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CONWAY MACMILLAN, 5/0^? Botanist

Minnesota
Botanical Studies

IRLF

3 313 fifl3

Contents

XXIII. The embryogeny of Ginkgo, Harold L. Lyon - - - 275

XXIV. Observations on Callymenia phyllophora J. Ag.,
Clara K. Leavitt 291

XXV. Observations on Endocladia muricata (P. and R. ) J.

Ag., Florence J/". Warner - - - 297

XXVI. Observations on Laminaria bullata Kjellm., Olga

Mueller ------ ..._ ^03

XXVII. Minnesota Helvellineoe, Daisy S. Hone - - - 309

XXVIII. Observations '.n Physalacria inflata (S.) Peck, J

^f, Policy - 323

XXIX. Symbiosis in the ^enus Lolium, E. E. Freeman - - 329

Third Series

Part III

October 18, 1904

MINNEAPOLIS, MINX.
1904

UNIVERSITY FARM

LIBRARY

UNIVERSITY OF CALIFORNIA
DAVIS

XXIII. THE EMBRYOGENY OF GINKGO.

HAROLD L. LYON.

INTRODUCTION.

The account published by Coulter and Chamberlain in 1901
is the most recent work on the embryogeny of Ginkgo and may
be taken as a summation of our knowledge on the subject up to
the present time. They wrote as follows, " Although the
embryo of Ginkgo is exceptional among Gymnosperms, and of
great interest, the details of its development are not sufficiently
known. We have been able to secure almost a complete series
showing the general outlines of the development, which merely
confirms the facts already published. Germination of the
oospore begins, as is usual, among Gymnosperms, with repeated
nuclear divisions. These nuclei, however, instead of organiz-
ing a parietal tissue as in the Cycads, or a basa1 group as in the
Conifers, proceed to fill the whole cavity of the enlarging
oospore with free nuclei, which is followed by the organization
of a compact tissue. In a certain sense this structure would
seem to represent the proembryo of Cycads, but it really repre-
sents the whole product of the oospore, in which proembryo,
suspensor, and embryo proper are not differentiated. The com-
plete filling of the spore with tissue, and the lack of early dif-
ferentiation into the great embryonic regions, would suggest
a habit more primitive than in either Cycads or Conifers. At
the same time, it may be merely a derived character. In any
event, the tissue near the base of the spore, which in the other
groups develops both suspensor and embryo, shows far greater
vigor than the remaining tissue. In the organization of the
embryo the whole mass of tissue is involved, and in the absence of
a suspensor the embryo invades the endosperm by direct growth.

The two cotyledons are differentiated early in October, and
are quite unequal in length. The larger one is two lobed, while
the shorter one is cleft halfway down, thus early indicating the
bilobed character of the leaf. The two cqtyledons are also
united at the apex, but the epidermal layers of the two are dis-

275

276 MINNESOTA BOTANICAL STUDIES

tinct where they are in contact. The plumule is very conspic-
uous between the elongated cotyledons, three or more leaves
appearing just behind the stem apex."

In 1872 Strasburger published the first paper dealing with the
embryogeny of Ginkgo. He determined the general course of
events in the formation of the spherical embryo but insufficient
material prevented the working out of details. Hirase ('95)
studied fecundation and the origin of the spherical embryo. He
found that there were eight simultaneous nuclear divisions in the
oosperm preceding free-cell-formation. More recently Ikeno
(1901) has made a very careful study ot the process of
fecundation.

The embryo of the seed has been frequently described, but
little however seems to be known of its histology. Endlicher
('47) was the first to mention the occurrence of two or more
embryos in one seed and recently Cook (1902, 1903) has called
attention to polyembryony in Ginkgo. Miss Wigglesworth
(1903) notes the presence of stomata on the cotyledons.
LeMaout and Descaisne ('76) and Masters ('91) figure seedlings
of Ginkgo. Van Tieghem ('70, '87) and Van Tieghem and
Douliot ('88) record observations on the anatomy of the root
system. Worsdell ('97) describes the vascular bundle and trans-
fusion tissue of the cotyledon, and Seward and Gowan (1900)
contribute some further observations on the anatomy of the
seedling.

The remarkable ciliated spermatozoids of Ginkgo have
received such critical attention from Japanese botanists l that no
observations on these need be offered here. . With the corrobo-
rative evidence from the cycads z our knowledge of the occur-
rence and development of ciliated spermatozoids in these Gym-
nosperms may be considered well established. In the present
paper the subjects of oogenesis, spermatogenesis and fecunda-
tion will be avoided, the recorded observations dealing solely
with the embryogeny. It is purposed to take up and continue the
subject where Strasburger left it some thirty years ago.

MATERIAL.

The investigations, which are recorded in the following pages,
were begun in 1901 on material received from the Botanical

'Hirase ('96, '97, '98), Fujii ('98, '99), Miyake ('98, 1902).

2Ikeno('96, '97), Ikeno and Hirase ('97), Webber ('971, '972, 1901), Lang (1900).

Lyon: THE EMBRYOGENY OF GINKGO. 277

Gardens of the Imperial University of Tokyo through the kind-
ness of Mr. K. Yendo. Later large quantities of excellent
material were obtained from the Missouri Botanical Garden,
and it is principally from this material that embryological data
have been obtained. The writer wishes to express his sincere
thanks to Dr. William Trelease who has done every thing pos-
sible to facilitate the work. Some valuable preparations have
also been obtained from a small number of seeds purchased from
Thorburn & Co., seed dealers of New York. The location of
the trees which bore these seeds was not determined. The
material sent from Japan by Mr. Yendo, amounted to several
quarts of seeds containing nearly mature embryos. The embryos
were removed from a large number for examination, those
shown in Plate XXXV. being some of the number, while many
of the seedlings studied were grown from these seeds. Under
the directions of Dr. Trelease, cuttings bearing megasporangia
were sent from the Missouri Botanical Garden at intervals of a
few days throughout an entire season. After the cut ends of
the branches had been sealed over with paraffin, they were
packed in moist sphagnum, wrapped first with oiled, then with
heavy paper and sent by parcel-post. These excellent precau-
tions brought the material to this laboratory in a perfectly fresh
condition. Upon arrival it was immediately placed in the
various fixing fluids.

RESEARCH.

A. The Embryo.

i. The Protocorm. — At the completion of free-division the
nuclei are quite evenly distributed through the cytoplasm of the
oosperm (Jig. /), and when the formation of walls between
these nuclei is first completed the resulting cells show no marked
dissimilarity in shape, size or contents (fig. 2). The cells, in
the upper two thirds or more of this spherical protocorm, divide
only a few times or not at all. Their protoplasmic contents be-
come thin and watery, and they take no part in the organiza-
tion of the metacormal bud or blastema. The cells in the lower
portion of the protocorm divide repeatedly, the relative activity
increasing towards the base so that in this region there is organ-
ized a small-celled tissue (figs. 3, 4). This basal tissue passes
over directly into the small-celled meristem of the blastema.
With the advance of the metacorm into the body of the gameto-
phyte, the protocormal tissue is forced back through the neck

278 MINNESOTA BOTANICAL STUDIES.

of the archegonium until it comes in contact with the firm nu-
cellar tissue (figs. 4-9)- Many of its cells are often crushed by
this backward pressure, but in the mature embryo of the seed
the protocormal region is still evident (figs. H, 34* 36)-

2. The Blastema. — The blastema arises directly from the
small-celled, basal tissue of the protocorm, and invades the
gametophyte as abroad, blunt cylinder (figs. 4, 5, 6, <?). The
central region of the gametophyte seems to be the least resistant,
and from the latterly disposed protocorm, the blastema directs
its growth into this central tissue (figs. 5, <?). The path, which
the embryo is to follow, is marked out for a considerable dis-
tance ahead of it by disorganized cells (fig. 8}.

At first the metacormal bud is throughout meristematic ; but
very soon there can be distinguished two growth-foci, one directly
behind the other, in the axis of the blastema, and only separated
from each other by a very few cells (fig. 5). The apical growth-
focus is the growing point of the stem, and includes the entire
apical region of the young metacorm. The second growth-
focus is the growing point of the root. It first becomes notice-
able through the apparent diverging from this area of the indefi-
nite cell-rows which extend forwards towards the apex of the
blastema (figs. 5, 6). It also immediately begins to cut off
rectangular cells from its end towards the protocorm, which
form the characteristic, parallel cell-rows of the central root
cap region (figs. 5-7). Both growing points arise through a
localization of growth activity out of one general meristematic
tissue ; and hence, from the first are many-celled meristems.

3. Cotyledons and Leaves. — The cotyledon primordia origi-
nate through a localization of growth-activity in the marginal
region of the broad apical meristem (fig. 6). They first appear
as crescent-shaped mounds of tissue, which push rapidly ahead
of the stem apex (fig. 7). Each cotyledon has an apical
meristem. Of the many embryos from Japan examined, all
were dicotyledonous with the exception of two ; but among the
hundreds of embryos from the Missouri Botanical Garden, which
have been studied, over fifteen per cent, were tricotyledons.
The more common number of cotyledons is therefore two ; but
it is by no means constant as is usually stated to be the case.
The plumular leaves, whose origin directly follows that of the
cotyledons, arise in the same manner as the latter, but have a
much more restricted intraseminal growth (fig. 10).

Lyon: THE EMBRYOGENY OF GINKGO. 279

4. Embryonic Tissue-Systems. — As the blastema grows out
from the protocorm, its superficial cells are irregular, and divide
by periclinal as well as anticlinal walls (Jig- 4). This condi-
tion prevails even after the stem apex has become quite distinct
(jig. 5). Later, with the broadening of the apex, which causes
a rapid increase of surface, periclinal divisions become less fre-
quent. They, however, continue to occur for some time in the
most central region of the stem apex, and can often be detected
here in nearly mature embryos. Superficial cells of the coty-
ledon and leaf-meristems have been found dividing periclinally.
In regions, other than meristems, the dermatogen appears to be
morphologically distinct from the subjacent tissue.

The plerome and periblem can only be recognized as regional
in the embryo, for they are not sharply marked off from each
other. The first procambial tissue is differentiated in connec-
tion with the cotyledons, passing from these straight back into
the body of the embryo (Jig. j)- The next procambium-strands
arise, in like manner, in connection wtth the first plumular
leaves ; but since these are closer together than the cotyledons,
their procambium-strands are closer together. The shape of
an embryo's stele is determined by these two sets of procambial
strands. Hence, if there are two cotyledons, the stele will be
ellipsoidal (figs. 13-15) ; but if there are three cotyledons, it
will be triangularly prismoidal (fig. 18). Attheroot-meristem,
an ellipsoidal plerome narrows down into a broad wedge (Jigs.
n, 12) ; while in the same region a prismoidal plerome narrows
down into a blunt pyramid. A single procambium-strand enters
each cotyledon. As it passes upward in the cotyledon, this
strand, usually but not always, divides once. A few pro-
toxylem-elements are often differentiated in the cotyledon-
bundles before intraseminal growth stops.

In both plerome and periblem numerous secretory vessels are
formed through the breaking down of cells in longitudinal
rows (Jigs. 10-12). Many spherical resin-reservoirs also arise
in the cortex of the stem, cotyledons and leaves by the disor-
ganization of masses of cells (Jig's. 10-20). In mature embryos
the cells of the cortex are packed with starch.

5. Polyembryony. — It often happens that when the oosphere
of one archegonium of a gametophyte is fertilized, the oosphere
of the other is also fertilized and two protocorms develop. Occa-
sionally gametophytes will be found with three archegonia ;

280 MINNESOTA BOTANICAL STUDIES.

and in one such exceptional case three young embryos were
found. Of the two embryos often formed in one seed, the
more vigorous soon occupies the central, non-resistant tissue of
the ganietophyte, while the other aborts. In the many seeds
examined, only one case has been found (fig. 29) where two
embryos from different oosperms have developed to maturity
in the same gametophyte. Several cases, however, have been
met with, where two metacorms have resulted through the
production of two blastemata by one protocorm. Figs. 36 and
37 show two such cases ; while fig. 9 shows an instance where
two blastemata have arisen on one protocorm, and then one has
aborted.

6. The Mature Embryo of the Seed. — The single embryo
figured and described by Seward and Gowan (1900) was, with-
out question, a freak ; for it bears little resemblance to a typical
embryo. The general description of Ginkgo embryos given
by Coulter and Chamberlain, as quoted above, was undoubtedly
constructed from that of Seward and Gowan ; for they describe
the same anomalies as constant characters of the embryo.

