12 to 18 per ovary. Fruit linear, 1-2(-3) 
cm X ca. 2 mm, slightly flattened to subterete; 
valves pilose, distinctly veined; septum perforate or 
reduced to a narrow rim, membranous; style obso- 
lete to 0.7 mm long; stigma 2-lobed. Seeds oblong, 
1.5-1.8 х 0.8-1 mm, papillate. 


ebracteate. 


Phenology. Flowering July and August. Fruit- 
ing August and early September. 

Habitat and distribution. Gravelly slopes; 
4000-5000 m. China (Xinjiang), Kashmir, Tajikis- 
tan. 


Selected specimens examined. KASHMIR. Karako- 
rum: above Caracash Valley, Chisil Gilgha Pass, 5300 m, 
21 ie 1914, Dainelli & Marinelli 2 (photo, FI). TAJI- 

KISTAN. Pamir: Badakhshan Mt., close to river Mald- 
zhuran, Јаја 700 (LE); Ва яа Mt., 7 km N of Pass 
Takhta-Korum, Tzvelev 1060a (LE); Badakhshan Mt., ba- 
sin of 2 Pshart, Tzvelev 535 (LE); valley of river Chun- 
jabay, Kuzmina 6060 (LE); Checkekty slope. 19 Aug. 
1965, Ikonnikov s.n. (LE) 


Although Pampanini (1926) did not cite any col- 
lections within the original description of Desideria 
mirabilis, he listed in the preceding discussion 


562 


Annals of th 
Missouri ра Garden 


three localities from which the species was collect- 
ed, and he (Pampanini, 1930) gave the details of 
these three syntypes. 

y their reduction of Desideria mirabilis to syn- 
onymy of Cheiranthus himalayensis (as Ermania or 
Christolea), Schulz (1927a, 1936), Botschantsev 
(1955, 1956), and Jafri (1955) overlooked the fact 
that the latter species has bracteate instead of 
ebracteate racemes, septate instead of eseptate 
fruits, free instead of united sepals, and fruits 3— 
4.5 mm instead of ca. 2 mm wide. The differences 
between the two species are so significant that it is 
hard to imagine they are conspecific. 

Although they correctly noticed that the sepals 
in Christolea karakorumensis are united, Wu and 
Ап (1994) were probably unaware of Desideria mir- 
abilis, a species endemic to the Karakorum Moun- 
tains and indistinguishable in every aspect from 
their novelty. 

Jafri (1955) did not mention the connation of se- 
pals in his original description of Christolea sca- 
posa, though the illustration clearly shows gamo- 
sepalous calyces. - contrast, his (Jafri, 1973) 
illustration did not show gamosepaly accurately, 
though both dedi were based on the same 
specimen. However, he mentioned that the sepals 
are "rarely connate below." Jafri (1955) suggested 
that C. scaposa is related to C. prolifera, while he 
(Jafn, 1973: 158) indicated that the species is 
closely related to Desideria suslovaeana except for 

"the absence of septum and slight difference in 
leaves." In my opinion, C. scaposa is indistinguish- 
able from Desideria mirabilis and is unrelated to D. 
prolifera. The latter has solitary flowers and decid- 
uous polysepalous calyces, whereas D. mirabilis 
has the flowers in racemes and persistent gamosep- 
alous calyces. 


11. upside к Н. Нага, Ј. Јар. Вог. 

YPE: Nepal. Barum Valley, 

Pe ft., a May 1954, L. W. Swan 71-72 
(holotype, ВМ!). 


Plants 2-3 cm tall. Trichomes simple, straight, 
to 1 mm long. Stems minute, simple, glabrous. Bas- 
al leaves subfleshy; petiole 2-5 mm long, sparsely 
pilose with simple trichomes, ciliate at base, not 
expanded or papery at base; leaf blade broadly ob- 
ovate to subflabellate, 2-3 х 1-3 mm, densely pu- 
bescent, base cuneate, margins 3- to 5-toothed, 
apex acute. Stem leaves absent. Flowers 2—4, 
ebracteate. Pedicel ascending, straight, 3-5 mm 
long, solitary from basal rosette, spreading pilose. 
Sepals united, 5-6 X 3—4 mm, densely pilose, base 
not saccate; calyx lobes ovate, 1.5-2 mm long, mar- 


gins membranous. Petals ?purplish, obovate, 11— 

x mm, apex obtuse; claw 6-7 mm long. 
Filaments slightly dilated at base, median pairs 
4.5—5.5 mm long, lateral pair 3—4 mm long; anthers 
oblong, 0.9—1.1 mm long. Ovule number, fruits, and 
seeds unknown. 


Desideria nepalensis is known only from the type 
collection made at an altitude of about 5400 m. It 
is readily distinguished from D. mirabilis by its 
much larger flowers (see key). 


Literature Cited 


Al-Shehbaz, 1. A. 1987. The genera of Alysseae (Cruci 
ferae; Brassicaceae) in the southeastern United n 
J. iip god 68: 185-240. 

The genera of Sisymbrieae (Cruciferae; 

inpr in the southeastern United States. J. Ar- 

nold is 69: 213-237 

eneric limits and taxonomy of Brayopsis 

and s Вы, J. Arnold Arbor. 71: 93- 

109. 


1990b. Brayopsis gamosepala (Brassicaceae), a 
remarkable new species with gamosepalous calyx. Ann. 
Missouri Bot. Gard. 77: 843—844. 

. 2000a. A revision | un (Brassica- 
dad. Edinburgh J. Bot. 57: 157-170. 

. 2000b. A revision af "n genus Phaeonychium 
(Brassicaceae). Nordic J. Bot. 20: 

& 5. L. О'Капе, Jr. 1997. A Aron gamosepala 
and A. tuemurnica Е to Neotorularia (Brassica- 
ceae). Novon 7: 93—¢ 

& H. Ohba. 2000. The status of тие 
and two new M in Dontostemon (Brassi 
сеае). Novon 10: 95-98. 

г С. Yang. er A reconsideration of the genus 
Еигусагрих (Brassicac wi Novon 10: 346—348 

& —. 2000b. A revision of Solmslaubachia 

(Brassicaceae). Harva | Pa ap. Bot. (in press). 
у thr nin In: T. Y. Cheo (editor), Fl. 
Reipubl. Popularis Sin. 33: 289-299, Science Press, 
eijing. 
. 1995. erp In: Z. М. Mao (editor), Fl. Xin- 
jiangensis 2(2): 3 8-229. Xinjiang Science & Technol- 
ору ygiene Publishing House, Urumqi. Science 


88. Brassicaceae. /n: S. S. Charkev- 
icz [khasi h] (edito Plantae vasculares orientis ex- 
tremi Sovietici. 3: 38-1 V ,eningrad. 

> a V. 1955. De Cruciferis notae criticae. ma 

er. Em Inst. У арх Akad. Nauk S.S.S.R. 
pnt 


956. [A review of] S.M.H. Jafri. Christolea: With 
spec ai reference to the species in N.W. Himalayas, W. 
Pakistan = n Bot. Zhurn. (Moscow & Len- 
ingrad) 41: 

Cambessédes, s a Plantae rariores, qe in India or- 
ientali collegit Victor Jacquemont. Pp. 1-56 т V. Jac 
quemond (editor), Voyage dus TInde ес. "1828-1832. 
Paris. 

Chamisso, L. 1831. De plantis in expeditione spe- 
culatoria Romanzoffians observatis disserer pergunt: 

rcticae, quae supersunt. Linnaea 6: 528—544. 
(o repanov, S. K. 1995. Vascular Plants of Russia and 


Volume 87, Number 4 
2000 


Al-Shehbaz 
Gamosepaly in Brassicaceae 


563 


Adjacent States (the Former USSR). Cambridge Univ. 

Press, ew. 
Dar, G. H. oR, 1990. New plant taxa es 
the Sind Valley, Kashmir. n Bombay Nat. Hist. Soc. 8 
279. 


Drury, W. H., Jr. & R. С. Rollins. 1952. The North Amer- 
ican representatives of Smelowskia (Cruciferae). Rho- 
dora 54: 85-119. 

Dudley, T. R. 1964. Synopsis of the genus Alyssum. J. 

Arnold Arbor. 45: 358-373. 

Сане. У. Р 1976. De Genere Dimorphostemon Kitag. 
(Cruciferae). Novosti Sist. Vyssh. Rast. 13: 120-130. 
Greuter, W, R. K. Brummitt, E. Farr, N. Kilian, P. M. 
Kirk & P. C. Silva (Editors). 1993. Names in Current 

Use for Extant Plant Genera. Regnum Veg. 129. 

McNeill, F. R. Barrie, H. M. Burdet, W. G. 

Chal oner, V. Demoulin, T. S. Filgueiras, D. H. Nicolson, 

P. C. Silva, J. E. Skog, P. Trehane, N. J. Turland & D 


Nomenclature (Saint Louis Code). Regnum Veg. 138. 

j ~ sei & G. H. preis 1993. 
Hesperideae. /n: Sharma & N. P. Balakrishnan 
(editors), d i 2: purs и Survey of In- 
dia, Calcut 

Hara, H. 1974. New or noteworthy flowering plants from 
eastern Himalaya (15). J. Jap. Bot. 49: 193-205. 

New or noteworthy flowering plants from 
eastern Himalaya (16). J. Jap. Bot. 50: 263-271. 

— ———. 1979. Cruciferae. In: H. Hara & L. H. J. 
liie (editors), An enumeration of the flow 
of Nep 


Wil- 
wering plants 


al. Bull. Brit. Mus. (Nat. Hist.), Bot. Publ. 810: 


38—46. 
ve I. C. 1968a. Arabidieae. /n: K. H. Rechinger (ed- 
tor, Fl. Iran. 57: 193-218. Akademische Druck-u. 
та авс Graz. 
.1 . Sisymbrieae. /n: К. H. Rechinger ferto. 
Fl. Iran. 57: 309-342. Akademische Druck-u. Verlag 
sanstalt, Graz. 

Hooker, J. D. & T. Thomson. 1861. Precursores ad floram 
Indicam, Cruciferae. J. Linn. Soc., Bot. 5: 128- ки 

. Anderson. 1872. Cru сене In: J. D. Hooker 
(editor), Flora of British India, 1: 128—167. L. LT 
London. 

Huang, R. F. 1997a. New plants of Cruciferae from Qing- 
hai, onem Acta Phytotax. Sin. 35: 556-561. 

997b. Cruciferae. In: S. W. Liu (editor), Fl. Qin- 
Mia бА 410-505. Qinghai People’s Publishing 
House, Xining. 

Hultén, E. 1945. Flora of ста and Yukon. Lunds. Univ. 
Атай М. Е. Avd. 2, 41: 799-978. 

Jafri, S. M. H. 1955. B cae. With special РЕНЕ [о 
the species in N.W. Himalayas, W. Pakistan and Af- 
sii Notes Roy. Bot. Gard. Edinburgh 22: 49-59. 