A very good idea of the embryo of the seed can be obtained
through an examination of the photographs of Plates XXXIV-
XXXVI. Figs. 23 and 25-28 are typical dicotyledonous, and
figs. 22 and 30-34 typical tricotyledonous embryos. Figs. 13-
17 are sections of a dicotyledonous, and figs. 18 and 19 sections
of a tricotyledonous embryo. Occasionally freakish, deformed
embryos are met with, but are no more numerous than is usual
in other plants. The one portrayed in fig. 35 is the only one
among the hundreds examined which approximated the one
described by Seward and Gowan ; and the shorter cotyledon of
this one was quite entire. The embryo, portrayed in fig. 24,
is one of several found showing a condition intermediate
between typical dicotyledonous, and tricotyledonous embryos.
Fig. 20 is a section of the same embryo. In this case a third
leaf was produced at the base of the plumule on the same level
as the cotyledons. It was, however, partially enclosed by the
functional cotyledons (fig. 26), and in length only equalled the
plumule. The stele of this embryo was triangularly prismoidal
as in the case of typical tricotyledons. Fig. 21 is a section
through the cotyledons of an apparently dicotyledonous embryo,
which was morphologically tricotyledonous ; two cotyledon-
primordia having developed conjointly, producing a single

Lyon: THE EMBRYOGENY OF GINKGO. 281

large member. This embryo possessed the stele of a typical
tricotyledon. In several similar cases noted, the primordia had
assumed independent growth after a period of conjoint develop-
ment, so that the cotyledons were not united throughout their
entire length (fig. jj).

Five leaf-primordia are usually to be distinguished on the
plumule of a mature embryo. In a dicotyledonous embryo,
the cotyledons are often not so truly diametrically opposite each
other, as are the two following leaves ; one of the latter being
regularly larger than the other, and occupying more space
between the cotyledons (Jigs. 75-77). Beyond the cotyledons
and first pair of plumular leaves, a decussate arrangement does
not obtain in the embryo (fig. //). In a tricotyledonous em-
bryo, the leaves are arranged spirally and have an indeterminate
divergence (fig. /p).

In a mature seed the embryo extends through one half to
two thirds the length of the gametophyte. The cotyledons are
of equal length ; often slightly reflexed at their tips (figs. 24.-
2j), and when the embryo is removed from the seed, they
usually separate of themselves. Frequently, a considerable
mass of disorganized gametophytic tissue is included between
the cotyledons. The stem and root regions and also the prox-
imal halves of the cotyledons are white ; the ends of the coty-
ledons being yellowish or light green in color. The resin-
reservoirs are most numerous in the stem and basal portions of
the cotyledons, and in fresh embryos stand out as prominent
greenish or amber-colored pustules.

When an embryo is placed in a chromic acid fixing solution,
the root-cap region turns brown. A section (figs. 7, 77, 12)
shows this color-change to be limited to the peripheral layer of
cells; the change being due to a change in their contents. The
fixing-fluid has a similar effect on the contents of the secretory
vessels, and also darkens the resin-globules.

B. The Seedling.

i. Morphogenesis. — When seeds are placed under favorable
conditions for germinating, the hard shell is cracked, at the
micropylar end, by the swelling of the gametophyte. Through
this opening the body of the embryo is thrust out by the elon-
gation of the cotyledons. The root immediately turns down
into the soil (fig. jp), and as soon as the stem is free from the

282 MINNESOTA BOTANICAL STUDIES.

gametophyte it begins to grow in the direction opposite that fol-
lowed by the root (fig. 41). With the elongation of the stem,
the plumular leaves are quickly displaced from their original
relative positions (figs. 42~47)- The first two or three leaves
directly following the cotyledons do not develop the character-
istic blades, but remain small and scale-like (fig. 47). The stem
soon stops its rapid elongation, and having expanded a rather
close crown of leaves at its apex (figs. 47, 48), spends the
remainder of the first season in strengthening and protecting
itself by secondary growth in its tissues.

As is the rule among gymnosperms, the primary root of
Ginkgo persists as the tap-root. During the first season it
develops numerous short secondary roots (figs. 47, 48}.

Upon the germination of the seed, the proximal- portions of
the cotyledons, which, as we have already noted, are well sup-
plied with resin-reservoirs, protrude from the gametophyte as
large, arching petioles (figs. 41-44). The portion of the coty-
ledon, remaining within the gametophyte, enlarges considerable,
(fig. 48), its greatest diameter being about twice that of a coty-
ledon of a mature embryo of the seed. The cotyledons persist
throughout the first season. Fig. 48 is a photograph of a liv-
ing seedling at the end of its first summer's growth. It was
taken in October, after the seedling had experienced several
quite heavy frosts. The remains of the seed were removed
exposing the large fleshy cotyledons, which sprang apart of
themselves as soon as freed from the tissue of the gametophyte.

A large terminal bud is the only prominent one on the stem
during its first winter ; and, if uninjured, is the only one to
become active during the second season. Fig. 49 is a photo-
graph of a young plant which has made a considerable portion
of its second year's growth. The leaves all possess fully
expanded blades, and are well distributed along the stem.
During its second season the plant develops a much-branched
root-system.

2. Histogenests. — As has already been mentioned in discuss-
ing the embryo of the seed, the first permanent vascular tissue
is differentiated in the cotyledons. As the embryo begins its
extraseminal development, differentiation progresses from the
cotyledons towards the root- apex. The two bundles of a coty-
ledon unite eventually to form one of the persistent bundles
which extends throughout the length of the root. If the seed-

Lyon : THE EMBRYOGENY OF GINKGO.

283

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284 MINNESOTA BOTANICAL STUDIES.

ling has two cotyledons, the primary root will be diarch ; if it
has three cotyledons, the root will be triarch, a condition which
Dangeard ('90) has noted in other gymnosperms. In the transi-
tion-region between root and stem, considerable diversity of
structure prevails among Ginkgo seedlings. Yet the numerous,
and seemingly very different types can be readily explained as
modifications of the one general plan illustrated conventionally
in the adjacent diagrams.

Examining serial sections of a seedling of about the age of
those shown in figs. 42-46 we find, a short distance above the
root-meristem, two xylem-bundles appearing at the ends of the
elongated oval stele ; and in the sides of the latter, two long
arching bundles of protophloem (Diagram A and Jig. 50). In
each xylem-bundle the protoxylem lies away from, and the
metaxylem towards, the center of the stele (Diagram A). En-
tering the transition-region the protoxylem broadens and finally
divides into two masses which continue to separate as they pass
upward (Diagrams B and C). With the separation of the two
protoxylem groups the metaxylem is drawn out between them
into a broad plate (Diagram D), and in this condition the xylem-
mass passes out to the cotyledon (Jigs. 55, 56, 61). In the
petiole the metaxylem plate divides, each portion continuing to
rotate about its contiguous protoxylem until it reaches the posi-
tion shown in Diagram E. As the xylem-bundles pass on into
the blade of the cotyledon they continue to separate. The
metaxylem broadens out, appearing fan-shaped in section (Jig.
5(5?), and in the center of the cotyledon it nearly surrounds the
protoxylem, thus forming a mesarch bundle (Jig. jp) as de-
scribed by Worsdell ('97). With the first splitting of the pro-
toxylem-bundle, as it enters the transition-region, a proto-
phloem-group appears between and beyond its two points
(Diagram B). This protophloem-bundle increases in bulk as it
passes upward, and enters the cotyledon as a single bundle
(Diagram D and E). Later it divides to contribute a proto-
phloem-group to each bundle of the cotyledon (Jig. jp).

In the lower portion of the transition-region, additional protox-
ylem-elements appear at various points between the cotyledon-
traces, and just inside the prospective cambium-zone. Farther
up, these scattered elements give place to usually six rather def-
inite strands, which continue upward as the protoxylem-groups
of the six primary bundles of the stem (Diagrams B to D and

Lyon: THE EMBRYOGENY OF GINKGO. 285

Jigs. 52-57). Usually, the four secondary protoxylem-bundles,
adjacent to those of the cotyledon-traces, contribute to the latter
small groups of elements which unite with them just before they
pass out to the cotyledons (Diagram C. and Jigs. 55-61).

While the plan of transition outlined above is often followed
to a nicety in a seedling, exceptions are very numerous. The
length of the transition-region varies considerably in different
seedlings. As a rule, the cotyledon-traces do not leave the stele
at the same height (Jigs. 51, 61), and the reduction of the traces
into the two root-bundles may take place at very different levels.
It is often impossible to recognize distinct bundles in the traces
through the absence of protoxylem, or the indiscriminate ming-
ling of protoxylem and metaxylem elements. The sharp limi-
tations of protoxylem and metaxylem indicated in the diagrams
are rarelv met with in a series of sections.

The origin of the stem-bundles in the transition-region, is
usually after the manner already described (Jigs. 50-57, 60,
61). Occasionally, seedlings are met with, however, in which
the traces of the first two leaves are prominent in the transition-
region, and, with the four bundles of the cotyledon-traces, form
a hexarch stele, which may become resolved, lower down, into
such a tetrarch stele as seen in fig. 63. Such leaf-traces are
often distinctly mesarch. Then again, the trace of a single
leaf may continue prominent through the transition-region, and
produce a triarch stele for a considerable distance in the root.
In a tricotyledonous seedling, the primary root is persistently
triarch. Secondary roots are always diarch (Jig. 62).

Secondary thickening in the stem and root of Gtnkgo takes
place in the ordinary manner. The position of the cambium in
the root is shown in the diagrams and in figs. 50-57, 60-64.
Below the transition-region, -the pith of the root may be partially
or almost wholly replaced by metaxylem (Jigs. 62-64). The
centrifugal xylem becomes continuous in places with the metax-
ylem, but it rarely if ever comes in contact with the protoxylem.
It is often separated from the latter by only a single layer of
pith-cells (Jig. 62). The growing-point of the root ( fig. 65)
presents essentially the same structure as those of the cycads
and conifers which have been described by De Bary ('84). The
cork-cambium of the stem arises directly beneath the dermato-
gen ; in the root its origin is deeper seated.

The structure of the cotyledons of the seedling is illustrated

286 MINNESOTA BOTANICAL STUDIES.

in figs. 58 and 59. The tissues of the petiole are firm, while the
portion within the seed is fleshy. The cells of the region which
comes in contact with the tissue of the gametophyte are densely
packed with starch (Jig. 59). The rest have fluid contents.
Stomata occur on both surfaces of the cotyledon.

SUMMARY.

The essential features of the embryogeny of Ginkgo can be
summarized as follows.

1. By free-cell-formation, following free-nuclear-division, a
spherical protocorm is organized which completely fills the
venter of the archegonium.

2. The basal cells of the protocorm, through continued activ-
ity, pass over into the blastema or metacormal bud.

3. The meristems of the stem and root are localized out of
the one general meristem of the blastema.

4. Cotyledons and leaves arise as exogenous outgrowths
upon the growing-point of the stem and are here morphologic-
ally homologous structures.

5. Cases are infrequently met with, where two embryos from
different oosperms have developed to maturity in the same seed.

6. Polyembryony occurs, occasionally, through the produc-
tion of two blastemata by one protocorm.

BIBLIOGRAPHY.

Cook, Mel T. 1902. Polyembryony in Ginkgo. Bot. Gazette, 34 :
64-65.

Cook, Mel T. 1903. Polyembryony in Ginkgo. Bot. Gazette, 36:
142.

Coulter, J. M., and Chamberlain, C. J. 1901. Morphology of

Spermatophytes. Part i. Gymnosperms.
Dangeard, P. A. '90. Le mode d'union de la tige et de la racine

chez les Gymnosperms. Comptes Rendus, HO: 253-254.
Endlicher, S. '47. Synopsis Coniferarum.
Fujii, K. '98. Has the spermatozoid of Ginkgo a tail or not ? Bot.

Mag. Tokyo, 12 • 287-290.
Fujii, K. '99.' On the Morphology of the spermatozoid of Ginkgo

biloba. Bot. Mag. Tokyo, 13 : 260-266.
Hirase, S. '95. Etudes sur la fecondation et I'embryogenie clu

Ginkgo biloba. Journ. Coll. Sci. Imp. Univ. Tokyo, 8: 307-322.
Hirase, S. '96. On the spermatozoid of Ginkgo biloba. Bot. Mag.

Tokyo, 10: 325-328.

Lyon : THE EMBRYOGENY OF GINGKO. 287

Hirase, S. '97. Untersuehungen iiber das Verhalten des Pollens von

Ginkgo biloba. Bot. Cent., 69: 33-35.
Hirase, S. '98. Etudes sur la fecondation et 1'embryogenie du

Ginkgo biloba (second memoire). Journ. Coll. Sci. Imp. Univ.

Tokyo, 12 : 103-149.
Ikeno, S. '96. The spermatozoid of Cycas revoluta. Bot. Mag.

Tokyo, IO: 367-368.
Ikeno, S. '97 Vorlaufige Mittheilung iiber die Spermatozoiden bei

Cycas revoluta. Bot. Cent. 69: 1-3.
Ikeno, S. 1901. Contribution a 1'etude tie la fecondation chez le

Ginkgo biloba. Ann. Sci. Nat. Bot., VIII., 13: 305-318.
Ikeno and Hirase. '97. Spermatozoids in Gymnosperms. Ann. of

Bot. ii : 344-345-
Lang, W. H. 1900. Studies in the development and morphology of

cycadean sporangia. The ovule of Stangeria paradoxa. Ann.

of Bot., 14: 281-306.

Le Maout and Descaisne. '76. Traite de Botanique.
Masters, M. T. '91. Review of some points in the comparative

morphology, anatomy and life-history of the coniferae. Journ.

Linn. Soc. 27: 226-332.
Miyake, K. '98. On the spermatozoid of Ginkgo. Bot. Mag.

Tokyo, 12: 333-339.
Miyake, K. 1902. The spermatozoid of Ginkgo. Journ. Applied

Micros., 5 : 1773-1780.
Seward, A. C. and Gowan, Miss J. 1900. The maidenhair tree

(Ginkgo biloba}. Ann. of Bot. 14: 109-154.
Strasburger, E. '72. Die Coniferen und die Gnetaceen.
Van Tieghem, Ph. '70. Recherches sur la symmetric de structure des

plantes vasculaires. Ann. Sci. Nat. Bot. V, 13: 15-314.
Van Tieghem, Ph. '87. Sur le second bois primare de la racine.

Bull. Soc. Bot. de France, 34: 101-105.
Van Tieghem, Ph., and Douliot, H. '88. Recherches comparatives

sur 1'origine des membres endogenes dans les plantes vasculaires.

Ann. Sci. Nat. Bot. VII., 8: 1-660.
Webber, H. J. '97. * Peculiar structures occuring in the pollen-tube

of Zamia. Bot. Gazette, 23 : 453-458.
Webber, H. J. '97. 2 The development of the antherozoids of Zamia.

Bot. Gazette, 24: 16-22.
Webber, H. J. '97. 3 Notes on the fecundation of Zamia and the

pollen-tube apparatus of Ginkgo. Bot. Gazette, 24: 225-235.
Webber, H. J. 1901. Spermatogenesis and fecundation of Zamia.
Bureau of Plant Industry, Bulletin No. 2.

288 MINNESOTA BOTANICAL STUDIES.

Wiggles worth, Miss Grace. 1903. The cotyledons of Ginkgo biloba
and Cycas revoluta. Ann. of Bot., 17: 789-791.

Worsdell, W. C. '97. On " Transfusion-tissue " its origin and func-
tion in the leaves of gymnospermous plants. Trans. Linn. Soc.,
5: 301-319.

DESCRIPTION OF PLATES.
Plates 3^, 36, 37 and 39 are after photographs by Mr. C. J. Hibbard.

PLATE 29.

1. Sectional view of a protocorm in which free-nuclear-division
has ceased. From a section 20 thick ( x 160).

2. Section of protocorm just after completion of free-cell-formation.
Section 10 thick (x 160).

3. Surface view of a protocorm (x 160).

4. Section of a protocorm in the basal portion of which a blastema
is being organized (x 160).

The outlines for figs, i, 2 and 4 were traced from photomicrographs.

PLATE 30.

5. Longitudinal section of a young embryo in which the position
of the root-apex is just distinguishable (x 130).

PLATE 31.

6. A similar section of an older embryo showing first indication
of cotyledon-primordia (x 130).

PLATE 32.

7. Shows relation of metacorm to protocorm. The cotyledons
are well advanced and secretory canals are appearing in both plerome
and periblem (x 57).

PLATE 33.

8. From a section of a gametophyte containing a young embryo
(X 39). To the right can be seen the empty venter of the second
archegonium. Disorganization of the tissue of the gametophyte is
evident for some distance ahead of the embryo.

9. An instance where two blastemata were organized in one proto-
corm, but only one developed into a metacorm (x 36).

10. Section of a young embryo in a plane parallel to the faces of
the cotyledons, passing through the stem-apex and primordia of the
first two plumular leaves (x 57).

11. 12. Two sections of mature embryos cut, the one parallel, the
T perpendicular to the plane of contact of the cotyledons ( x 12).

These sections show, in a general way, the relation of parts, differ-

Lyon : THE EMBRYOGENY OF GINKGO. 289

entiation of tissues and shape of meristems. The secretory canals are
black. The openings in the cortex are resin-reservoirs.

PLATE 34.

13-17. Sections taken at intervals from a series of cross-sections
of a dicotyledonous embryo (x 28). 13, through root-meristem ;
14, through transition-region; 15, through bases of cotyledons; 16,
through cotyledons and stem of plumule; 17, through cotyledons and
plumular leaves.

18, 19. Sections of a tricotyledonous embryo (X 28).

20. From an embryo showing a condition intermediate between a
typical dicotyledon and tricotyledon ( X 28).

21. A section near the tips of the cotyledons of a tricotyledonous
embryo, two cotyledons of which developed conjointly (x 28).

PLATE 35.

22-29. Embryos removed from seeds grown in Japan (x 5.5).
The dark knobs, very prominent on some of the embryos, are pro-
truding resin-reservoirs.

PLATE 36.

30-37. Embryos removed from seeds grown in the Missouri Bot-
anical Garden (x 5.5) • Figs. 36 and 37 are cases of true twinning;
two metacorms having developed from one protocorm in each case.

PLATE 37.
38-46. Young Ginkgo seedlings about natural size.

PLATE 38.

47. A young seedling which has just completed its growth in
length for the first season, slightly reduced.

48. A seedling at the end of its first season's growth, somewhat
reduced.

PLATE 39.

49. A young plant early in its second year, somewhat reduced.

PLATES 40 AND 41.

50-57. Sections, selected at intervals from a series, passing from
the root through the transition-region to the stem of a young seedling
(X 92).

PLATE 42.

58, 59. Sections of a cotyledon of the seedling shown in Fig. 48
(X 28). Fig. 58 is through the petiole, and Fig. 59 through the
thickest portion of the cotyledon. The circular openings in Fig. 58
are resin-reservoirs.

290 MINNESOTA BOTANICAL STUDIES.

60, 61. Sections through the transition-region of a young seedling

(x 60).

PLATE 43.

62. Cross section of a secondary root (x 62).

63. Cross section of a primary root in which the traces of the third
and fourth leaves continued distinct through the transition-region, and
at the point of this section, formed a tetrarch arrangement with the
two cotyledon-traces (x 57)-

64. Cross section of a primary root in the lower portion of the
transition-region (x 25). A central mass of pith is completely sur-
rounded by metaxylem.

65. Longitudinal section through the tip of a secondary root
( x no). A flake of tissue, torn from the primary root, adhered to
the tip of the secondary root and appears in the section.

VOL. in.

MINNESOTA

3AL STUDIES

PART III.

DL. HI-

MINNESOTA BOTANICAL STUDIES.

PLATE XXX.

VOL. III.

MINNESOTA BOTANICAL STUDIES.

PART I

PLATE XXXI,

VOL. III.

MINNESOTA BOTANICAL STUDIES.

PLATE XXXII.

VOL III.

MINNESOTA EOT

PLA

'CAL STUDIES.

PART III.

iikrm

XXXIII.

VOL. III.

MINNESOTA BC

I

iUCAL STUDIES.

PART III.

20

18

VOL III.

MINNESOTA BOTANICAL STUDIES.

PART III.

27

PLATE XXXV.

VOL III.

MINNESOTA BOTANICAL STUDIES.

PART III.

34

PLATE XXXVI.

VOL III. MINNESOTA BOTANICAL STUDIES. PART III.

PLATE XXXVII.

VOL. III. MINNESOTA BOTANICAL STUDIES.

PART III.

48 47

PLATE XXXVIII.

VOL. III. MINNESOTA BOTANICAL STUDIES. PART III.

49

PLATE XXXIX.

VOL. III.

MINNESOTA BCJ

•H

STUDIES.

PART III.

52

^^ ' I

53

XL.

VOL. III.

MINNESOTA BC

54

ICAL STUDIES.

PART III.

XLI.

VOL. III.

MINNESOTA

• «•' * *

XH!

t.v;

S&tSb

~N X

60

;AL STUDIES.

PART III.

59

VOL. III.

MINNESOTA B(

CAL STUDIES.

PART III.

XXIV. OBSERVATIONS ON CALLYMENIA PHYL-
LOPHORA J. AG.

CLARA K. LEAVITT.

MATERIAL.

This study was made from plants collected by the writer in
July and August, 1901 and 1902, and by Miss J. E. Tilden in
December, 1901 at the Minnesota Seaside Station, Port Ren-
frew, B. C. All the older plants were washed up on shore.
Only very small young plants were collected on the rocks at
low tide.

HABITAT.

The plant is elittoral. It occurs in crevices in the rocky cav-
erns where the surge is strong as the tide runs in and out. It
grows in large quantities on the rocks running out from John-
son's Cove and is always to be found in the wash in the cove.
It is also attached to rocks on the sea bottom and was sometimes
washed in attached to the holdfast of a Nereocystis. Only
young plants were uncovered by the low tides ( — 1.2 ft.) of
August 2-4, 1902. The mature plants were well out beyond
the low tide line, indicating that the conditions were more
favorable at the lower points.

GROSS STRUCTURE.

The young plant consists of a small disk-shaped holdfast, a
short cylindrical stipe and one or sometimes two, entire some-
what circular laminae (jig. /). As the plant grows older pro-
liferations appear on the margin of the lamina, each consisting
of a short cylindrical stipe and a broad, flat, usually oval blade.
The main central lamina later becomes thickened longitudinally
from the secondary stipes on the margin to the main stipe, form-
ing a sort of palmate veining (fig. 2). A tearing of the lamina
between the " veins " results, and the stipe appears to have sev-
eral laminae arising from it, where originally there was but one.
Often several laminae arise from one holdfast each on a dis-
tinct stipe. The older primary fronds are dark red in color,
thick, coarse and leathery, while the younger laminae are much

291

292 MINNESOTA BOTANICAL STUDIES.

finer and thinner and of the same dark red color. The second-
ary laminae not infrequently arise from the surface instead of
the margin of the primary frond (Jig. 2). A third series of
laminae rising from the margin of the secondary laminae was
common, but a series of four as in fig. 3 was comparatively
rare. A plant often reaches twenty-five and thirty centimeters
from stipe to margin of outermost lamina. The dried specimen
does not adhere well to the paper.

MINUTE STRUCTURE.

The material used consisted chiefly of free-hand razor sec-
tions of fresh plants gathered in the summer of 1902. The
mature cystocarps were studied from formalin material collected
in December, 1901, at the same point.

1. Holdfast. — The holdfast is comparatively small and fits
like a sucker to the surface of a flat rock or curves over a pro-
jection or barnacle (figs. 4., 5). A thick layer of gelatine lying
close to the rock fits into all uneven places and takes the im-
pression of the rock (fig. 6). Above this, thick at the center
and thinner at the margin, lies a layer of cortical cells covered
by an epidermal layer several cells in thickness (fig. 7). Several
holdfasts were sectioned in which a second cortical and epi-
dermal layer appeared to have grown out over the first, so that
a layer of gelatine lay within the tissues of the holdfast.

2. Stipe. — The main stipe was about five millimeters in
length, cylindrical in form, becoming oval in cross section where
it passes into the lamina. The older stipes are thicker but not
longer than the younger ones.

The epidermis consists of deeply colored, thick walled, some-
what rectangular cells arranged in several rows. The cells of
the true epidermis are rather larger than those of the subepi-
dermis. Dimensions, 6.5 to 16.5 mic. in length and 6.5 mic. in
width (Plate XLIV.,fig. 8}.

The cortex is made up of large, clear, colorless cells. The
cells of the inner cortex are much larger than those of any other
part of the plant. Dimensions, diameter 16 to 50 mic. (Plate
XLIV.,fig. 9).

In the center lie the long, narrow, thick-walled cells of the
pith strand. These run mainly longitudinally, very few running
transversely. Dimensions, diameter 20 mic. (Plate XL IV.,
figs. 10, n).

Leavitt: OBSERVATIONS ON CALLYMENIA PHYLLOPHORA. 293

Peculiar layers of tissue appeared in some of the older stipes.
These remind one of annual rings. Similar structures are
reported by Jonsson in such related forms as Ahnfeltia -plicata
and Phyllophora membranifolia. In these Jonsson found a very
clear layering of the cortex marked by a difference in color and
diameter of cells and formed probably by a division of the cells
of the outer cortical layer.

The layers of stipe in Callymenia -phyllofhora as observed in
the few older stipes collected, showed no such origin. They
are irregular as to width and position, sometimes encircling the
stipe, sometimes appearing on one side only. Some partial
annulations were apparently due to a change of direction of the
filaments of the medullary layer. Most of the filaments of one
layer ran in a direction perpendicular to those of the next layer.

In other instances the annulations were apparently due to an
overgrowth covering a primary cortical layer. The epidermis
of the inclosed layer was somewhat disorganized and the gela-
tine was filled with diatoms which thus became imbedded some
distance in the stem. Several layers of cortex and epidermis,
distinctly marked by their imbedded parasites, appeared on one
side of some stipes. In these instances the buried epidermal
cells were very distinct.

LAMINA.