3. Brassicaceae. In: E. Nasir & S. I. Ali (ed- 
jii E T Pakistan 55: 1-308. Ferozsons, Karachi. 

Jurtsev, B. A. 1975. Ermania. In: А. 1. Tolmatchev (edi- 
tor), Flora Arctica URSS. 7: 61-64. Leningrad. 


Kuan, K. C. 1985. pcs In: C. Y. E. = Fl. 
Xiz zangica 2: 323-411. Science то 

Нез ње rr In: E (editor), Fl. 
Reipubl. Populate Sin. 33: TR 396. Science Press, 
Beijing 

Kurz, i: 1872. Drei neue Tibetische Pflanzen. Flora 55: 
284— 


uod C. J. 1889. Historia naturalis itinerum N 
Przewalski per Asiam centralum. Pars botanica, Vol. 2 
Fl. Tangutica. Imperial Academy of Sciences Press, St. 
кел рр. 1-1 

ш; р. 1893. Вгауа uniflora. Hooker's Icon. Pl. 23: РІ. 


ео. P. М. & S. Yunussov. 1978. Ermania. Fl. 
Tadzhitskoi SSR. 5: 170-176. Academy of Sciences, 
Leningrad. 
Pachomova, M. С. 1974. Desideria. In: А. 1. Vvedensky 
(editor), Conspectus florae Asia Mediae. 4: 157-156. 
of Sciences of UzSSR, Tashkent. 


eic R. baee p mirabilis Pamp., gen. et 


a Cr cifera anomala del Dum (Asia 
Cental Boll. Soc. Bot. Ital. 1926: 107- ri 
. La flora del Caracorüm. Pp. EA ) in G. 


о rd ey Speditione italiana de Filippi nell 
Himalaya, Caracorüm et Turchestàn Cinese (1913- 
1914). Ser. 2, vol. 10. Nicola Zanichelli, Bologna 

Rollins, R. C. 1993. The Cruciferae of qae North 
America. Santor Univ. Press, Stan 

Schulz, O 4. Cru кета ем ae. e. In: A. Bugler 
(editor), ле: IV. poni eft 86): 1-388. Verlag von 
Wilhelm Engelmann, Leip 


3 2 Deiode zur M der Cruciferen des 
мрн ын Himalaya-Gebirges. Notizbl. Вог. Сап 
Berlin- pera 9: 1057-1095. 

27b. Über die Gattung Gamosepalum Hausskn. 
ERE Bot Gart. Berlin-Dahlem 10: 109-111. 

. Amerikanische Cruciferen verschiedener 


Небо. a Bot. Gart. Berlin-Dahlem 10: 558- 


1931. Seltene P ries aus Kashmir. Repert. 

Sp. Nov. Fat Veg. 33: 330-334. 

— ———. 1933a. Kurze на iiber neue Gattungen, Sek- 
tionen a noe der Cruciferen. Bot. Jahrb. Syst. 66: 
bg 


: Über verschiedene Cruciferen. Repert. 
Sp. NE p Veg. 33: 183-191. 
—— ———. 1933c. Seltene Cruciferen aus Kashmir. Керегі. 
Spec. Nov. Regni Veg. 31: 330—334. 
. 1936. Cruciferae. /n: A. Engler & K. Prantl (ed- 
itors), Nat. Pflanzenfam., ed. E т 227-658. Verlag 


>= Ка ће RÀ »* d 

Zhurn. (Mesum & Leningrad) 57: 647-653 

973. Species nova generis Desideria Pup. in 

Pamir. Novosti Sist. Vyssh. Rast. 10: 160-165. 

Wu, Y. H. & Z. X. wo new species of Chinese 

Chr. istolea. Acta ТЕР" з = h 7- ih 

Yunusso esideria . Ovezinnikov & S. 
VM eio "FL. Todihitskol SSR. 5: 176-177. 

ad. 


Academy of Sciences, Leningr: 


Peter Goldblatt,? Peter Bernhardt,? and 


ohn C. Manning* 


ADAPTIVE RADIATION OF 
POLLINATION MECHANISMS 
IN IXIA (IRIDACEAE: 
CROCOIDEAE)! 


ABSTRACT 


Field observations, floral dissections, and pollen load analyses of insects captured on 20 species of /xia (Iridac eae), 
this southern African genus of : 


atrandra, 1. curta, 1. ! I. maculata, 1. metelerkampiae, and I. versicolor have salver-shaped, nectarless flowers, in 
bright colors contrasting with dark “beetle marks” and are ти и, by hopliine scarab beetles. Four /xia 
species with narrowly tubular flowers, spreading tepals, and ample nectar are pollinated by long-proboscid flies (Moe- 


p longirostris and Philoliche species). Three additional species with tubular flowers, and modest nectar 

mes, appear to be pollinated by the pieriid парна Colias electo (Ixia orientalis), or by a combination of hopliine 
га and tabanid flies with short probosces (/. aurea, 1. esterhuyseniae, [. tenu ifol ia). The remaining species are 
largely и by anthophorine bees ог Apis mellifera, but bee pollination comprises three discrete systems. Spec лев 


pollina y Ant 
ave с apt 

by pollen-collecting xg il 

presentation, nectar 


the one in which flowers are cup-sha 
of e Bebo" into an elongate tub 
or the 


и strategies 
Key 


short, 
А incompletely from the base. Vien пне и suggests that the ancestral pollination system 


closed perianth tube е, ee stub 


e development of hen anther dehiscense must be regarded as specialized adaptations related to their derived 


ords: Ados Hopliini, Iridaceae, /xia, Nemestrinidae, pollination biology, Tabanidae. 


Macroevolution of the African Iridaceae depends 
in part on the plasticity of pollination mechanisms. 
Nivenia (10 spp.) appears to be one of the few gen- 
era of any size in which species are pollinated pri- 
marily by two pollinator groups, long-proboscid 
flies (Nemestrinidae) or long-tongued anthophorine 
bees (Goldblatt & Bernhardt, ). In contrast, 
the larger genera of African Iridaceae are сћагас- 
terized by a diversity of pollination systems. For 
example, Lapeirousia consists of species pollinated 
by long-proboscid flies, bees and butterflies com- 
bined, or moths (Goldblatt et al., 1995). Romulea 
species exploit scarab beetles, pollen-collecting 
bees x in one case, nemestrinid flies (Goldblatt et 

998a; Manning & Goldblatt, 1996, and un- 
ране), The majority of species of Gladiolus ар- 
pear to be pollinated primarily by nectar-feeding 
anthophorine bees (Goldblatt et al., 1998b), but re- 
cent fieldwork also indicates that some red-flowered 


species are pollinated by the large butterfly Aero- 
petes (Johnson & Bond, 1994), while other species 
are dependent on andrenid bees, a combination of 
these bees and ћоршпе beetles (Goldblatt et al., 
1998a), long-proboscid flies, moths, or birds (Gold- 
blatt & Manning, 1998). The adaptive radiation of 
floral characters thus appears to have played a 
prominent role in evolution and speciation within 
African Iridaceae. 

Ixia, a moderate-sized genus of Iridaceae sub- 
family Crocoideae Burnett, Outl. Bot.: 451 Jun. 
1835. (syn. Ixioideae Klatt, 1866 аз subordo 
Ixieae), comprises some 52 species (Lewis, 1962; 
de Vos, 1999; Goldblatt & И 1999, 2000). 
The genus extends from Namaqualand in the north- 
west of the subcontinent to Eastern Cape Province 
near Grahamstown in the east, a range that coin- 
cides almost exactly with the southern African win- 
ter-rainfall zone. Species diversity is highest in the 


! Support for this study by grants 5408-95 and 5994-97 from the National Geographic d is gratefully acknowl- 


РЧ hi Sus Robert 


Brooks, University of Kansas, and Holger Dombrow, 


insect identifications. 


rukoff Curator of African Botany, Missouri Botanical Garden, P.O. Box 299, St. qu Missouri 63166, 


USA. 
* Department of Biology, St. Louis University, St. Lou 


, uis, Missouri 63103, U.S.A. 
* Compton Herbarium, National Botanical Institute, P. Bag 


7, Claremont 7735, South Africa. 


ANN. Missour! Вот. GARD. 87: 564—577. 2000. 


Volume 87, Number 4 
2000 


Goldblatt e 


al. 565 
Pollination АНИ in Ixia 


eographical center of this zone, the Western Cape 
Province. At first glance, [xia appears to exhibit 
only moderate floral diversity compared, for exam- 
ple, with the ca. 245 sub-Saharan African species 
of Gladiolus, the 52 species of Watsonia, or the ca. 
40 species of Lapeirousia. All Ixia species have 
radially symmetric perianths and relatively small 
flowers 1.5—3 cm in diameter, typically arranged in 
a dense spike at the apex of a slender flowering 
stem. The androecium is also symmetric in the ma- 
jority of species, and the style diverges into three 
branches near the mouth of the floral tube, at or 
below the level of the anthers. On the other hand, 
Ixia flowers vary remarkably in the range of peri- 
anth color and pigmentation patterning, perianth 
tube length and structure, staminal coalescence 
and mode of anther dehiscence, and nectar pro- 
duction. A particularly unusual aspect of floral var- 
iation within this genus is that while the anthers of 
most Iridaceae are longitudinally dehiscent and 
mounted on long filaments, species of Ixia subg. 
Dichone have short, stubby filaments and short, in- 
flated anthers with a narrow connective. The an- 
thers of all but one of the nine species of this sub- 
genus dehisce incompletely, and are more or less 
basally porose in some species (Lewis, 1962). 
Field study of the pollination systems of a range 
of [xia species was undertaken to obtain additional 
information about the evolution of adaptive mech- 
anisms, to provide insights into the way the flowers 
of Ixia species function, and to help understand the 
seeming contradiction in apparently simple and not 
particularly diverse floral morphology and a range 
of quite distinct pollination systems. 


METHODS 


FLORAL PHENOLOGY, LIFE SPAN, AND FLORAL 
PRESENTATION 


Direct observations are presented on 20 /xia spe- 
cies made during the years 1993 to 1999 in the 
field and in living а а! Kirstenbosch Bo- 
tanic Gardens, Cape Town. Observations on the 
pollination biology of Ixia were made in the course 
of other field research in the southern spring at 
various sites (Table 1) in the southwestern Cape 
and the western Karoo, South Africa, areas of Med- 
iterranean climate with wet winters and dry sum- 
mers. Observations of insect foraging involved 4— 
10 hours per plant species and included recording 
of both floral attractants (pigment patterns, scent), 
the mode and timing of anthesis (opening of indi- 
vidual buds), anther dehiscence, expansion of stig- 
matic lobes, the behavior of insects on the flower, 
and the taxonomic diversity of floral foragers. The 


range of species studied includes examples from all 
infrageneric taxa (de Vos, 1999) and all the major 
floral types except 1. acaulis, a rare limestone en- 
demic, which has long-tubed yellow flowers borne 
at ground level and a subterranean ovary (Goldblatt 
& Manning, 1993 

Floral scent was noted in the field and in culti- 
vated plants. Scents too weak to be discerned by 
the human nose were recorded after individual 
flowers were picked and placed in clean, lidded 
glass jars and stored in a warm place. The contents 
of each jar were smelled after a minimum of 60 
minutes (Buchmann, 1983). 