The lamina is made up of the same three tissues, an epider-
mis from three to five cells in thickness, a cortex two or three
cells deep and a pith strand occupying the main cross section

(figS. 12, 13).

The leaf is abruptly thickened at the margin, due mainly to
the greater number of cells in the pith strand and cortical layer

(fig- **)•

FRUIT.

The cystocarps form dark dots showing through the surface
of the thallus when held to the light. The mature cystocarp
increases only slightly the thickness of the lamina in the center
of which it lies. The spores are of rounded but rather irregular
form, many of them enclosed together in a compartment-like
portion of the cystocarp formed by a single row of long clear
cells (Jig. 75). The cystocarp had no well defined wall as
described by Carruthers for Rhodymenia. The spores are dis-
charged by a rupture of the epidermal layers of the lamina just

294 MINNESOTA BOTANICAL STUDIES.

over the cystocarp. Dimensions of spore, length 20 to 35 mic.
width 13 to 20 mic.

Young cystocarp material was collected late August, 1902,
showing the oogonium and accessory cells in figs. 16 and 17.

The stages of development described in Callymenia J. Ag.
by Bornet could not be made out.

PARASITES.

Microcladia coulteri. — This plant occurred parasitic on
nearly every plant collected. The largest found was six cen-
timeters in length. It is usually found on the margins of the
fronds but sometimes occurs on the surface. A section through
the lamina of the host shows the rounded base of the parasite
standing in a cup-like depression of the host thallus formed by
the disappearance of the epidermal cells. Long rhizoid-like
cells project from the short rounded cells at the base of the
parasite down between the cortical cells of the host {Plate
XLV., fig. i).

Callithamnion sp. — Almost all laminae showed at some
point traces of this parasite. The primary frond is frequently
covered with a fine short downy mat of it. Plate XLV., fig. 2,
shows a young filament extending up from the surface of the
host thallus. Beneath the surface the parasite sends down a
long rhizoid to the pith strand of the host through which it
ramifies. This rhizoid sometimes branches in the pith strand.
Above the surface the Callithamnion shows its characteristic
branching. Plate XLV., fig. 3 shows this parasite in the tetra-
gonidial condition.

Comparison of Microcladia and Callithamnion. — The former
is comparatively large, is borne on the margin of the thallus and
its penetration into the pith strand was slight, not branching in
any direction. The latter was either invisible to the naked eye
or appeared as minute down on both surfaces of the host thal-
lus. It penetrated through all parts of the host as a single
branching filament.

Porphyra sp. — Young Porphyra plants were found growing
epiphytically upon this parasitic Callithamnion. Plate XLV.,
fig. 4, shows such a plant consisting of a filament of four cells
which broadens out to form the characteristic flat thallus of Por-
phyra. At the base the filament forks, forming two branches
of three cells each, which served as a holdfast.

Leavitt : OBSERVATIONS ON CALLYMENIA PHYLLOPHORA. 295

Chlorochytrium inclusum. — A section cut in fresh material
August 12, 1902, showed a mature vegetative stage of this
parasite, Plate XLV., fig. 5. It corresponded very closely to
the parasite on Constantinca rosa marina found and described
by Mr. E. M. Freeman. Dimensions: 150 mic. x 75 mic.

It was oval in form and was inclosed in a thick cell wall
which protruded beyond the epidermis of the host. The parasite
lay mainly in the epidermis scarcely extending into the cortex
of the host. Several distinct pyrenoids could be distinguished
in the bright green chlorophyll.

Endophyte, genus unknown. — This parasite was to be found in
the great majority of the sections cut in the laminae. It occurs
but rarely on stipe or holdfast. It is usually distinctly vase
shaped with the interior of the vase toward the interior of the
lamina. It often lies entirely in the subepidermis and never
extends beyond the surface of the host. The larger ones extend
down into the outer cortex. Each is enclosed in a thick wall.
The contents are homogenous, granular and vary from an almost
colorless to a yellow green in color. Often what appears to be
a large oil drop lies in the base of the vase. The parasite con-
sists often of two and sometimes of three or four cells (Plate
XLV., fig. 6), one large cell in the base of the vase and one or
two in the narrow neck. The larger plants are often one celled,
Plate XLV., fig. 7, and the small ones are sometimes two.

Dimensions: length 35 mic. to 140 mic. width 25 to 45 mic.
Plate XVL., fig. 8, shows diagramatically the relative positions
of the cystocarp and the two parasites Callithamnion and .

EXPLANATION OF PLATES.

Plate XLIV. figures Callemenia phyllophora f. orbicularis as to
general habit and structure. Plate XLV. figures various parasites
upon the plant.

PLATE XLIV.

1. A young plant about half natural size.

2. Older plant with secondary lamina? rising from margin and
surface, about one half natural size.

3. Mature plant consisting of series of four lamina?, about one
half natural size.

4. Joint holdfast of young and mature frond attached to clam shell,
about natural size.

5. Side view holdfast, natural size.

6. Sucker-like under surface of holdfast, natural size.

296

MINNESOTA BOTANICAL STUDIES.

7. Cross section of epidermis and cortex of holdfast, x 250.

8. Cross section through epidermis and cortex of stipe, x 250.

9. Detail of cortex eel Is of stipe, x 250.

10. Cross section through pith strand of stipe, x 250.

11. Longitudinal section through pith strand of stipe, x 250.

12. Cross section through epidermis of lamina, x 250.

13. Cross section through cortex and pith strand of lamina, x 250.

14. Diagram of cross section of leaf showing thickening of margin
due to increase of cortex and pith strand, x 30.

15. Section of cystocarp showing spores inclosed by row of clear
cells, x 250.

1 6. Oogonium and accessory cells among cortical cells of lamina
X 250.

17. Another oogonium and accessory cell, x 250.

PLATE XLV.

1. Outline of young thalius of Microcladia coulteri from the base
of which project long rhizoid-like cells into the cortex of the host
plant, x 170.

2. Young filament of Callithamnion among cells of host, x 170.

3. Filament of Callithamnion bearing tetragonidia, x 170.

4. Young frond of Porphyra epiphytic on parasitic Callithamnion,
X 170.

5. Chlorochytrium inclusum lying in epidermis of host, x J7o.

6. Endophytic parasite of unknown genus consisting of four cells,
X 170.

7. Same parasite surrounded by epidermal cells, x 170.

8. A diagram of cross section of lamina showing relative positions
of cystocarp, parasitic Callithamnion and endophyte, x 20.

VOL. III.

MINNESOTA BOTANICAL STUDIES.

PART III.

9

VOL. III.

MINNESOTA BOTANICAL STUDIES.

XXV. OBSERVATIONS ON ENDOCLADIA MURI-
CATA (P. AND R.) J. AG.

FLORENCE M. WARNER.

The collections upon which this paper is based were made at
the Minnesota Seaside Station in August, 1902.

This species was given the same name by two different stu-
dents of algag at about the same time. Harvey gave the
name Gigartina muricata to a form from San Francisco in 1839
or early in 1840 while Postels and Ruprecht gave the same
name to a form of the same species in 1840. Harvey describes
the plant (Ner. Bor. Am., p. 182, pi. 27, B) as follows : " The
frond is formed of a simple, jointed axial filament of large di-
ameter, with internodes containing endochrome and about thrice
as long as broad, coated externally by a thin stratum of minute
cellules, from which radiate to all sides numerous, dichoto-
mous, moniliform, horizontal filaments, whose apices, strongly
soldered together, unite to form the periphery. The substance
is firmly cartilaginous, rigid when dry, Color a very dark
brown. Conceptacles spherical, sessile on the ramuli."

Schmitz and Hauptfleisch in Engler and Prantl's Die natiir-
lichen Pflanzenfamilien, describe Endocladia and figure the
three-celled carpogonial branch with the auxiliary cell which
form the procarp of E. vernicata J. Ag. The thallus, accord-
ing to the above, is cylindrical, very much branched on all sides
with small hooked spines, and has a distinct filamentous struc-
ture. There is a rather thick, long-jointed central axis, with
an alternating, inclined, jointed apical cell. This sends off in
alternate order dichotomously branched filaments which grow
diagonally upwards. These branches are more loosely con-
structed and longer jointed toward the center, but toward the
cortex become smaller celled and closer, lying finally side by
side. The inner layer is more or less quickly traversed with
dichotomously branched short-celled rhizoids. The central
axis is surrounded by numerous analogous filaments running
lengthwise. Gonidia are found in great numbers in the thick-
ened branches of certain sections of the thallus. Procarps are

297

298 MINNESOTA BOTANICAL STUDIES.

found in the somewhat loosened, fruit-bearing section of the
thallus appearing in great numbers in the central part of the
cortex. A short, two or more celled, many forked, small second-
ary side-branch of a filament forms an auxiliary cell from an
end cell. Near the auxiliary cell is developed a three-celled
carpogonial branch bent in shape of a hook. The gonimoblast
apparently arises from the fecundated auxiliary cell, branches
profusely, at times towards the center, into the somewhat loos-
ened tissue of the inner layer. The branches of the gonimo-
blast creep between the rows of cells of the sterile tissue, often
fusing with these cells, and finally the end cells develop into
spores.

The fruit-body is an irregular mass of interlacing fibers of
which the lower, stronger sections of the branches stand out
plainly, with numerous spores irregularly massed in the inter-
stices. The cystocarp, without a special protective layer, is
sunk in the locally, slightly thickened thallus. It protrudes
slightly to one side of the thallus near the short spiny point of
a branch. The fruit wall formed by the local thickening of the
cortex of the thallus does not show a pore.

The same authors describe the reproductive organs of the
Gigartinaceae as follows : Reproduction occurs both sexually
and non-sexually. The tetraspores are strewn over the surface
under the outer cortex or in many irregular groups and then
sunk in the inner cortex of the thallus, or arranged in project-
ing nemathecia. The sporangia usually divide transversely but
they also divide obliquely (Endocladid).

Antheridia are spread over the upper surface of the thallus,
sometimes in the form of small, cup-shaped capsules, opening
outwards, and sunk in the outer cortex of the thallus.

The carpogonial branch develops from a lateral branchlet of
a primary branch. The carpogonial branch is three-celled,
bent inwards like a hook, and connected with the swollen auxil-
iary cell, rich in contents. The fecundated auxiliary cell grows
inwards and develops the gonimoblast branches. The end cells
of these branches are transformed into spores.

Endocladia hamulosa (Ruprecht) J. Ag., described in De-
Toni's Sylloge Algarum, seems to differ from E. muricata only
in the position of the cystocarps. " E. hamulosa seems to
differ from E. muricata only in having the cystocarps at the
bases of the ramuli, while in the latter species they are simply

Warner: OBSERVATIONS ON ENDOCLADIA MURICATA. 299

lateral. We have found both sorts on the same plant so it has
seemed best to include both under the same name" — Setchell and
Gardner. This is true also of the specimens examined by me.

Endocladia muricata (P. and R.) J. Ag. is a red alga belong-
ing to the genus Gigartinaceae. The plants studied at the
Minnesota Seaside Station, Vancouver island, seem to be the
typical form. Setchell and Gardner describe two other forms,
E. muricata forma compressa and forma inermis, but as the
specimens in hand have not a particularly flattened frond and
are not destitute of spines, they are probably neither of these
forms.

The plants were found growing on rocks and boulders in the
upper portion of the littoral zone very near high water mark.
They were fastened quite firmly to the substratum. The fronds
are low, from 2-4 cm. in height, shrubby in appearance, and
very dark red or brown in color. The branching is dense and
irregular and the branches are profusely covered with spines.
The frond seems to proceed from a branch which runs hori-
zontally along the surface of the substratum. This horizontal
branch sends off downward branches at the ends of which hold-
fasts are developed. Upright branches develop into the
frond.

Frond. — Examining a longitudinal section (Plate XL VI., fig.
4) of the frond a conspicuous central cylinder is seen surrounded
by a mucilaginous sheath. This axis is divided into cells about
three times as long as broad. There appear to be protoplasmic
connections running through the dividing cell walls of the axis
cylinder. Branches are given off quite regularly from this
central axis, the branches arising just below the cross walls of
the central axis, and often from two sides of these cells. These
branching filaments do not extend radially out to the cortex as
described by Harvey, but rather diagonally upward and outward,
terminating in the cortex opposite the lower part of the third cell
of the central axis from which they started (Plate XL VI., fig. 4).
The branching seems to be more or less regular (Plate XL VI. ,
fig. 2]. Two branches are given off from the upper third
of the cell, following somewhat the method of branching of the
central cylinder. The branching, in this way, seems to be quite
regular for about eight cells, when it sends off two branches,
each of these branching dichotomously until the cortex is
reached. There are, however, exceptions to this rule.

300 MINNESOTA BOTANICAL STUDIES.

Massed around the central axis are small round cells. These
seem to have developed, at least in some cases, from the upper
branch (Plate XL VI., figs. 2, a, and j, a) of the original branch
coming from the central axis. Schmitz and Hauptfleisch
speak of these rounded cells as rhizoids, while Harvey does
not discuss their origin, but speaks of them as coating the axis.
In the material studied the branching was not always dichoto-
mous nor did the branches run radially outwards. The fila-
ments in the center of the frond are loosely scattered, being
massed together closely to form the cortex. Examining a cross
section of the frond we find a large round central cylinder
(Plate XL VI., fig. i). Massed around this cylinder are small
round cells. The cells in the center of the section are not con-
nected with filaments showing that the filaments of which these
cells are cross sections run parallel to the central axis. The
tissues are loosely arranged, but towards the periphery dichoto-
mous branching can be observed and these branches held to-
gether by a gelatinous secretion form the periphery.

Holdfast. — The holdfast is strong, although quite incon-
spicuous. It does not appear to be disc-like, but rather to be
composed of branches (Plate XL VI., fig. 7). There is a brown
cellular substance which is developed beneath the holdfast.
Apparently the holdfast can be developed at any point where
the horizontal branch may come in contact with the substratum
or at the ends of the small branches which radiate downward
from the horizontal branch.

Asexual Reproduction. — Some branches appear slightly
fleshier than others. When sectioned there are found distributed
around them, the tetragonidia. These are developed from the
peripheral cells. In one section (Plate XL VI., fig. 8) we may
find the younger gonidangia the contents of which are not yet
divided, those which have divided obliquely forming two masses,
and the mature tetragonidangium containing four gonidia. As
has been previously stated they do not divide perpendicularly but
obliquely. Plate XL VI., Jig. 8, illustrates a portion of a longi-
tudinal section drawn from the side of the axis cylinder to the
periphery. The same structures would be seen on the other
side of the axis cylinder. Long narrow paraphyses extending as
far again as the tetragonidangia are found which function as a
protection for the gonidia. They are developed from certain
peripheral cells and are made up from fourteen to sixteen cells.

Warner: OBSERVATIONS ON ENDOCLADIA MURICATA. 301

Sexual reproduction. — The cystocarps are found on the
branches, sometimes singly, often two on one branch, in which
respect this species differs, as has been stated, from E. hamulosa.
Beyond the cystocarps, sterile tissue extends in the form of a
projection or spine. In a cross section of the cystocarp (Plate
XL VI., Jig. 5) slender branching filaments pass in and out
among the carpospores. The gelatinous sheath of these filaments
is not plainly seen here, the protoplasmic contents alone being
visible, and the cells of the filament have become very much
distorted. The carpospores are uninucleate and vary in shape,
some being spherical, some oblong and others oval. The
wall of the cystocarp has the characteristic structure of
the wall of the frond. Distorted branches connected at
times with the cortical cells are found ramifying through
the structure. The development of the cystocarp has been
given previously in the paper as described by Schmitz and
Hauptfleisch.

The author desires to thank Professor Conway MacMillan
for suggesting the subject for study, Miss Josephine E. Tilden
for a detailed outline of the work, and Professor R. A. Harper
for encouragement and helpful suggestions during the progress
of the work.

BIBLIOGRAPHY.

Postels and Ruprecht. Illustrationes Algarum, 16. 1840.
Harvey, W. H. Nereis Borealis Americana. Part 2. 182. 1852.
Saunders, De A. The Algae. Papers from the Harriman Alaska

Expedition. Proc. Wash. Acad. 3: Part i. 1901.
Agardh, J. G. Species, Genera et Ordines Algarum, 3 : 558. 1876.
Setchell and Gardner. Algae of Northwestern America. Univ. of

Calif. Pub. Botany, 296. 1903.
DeToni, J. B. Sylloge Algarum, 4: 175. 1897.
Engler and Prantl. Die Natiirlichen Pflanzenfamilien. 353. 1897.

EXPLANATION OF PLATE XLVI.

All drawings were made by the aid of a camera lucida.

1. Cross section of the frond. Central axis; small cells surround-
ing central axis ; periphery.

2. Branching of one of the filaments taken about three cells from
the central cylinder. Shows somewhat regular branching, similar to
the branching of the main axis. At the periphery dichotomous branch-
ing is illustrated.

302 MINNESOTA BOTANICAL STUDIES.

3. A portion of the central axis showing a branch which is leaving
it. Protoplasmic axis of the central cylinder. The protoplasm
appears to separate into threads to pass] through the thickened plate.

4. Longitudinal section of frond. Axis cylinder surrounded by
mucilaginous sheath. This figure illustrates the point that the main
branches from the axis cylinder do not run out radially, but diagonally
upward, reaching the periphery opposite the lower end of the third
cell of the central axis from which they started.

5. Cystocarp containing carpospores; filaments ramifying among
he spores.

6. Carpospores and branching filaments.

7. Holdfast. #, horizontal branch; ^, stipe; c, brown cellular
substance developed beneath the holdfast; d, vertical branch.

8. Section showing tetragonidangia and paraphyses in different
stages.

MINNESOTA BOTANICAL STUDIES.

PART III.

PLATE XLVI,

XXVI. OBSERVATIONS ON LAMINARIA BULLATA

KJELLM.

OLGA MUELLER.

The plant, Laminar ia bullata, was first studied by Kjellman
in 1889 from material found by him in St. Lawrence Bay of St.
Lawrence island in Behring Sea. Some of the material used
for these observations was found by Miss Josephine Tilden at
Tracyton, Washington, but the greater part was collected
by the writer near Port Renfrew, on Vancouver island, on the
straits of Juan de Fuca, in August, 1902. From this it appears
that the plant has a wide distribution on the western coast of
North America, extending from Alaska to Puget sound, and
possibly to California.

The writer is deeply indebted to Professor Conwray MacMillan
and to Miss Josephine Tilden for helpful suggestions.

The plants collected at Port Renfrew were found growing
attached to rocks in a narrow arm of the sea leading into a
cave. Here the tidal currents were very strong, moving the
plants constantly to and fro, and bringing to them the food and
oxygen necessary for their life. They were found growing in
the sublittoral zone and could be collected only at low tide and
then with difficulty.

EXTERNAL MORPHOLOGY.

The plant, like other members of the Laminariaceae, consists
of three portions ; the holdfast, the stipe, and the lamina. The
distinguishing feature of the plant is its rows of undulations,
or of alternating elevations and depressions, called bullations,
which run nearly parallel to the margin of the lamina, but at
some distance from it. The color of the plant is dark brown.
The margin of the lamina is straight and not undulated. Its tex-
ture is like that of strong, firm, sheet rubber, and its smooth glisten-
ing surface offers but little resistance to the action of the waves.

Plants of various ages were examined. The youngest was a
tiny individual 2.5 cm. in length. The primitive disc showed
to good advantage ; its lower surface was almost flat, while the
margin was slightly irregular, with indications of where the first
hapteric branches would arise.

303

304 MINNESOTA BOTANICAL STUDIES.

The stipe was .8 cm. long — cylindrical below and slightly
flattened above where it merged into the lamina. The lamina
was .17 cm. long, rather oblong in shape, showing as yet no
traces of the bullations which are so characteristic of the older
specimens, and the free end of the lamina even in so young a
specimen was not perfect, but slightly notched and irregular —
due no doubt to the action of the waves.

The largest specimen studied was found growing at Tracyton,
in quiet water. Its length was 143 cm. and the greatest width
of the lamina was 30 cm.

The holdfast by means of which the plant attached itself to
the rocks on which it grew, consisted of a mass of dichotomously
branched hapteres, which had arisen from above the primitive
disc. These hapteres were brown in color, being of a lighter
shade and of a more delicate texture towards the apex. In this
largest specimen the stipe was unusually short, being but 2.5
cm. in length. Most of the specimens examined showed larger
stipes, as in one whose lamina was 52 cm. long, the stipe had a
length of 8 cm.

The stipe is strong and tough in texture. It retains the
characteristic of the earlier stage of being cylindrical at the
base and flattened where it merges into the lamina.

The laminae of different specimens varied considerably in
outline, some being almost oblong, others broad ovate and
others elliptical.

The texture varies also, the laminae of some plants being
much thicker and firmer than of others. These differences
of form and texture are due, no doubt, to differences in the in-
tensity of the light, and the strength of the tidal currents.

The margins are in every case straight but the apex is almost
always frayed and split. There is often one long split extend-
ing nearly to the base of the lamina, a second one not so deep,
and several minor indentations besides. This splitting takes
place in a direction parallel to the margin of the lamina and
usually near the rows of bullations.

Although splitting seems to be the rule, yet our largest speci-
men showed scarcely a trace of it. This was no doubt due to
the fact that it was found growing in quiet water. Its apex
however was frayed somewhat by the action of the waves.

The region of growth is at the point where the stipe joins the
lamina, and here, in the younger portion of the lamina, the bul-

MUELLER : OBSERVATIONS ON LAMINARIA BULLATA KJELLM. 305

lations are most perfect, being more greatly elevated and de-
pressed, though not so large in diameter as in portions near the
apex.

The plant is a perennial, and towards the older portion of the
lamina the bullations become broader and more shallow until
they finally disappear, leaving the older portion of the lamina
with an even surface (Jig. /).

The greater part of the material was preserved in a five per
cent, solution of formaline, a little alcoholic material was used.
The former proved the more satisfactory. Free hand sections
mounted in glycerine jelly were used for study.

ANATOMY.

Laminaria bullata, like the other members of the Laminari-
aceae consists of three tissues, viz., the epidermal, the cortical,
and the pith. Only the first two are found in the hapteres,
while the stipe and lamina contain them all.

The surface of the plant is covered by a thick structureless
cuticle ; below this are the epidermal cells, prismatic in form,
about one and one-half times as long as broad, but with the
two shorter diameters equal, so that when the cells are seen
from the surface, they appear as cubes or pentagons.

These epidermal cells are densely crowded with chromato-
phores, their chlorophyll being masked by the brown coloring
matter characteristic of the kelps. The outer wall is compara-
tively thick while the lateral and inner walls are rather
thin.

Below the single layer of epidermal cells are found from two
to four layers of cells which are shorter and broader than the
epidermal cells, and not so densely crowded with chloroplasts.
These are the hypodermal cells.

Next to the hypodermal cells are found the cortical cells,
which are more irregular in form, though still prismatic. They
increase in size towards the center of the plant, and are followed
by strengthening cells which are smaller in diameter and longer
than the cortical cells. They also have thicker walls and some
of them are imbedded in the mucilaginous material of the central
part of the plant. These cells are devoid of chromoplasts, but
contain granular protoplasm.

The pith web consists of numerous colorless, interlacing
and anastomosing hyphae embedded in mucilage.

306 MINNESOTA BOTANICAL STUDIES.

HAPTERE.

A longitudinal section of the haptere shows that it is composed
of a layer of cuticle on its surface and below this are the pris-
matic epidermal cells with their numerous chromatophores.
Next to these are two layers of hypodermal cells, and just
within are the cells of the cortex. These are much larger and
more irregular in form than the epidermal cells (fig- j). In the
center of the haptere are found rather thick walled elongated
cells arranged in rows.

These rows run in a straight course to the apex of the hap-
tere (Jig- 4)9 while cells at the sides of the haptere bend in
curving rows from the circumference towards the center ( figs.
5, 2.) The cells near the end of each row have the power of
dividing and it is here that the haptere increases in length and
in thickness. Both cross and longitudinal sections reveal
numerous circular openings in the hypodermis. These are the
mucilage ducts which in the haptere seems to take the form of
spherical pits. Faint traces of branches may sometimes be seen,
so that it is possible that these pits are in communication with
each other. Each pit is surrounded by little granular secreting
cells.

There is no pith in the haptere, the thick-walled, elongated
cells of the cortex occupying the interior of the haptere.

In the stipe is found much the same arrangement of tissues
as in the haptere, but with this difference, that a pith web occu-
pies a considerable portion of the interior of the stipe.

The lower cylindrical portion of the stipe is hollow with only
traces of the pith web left. In the upper, younger, more flat-
tened portion, the pith web fills the center of the stipe
(fig. 6, A, B).

The cuticle is thicker on the surface of the stipe than on the
haptere ; the epidermal cells are slightly more elongated but
otherwise much like those of the haptere (Jig. p). The hypo-
dermal cells consist of several rows and among them are found
the mucilage ducts. A cross section shows them to be large
elliptical openings, very close together, in fact with only the
bounding cells of each duct between. Each duct appears to
have been formed as a fissure between four or five adjoining
cells (fig. 8). These cells have very granular contents, are
enlarged at the ends of the ducts but compressed where the
ducts are broadest. The longitudinal section shows the ducts

MUELLER: OBSERVATIONS ox LAMIXARIA BULLATA KJELLM. 307

as long cylindrical tubes — with the inner side crenate and the
outer straight, and with the secreting cells very granular (jig. J).

The cortex cells of the stipe are not so large as those of the
haptere (Jigs. 10, //)• The strengthening cells found near the
pith web, are in general cylindrical in form ; the innermost are
imbedded in mucilage. These cells are characterized by each
possessing an unusually large nucleus. The granular proto-
plasm passes through the center of the cell and communicates
with that of adjoining cells through the end partition walls.