~ 
" 


NECTAR ANALYSIS 


Nectar volume measurements were taken pri- 
marily from unbagged flowers in the field, reflecting 
both rates of secretion and depletion, but some spe- 
cies were sampled in the laboratory where no in- 
sects visited the flowers. Nectar sugar chemistry 
and concentration are unlikely to be affected sig- 
nificantly, as opposed to sampling bagged flowers. 
Ideally both bagged and unbagged flowers should 
be sampled. Studies on nectar characteristics of 
Lapeirousia (another southern African genus of Ir- 
idaceae—Crocoideae) suggest that nectar concentra- 
tion is not affected using unbagged flowers versus 
those examined in the laboratory where insects 
were excluded (Goldblatt et al., 1995). Nectar vol- 
ume is expected to be lower in unbagged versus 
bagged flowers, but sampling of nectar of unbagged 
flowers in populations being visited by pollinators 
reflects a realistic situation that confronts a partic- 
ular pollinator in the field. To collect nectar whole 
flowers were picked and nectar was withdrawn from 
the base of the perianth tube with 3 pl capillary 
tubes after separating the ovary from the perianth 
base. The percentage of sucrose equivalents in 
fresh nectar was measured in the field or laboratory 
on a Bellingham and Stanley hand-held refractom- 
eter (0–50%) from five or more individuals per pop- 
ulation, unless fewer individuals were available. 
Additional nectar samples were dried on Whatmans 
filter paper no. 1 and sent to B.-E. van Wyk, Rand 
Afrikaans University, Johannesburg, for HPLC nec- 
tar chemistry analysis. 


INSECT OBSERVATION, POLLINATION MECHANISMS, 
AND POLLEN LOAD ANALYSIS 


Observations of insects on [а flowers included 
whether insects contacted anthers and stigmas 
while foraging. Insects observed probing the floral 
tube or brushing the anthers or stigmas were cap- 
tured and killed in a jar using ethyl acetate fumes. 


566 Annals of the 


Missouri Botanical Garden 


Table 1. 


Study sites and voucher information for species studied. Vouchers are housed at MO (Goldblatt) or at 


NBG (other collectors). All study sites are in South Africa. 


Species 


Study site 


Voucher 


Subgenus Ixia 
Section Morphixia 
aurea Goldblatt & J. C. Manning 
capillaris L. f. 
нан М. Р. de Vos 


poe D. Delaroche (site 1) 
2) 


_ ~ гм 


a Ker Gawl. 
orientalis L. Bolus 
rapunculoides Delile 
tenuifolia Vent. 
thomasiae Goldblatt 


~ ~ ~ ~ ~ 


Section Hyalis 

1. longituba N. Е. Br. 

I. paniculata D. Delaroche (site 1) 
(site 2) 


1. paucifolia G. J. Lewis (site 1) 
ite 2) 


Section Ixia 


1. atrandra Goldblatt & J. С. Man- 
ing 
1. сипа Andrews 


1. flexuosa L. 

1. lutea Eckl. (site 1) 
(site 2) 
(site 3) 

. maculata L. (site 1) 
(site 2) 


M 


(site 3 
. metelerkampiae L. Bolus 
1. versicolor G. J. Lewis 
Subgenus Dichone 
I. scillaris L. (site 1) 


Western Cape, near Darling 
Cape, near Bot River 
Western Cape, Landrostko 
Western Cape, Cold Bokkeveld 
Wee tern Cape, near Touwsrivier 
estern Cape, Darling, Wa ylands 
Western Cape, near Caledon 
Northern Cape, Bokkeveld Plateau 
Western Cape, Camphill road 


estern 


Western Cape, near Middelpos 


Western Cape, near pii 
Western Cape, Ronde 
Northern Cape, Nieuwoudtville Water- 
fall 
Western Саре, Burger’s Pass 
Western Cape, Worcester district, near 
e We 


Western Cape, Brandvlei Hills 
Western Cape, Versveld Reserve, Dar- 


ing 

Western Cape, Bot River 

Western Cape, Darling Nature Reserve 

Western Cape, Bur Far 

Western Cape, Piketber, 

Western Cape, Waylands, Darling 

Western Cape, Clanwilliam, Farm Yst- 
erfontein 

Western Cape, near Leipoldtville 

Western Cape, Bainskloo 

Western Cape, Strand 


Western Cape, Lion’s Head, above 
Camps Bay, Cape Town 


Goldblatt & SN 10338 
Goldblatt 10674 

Oliver & ue 11461 
Thomps 550 

Goldblatt & ш 11202 
Burgers 

Goldblatt & ETE 9789 
Pretorius 7. 

Goldblatt & Manning 10333 
Goldblatt & Manning 10368 


Goldblatt & Manning 10341 
Goldblatt & Manning 10051 
Goldblatt & Manning 10429 


Goldblatt & Manning 10012 
Goldblatt & Manning 9690 


Goldblatt 10568 
Goldblatt & Manning 10358 


Goldblatt 10671 

Goldblatt 11151 

Goldblatt no voucher 

Goldblatt & Manning no voucher 
blatt & aah ng 10: 


Goldblatt & Manning 10349 
Goldblatt & Manning 10324 


"Wis ‹ 
Goldblatt & Manning 11031 


Barker 2214 


(site 2) Western Cape, Darling Nature Reserve Goldblatt 1196A 


Pollen was removed from insects after specimens 
were pinned. To prevent contamination of the body 
of an insect with pollen carried by another in the 
same jar, each insect was wrapped in tissue as soon 
as it was immobilized by jar fumes. Body length 
and proboscis length of insects were recorded from 
captured specimens. Capturing a bee or fly at any 
site appeared to reduce the insect population sig- 
nificantly. We therefore killed as few insects as nec- 
essary to obtain specimens for identification and 
pollen load analysis. Removal of pollen from insect 


bodies involved gently scraping pollen off the body, 
including the scopae or corbiculae of bees, with a 
dissecting needle (see Goldblatt et al., 1998a, b). 
The residue from needle probes was collected on 
glass slides and mounted in 1-2 drops of Calberla’s 
fluid (Ogden et al., 1974). In the case of flies, which 
are comparatively large insects, sites of pollen de- 
position are quite discrete for a particular plant vis- 
ited, and pollen species can usually be identified 
without recourse to microscopic examination. Pol- 
len grains were identified microscopically by com- 


Volume 87, Number 4 
2000 


Goldblatt et al. 
Pollination Mechanisms in Ixia 


567 


parison with a reference set of pollen grain prepa- 
rations made from plants flowering at the study 
sites. Ixia pollen grains are recognized by their 
large size, perforate-scabrate exine, and monosul- 
cate aperture with prominent 1-banded operculum, 
the latter feature unique as far as is known (Gold- 
blatt et al., 1991). 

Insect specimens were identified by В. У. 
Brooks (Apidae), University of Kansas, H. Dom- 
brow, Worms, Germany (Scarabaeidae), and J. C. 
Manning (Diptera, Lepidoptera). Voucher speci- 
mens are deposited at the Natal Museum, Pieter- 
maritzburg, South Africa, or the Snow Entomolog- 
ical Museum, Lawrence, Kansas. 


RESULTS 


FLORAL PHENOLOGY, LIFE SPAN, FLORAL 
PRESENTATION, AND ATTRACTANTS 


Floral phenology and life span. Species of Ixia 
are seasonal, corm-bearing geophytes of small to 
moderate size, typically 15—40 cm high (Fig. 1). 
Individuals produce a single, simple or branched 
flowering stem annually, and flowering is closely 
synchronized in a population. Inflorescences are 
spikes with helically arranged flowers (Lewis, 
1962). With a few exceptions, flowering occurs in 
late winter and spring (August to October) (Table 
2). This coincides with the period of optimal plant 
growth, during or soon after the main rainy season. 

The pattern of flower buds opening on an inflo- 
rescence is acropetal. In all species a mature bud 
expands in the early to late morning, and the open 
flower typically lasts three or four days. Flowers 
open sequentially, usually one day apart, hence 
there are often three or four flowers open at any 
time on an inflorescence. At sunset the tepals 
loosely close to enfold the anthers and stigmas. Te- 
pals unfold again the next day, between 08.00 and 
12.00 hours, depending on species and to some ex- 
tent on ambient temperature. On cold (« 15?C) or 
overcast days they may unfold partially or not at 

| 


Flowers of all species studied show weak me- 
chanical protandry. In most species the anthers de- 
hisce longitudinally within one to four hours after 
the tepals first unfold (Fig. 2A, B, C). In these spe- 
cies pollen remains in place and exposed in the 
anthers. In contrast, in most species of section Di- 


— 
Figure 1. /xia paucifolia, a typical member of the ge- 
nus, showing general habit, inflorescence, and flowers 


(scale bar 1 ст 


568 


Annals of the 
Missouri Botanical Garden 


Table 2. 


= trace amount too little 


Floral characteristics of /xia species arranged according to flower type. + = presence, — = absence, tr 
to measure volumetrically. Conventional perianth tubes are cylindrical Mid hollow (indicated 


by +), but in closed tubes the perianth tissue tightly envelops the style so that the interior of the tube is filled by 
stylar tissue (indicated by —); additionally, in these species the filaments are contiguous and sometimes fused, thus 


closing the mouth of the tube. 


Lower Perianth ibe Beetle Nectar Flowering 
Species shape color length mm open marks presence time 

Subgenus Ixia: section Morphixia 

1. aurea salver orange ca. 8 + = + Sep.-Oct. 

1. capillaris up blue ca. 7 + = + Aug.—Sep. 

I. esterhuyseniae cup yellow 5 + = + Nov. 

I. latifolia cup pink to mauve 12-14 + - + Aug.—Sep 

1. о сир yellow ca. 8 + = + Зер 

I. orientalis salver ca. 8 + – + Oct. 

1. rapunculoides up blue-mauve ca. 10 + - + Aug.-Sep 

I. tenuifolia salver orange ca. 12 + + tr Sep. 

1. thomasiae cup pink ca. 7 + = Oct 
Section Hyalis 

1. longituba tube pink 30-32 + = Oct. 