The pith web is formed of interlacing, colorless hyphae. In both
cross and longitudinal sections can be seen some hyphae which
have been cut obliquely and others which have not been cut at
all, showing that they run in various directions. The majority
of them take a lengthwise course, however. The hyphae (figs.
12, /j) have comparatively thick walls, are imbedded in mucilage
and contain protoplasm and some starch grains. Trumpet
hyphae are numerous and are found running lengthwise more
often than crosswise.

In the lamina we see again the same tissues as in the stipe.
The chromoplasts of epidermal and hypodermal cells are more
numerous than in the corresponding cells of the stipe. The
mucilage ducts are not so numerous nor so compressed, being
circular rather than elliptical in outline (Jig. if). The cells of
the cortex are large and cuboidal in form (Jig. 14) and follow-
ing these are the thick-walled strengthening cells, circular or
oval in cross section (Jig. 75), and often arranged in pairs as
though recently divided ; elongated and of uniform diameter in
longitudinal section (Jig. 16). In these cells the hyphae of the
pith web take their origin, as branches from the sides of the
cells, or as a prolongation of the cell proper (Jigs. 15, 16).

In some cases the hyphae can be traced from their origin in
one cell, across the pith web to their termination in another cell
on the other side. The pith web is richly supplied with mucilage,
and imbedded in this are numerous trumpet hyphae — which do
not differ from those of the stipe (Jig. //).

The bullations, which are so striking a feature of the plant,
are not due to a thickening of any of the tissues of the lamina,
but rather to a bending in and out of these tissues, the epi-
dermis, cortex and pith web following each other in the same
order and proportion as they do in the even portions of the
lamina.

308 MINNESOTA BOTANICAL STUDIES.

Material in the fruiting condition was not available, and there-
fore observations on the reproduction will be made at a future
time. According to Kjellman the sori are found as oval areas
spread over that portion of the lamina where the old lamina
merges into the new. The sporangia do not differ from those
of other Laminariaceae.

BIBLIOGRAPHY.
Kjellman, F. J. Beringhafvets Algflora, page 46, 1889.

EXPLANATION OF PLATE XLVII.

1. Photograph of Laminaria bullata.

2. Diagram of haptere showing the direction taken by the cells of
the cortex. They run in straight lines towards the apex of the haptere,
but bend outward at the sides.

3. Longitudinal section of haptere showing epidermal cells, hypo-
dermal cells, cortex and a mucilage duct.

4. Cells near apex of haptere, as seen in longitudinal section.

5. Cells from side of haptere showing curving direction of cortex
cells. Longitudinal section.

6. a. Diagram of lower hollow portion of stipe.

6. Diagram of upper portion of stipe.

7. Diagram of longitudinal section of stipe. #, epidermal region;
<5, mucilage duct; c, cortex; d, strengthening cells; <?, pith web.

8. Cross section stipe, showing cuticle, epidermal and hypodermal
cells, mucilage ducts and cortex cells.

9. Epidermal and hypodermal cells of stipe. Longitudinal section.

10. Strengthening cells of cortex of stipe. Cross section.

11. Strengthening cells of cortex of stipe. Longitudinal section.

12. Pith web of stipe. Longitudinal section.

13. Pith web of stipe. Cross section.

14. Cross section of lamina showing cuticle, epidermal cells, hypo-
dermal cells, mucilage duct and cortex cells.

15. Cross section of lamina showing strengthening cells and origin
of hyphae of pith web.

1 6. Longitudinal section of lamina showing same points as 15.

17. Pith web of lamina in longitudinal section.

VOL. III.

MINNESOTA BG

11

10

«

ICAL STUDIES.

PART III.

XXVII. MINNESOTA HELVELLINE^.

DAISY S. HONE.

The following is a list of Minnesota Helvellineas collected at
various times since 1886.

In the determination of these species, Dr. H. Rerun's work
has been carefully consulted, as have also among many others
the British works of Phillips and Massee and the American
papers of Peck and Morgan. These works have been used
freely in the descriptions, though only where they agree with
my observations on the Minnesota plants. Krombholz's plates
have been compared and cited under each species. Compari-
sons have also been made with the Exsiccatae in the University
Herbarium and the results of such comparisons have been noted
in each case. Schroeter's generic classification as outlined in
Engler and Prantl, Die Naturlichen Pflanzenfamilien, has been
closely followed.

All drawings were made with the aid of an Abbe camera
lucida, usually from material preserved in formalin or from
dried material soaked in water. In a few cases fresh material
was used. The photographs were taken by Mr. C. J. Hibbard,
photographer on the Minnesota Geological and Natural History
Survey. All species, described are now in the collection of the
Museum and Herbarium of the University. They are pre-
served in a mixture of two per cent, formaline and seventy per
cent, alcohol and also in the dried condition.

Eight genera with fourteen species and two varieties here
reported are from Minnesota. Five species belong to the Geo-
glossaceae and nine to the Helvellaceae. No member of the
Rhizinaceae has yet been reported, but the common species of
Rhizinia probably occurs in the State. Specimens marked * are
those from which the photographs have been taken. The
reports of localities are given in counties.

I wish to express by thanks for the aid and supervision of
Professor E. M. Freeman, of the University of Minnesota,
under whose direction the work was done. I also wish to

309

310 MINNESOTA BOTANICAL STUDIES.

HELVELLACE^E.

thank Mr. C. J. Hibbard who kindly assisted by making the
photographs.

1. Helvella lacunosa AFZEL, Act. Holm. 304. 1783. (Plate
II., Jigs. n, 12, 13; Plate IV., figs. 11-16.)

Solitary or gregarious ; stipe lacunose, fistulose, slender,
grayish to mouse-colored, up to 3.25 in. high, 1.25 in. wide;
pileus saddle-shaped or three lobed, slightly wrinkled, mouse-
colored, up to 2 in. in diameter; spores elliptical, obtuse,
smooth, containing one large oil drop, 16-18 mic. long, 9-12
mic. wide ; paraphyses filiform, branched, clavate, about 6 mic.
wide at the tip.

On ground in moist, soft woods.

Ramsey, Sept. 1898, Freeman 229 ; Cass, Sept. 1898, Free-
man, 186 ; * Hennepin, Oct. 1900, Butters 63; Hennepin,
Sept. 1900, Freeman 760; Hennepin, Oct. 1900, Hibbard.

According to Phillips : " Differs from H. crispa, for a variety
of which it may easily be taken, by the more regular pileus,
2-4~lobed, scarcely laciniate, lobes later becoming free, and
especially by the colour. The stature generally smaller. Acute
characters are wanting in nature, therefore it is constant." The
stipe is more slender than that of H. crispa.

The specimens agree with Thuemen Mycoth. Univ. 809,
spores of which are up to 16 x 12 mic. ; Sydow Mycoth.
March, 182, spores of which are 16-18 mic. x 8— 10 mic. ;
Sydow Mycoth. March 2845, spores are 14-16 mic. x 8-10
mic.

Krombholz Schwamme III., pi. 19, Jigs. 18—21 (lacunosa),
Jigs. 22-26 (sulcatd)-, -pi. 21, jigs. 22-24.. I834-

2. Helvella crispa (Scop.) FR. Syst. Myc. 2: 14. 1822.
(Plate II., fig. 10.)

Generally solitary ; stipe lacunated, fistulose, slightly bulbous
at the base, stout, grayish, measures up to 4 in. high and 2 in.
wide ; pileus very much lobed and folded or wrinkled, white,
up to 2.5 in. across; spores elliptical, obtuse, smooth, contain-
ing one very large oil drop, 10-16 mic. long by 8-10 mic.
wide; paraphyses filiform, clavate, septate, branched, about 4
mic. wide at the tip.

On ground in hard woods, chiefly oak.

Hone: MINNESOTA HELVELLINE^:.

311

Hennepin, Oct. and Sept. 1900, Freeman 895, 734, 804;
Becker, Aug. 1901, Freeman 1053; * Chisago, Sept. 1900,
Butters 15; Ramsey, Sept. 1903, Cuzner ; Ramsey, Sept.
1903, Wilcockson.

H. crispa is closely related to H. lacunosa ; for differences
see H. lacunosa.

The specimens agree with Sydow Mycoth. March 181, spores
of which measure up to 12 mic. x 16 mic.

Krombholz Schwamme \\\.,pl. 19, fig. 27-29. 1834.

3. Helvella elastica BULL. Champ, franc. 299,^7. 24.2. 1785.
(Plate I I., figs. 14, 75.)

Stipe cylindrical, not lacunated, hollow, smooth, or pruinose,
grayish, up to 2.25 in. high by i in. across; pileus saddle-
shaped, bilobed, mouse-colored above, grayish beneath, up to
1.25 in. across; spores elliptical, obtuse, smooth, containing
one large, central oil drop, up to 16-20 mic. long by 10-12
mic. wide; paraphyses filiform, septate, branched, clavate,
about 6 mic. wide at the tip.

On ground in moist places and among grass.

* Hennepin, Sept. 1900, Freeman 877 ; Hennepin, Oct.
1900, Butters 66 ; Becker, Aug. 1901, Freeman 1040; Cook,
Aug. 1903, Freeman and Ballard 3.

The specimens agree with D. Saccardo Myco. Ital., spores
of which average 14 mic. x 9 mic.

Krombholz Schwamme \\\.*pl. 21, fig. 21, 1834.

4. Helvella infula SCHAFFER, Icon. Fungi, -pi. 159. 1763.
(Plate //., figs. /, 2, j; Plate IV., figs. 24-29.) "
Gregarious or solitary ; small or large ; stipe cylindrical,

tapering toward top, at first solid later hollow, cream to flesh-
colored, densely pubescent at the base, up to 2.25 in. high by
i in. wide ; pileus more or less saddle-shaped or irregularly
undulated, margin attached in places to the stipe, yellow to
cinnamon brown or chestnut brown above, cream to flesh-col-
ored beneath, finely pubescent beneath, up to 2.25 in. deep;
spores elliptical, obtuse, smooth, with two equal oil drops, 18-
24 mic. by 8-12 mic. ; paraphyses clavate, septate, branched,
brownish, about 4 mic. wide.

On the ground along paths and trails or among moss. It
usually prefers clayey ground but occasionally is found even
on decayed logs.

312 MINNESOTA BOTANICAL STUDIES.

* Cook, Aug., 1902, Fink; Cook, Aug., 1903, Freeman and
Ballard 5.

This is Gyromitra infida (Schaff.) Buel. Schroter in Die
Naturlichen Pflanzenfamilien, has defined Gyromitra as pos-
sessing an inflated cap. In the above species no true inflation
of the cap is found, *". £., the cavity of the stipe is not prolonged
into a cavity in the cap as in the morels. Hence the condition
here is similar to that of Helvella. It approaches the true
Gyromitras however in the tendency of the cap to become
irregularly convoluted and especially in the fusion of the cap
with the stipe. Helvella infula Schaff. therefore stands be-
tween the Gyromitras with truly inflated cap and the typical
Helvellas without fusion of cap and stipe. There is moreover
often a very marked saddle form to specimens of this species.
Schroter's conception of the genus Gyromitra accords with
Fries' original description " discus bullato-inflatus, costis ela-
vatis gyrosus." (Summa Veg. Scand. 346. 1849.)

Our specimens agree with Krombholz Schwamme III., pi.
/p, Jigs. 11-13 (•&• rhodopodq which is clearly a true Helvella
and is included by Rehm as a synonym under H. infnla.
They also agree with Roumeguere Fungi Gall. Exsicc. 1208,
in which the spores measure 18-22 mic. x 9-12 mic. ; Kromb-
holz Schwamme HI., pi. 21, figs. 12-17. Helvella infula also
agrees as far as exterior views are concerned but the sectional
views do not clearly show the relation of stipe and cap cavities
as seen in the Minnesota specimens.

Certain specimens of these Helvellas contained an abundance
of Sphceronamcemella helvella Karst. It differs from Karsten's
descriptions (Hedw. 23; 18. 1884) in its possession of ultimately
uniseptate spores. These are at first continuous and later uni-
septate. In all other points the agreement is complete.
5. Verpa conica (MILL.) SWARTZ. Vet. Ak. Handl. 136.

1815. (Plate //., figs. 4, 5, 6; Plate IV., figs. 30-33.)

Stipe cylindrical, tapering at the apex, hollow, white, even
or slightly wrinkled, floccose, up to 2.5 in. high and 1.5 in.
wide ; pileus conical, even or slightly wrinkled, brownish
above, white beneath, up to .5 in. deep by i in. across at the
base.; spores elliptical, obtuse, smooth, containing one large
central oil drop, 20-22 mic. long by 10-14 mic. wide; para-
physes filiform, septate, branched, 6 mic. wide.

On moist ground.

Hone: MINNESOTA HELVELLINK^E.

313

Ramsey, May 1899, Freeman 313 ; * Wright, May 1900,
Freeman 588 ; Beltrami, May 1902, L. R. B. W. 52 ; Ramsey,
May 1903, Policy; Hennepin, May 1903, Ramsey and Hone;
Chisago, May 1903, Nelson.