I. paniculata tube cream 65-75 + = + Oct.-Nov. 

I. paucifolia tube cream 16-28 + = + Aug.-Sep 
Section Ixia 

1. atrandra salver cream ca. 10 - + - Oct.-Nov 

I. curta salver orange ca. 12 = + m Sep.-Oct 

1. flexuosa salver white/pink ca. 6 = = = Аир.—бер 

I. lutea salver cream 6-12 = + — ug.-Sep 

. maculata salver orange ca. 8 = + Sep.-Oct 

I. metelerkampiae salver mauve ca. 4 Е + = Oct.—Nov 

I. versicolor salver white/pink ca. 10 - + = Sep.-Oct 
Subgenus Dichone 

I. scillaris salver pink 3-4 = - = Oct 


chone the anther dehiscence is virtually porose, 
starting from the base (Fig. 2D). Pollen in these 
species is not shed but remains within the anthers 
unless actively removed by an insect. The time of 
anther dehiscence depends to some extent on am- 
bient temperature and humidity, and anthers de- 
hisce later in wet cool conditions. The three stylar 
lobes, the distal adaxial surfaces of which comprise 
the stigmas, are held together when the flower first 
opens, but they diverge later during the same day. 
e did not determine pollen-stigma compatibil- 
ity ourselves, but Horn (1962) showed that a range 
of species аге self-compatible, including Ixia ma- 
culata and 1. polystachya, which were studied here, 
but have reduced fruit and seed production when 
selfed by hand compared with xenogamous crosses. 
Horn also reported that 1. odorata was self-incom- 
patible. Mechanical selfing is restricted in [xia scil- 
laris by the spatial separation of anthers and stig- 
matic surfaces 
Floral presentation and attractants. Open flow- 


ers are typically held erect to suberect in a fairly 
congested spike (Fig. 1). [xia viridiflora (sect. Mor- 
phixia), not studied here, has elongate, lax spikes, 


and relatively widely spaced on the spike axis. [ма 
odorata is one of two species examined that pro- 
duces an odor, and the flowers have a sweet scent 
reminiscent of commercial cultivars of Viola odor- 
ata. In the other scented species, 1. flexuosa, the 
flowers have a sour, acrid, musky odor, unpleasant 
to the human nose. 

Floral presentation (including shape, pigmenta- 
tion, attractants, and rewards) in /xia falls into four 
main categories (Table 2), although the basic peri- 
anth plan is always symmetrical and actinomorphic. 
The first category, of which 1. latifolia and 1. ra- 
punculoides (sect. Morphixia) are typical examples, 
have cup- or salver-shaped flowers (Fig. 3). The 
floral tube is funnel-shaped with a hollow, narrow 
cylindric basal half and a flared upper half, this 


Volume 87, Number 4 Goldblatt et al. 569 
2000 Pollination {эй ан in Ixia 


enclosing the style and prom os Dé wp dehiscent anthers. —C. 1. вија 25 (sect. Hyalis), with elongate, 
hollow tube with included stamens. —D. 1. scillaris (sect. Dichone), with unilateral anthers dehiscing from the base. 
Whole flowers full size, deis variously eer (drawn by John Manning). 


ranging from fairly wide and 7-10 mm wide at the and located around the center of the flower and 
mouth or narrow and only 2-4 mm wide at the either within the wide part of the floral tube or are 
mouth. The cylindrical part of the tube contains shortly to fully exserted. Anthers are extrorse with 
nectar, which may extend upward into the base of — loculicidal dehiscence, and the pollen adheres to 
the flared portion. The stamens are symmetrical the dehisced anther locules (Lewis, 1962). The 


Annals of the 
Missouri Botanical Garden 


Volume 87, Number 4 
2000 


Goldblatt et al. 
Pollination Ме in Ixia 


style is central in all species and divides apically 
into three lobes that extend between the filaments 
or the lower half of the anthers, depending on the 
level at which the style divides. Perianth coloration 
in flowers of this group is usually pink to mauve or 
blue but may sometimes be deep pink or magenta 
in forms of 1. latifolia. The fragrant flowers of I. 
odorata are yellow and unusually small and crowd- 
ed together, collectively forming the prominent sig- 
nal to pollinators. /xia aurea and I. tenuifolia are 
exceptional among the species with tubular flowers 
in having bright yellow to orange perianths, marked 
in the latter with a dark reddish center. 

A second flower category is exemplified by [xia 
paucifolia (Fig. 1) and I. paniculata (Figs. 2C, 4) 
and is found in all species of section Hyalis (Table 
2). The floral tube is hollow, cylindric, much ex- 
ceeds the tepals, and has a narrow diameter ca. 

5-2 mm wide. The style and stamens are cen- 
trally placed. The anthers may be included in the 
tube (1. paniculata) or, more commonly, are exsert- 
ed (1. longituba, I. paucifolia). Perianth color in 
these tubular flowers is white, cream, or pink, 
sometimes more darkly pigmented around the 
throat. All the tubular flowers are unscented. 

In the third floral category, typified by Ixia ma- 
culata (sect. Ixia), the perianth forms a broad, flat 
salver, typically 2-3 cm in diameter, with the tepals 
extended horizontally at the apex of a filiform floral 
tube (Fig. 2B, 5). The tube is narrow and complete- 
ly filled by the style and closed off at the mouth by 
coherent or coalescent filaments, contains no nec- 
tar, and functions as a pedicel, supporting the te- 
pals and stamens. The perianth color is remarkably 
diverse among species of this group, ranging from 
white or cream to pink or mauve, or dark red, pur- 
ple, turquoise, or intense orange or yellow, but flow- 
ers are usually brilliantly pigmented. The flowers 
are unscented and usually have a dark, contrasting 
central marking, sometimes including the filaments 
and occasionally the anthers, i.e., beetle marks sen- 
su Goldblatt et al. (1998a). Other authors (Steiner, 
1998a, b) have restricted the term beetle mark to 
describe markings that more closely resemble a 
beetle, but our broader definition seems useful. The 
longitudinally dehiscent anthers are centrally 
placed and the pollen is retained on the dehisced 


anthers. Ixia flexuosa has unusually small flowers 
mm in diameter, and they 
are fragrant and borne on particularly slender, wiry 
peduncles. Its crowded inflorescences tend to droop 
and wave in the wind, unlike the erect spikes of 
other species in the group. 

The last floral category, represented by /xia scil- 
laris (Fig. 6), is restricted to species of subgenus 
Dichone (Lewis, 1962). The perianth is salver- 
shaped and pale to deep pink, often darker pink 
toward the center of the flower. The flowers are se- 
cund with tepals vertically oriented. The perianth 
tube is filiform with the tube wall enveloping the 
style in a tight, continuous sheath. The tube con- 
tains no detectable nectar. The stamens are fully 
exserted, extending horizontally on short, somewhat 
stubby filaments. The anthers are 3—4 mm long, 
usually as long as the filaments, and inflated and 
basally porose. Dehiscence is delayed and begins 
from the base and does not reach the anther apices 


for this group, ca. 1. 


(Fig. 2D). In Z. scillaris the anthers are more or less 
unilateral, and lie slightly below horizontal, per- 
haps due only to gravity. Pollen is retained within 
anther locules and not exposed as it is in anthers 
of most other Iridaceae. The anthers have been de- 
scribed as subdidymous (Lewis, 1962), indicating 
that they have virtually no connective tissue. 

We were unable to include [xia acaulis in the 
study, and the species may represent an additional 
floral category. Plants are unique in the genus in 
being acaulescent, and the long-tubed, yellow flow- 
ers are borne at ground level with the ovary held 
below ground (Goldblatt & Manning, 1993). In- 
cluded in section Hyalis by de Vos (1999), I. acau- 
lis is unlikely to be pollinated by the long-probos- 
cid flies that visit other species in the section for 
the flowers are produced in May when no known 
species of this pollinator group are on the wing. 


NECTAR PROFILE 


Nectar, when present (Table 2), is goin in 
septal nectaries in Ixia species, as in the entire 
subfamily Crocoideae (Goldblatt, 1990, 1991). 
Nectar is secreted from minute, circular pores at 
the top of the ovary, flowing directly into the base 
of the perianth tube, and is typically retained in 


€— 


Figures 3-6. Representative species of /xia showing general features of habit and floral form and pigmentation. 


—3 (tor 


curta, with 


> left). Ixia rapunculoides, with nodding flowers with a wide tube. 
colored ped lacking markings, an elongate tube, and the anthers included in the floral tube. —5 (bottom 
flowers crowded and with prominent darker, central markings and о anthers. —6 (bottom right). fiis 


—4 (top right). /. paniculata, with pale- 
left). Z 


scillaris, with vertically oriented flowers, secund on the spikes, and nodding anthers 


572 


Annals of the 
Missouri Botanical Garden 


able 3. 


Nectar properties of selected /xia flowers. Fru = tue tose, Glu = glucose, Suc = sucrose. Nectar sugars 


T 
analyzed by B.-E. van Wyk. Sample size indicates number of flowers (of different dn a. examined at study site. 


Nectar % of different sugars 
Sample volume % sugar Mean Suc/Glu 
Species size ul (n) (+SD) Fru Glu Suc + Fru (n) 

I. longituba 6 2.0–2.4 23.7 (3.3) 14-19 12-22 60—75 1.94 (3) 
1. latifolia 5 1.7-2.9 21.6 (2.5) — — 

10 0.4-1.3 38.5 (3.6) = = == = 
I. paucifolia 5 1.1-1.6 29.8 (1.8) 17-21 20-25 54—63 1.41 (2) 
1. paniculata 8 3.9-5.7 26.1 (2.7) 16-19 17-23 58—66 1.66 (3) 
1. rapunculoides 3 1.1-1.5 28.0 (1.0) 21-28 29-37 35-44 0.69 (3) 


(1. curta, 1. flexuosa, I. maculata, I. metelerkampiae, 1. monadelpha, 1. polystachya, I. scillaris: tubes filiform, closed at 
le) 


the mouth, and nectar not detectable 


the lower part of the tube. The length of the peri- 
anth tube varies among the Ixia species (Table 2 
examined, ranging from 4 mm in I. scillaris to 60 
mm in 1. paniculata. In species with a funnel- 
shaped tube, the lower, slender part is mostly 4—8 
mm long. Nectar is present in flowers of groups 1 
and 2 but is not produced in the та maculata 
group or in subgenus Dichone. Nectar volumes are 
modest in the /xia latifolia group, mostly 1.5—3 ш, 
and rarely exceed 5 ш (Table 3). In the long-tubed 
I. paniculata group nectar secretion ranges from 
1.1-1.6 pl (in 7. paucifolia) to 3.9-5.7 yl (in Z. 
paniculata). The nectar is sucrose-rich to sucrose- 
dominant with sugar solute making up 25-35% o 
the total volume of fluid (Table 3). 


N 


INSECT OBSERVATION, POLLINATION MECHANISMS, 
AND POLLEN LOAD ANALYSIS 


Pollination strategies vary among [ма species 
and largely correlate with the mode of floral pre- 
sentation. There appears to be a correlation be- 
tween pollinators of species in category 1 and the 
width of the upper part of the perianth tube. Spe- 
cies with a wide, cup-shaped upper tube are pol- 
linated primarily by female Anthophora diversipes, 
a large bee (body 14-17 mm long) with a relatively 
long proboscis, 6.5-8 mm long (Goldblatt et al., 
1998b). These bees land on the flower and brush 
both the anthers and stigma lobes as they push 
their heads into the floral cup. Anthophora diversi- 
pes is a polylectic forager (Table 5), and different 
individuals were found to carry the pollen of co- 
blooming Fabaceae, Salvia sp. (Lamiaceae), Lobos- 
temon (Boraginaceae), and Iridaceae (including 
Gladiolus, Hesperantha, Moraea) in their scopae. 
One bee, captured on 1. latifolia, carried three pol- 
linaria of Satyrium humile Lindl. (Orchidaceae) on 
its frons. We have also captured the nemestrinid 


fly, Prosoeca sp. (mouth parts 12 mm long), foraging 
for nectar on 1. rapunculoides. The fly grasps the 
extended tepals with its tarsi while continuing to 
vibrate its wings. 

Among the species of category 1, those with a 
narrow upper floral tube have a different range of 
insect visitors. Ixia orientalis is visited by the but- 
terfly Colias electo (Pieriidae). This insect lands on 
the tepals and its head contacts anthers and stigma 
lobes while it extends its proboscis into the floral 
tube. The small-flowered and short-tubed [xia or- 
ientalis has a particularly narrow floral tube, and it 
appears to be visited only by this insect. 

Ixia aurea and 1. tenuifolia are visited by a com- 
bination of short-proboscid tabanid flies, Mesomyia 
edentula or Philoliche atricornis (proboscis 3—5 mm 
long), respectively, and hopliine beetles (Scara- 
baeidae) (Table 5), while Р. atricornis alone was 
captured on /. esterhuyseniae. The tabanid flies for- 
aged for nectar on, and carried mixed loads of pol- 
len of, Ixia, Ornithogalum thyrsiflora Jacq. (Hy- 
acinthaceae), and Asteraceae. Beetles ignored the 
nectar in the floral tube, feeding exclusively on pol- 
len, and contacted both dehisced anthers and stig- 
ma lobes while foraging, copulating, or engaging in 
agonistic behavior (see Goldblatt et al., 19984). 
These beetles also carried mixed loads of /xia, Or- 
nithogalum, and asteraceous pollen. Both these 
short-proboscid tabanid flies and the hopliine bee- 
tles carried pollen in the dorsal and ventral parts 
of their bodies as well as the frons. Pollen of Ixia 
species was, however, located on the frons and dor- 
sal part of the thorax in the tabanids, reflecting 
their body orientation as they foraged for nectar on 
Ixia flowers. 

Tubular flowers (category 2) were visited by long- 
proboscid flies, either Nemestrinidae or Tabanidae, 
with probosces between 18 and 70 mm long (Table 


Моште 87, Митрег 4 
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Goldblatt et 
Pollination Фей ЖЕЙ їп /хїа 


573 


Table 4. Comparison of the length of perianth tube 
and mouth parts of long-proboscid flies collected on /xia 
species. Measurements are from study sites and represent 
the ranges found by sampling flowers of 10 individuals. 
For insects only three individuals were captured and mea- 
sured. 


Peri- 
anth  Probos- 
tube cis 
Plant length length 
species (mm) (mm) Insect species 
I. paniculata 65-75 67-70 Moegistorhynchus 
longirostris 
I. longituba 30-32 22-24 Philoliche gulosa 
I. paucifolia 16-28 18-22 P. gulosa 


4). These flies grasp the tepals with their tarsi and 
probe for nectar while continuing to vibrate their 
wings. Field observations and pollen load analyses 
show that long-proboscid flies visit [ма species 
during the same foraging bouts in which they visit 
open flowers of Pelargonium species (Geraniaceae) 
and species of several genera of Iridaceae (includ- 
ing Babiana, Gladiolus, Geissorhiza, and Lapeirou- 
sia species). One specimen of Moegistorhynchus 
also carried one pollinarium of Disa draconis (L. f.) 
Sw. (Orchidaceae), also a member of the guild of 
plant species that depend on this particular fly for 
pollen dispersal (Manning & Goldblatt, 1997; John- 
son & Steiner, 1997). 

Flowers of category З, section [xia (with a salver- 
shaped perianth and longitudinally dehiscent an- 
thers), are visited primarily by hopliine beetles (Ta- 
ble 5), but one species is visited primarily by honey 
bees (Apis mellifera). Observation and pollen load 
analyses of beetles captured on species of the [xia 
maculata group show unusually diverse foraging 
behavior, visiting flowers of a range of species in- 
cluding other lridaceae (e.g., Gladiolus, Moraea, 
Romulea), Asteraceae, Monsonia (Geraniaceae), 
Prismatocarpus pedunculatus (P. J. Bergius) A. DC. 
(Campanulaceae), Spiloxene capensis (L.) Garside 
(Hypoxidaceae), and Ornithogalum spp. When Apis 
mellifera visited І. flexuosa it contacted the stig- 
matic lobes while scraping anthers for pollen. Cap- 
tured individuals of Apis mellifera carried pure 
loads of Ixia pollen on their bodies and in their 
corbiculae (Table 

t one study site, Bainskloof, we noted a possi- 
ble example of Batesian mimicry. [ма metelerkam- 
piae, which has pale mauve flowers with a darker 
central area, and co-blooming Thereianthus ixioides 
G. J. Lewis (also Iridaceae), with whitish flowers 
also with purple markings in the center, closely re- 


Table 5. Pollen load analysis of pois insects on /xia 
ies. Scarabaeidae: Anisochelus ‚ Heterochelus, 


idea: Apidae: Amegilla, Anthophora, Apis, Pachymelus. Mel- 
ittidae: Rediviva. Nemestrinidae: Moegistrohynchus, Prosoeca. 
Tabanidae: -— тын. In addition to the insects 
listed below. i ls of Rediviva aurata, and one each 
of Andrena еа and Colletes sp. captured on 1. scillaris carried 
pollen only of non-host species, and one Lepithrix individual 
captured on 1. tenuifolia carried no pollen at all. 


Number of 
insects carryin 
pollen loaded 


Host 
flr 
Host plus 
r other 
Plant and [insect] taxon only sp. 
Subgenus Ixia: Section Morphixia 
1. aurea 
[Heterochelus arthriticus] 3 2 
[Lepithrix ornatella 0 1 
[Pachycnema crassipes] 0 2 
[Mesomyia edentula] 2 2 
1. capillaris 
[Anthophora diversipes 9 | 0 2 
1. esterhuyseniae 
[Philoliche atricornis] 0 1 
1. latifolia 1 
[Anthophora diversipes $ | 0 2 
popln 2 
Jean diversipes 2 | 0 2 
[A. schulzei $ | 0 2 
[Amegilla ie m 9] 0 1 
I. odorata 
[Anthophora diversipes 9] 0 2 
І. orientalis 
[Colias electo] 3 0 
1. rapunculoides 
[Anthophora schulzei 9] 0 1 
[Pachymelus peringueyi $ | 0 2 
I. tenuifolia 
[Lepithrix ornatella| 0 5 
[Philoliche atricornis] 3 3 
I. thomasiae 
[Anthophora diversipes 9 | 0 2 
Section Hyalis 
I. longtuba 
[Philoliche gulosa] 0 2 
I. paniculata 
[Moegistorhynchus longirostris] 0 2 
I. paucifolia (site 1) 
[Philoliche gulosa] 0 3 
[Philoliche gulosa] 0 2 


574 


Annals of the 
Missouri Botanical Garden 


Table 5. Continued. 


umber 


of 


insects carryi 


ng 
pollen load(s) 


Host 
flr 
Host plus 
flr other 
Plant and [insect] taxon only sp. 
Section Ixia 
1. atrandra 
Peritrichia subsquamosa 0 3 
І. curta 
[Lepisia rupicola] 0 3 
[Lepithrix fulvipes] 0 5 
[Pachycnema crassipes] 0 l 
1. flexuosa 
[Apix 2 3 0 
1. lutea (site 
[Lepithrix din 0 5 
site 2 
[Anisochelus inornatus] 1 4 
site : 
[Anisochelus "o E 1 3 
|Сћазте dec 0 1 
[Peritrichia ral 1 1 
1. maculata 
(site 1) 
[Pachynema crassipes] 0 2 
ite 
[Pachycnema crassipes] 0 2 
[Lepithrix fulvipes | 0 3 
(site 3) 
[Heterochelis sexlineatus] 0 1 
[Scelophysa militaris] 0 1 
[Pachycnema crassipes] 0 | 
[Lepithrix ornatella| 0 5 
1. metelerkampiae 
[Peritrichia sp.] 1 2 
1. monadelpha 
mee fulvipes] 0 l 
І. versic 
[Peritrichia рѕеийигѕа | 0 4 
Subgenus Dichone 
I. scillaris (site 1) 
[Amegilla spilostoma 9 | 0 4 
site 2 
Laci aurata. 9 | 0 0 
[C sp. 8] 0 0 
dd sp. 9] 0 0 
Totals 18 93 


semble one another as regards floral presentation, 
and they appear to be visited indiscriminately by 
the same beetle, Peritrichia sp. Both species have 
a filiform floral tube in which the style is tightly 
enclosed by the walls of the tube. In T. ixioides 
small amounts of nectar are produced and are vis- 
ible at the mouth of the tube. This suggests that 
the species may offer a secondary reward and may 
ave a more generalist pollination system than 1. 
metelerkampiae, which does not offer nectar. 
Category 4 flowers of subgenus Dichone (with se- 
cund, salver-shaped flowers and basally dehiscent 
anthers) appear to be pollinated by female Amegilla 
(Apidae). These bees have bodies 10-12 mm long, 
thus smaller than Anthophora diversipes, although 
they also have an elongated proboscis (Goldblatt et 
al., 1998b). These bees cling to the anthers of Ixia 
scillaris while foraging exclusively for pollen. Pol- 
len removal is accomplished by thoracic vibration 
in which the foraging female produces the charac- 
teristic high-pitched whine (sensu Buchmann, 
198 bee did not remain on an individual flower 
of I. scillaris for longer than 1.5 seconds. The head 
of a bee appeared to contact one of the three ex- 
tended stylar lobes while it grasped the anthers. 
Pollen load analyses of Amegilla captured on I. scil- 
laris (Table 5) showed that the bees also foraged on 
nectariferous flowers of Aspalathus (Fabaceae) and 
Muraltia (Polygalaceae). An interesting aspect of 
the pollination of /. scillaris is that although the 
anthers are prominently displayed and evidently at- 
tractive to bees other than Amegilla, the melittid, 
Rediviva aurata, as well as Andrena sp. and Colletes 
sp. seen visiting flowers and clasping the anthers, 
did not carry pollen of the species. This is evidently 
a reflection of these bees’ inability to vibrate their 
wings at the necessary frequency to dislodge pollen 
from the anthers. 