The specimens agree with Sydow Mycoth. March 3166, of
which the average spore measures 20 mic. x 12 mic. ; Roume-
guere Fungi Selecti Exsicc. No. 4554, of which the average
spore measures 20-22 mic, x 12-14 mic.

Krombholz Schwamme III., pi. 5, figs. 29-31. 1834.

6. Verpa bohemica (KROMBH.) SCHROT. "Schles. Krypt 32: 25.
1893. (Plate //., figs. 7, <?, p; Plate IV., figs. 17-23.)
Stipe cylindrical, stuffed or hollow, floccose, white, even, not

wrinkled, up to 4 in. high by i in. across ; pileus conical obtuse,
reticulated or longitudinally ribbed, edge sometimes inflexed so
as to form a white border, brownish above and white beneath,
up to 1.25 in. deep; spores elliptical, obtuse, straight, or bent,
one-celled, hyaline, only two in an ascus, up to 60 mic. long by
16 mic. wide; paraphyses filiform, branched, septate, up to 4
mic. wide at the tip.

On damp ground, often among broken limestone.

* Hennepin, May 1899, Freeman 302 ; Hennepin, April 1901,
Lyon ; Hennepin, April 1903, Butters and Ramsey; Ramsey,
May 1903, Policy.

Verpa bohemica is easily distinguished by the two large
spores in each ascus.

The specimens agree with Rathay Fl. Exsicc. Austro-Hung.
No. 1573, of which the spores measure 60 x 14 mic. ; Thuemen
Mycoth. Univ. 609, of which the spores measure 60-80 mic. x
16-18 mic.

Krombholz Schwamme III.,^>/. 15, figs. 1-13. 1834.

7. Morchella hybrida (SON.) PERS. Syn. Meth. Fungi. 620.
1801. Plate I., figs. 7, 8; Plate IV., figs. 8-n.

Stipe cylindrical, bulbous at base, tapering toward apex,
whitish, granulose, I— 6 in. high : pileus brownish to tan, coni-
cal, acute, lower half free from stipe, longitudinally pitted and
ribbed, about 1.5 in. deep; spores elliptical, obtuse, smooth,
hyaline, 18-20 mic. long by 10-14 m^c* wide ; paraphyses
slightly clavate, vacuolated, septate, branched, about 12 mic.
wide at the tip.

On ground in shady places, sometimes in gravel and also
along roadsides.

314 MINNESOTA BOTANICAL STUDIES.

* Hennepin, May 1902, Butters 201 ; * Hennepin, May 1903,
Policy ; * Ramsey, May 1903, Freeman ; Hennepin, May 1903,
Lyon and Rosendahl.

Morgan states (Morchellae — The Morels, Journ. Myc., Vol.
VIII., June, 1902) that no paraphyses are present in M. hybrida
or in M. esculenta. M. hybrida as generally accepted however
has paraphyses. They are large and are therefore easily mis-
taken for young asci, but are septate and sometimes branched.
Collections marked with a * contain small forms, one inch high
or less.

The specimens agree with Thuemen Mycoth. Univ. No. 412,
the spores of which measure 20-24 mic. x 14 mic. ; Ellis and
Everhart N. A. Fungi No. 2628, the spores of which measure
20-22 mic. x 12-14 m^c-

Krombholz Schwamme III. 9 pi. 75, figs. 14-21. 1834.
8. Morchella esculenta (L.) PERS. Syn. Fungi. 618. 1801.

(Plate I., figs. 3-9 ; Plate IV., figs. 1-6.)

Solitary or gregarious; stipe cylindrical, hollow, sometimes
bulbous, granulose or glabrous, white, entire, .5 to 2 in. high by
.5 to i in. thick; pileus is very varied from conical to obtuse,
irregularly or longitudinally pitted, olive brown to grayish
brown ; ribs of the pileus are thick and obtuse at the edge ; the
surface being even, pileus is about 2.5 in. long; spores are
elliptical, obtuse, smooth, one large oil drop in center, sometimes
yellowish, 14-22 mic. long by 8-14 mic. wide ; paraphyses
filiform, clavate, septate, branched, in some specimens very
abundant.

On the ground in shady woods. Very common in oak woods.

* Hennepin, May 1891, Sheldon 20; Wright, May 1900,
Freeman 675; Hennepin, May 1901, Polley; Hennepin, May
1903, Rosejidahl ; Wright, May 1903, Polley; *Hennepin,
May 1903, Hone 218; * Hennepin, May 1903, Polley.

Like M. hybrida, M. esculenta is generally accepted as hav-
ing paraphyses. The above cited material certainly possesses
structures which resemble paraphyses in all essentials. They
are septate, often branched, and are smaller than the asci,
growing among the latter in the hymenium. The collections
marked with a * have rather conical caps and the ribs are longi-
tudinal and regular. In all other characteristics they are true
esculenta forms and I have no doubt that these two collections
contain the forms which have been described as M. conica. As

Hone: MINNESOTA HELVELLINE^E. 315

M. esculenta occurs in such varied sizes and forms in all of the
material which I have examined, I consider that if these are M.
conicq, that they are only a form or variety of M. esculenta and
not a distinct species. Plate I. , figs. J-6, show photographs of
both forms.

The specimens agree with Thuemen Fungi Austr. 12, 13,
where the spores measure 14—22 mic. x 6—12 mic (12 contains
paraphyses measuring 10 mic. at tip and branched) ; Rathay
Fl. Exsicc. Austro-Hung. 1572, in which the spores measure
20 X 12 mic. ; D. Saccardo Mycoth, Italica, 507, in which the
spores measure 14—18 x 10—12 mic.

Krombholz Schwamme III., pi. i6,Jigs. j, 4 (Morch. escul.
rotunda Fr.), 5, 6 (Morch. escul. vulgaris F.), pi. ij, fig's.
3-4. (Morch. escul. fulva Fr.), 9-16; pi. 19, figs. 6-j.

9. Morchella crassipes (VENT.) PERS. Syn. Meth. Fungi 620.
1801. (Plate I., figs. /, 2; Plate IV., fig. 7.)

Solitary or gregarious ; stipe cylindrical, hollow, bulbous,
very much furrowed, granulose, white, .5—4.5 in. long by -75—
1.75 in. wide ; pileus conical or subconical, ribs very irregular,
undulating, thick, acute at the edge, pits deep, surface very
ragged and uneven, yellowish, 1.5-3 m- deep ; spores elliptical,
obtuse, smooth, one large central oil drop, 20-22 mic. long by
10-12 mic. wide ; paraphyses broad, as wide as the asci, septate
and vacuolated, 10-18 mic. wide.

On ground in most shady woods or open places.

LeSueur, June 1891, Taylor 49; Ramsey, June 1899, Free-
man 359; Hennepin, May 1899, Buell ; Hennepin, May 1901,
Freeman 1000^ ; * Hennepin, May 1903, Hone 217.

M. crassipes differs from M. esculenta in its greater size, up
to 7.5 in. high, yellowish color and very uneven surface. The
ribs also are very acute at the edge and thick at the base while
in M. esculenta they are even and very obtuse at the edge.

Krombholz Schwamme \\\.,pl. 1 6, figs. 1—2. 1834.

GEOGLOSSACE^E .

10. Spathularia clavata (SCHAEFF.) SACC. Michelia 2: 77.
1880. (Plate II I., fig. i; Plate V.,figs. 13-15, 20.)
Gregarious ; stipe cylindrical slightly compressed, hollow,

erect, single or caespitose, fleshy, yellowish brown, up to 3 in.
long by .5 in. wide; pileus spatulate or broadly clavate, gen-

316 MINNESOTA BOTANICAL STUDIES.

erally obtuse, much compressed, running down the stipe for
some distance on opposite sides, hollow, glabrous, margin un-
dulated, surface wavy or slightly lacunose, yellowish, up to
1.25 in. wide by 1.5 in. long; spores parallel in fascicles, hya-
line, linear-clavate, slightly bent, multiseptate, containing small
oil drops, up to 45 mic. long by 2 mic. wide ; paraphyses fili-
form, septate, branched, tips not thickened but wavy, up to 2
mic. wide, numerous.

On rotten wood among moss.

St. Louis, July 1886, Arthur 194 ; * Cook, Aug. 1903, Free-
man and Ballard 20.

These specimens agree with Thuemen Fungi Austr. 925, in
which the spores measure up to 50 mic. x 2 mic. ; Sydow My-
coth. March 2516, in which the spores measure 45-60 mic. x
2 mic. ; A. Kerner Fl. Exsicc. Austro-Hung. 1874, m which
the spores measure 40-50 mic. ; Ellis N. A. Fungi 1268, in
which the spores measure 40-60 mic. x 2 mic.

Krombholz Schwamme III., -pi. 5>>fig. 22. 1834.

ii. Geoglossum hirsutum PERS. Comm. Schaff. Icon. Fungi
Bav. 37. 1800. (Plate III., fig. 5; Plate V.,figs. 1-4.}
Caespitose, erect, black : stipe cylindrical, solid, even, black,
hirsute, up to 2.25 in. long by i in. wide; pileus club shaped,
compressed or plicate, distinct from stipe, hirsute, up to i in.
long and .75 in. wide; spores linear, slightly curved, septate
into about 16 cells, multiguttulate, brown, obtuse, 100-120
mic. long by 4-7 mic. wide ; paraphyses filiform, septate, curved
or arched at the tip, much enlarged, brownish, 3 mic. wide, tip
4-6 mic. wide; setae rigid, simple, brown or black, intermingled
with the asci, sometimes twice as long as the asci, projecting
beyond.

On moist ground among grass.

* Washington, July and Sept. 1903, Lyon ; Washington,
July 1903, Wheeler.

The specimens agree with Jaczewski, Komarov, Franzchel.
Fungi Rossiae Exsicc. 245, in which the spores measure 100-
135 x 4 mic. ; Sydow Mycoth. March 1069, in which the spores
measure 100-120 x 4 mic.

They agree with the figures and description of Massee,
Monograph of Geoglossaceae, Ann. Bot., Vol. n, PL XIII.,
figs- 78^ 79 5 PI- XII., figs, j/, j/, a, 32. 1897 ; Krombholz
Schwamme III., PL 5, figs. 20-21. 1834.

Hone: MINNESOTA HELVELLINEJE. 317

Geoglossum hirsutum americanum COOKE, Mycogr. 3. fig.

1875-

Hymenium appears glabrous when examined under a hand
lens as the setae project but slightly beyond the asci ; spores
light brown and only 7-10 septate. It agrees with G. hirsutum
in all other characters.

On ground among moss in an alder swamp.
Sherburne, Aug. 1901, Policy.

12. Leptoglossum luteum (PK.) SACC. Syll. Fungi 8 : 48. 1889.
(Plate III., fig. 2; Plate V., figs. 16-19.)

vStipe cylindrical, minutely scaly, whitish, stuffed, even, up
to i in. high; pileus club shaped, slightly compressed, grooved
on one side, yellowish, smooth, up to .5 in. high ; spores oblong,
slightly curved, hyaline, obtuse, 28-38 mic. long by 5-6 mic.
wide (most of the spores were immature but some show distinct
indications of septations) ; paraphyses filiform, clavate, curved
at the tip, branched, numerous, about 2 mic. wide.

On ground in swamp.

* St. Louis, July, 1886, Arthur 195.

The specimens agree with a specimen of unknown origin No.
82 collected at Kirkville, N. Y., June, 1889, the spores of which
measure 26-34 x 4~5 m^c- an<^ each cell often contains one large
oil drop.

They agree with the plates and descriptions of Peck, Rep.
N. Y. St. Mus. Nat. Hist. 24: 94, pi. 3,^*5. 20-24. I^72 anc*
Massee Monograph of Geoglossaceae, Ann. Bot., Vol. u,^>/.
XII., figs. 28, 29, jo. 1897.

13. Leotia lubrica (Scop.) PERS. Syn. Meth. Fungi 613. 1801.
(Plate III., fig. 4; Plate V., figs. 5-8).

Gregarious, caespitose, usually gelatinous especially when
old ; stipe cylindrical or inflated at the base, compressed and
often bent, hollow or pulpy within, pruinose, yellowish to green-
ish, up to 2 in. long by .5 in. wide; pileus irregularly hemi-
spherical, inflated, wavy margin obtuse and folded, yellow to
yellowish olive-green and very dark green, up to i in. across ;
spores elliptical slightly acute, straight or bent, smooth, with sev-
eral large oil drops, when mature 2~4-celled, colorless or slightly
greenish, often in two rows, 4-5 mic. wide by 18-22 mic. long ;
paraphyses filiform, septate, branched, slightly clavate, hyaline
or greenish, up to 2 mic. wide, 4 mic. wide at tip.

3] 8 MINNESOTA BOTANICAL STUDIES.

On ground in woods often among moss. Usually on sandy
soil.