DISCUSSION 


The adaptive radiation of pollination mecha- 
nisms in /xia is consistent with aspects of polli- 
nation systems described in Lapeirousia (Goldblatt 
et al., 1995), Moraea (Goldblatt et al., 1989; Gold- 
blatt & Bernhardt, 1999), and Gladiolus (Goldblatt 
& Manning, 1998; Goldblatt et al., 1998b). Specif- 
ically, pollination systems in moderate to Eo 
sized genera of Iridaceae in Africa diverge as dif- 
ferent species in the same genus exploit different 

pollen vectors. Lapeirousia (Goldblatt, 1990b; 
Goldblatt & Manning, 1992) has specialized spe- 
cies that utilize long-proboscid Tabanidae, Nemes- 
trinidae, or sphinx moths, as well as species with 
generalist systems that include various long- 


Volume 87, Number 4 
2000 


Goldblatt et al. 
Pollination Mechanisms in Ixia 


575 


tongued bees, bee flies (Bombyliidae), and butter- 
flies. Pollination in the much larger genus Gladio- 
lus, with some 245 sub-Saharan African species, is 
dominated by long-tongued anthophorine bees, but 
pollination by long-proboscid Nemestrinidae and 
Tabanidae, the large butterfly, Aeropetes tulbaghia, 
night-flying moths, and sunbirds (Nectarinia spp.) 
has also been observed in several species. Moraea 
species appear to be pollinated primarily by bees 
with elongated or extended mouth parts (Goldblatt 
et al., 1989). However, recent work by Steiner 
(1998a) and Goldblatt et al. (1998a) shows that sev- 
eral southwestern Cape species are pollinated by 
hopliine beetles, while other species are pollinated 
only by pollen-collecting bees, or a combination of 
bees, muscid and scathophagid flies, and hopliine 
beetles (Goldblatt & Bernhardt, 1999). Like these 
other genera of the Iridaceae, [xia also has species 
adapted to a range of pollination systems. The flow- 
ers of [xia species thus show different sets of cor- 
related morphological specializations that represent 
co-adaptations with different floral foragers. 

Pollination by hopliine beetles in [xia parallels 
pollination by hopliine beetles in other genera of 
the Iridaceae. As in Moraea insolens Goldblatt, M. 
cf. lurida Ker-Gawl., M. tulbaghensis L. Bolus, and 
M. villosa (Ker-Gawl.) Ker-Gawl. (Goldblatt et al., 
1998a; Steiner, 1998a), the flowers of the [xia ma- 
culata group have darkened floral markings and 
large areas of flat surface, brightly colored pollen, 
and little or no nectar. Pollination by nectar-for- 
aging anthophorine bees in some species of Ixia 
parallels that in Moraea inclinata, many Gladiolus 
species, and Lapeirousia divaricata N. E. Br. (Gold- 
blatt et al., 1989, 1995, 1998b). These [xia species 
also have flowers broad enough to offer a landing 
platform, pigmentation patterns mostly in the pink 
to blue range, inconspicuous stamens, and nectar 
retained at the base of a floral tube. 

The adaptive radiation of pollination mecha- 
nisms in Ixia, however, appears less diverse than 
in Lapeirousia or Gladiolus. Neither moth nor bird 
pollination appears to occur in the genus. While 
butterfly pollination is described here in Ixia, 1. 
orientalis is definitely not part of the southern Af- 
rican red-flowered, summer-blooming Aeropetes 
butterfly guild described by Johnson and Bond 
(1994). 

The unique feature of pollination systems in Ixia 
is represented by the unusual buzz-pollination sys- 
tem in subgenus Dichone. In other genera of Iri- 
daceae in southern Africa there is a divergence be- 
tween bee pollination 
nectariferous flowers, correlated with either pollen 
or nectar as the reward. For example, most species 


in nectarless versus 


of Gladiolus are nectariferous, and large antho- 
phorines forage on gullet- or flag-shaped flowers 
(sensu Faegri & van der Pijl, 1979). Gladiolus 
quadrangulus (D. Delaroche) Barnard and С. stel- 
latus G. J. Lewis have stellate flowers, and Andrena 
sp. or Apis mellifera scrape pollen from prominent, 
exserted anthers (Goldblatt et al., 1998b). A similar 
divergence is also found in bee-pollinated species 
of Moraea (Goldblatt & Bernhardt, 1999). 

The genus /xia actually exhibits three different 
modes of bee pollination. The nectar-bearing tube 
and cup-shaped floral system of section Morphixia 
and the pollen-rich, nectarless, salver-shaped floral 
system of I. flexuosa parallel systems described in 
Gladiolus (Goldblatt et al., 1998b). However, buzz 
pollination described in subgenus Dichone has not 
been observed in any other genus of the Iridaceae 
to date. Harris (1905) noted that porose or porate 
anthers are found in the Iridaceae, but Buchmann 
(1983) did not mention buzz-pollinated flowers in 
the family in his review of the subject. Buchmann 
showed how varied pore position may be on a sol- 
itary anther but did not mention basal pores or bas- 
al dehiscence. Buzz pollination in subgenus Dicho- 
ne has evolved independently adding another 
example to the widespread convergence within the 
monocots for buzz pollination. Buzz pollination has 
been reported within the petaloid monocots in Cy- 
anella (Tecophilaeaceae—Dulberger & Ornduff, 
1980), Echeandia (Anthericaceae—Bernhardt & 
Montalvo, 1979), Dianella (Hemerocallidaceae— 
Bernhardt, 1995), Dichopogon (Lomandraceae— 
Bernhardt & Burns-Balogh, 1986; see Bernhardt, 
1996, fig. 3), and Xiphidium (Haemodoraceae— 
Buchmann, 1980). In these taxa filaments are rel- 
atively short and anthers are inflated. Buzz-polli- 
nated species of Ixia, however, have one unusual 
character found only in a few buzz-pollinated an- 
giosperms such as the Australian Calectesia (Da- 
sypogonaceae) and Hibbertia (Dilleniaceae). In 
these last two taxa the stylar lobes diverge, forming 
a triangular perimeter outside the centrally placed 
anthers (Bernhardt, 1986). We presume that this 
alignment of sexual organs may encourage cross 
pollination, as a bee should contact stigmatic sur- 
faces before it vibrates the anthers. 

Flowers of Ixia flexuosa appear to represent a 
secondary shift to bee pollination within the pre- 
dominantly beetle-pollinated section /xia. The flow- 
ers have the morphological modifications of hopli- 
ine beetle-pollinated species of the section, but the 
flowers are relatively small, ca. 15 mm in diameter, 
too small to easily accommodate more than one 
beetle at a time. Moreover, they are scented and 
also often fairly dull-colored, either cream or mauve 


576 


Annals of the 
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(but sometimes pink) and without a prominent cen- 
tral mark, features not normally associated with ho- 
pliine pollination (Picker & Midgley, 1996; Steiner, 
1998b; Goldblatt et al., 1998b). The entire inflo- 
rescence often droops and waves about in the wind 
on a slender peduncle, making the flowers poor 
sites for beetle assembly. Instead, the flowers seem 
to offer pollen to foraging bees. Apis mellifera is a 
common visitor, which we have seen actively col- 
lecting pollen on the species. The corbiculae of 
bees captured on J. flexuosa contained pure loads 
of Ixia pollen. 

Outgroup comparison with the genera most 
closely allied to Ixia, Sparaxis, and Tritonia (Lewis, 
1962; Goldblatt et al., 2000), suggests that the an- 
cestral pollination system in /xia is the one in 
which flowers are cup-shaped, produce nectar, and 
are pollinated by large anthophorine bees. This is 
the plesiomorphic pollination strategy in Sparaxis 
and Tritonia according to current views on the evo- 
lution of these genera. In both genera a zygomor- 
phic, bilabiate flower that produces nectar is be- 
lieved to be ancestral, as it probably is for the 
entire subfamily Ixioiodeae (Goldblatt, 2000). All 
Ixia species have a radially symmetric perianth, a 
major synapomorphy for the genus. Flowers of spe- 
cies of sections Morphixia and Hyalis still produce 
nectar in a hollow perianth tube. Specialization for 
pollinators іп /xia has shifted in two major direc- 
tions. 

An emphasis on nectar as the sole reward ac- 
companied by an elongation of the floral tube has 
occurred in species of section Hyalis and is the 
only system known to be utilized by any species of 
the section. This pollination system is most highly 
developed in [xia paniculata, which has a floral 
tube up to 65(-75) mm long and is the only species 
of the section visited by Moegistorhynchus longi- 
rostris. Other members of the section utilize shorter- 
proboscid Tabanidae. In neither fly family is pollen 
consumed, and anthers of /. paniculata are actually 
concealed within the floral tube, a mode of presen- 
tation also found in 1. fucata Ker-Gawl. (Lewis, 
1962), on which we have not captured any insects. 

A second direction of morphological evolution 
has been the reduction of the perianth tube in width 
and sometimes length, accompanied by the sup- 
pression of nectar production. In most species of 
section /xia ample pollen is produced by longitu- 
dinally dehiscent anthers, but the flowers primarily 
appear to constitute sites for the assembly and mat- 
ing of species of hopliine beetles. While these bee- 
tles do consume pollen, they spend most of their 
time on flowers either crawling over the perianth or 
lying with their heads in the center of a flower. 


Beetle pollination is a strategy adopted by many 
southern African petaloid geophytes, as well as an- 
nual and perennial Asteraceae, and is known in the 
Iridaceae in Апчеа, Moraea, Hesperantha vaginata 
(Sweet) Goldblatt, and Romulea (Goldblatt & Man- 
ning, 1996; Steiner, 1998a; Goldblatt et al., 19983, 
2000). Pollination by hopliine beetles also occurs 
in other genera, either alone or in combination with 
other insects, usually nectarivorous, short-probos- 
cid Tabanidae (e.g., species of Sparaxis) or with 
pollen-collecting female halictid and andrenid bees 
and Apis mellifera (e.g., species of Romulea, Glad- 
iolus meliusculus (G. J. Lewis) Goldblatt & J. C. 
Manning). Hopliine beetle pollination, however, ap- 
pears to be particularly well developed in /xia sect. 
Ixia, a taxon that appears to comprise a clade, de- 
fined by the morphological adaptations associated 
with the pollination system. These include the re- 
duction in the diameter of the perianth tube, which 
is filiform and tightly sheaths the style, and the 
closure of the apex of the tube by broad-based fil- 
aments, inserted at the base of the tepals and often 
united below or entirely. Nectar production is also 
suppressed, and the floral tube, a plesiomorphic 
structure in Iridaceae subfamily Crocoideae, simply 
functions as a stalk for the petaloid, salver-shaped 
part of the flowers. Additional adaptations include 
the development of bright floral pigmentation, most- 
ly orange, yellow, red, or purple, often with dark 
centers that appear to represent beetle marks, a 
common feature of beetle flowers (Picker & Midg- 
ley, 1996). Ixia aurea and I. tenuifolia (sect. Mor- 
phixia) offer a remarkable example within the ge- 
nus of convergence for hopliine beetle pollination. 
Both species have a narrowly funnel-shaped floral 
tube that contains nectar, but the floral pigmenta- 
tion is like that of species of section Ixia, being 
deep yellow to orange, and with a dark center in |. 
tenuifolia. They are pollinated by a combination of 
hopliine beetles and the tabanid flies Mesomyia ed- 
entula and Philoliche atricornis. This system offers 
a striking parallel with the beetle pollination sys- 
tem in Sparaxis species (Goldblatt et al., 1998b, 
2000) in which small amounts of nectar are also 
offered. 

While there are far fewer species in Ixia com- 
pared to Gladiolus and Moraea, pollination systems 
of Ixia are relatively diverse. The diversity of pol- 
lination systems in Ixia reflects relatively minor 
changes in floral characters based on changes in 
pigmentation, the presence or absence of nectar, 
floral tube, and mode of anther dehiscence. With 
the exception of the mode of anther dehiscence, all 
other floral modifications in [ма parallel those 
found in allied genera of the family. 


Volume 87, Number 4 
2000 


Goldblatt e 
Pollination ace А in Ixia 


577 


Literature Cited 


Bernhardt, P. 1986. Bee pollination in Hibbertia ee 
lata gt ча Pl. Syst. Evol. 152: 2 
—— . Buzz- ан of Dianella Mies var. 
assera Mop ons Cunninghamia 4: 9-20. 
. Anther adaptation in animal pollination. 
Pp. oo in W. С. D’Arcy & R. С. Keating (editors), 
The Anther: Form, Function, and Phylogeny. Cambridge 
Univ. Press, Cambridge, Englan 
P. Burns-Balogh. 1986. Floral mimesis in The- 
[утига. Pl. Syst. Evol. 151: 97- 
& E. A. Montalvo. 1979. ia па аи ecology 
of Echeandia macrocarpa (Liliaceae). Brittonia 31: 64— 
71l. 


Buchmann, S. L. 1980. Preliminary anthecological obser- 

vations on Xiphidium caeruleum Aubl. (Monocotyledo- 

eae: Haemodoraceae) in Panama. J. Kansas Entomol. 
Soc. 53: peal 


. Buzz ee in angiosperms. Pp. 73— 
113 i in 3 x Jones & R. J. Little (editors), Handbook 


of Experimental Pollination. Van Nostrand Reinhold, 


ork. 

Dulberger, R. & R. Ornduff. 1980. Floral morphology and 
reproductive biology of four о of Cyanella (Teco- 
eters. New Phytol. 86 

Faegri, L. van der Pijl. 1979. The Principles of 

ollination Ecology, 3rd ed. Pergamon Press, New York. 

Goldblatt, P. 1990a. Phylogeny and classification of Iri- 
daceae. I Missouri Bot. Gard. 77: 607—627. 

—— ——. |990b. Systematics of Lapeirousia (Iridaceae— 
m | tropical Africa. Ann. Missouri Bot. Gard. 


97 overview of the systematics, phylogeny 
and deis of ci southern African Iridaceae. Contr. 
Bolus oe 13 
—— E of the bara and the rela- 
m v г Апп. Вог. (Ко 6. 
& P. Bernhardt. 1990. Pollination biology of Niv- 
ета Веи ae and the presence of heterostylous self- 
compatibility. ipi Bot. 39: 93-111. 
Pollination of Moraea species 
(Iridac ene with a к ы column. Ann. Missouri Bot. 
Gard. 86: 47-56. 
& J. ri Manning. 1992. Systematics of the south- 
ern African Lapeirousia corymbosa (lridaceae: Ixioi- 


deae) complex (sect. Peek ) and a new species of 
sect. Paniculata. S. A n J. Bot. 58: 326— 
& acaulis, a new ac јен еп! 


species of Iridaceae: Ixioideae from ч ips vlakte, 
Namaqualand, South Africa. Novon 3: 
& . 1996. Aristeas and iiis "om 
Veld & Flora 82: т 
& 998. Gladiolus in Southern Africa. 


Fernwood Press, ш Town. 
& ‚1 


w species of Sparaxis and 
Ixia apes pd К) from Western Саре, Sout 
Africa, and t omic notes on [ма and Gladiolus. 


Bothalia 29: M 
& 


000. Cape Plants: А conspectus of 
the vascular iud ofthe Cape region of South Africa. 
Strelitzia 2. National Botanical Institute of South Africa, 
Cape Town 


—— Bernhardt s у со 1989. Notes on 
the Бе > mechanisms of Moraea inclinata an 
к = жее). Pl. St Evol. 163: 201-209 

ning ari. 1991. Sulcus and oper- 
cu “ee structure in the pollen grains of Iridaceae sub- 
family Ixioideae. Ann. Missouri Bot. Gard. 78: 950- 
961. 


— = & P. Bernhardt. 1995. Pollination biol- 
ogy of Lapeirousia subgenus Lapeirousia (Iridaceae) in 
southern Africa; Floral divergence and adaptation for 
long-tongued fly pollination. Ann. Missouri Bot. Gard 
82: 517-534. 


& —. 1998a. Adaptive radiation of 

bee- piion Gladiolus species (Iridaceae) in southern 
Africa. Ann. Missouri Bot. Gard. 85: 492-517. 
, P. Bernhardt & J. C. Manning. 1998b. Pollination 
by каин beetles (Scarabaeidae—Hopliini) in petaloid 
geophytes in southern Africa. Ann. Missouri Bot. Gard. 
85: 215-230. 


. Manning & P. Bernhardt. 2000. Adaptive 

peius of P systems in Sparaxis Ker Gawler 

(Iridaceae: Ixioideae 

Harris, J. A. 1905 rs by Apical 

Pores. Ph.D Dissertation, Washington Beni St. 
Louis. 

Horn, W. 1962. ай а menos on South African 
us III. Intra- and inter patibility in /xia 

a, Sparaxis Ker., Watsonia n sid Zantedeschia 
TR J. : 2: Bot. 28: 269-277. 

Johnson, S. I W. J. Bond. 1994. Red flowers and but- 
terfly de l Ay in the fynbos of South Africa. Pp. 137- 
148 in M. Arianoutsou & R. Grooves (editors), Plant 
м Interactions in Mediterranean-Type Ecosystems. 
Kluwer Academic Press, Dordrecht. 

. Steiner. 1997, Long-tongued z пеге 
tion and evolution of floral spur length in the Disa dra- 
conis complex. Evolution 51: 45—53. 

ioo G 1962. South African lridaceae.The genus 

1. J. S. African Bot. 28: 45-195. 

ME J. С. & Р. Goldblatt. 1996. The Prosoeca per- 
ingueyi E e pollination syndrome 
in southern Afric ued flies and their tubular 
flowers. Ann. Missouri Bot. Card. 8 

he Молнии њен longiros- 

tris. (Diptera: Nomestisidad pollination guild: Long- 

tubed flowers and a specialized long-tongued fly-polli- 

nation system in southern Africa. Pl. Syst. Evol. 206: 

51-69. 


Ogden, E. . 5. Raynor, J. V. Hayers & D. M. Lewi 
1974. bd of Sampling Airborne Pollen. Hafner 


& J. J. Midgley. 1996. Pollination by mon- 
key жен (Coleoptera: Scarabaeidae: Hopliini): Flow- 
er and color preferences. African Entomol. 4: 7-14. 

Steiner, K. 5. 1998a. Beetle pollination of о то- 
raeas т South Africa. Pl. Syst. Evol. 209: 47-65. 

. 1998b. The Meno of ео sollination in a 

seu Afric n orchi r. J. Bot. 85: 1180-1193. 
os, M. P. de. 1999. s "Pp. 1-87 i n G. Germishuizen 
editor), a of Southern Africa, ái 7 Iridaceae, part 

2 Ixioideae, fasc. 1. National Botanical Institute, Pre- 

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VOLUME 87 


AL-SHEHBAZ, IHSAN A. A Review of Gamosepaly in the Brassicaceae and а 
Revision of Desideria, with a Critical Evaluation of Related Genera __ 


AL-SHEHBAZ, IHSAN A. (See Marcus Koch & Ihsan A. Al-Shehbaz) ______ 
ALVERSON, WILLIAM S. (See Michael J. Donoghue & William 5. Alverson) _ 
BASKIN, CAROL С. (See Jerry M. Baskin & Carol С. Baskin) 
BASKIN, JERRY M. & Canor С. BASKIN. Vegetation of Limestone and Dolomite 
Glades in the Ozarks and Midwest Regions of the United States ____ 
BEENTJE, HENK. (See Ghillean T. Prance, Henk Beentje, John Dransfield & 
Robert Johns) 
BERDUC, ALFREDO. (See Leonardo Galetto, Gabriel Bernardello, Irene C. Isele, 
José Vesprini, Gabriela Speroni & Alfredo Berduc) 
BERNARDELLO, GABRIEL. (See Leonardo Galetto, Gabriel Bernardello, Irene C. 
Isele, José Vesprini, Gabriela Speroni & Alfredo Berduc) 
BERNHARDT, PETER. (See Peter Goldblatt, Peter Bernhardt & John C. Man- 
ning) о 
BERRY, PAUL Е. Book Review. Flora da Reserva Ducke by J. 5. да 5. Ribeiro 
et al. 


Briccs, BARBARA. What is Significant—the Wollemi Pine or the Southern 
Rushes? 


Brusca, RICHARD C. Unraveling the History of Arthropod Biodiversification 


CARLQUIST, SHERWIN. Wood and Bark Anatomy of Takhtajania (Winteraceae); 
Phylogenetic and Ecological Implications 


CHATROU, LARS W., ЛЕКЕ KOEK-NOORMAN & PAUL J. M. Maas. Studies in 
Annonaceae XXXVI. The Duguetia Alliance: Where the Ways Part __ 
CHIANG, TZEN-YUH. (See Ching-I Peng & Tzen-Yuh Chiang) 
Скозву, MARSHALL R. Statistical Summary of Some of the Activities in the 
Missouri Botanical Garden Herbarium, 19 
DANIEL, THOMAS F. (See Lucinda A. McDade, Thomas F. Daniel, Susan E. 
Masta & Katherine M. Riley) 
DEROIN, THIERRY. Notes on the Vascular Anatomy of the Fruit of Takhtajania 
(Winteraceae) and Its Interpretation 


DONOGHUE, MICHAEL J. & WILLIAM S. ALVERSON. A New Age of Discovery 


Doust, ANDREW N. Comparative Floral Ontogeny in Winteraceae — 


DovLE, JAMES А. Paleobotany, Relationships, and Geographic History of Win- 
teraceae 


DRANSFIELD, JOHN. (See Ghillean T. Prance, Henk Beentje, John Dransfield 
& Robert Johns) 


2000 


549 
246 
10 
286 


286 


67 


127 


127 


264. 


EBINGER, JOHN E. (See Jennifer T. Jawad, David 5. Seigler & John E. Ebinger) 


EHRENDORFER, F. & M. LAMBROU. Chromosomes of Takhtajania, Other Win- 
teraceae, and Canellaceae: Phylogenetic Implications 


ENDRESS, PETER K., ANTON IGERSHEIM, F. B. SAMPSON & GEORGE E. SCHATZ. 
loral Structure of Takhtajania and Its Systematic Position in Wintera- 
ceae 


ERTTER, BARBARA. Floristic Surprises in North America North of Mexico — 


FEILD, TAYLOR S., MACIEJ А. ZWIENIECKI & N. M. HOLBROOK. Winteraceae 
Evolution: An Ecophysiological Perspective 


GALETTO, LEONARDO, GABRIEL BERNARDELLO, IRENE C. ISELE, JOSE VESPRINI, 
GABRIELA SPERONI & ALFREDO BERDUC. Reproductive Biology of Ery- 
thrina crista-galli (Fabaceae) 

GILL, ANTHONY С. (See John С. Lundberg, Maurice Kottelat, Gerald R. Smith, 
Melanie L. J. Stiassny & Anthony C. Gill) 

GIUSSANI, LILIANA MONICA. Phenetic Similarity Patterns of Dioecious Species 
of Poa from Argentina and Neighboring Соипігіеѕ 


GOLDBLATT, PETER, PETER BERNHARDT & JOHN С. MANNING. Adaptive Ra- 
diation of Pollination Mechanisms in /xia (Iridaceae: Crocoideae) ___ 
GOLDBLATT, PETER & JOHN C. MANNING. The Long-proboscid Fly Pollination 
ystem in Southern Africa 


GUTIERREZ, Плесо С. (See Liliana Katinas, Diego С. Gutiérrez & Silvia S. 
Torres ПОВИ Пн eee рана ee 
Ногвкоок, N. M. (See Taylor S. Feild, Maciej A. Zwieniecki & N. M. Hol- 


TOO 


IGERSHEIM, ANTON. (See Peter K. Endress, Anton Igersheim, F. B. Sampson 
& George E. Schatz) 
IsELE, IRENE C. (See Leonardo Galetto, Gabriel Bernardello, Irene C. Isele, 
José Vesprini, Gabriela Speroni & Alfredo Berduc) 
JAWAD, JENNIFER T., DAvip S. SEIGLER & JOHN E. EBINGER. A Systematic 
Treatment of Acacia coulteri (Fabaceae, Mimosoideae) and Similar Spe- 

cies in the New World 
JOHNS, RoBERT. (See Ghillean T. Prance, Henk Beentje, John Dransfield & 
obert Johns 


KAROL, KENNETH G., YOUNGBAE SUH, GEORGE E. SCHATZ & ELIZABETH А. 
ZIMMER. Molecular Evidence for the Phylogenetic Position of Takhtaja- 
та in the Winteraceae: Inference from Nuclear Ribosomal and Chloro- 
plast Gene Spacer Sequences 


KATINAS, LILIANA, DIEGO С. GUTIERREZ & SILVIA S. TORRES ROBLES. Carlos 
Spegazzini (1858-1926): Travels and Botanical Work on Vascular Plants 


KEATING, RICHARD С. Anatomy of the Young Vegetative Shoot of Takhtajania 
perrieri (Winteraceae) 


528 


127 


528 


67 


414 


183 


335 


KELCHNER, Scor A. The Evolution of Non-coding Chloroplast DNA and Its 
Application in Plant Systematics ____ 


Косн, MARCUS & IHSAN A. AL-SHEHBAZ. Molecular Systematics of the Chi- 
nese Yinshania (Brassicaceae): Evidence from Plastid and Nuclear ITS 
DNA Sequence Data _ 

KOEK-NOORMAN, ЛЕКЕ. (See Lars W. Chatrou, Jifke Koek-Noorman & Paul J. 
Me Maa) эз ee ne neo a rece әш 

KOTTELAT, MAURICE. (See John С. Lundberg, Maurice Kottelat, Gerald В. 
Smith, Melanie L. J. Stiassny & Anthony C. Gill) 

LAMBROU, M. (See F. Ehrendorfer & M. Lambrou) 

Lowry, Porter P., II. (See Alexei А. Oskolski & Porter P. Lowry II) 

LUNDBERG, JOHN G., MAURICE KOTTELAT, GERALD R. SMITH, MELANIE L. J. 
SrIASSNY & ANTHONY С. GILL. So Many Fishes, So Little Time: Ап 
Overview of Recent Ichthyological Discovery in Continental Waters __ 


МсрАре, LucinpA A., THOMAS Е. DANIEL, SUSAN E. MASTA & KATHERINE M. 
RILEY. Phylogenetic Relationships within the Tribe Justicieae (Acantha- 
ceae): Evidence from Molecular Sequences, Morphology, and Cytology 

Maas, PauL J. M. (See Lars W. Chatrou, Jifke Koek-Noorman & Paul J. M. 


Maas) 


MACKINNON, Jonn. New Mammals in the 21st Century? _ 


MADIGAN, MICHAEL Т. Extremophilic Bacteria and Microbial Diversity ____ 
MANNING, JOHN С. (See Peter Goldblatt & John C. Manning) . 
MANNING, JOHN C. (See Peter Goldblatt, Peter Bernhardt & John C. Manning) 


МАЅТА, SUSAN Е. (See Lucinda A. McDade, Thomas Е. Daniel, Susan E. Masta 
& Katherine M. Riley) вара г 
ORTIZ, SANTIAGO. A Phylogenetic Analysis of Dicoma Cass. and Related Gen- 
era (Asteraceae: Cichorioideae: Mutisieae) Based on Morphological and 
Anatomic Characters 


OSKOLSKI, ALEXEI А. & PORTER P. Lowry П. Wood Anatomy of Mackinlaya 
and Apiopetalum (Araliaceae) and Its Systematic Implications _______ 
PENG, CHING-I & TZEN-YUH CHIANG. Molecular Confirmation of Unidirection- 
al Hybridization in Begonia X taipeiensis Peng (Begoniaceae) from Tai- 
wan 
PHILLIPS, OLIVER L. (See Rodolfo Vásquez Martínez & Oliver L. Phillips) _ 
PRANCE, GHILLEAN T., HENK BEENTJE, JOHN DRANSFIELD & ROBERT JOHNS. 
The Tropical Flora Remains Undercollected 


RICHARDSON, P. Mick. Our Unknown Planet: Recent Discoveries and the Fu- 
ture, the 45^ Annual Systematics Symposium of the Missouri Botanical 
Garden. Introduction to the Symposium 


RILEY, KATHERINE M. (See Lucinda A. McDade, Thomas F. Daniel, Susan E. 
Masta & Katherine M. Riley |... 


482 


26 


459 


171 


435 


ROBLES, SILVIA S. TORRES. (See Liliana Katinas, Diego G. Gutiérrez & Silvia 
S. Torres Robles) 


SAMPSON, BRUCE. (See Hiroshi Tobe & Bruce Sampson) 


SAMPSON, Е. B. The Pollen of Takhtajania perrieri (Winteraceae) — 
SAMPSON, Е. B. (See Peter К. Endress, Anton Igersheim, Е. B. Sampson & 
George E. Schatz) 
ScHATZ, СЕОКСЕ E. The Rediscovery of a — Endemic: Takhtajania 
perrieri (Winteraceae) __________ 


SCHATZ, GEORGE E. (See Peter K. Endress, Anton Igersheim, F. B. Sampson 
& George E. Schatz) _ 


SCHATZ, GEORGE E. (See Kenneth G. Karol, Youngbae Suh, George E. Schatz 
& Elizabeth A. Zimmer) 


SEIGLER, DAvip S. (See Jennifer T. Jawad, David S. Seigler & John E. Ebinger) 


SMITH, GERALD R. (See John G. Lundberg, Maurice Kottelat, Gerald R. Smith, 
Melanie L. J. Stiassny & Anthony C. Gill) 
SPERONI, GABRIELA. (See Leonardo Galetto, Gabriel Bernardello, Irene C. Is- 
ele, José Vesprini, Gabriela Speroni & Alfredo Berduc) 
STIASSNY, MELANIE L. J. (See John G. Lundberg, Maurice Kottelat, Gerald R. 
Smith, Melanie L. J. Stiassny & Anthony C. Gill) 
SUH, YOUNGBAE. (See Kenneth G. Karol, Youngbae Suh, George E. Schatz & 
Elizabeth A. Zimmer) ааа 
TOBE, HIROSHI & BRUCE SAMPSON. Embryology of Takhtajania (Winteraceae) 
and a Summary Statement of Embryological Features for the Family — 


VASQUEZ MARTÍNEZ, RODOLFO & OLIVER L. PHILLIPS. Allpahuayo: Floristics, 
Structure, and Dynamics of a High Diversity Forest in Amazonian Peru 


VESPRINI, JOSE. (See Leonardo Galetto, Gabriel Bernardello, Irene C. Isele, 
José Vesprini, Gabriela Speroni & Alfredo Berduc) 
ZIMMER, ELIZABETH А. (See Kenneth С. Karol, Youngbae Suh, George E. 
Schatz & Elizabeth A. Zimmer) 
ZWIENIECKI, MACIEJ A. (See Taylor S. Feild, Maciej A. Zwieniecki & N. M. 
Holbrook) 


414 


323 


Missouri Botanical Garden Press March 2000 


Orchids 


Orchids of Guatemala: 
ARevised 
n 


Checklist 
Margaret A. Dix and 
Michael W. Dix, 
MBG Press. 
Available 

March 2000. 


his study үл 
represents а compilation of all currently 


recognized species of orchids known from 


с : Manli in bLahahnral A 


Guatemala 


together 

older treatments, and their бгр. дату 
and habitat distribution in the country аге 
described. Where identification of species new to 
Guatemala may be difficult, distinguishing 
characters are briefly described. The list, based 
on extensive field collections and herbarium 
material, includes 734 taxa, of which 207 are new 
records or recently described species not reported 
in earlier studies. 

ISBN 0-915279-66-5, iii + 62 pp. 


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CONTENTS 


Phylogenetic Relationships within the Tribe Justicieae (Acanthaceae): Evidence from 
Molecular Sequences, Morphology, and Cytology 
— Lucinda А. McDade, Thomas Е Daniel, Susan E. Masta & Katherine M. Riley 435 
A Phylogenetic Analysis of Dicoma Cass. and Related Genera (Asteraceae: Cichorio- 
ideae: Mutisieae) Based on Morphological and Anatomic Characters — 
— Santiago Ortiz 459 
The Evolution of Non-coding Chloroplast DNA and Its Application in Plant Systematics 
| Scot A. Kelchner 482 
Allpahuayo: Floristics, Structure, and Dynamics of a High Diversity Forest in Amazo- 
nian Peru Rodolfo Vasquez Martinez & Oliver L. Phillips 499 
__А Systematic Treatment of Acacia cin (Fabaceae, Mimosoideae) and Similar 
Species i in the New World 


Jennifer T Jawad, David S. Seigler & John E. Ebinger 528 
A Review of Gamosepaly in the Brassicaceae ànd a Revision of pride with a Crit- 
ical Evaluation of Related Genera n A. Al-Shehbaz 549 
Adaptive Radiation of Pollination Mechanisms т Ixia "UR Беше ме ы СА, 
Peter Goldblatt, Peter Bernhardt & John С. Manning 564 
Checklist for Authors | | 578 
Index to Volume 87 — E ; 


Cover кйин: бозы simplicifolia 1. 5. Miller, бени Ьу Barbara Along from the book: 
let for the 45th Annual Systematics Symposium, Missouri Botanical Garden. First ааа іп 
Novon 8: 167-169 ПУБ