Hennepin, July 1903, C. C. Conser ; Washington, Aug.
1903, Wheeler; Ramsey, Sept. 1903, Freeman 1380.

The collection 1380 Freeman agrees in color and form with
L. cMorocephala. The stipe is green and only slightly lighter
than the cap and the plants are more slender. As many grada-
tions may be found between this and typical L. lubrica forms
so all of the above specimens have been listed under L. lubrica.
The dried material from collection 1380 Freeman shows a yel-
lowish stipe.

The specimens agree with Thuemen Fungi Austr. 517, the
average spores of which measure 20 x 5 mic. ; Sydow Mycoth.
March 278, 667, the average spores of which measure 20 x 4
mic.; Thumen Mycoth. Univ. 1112, the average spores of
which measure 18-20 mic. x 4-5 mic.

They agree with the descriptions and figures of Rehm Krypt.
Fl. Vol. I. ubi.Jfg. 1-4. 1896 and of Massee Monograph of
Geoglossaceae, Ann. Bot., Vol. n, -pi. XIII., Jigs. 61, 65.
1897.
14. Cudonia circinans (PERS.) FR. Summa Veg. Scand. 348.

1849. (Plate III., fg. j; Plate V.,figs. 9-12).

Gregarious, erect, somewhat cagspitose ; stipe fistulose or solid,
even, twisted, expanding upward with the pileus, granulose or
powdery, darker colored than pileus, up to 2 in. long, .25 in.
across ; pileus fleshy, convex and undulated, margin free, invo-
lute, variable in color with age, tan to dingy yellow, sometimes
flesh tinted, up to .5 in. broad; spores linear, multiseptate,
often containing several small oil drops in each cell, hyaline,
curved slightly when free, obtuse, 35-50 mic. long by 2 mic.
wide ; paraphyses filiform, branched, septate, tip not enlarged
but curved, up to 2 mic. wide.

On decayed log in deep balsam fir woods.

* Cook, Aug. 1903, Freeman and Ballard 131.

The specimens agree with Thuemen Mycoth. Univ. 1809, the
spores of which measure 35-45 x 2 mic. ; Ellis and Everhart
N.A. Fungi 3533, the spores of which measure 35-40 x 2 mic. ;
D. Saccardo Mycoth. Italica 873, the spores of which measure
35-45 x 2 mic.

They also agree with the descriptions and figures of Rehm
Raben. Krypt. Fl. Vol. I., 1163,^. 1-4. 1896 and Massee

Hone: MINNESOTA HELVELLINE^E. 319

Monograph of Geoglossaceae, Ann. Bot., Vol. n, -pL XII. ,
figs, /j, 14. 1897.

BIBLIOGRAPHY.

1. Fries. Syst. Myc. II. 1822.

2. Schweinitz. Syn. Fung. Car. Sup. 1822.

3. Krombholz. Schwamme III. 1834.

4. Fries. Sum ma Veg. Scand. 1849.

5. Fuckel. Symb. Myc. 1869.

6. Peck, C. H. Rep. N. Y. State Mus. Nat. Hist. No. 24-68.
1872-95.

7. Cooke. Mycographia. 1879.

8. Saccardo. Michelia, 2. 1880.

9. Saccardo. Conspectus genera Discomycetum hucusque cogni-
torum. Bot. Centralb. 18 : 213-220, 247-256. 1884.

10. Saccardo. SyllogeFungorum, 8: 7-53; 10 : 1-3; n : 39i~393«
1887-95.

11. Arthur. Helvellaceae, Report. Bot. Work in Minn, for 1886.
3: 35- 1887.

12. Trelease, W. " Morels and Puff-Balis." Trans. Wise. Acad.
Sci. A. and L. 7: 105. 1888.

13. Phillips, W. British Disco mycetes. 1893.

14. Schrbter, J. Kryptogamen Fl. von Schlesien. 32 : 16-31.
1893.

15. Schroter, J. "• HelvellineaB" in Engler and Prantl, Die natur-
lichen Pflanzen Familien. I2: 162-172. 1894.

16. Massee, G. British Fungus-Flora, 4 : 455-497. 1895.

17. Rehm, H. Rabenhorst's Kryptogamen Flora. Pilze, I3: 1134-
1208. 1896.

1 8. Underwood, L. M. " On the Distribution of N. A. Helvellales."
Minn. Bot. St. I : 483. 1896.

19. Massee, G. A Monograph of the Geoglossacea. Ann. Bot.
II: 225-307. 1897.

20. Burt. List of Vermont Helvelleae with descriptive Notes. Rho-
dora, I: 59-67. pi. 4. 1899.

21. Atkinson, G. u Mushrooms edible and poisonous." 1901.

22. Morgan, A. P. "MorchellaB. The Morels." Journ. Myc. 8:
49. 1902.

33. Morgan, A. P. The Discomycetes of the Miami Valley, Ohio.
Journ. Myc. 8: 179. 1902.

320 MINNESOTA BOTANICAL STUDIES.

EXPLANATION OF PLATES.
All figures of plates are natural size.

PLATE XLVIII.

Figures 1,2. Morchella crassipes.
Figures 3, 4. Morchella esculenta.
Figures 5, 6. Morchella esculenta form conica.
Figures 7, 8. Morchella hybrida.

PLATE XLIX.

Figures i, 2, 3. Helvella infula.

Figures 4, 5, 6. Verpa conica.

Figures 7^ 8, 9. Verpa bohemica.

Figure 10. Helvella crispa.

Figures n, 12, 13. Helvella lacunosa.

Figures 14, 15. Helvella elastica.

Figure 16. See paper by Miss Jessie Policy on Physalacria.

PLATE L.

Figure i. Spathularia clavata.
Figure 2. Leptoglossum luteum.
Figure 3. Cudonia circinans.
Figure 4. Leotia lubrica.
Figure 5. Geoglossum hirsutum.

PLATE LI.

Figures 1-6. Morchella esculenta.

Figure i . Longitudinal section ^ nat. size.

Figure 2. Ascus, x 195.

Figure 3. Ascus with sixteen spores, x 195.

Figure 4. Spores, X 387.

Figure 5. Apex of an open ascus, X 387.

Figure 6, a. Normal mature paraphyses, X 195.

Figure 6, b. Young paraphyses, X 195.

Figure 7. Morchella crassipes^ longitudinal section, ^ nat. size,

Figures 8-n. Morchella hybrida.

Figure 8. Longitudinal section, ^ nat. size.

Figure 9. Ascus, X 195.

Figure 10. Spore, x 387.

Figure n. Paraphyses, X 195.

Figures 12-16. Helvella lacunosa.

Figure 12. Cross section of stipe, ^ nat. size.

Figure 13. Paraphyses, X 195.

Figure 14. Ascus, x 195.

Hone: MINNESOTA HELVELLINE^E.

321

Figure 15. Young ascus, X 195.

Figure 16. Spores, X 387.

Figures 17-23. Verpa bohemica.

Figure 17. Two asci, one emptied of spores, X

Figure 18. Ascus, X 195.

Figure 19. Paraphyses, X 195.

Figure 20. Young paraphyses, X 195.

Figure 21. Longitudinal section, ^ nat. size.

Figure 22. Spores, X 387.

Figure 23. Open ends of asci, x 195.

Figures 24—29. Helvetia infula.

Figure 24. Longitudinal section, yz nat. size.

Figures 25, 26. Paraphyses, X 195.

Figure 27. Spores, X 387.

Figures 28, 29. Asci, X 195.

Figures 30—33. Verpa conica.

Figure 30. Longitudinal section, *^ nat. size.

Figure 31. Ascus, X 195.

Figure 32. Spores, X 387.

Figure 33. Paraphyses, X 195.

PLATE LII.

Figures 1-4. Geoglossum hirsutum.

Figure i. Spore, x 387.

Figure 2. Ascus, X 195.

Figure 3. Hair, X 387.

Figure 4. Paraphyses, X 195.

Figures 5-8. Leotia lubrica.

Figure 5. Longitudinal section, ^ nat. size.

Figure 6. Ascus, x 387.

Figure 7. Spores, X 387.

Figure 8. Paraphyses, X 387.

Figures 9-12. Cudonia circinans.

Figure 9. Longitudinal section, ^ nat. size.

Figure 10. Ascus, X 387.

Figure n. Spores, x 387.

Figure 12. Paraphyses, X 387.

Figures 13-15, 20. Spathularia clavata.

Figure 13. Longitudinal section, ^ nat. size.

Figure 14. Ascus, x 387.

Figure 15. Spores, X 387.

'95

VOL. III.

MINNESOTA"!

UAL STUDIES.

PART III.

7

"TTT

VOL. III.

MINNESOTA

•

•ifc.

.L STUDIES.

PART III.

14

-13

VOL. III. MINNESOTA BOTANICAL STUDIES. PART III.

VOL. III.

CAL STUDIES.

PART III.

VOL. III.

MINNESOTA BOTANICAL STUDIES. PART III

XXVIII. OBSERVATIONS ON PKYSALACRIA
INFLATA (S.) PECK.

JESSIE M. POLLEY.

This rather rare and interesting club fungus was first
described by Schweinitz in 1822, under the name of Leotia
inflata (i). In 1828 Fries described it as Mitrula inflata (2),
and it was again described by Cooke in 1879 as S-pathularia
inflata (3).

The first accurate description however, and the first appre-
ciation of the true position of this fungus, was given by Peck
(ii)in 1882. His description is as follows:

"Physalacria gen. nov.

("From (puaaztz, a bladder and axpa, the top.)

" Club subglobose, inflated, thin, somewhat tenacious, every-
where covered by the hymenium, supported on a distinct,
slender stem.

" Distinguished from Pistillaria by the thin, inflated bladder-
like club and the distinct slender stem. The following is at
present the only species known :

" PHYSALACRIA INFLATA.

"White, becoming tinged with yellow; club subglobose,
submembranaceous, glabrous, flaccid, more or less uneven with
irregular depressions or wrinkles, two to four lines broad ; stem
slender, equal, firm, straight, solid, four to nine lines high, mi-
nutely hairy or subfurfuraceous, mostly cespitose ; spores minute,
narrowly elliptical, colorless, .00016— .0002 of an inch long
and about half as broad.

<TDecaying wood and bark in woods and shaded places.
It occurs especially in mountains or hilly districts in sum-
mer."

Two years later Farlow (8) collected a few specimens in the
White Mountains and identified them with Peck's description.
His account is as follows :

323

324 MINNESOTA BOTANICAL STUDIES.

" PHYSALACRIA INFLATA (S.) Peck.

" On logs in wet places. Shelburne, a small number of
specimens of this curious species were found on a log in a brook
which was nearly dry. My specimens were in fruit and I am
able to confirm the account of the fructification given by Peck
in Bull. Torr. Bot. Club, Jan. 1882. The species does not
belong to the genus Mitrula where it was placed by Schweinitz,
but it is one of the Hymenomycetes, closely related to Pistil-
laria, as correctly shown by Peck. I could find only two spores
to a basidium."

The material from which the present description is written
was collected by E. M. Freeman at Detroit, Minn., in August,
1901. A part of the material used had been preserved in 2^J
per cent, formaline and the rest had been dried. Permanent
slides were prepared by the ordinary paraffin section method
and gentian violet and bismark brown were employed in stain-
ing. The sections from which the greater part of the work was
done were cut with an ether freezing microtome and mounted
in water or in glycerine. All detail drawings were made with
the aid of an Abbe camera lucida.

The plant grows upon decaying wood in shaded places or in
deep woods, though it seems not to need as much moisture as
many of the club-fungi. It grows in clusters and is | to | in.
in height. Stem ^ and club \ inch thick (PI. XLIX., fig. 16).
Many stipes come up together and diverge toward the top. The
plants spread apart by the enlargement of the clubs which fuse
on the sides where they press against each other. In general
appearance they simulate some of the ascomycetes of the
Geoglossacae, e. g., Spathularia and Mttrula, which led to the
assignment of this species under those genera by earlier authors.
They are creamy white in color except the base of the stem
which is black.

Peck's description is very apt. In addition to the characters
pointed out by him, the following are worthy of note. The
stem is tough, at least in formaline material, and the central
portion is more stringy than the peripheral region, this central
portion is composed of two distinct kinds of hyphae, small even
threads with septa far apart (jigs. 7, 12) and larger hyphas which
are composed of flasked-shaped cells set end to end. Both
kinds of hyphae are very rarely branched and lie parallel. In

POLLEY : OBSERVATIONS ON PHYSALACR1A INFLATA. 325

cross section all of the cells appear circular and the largest are
ii /JL in diameter and four or five times as large as the smallest
ones. The latter are 2.5 to 4^ in diameter and are probably
stereome in function, while the former have perhaps to do
mainly with conduction. There are, even in the young stems,
fairly large air spaces between the hyphae. The outer portion
of the stipe consists of a looser weft of septate branching
hyphae which produce large flask-shaped cells, (7-9^ x 22-40
IJL ) pointing outward and giving to the surface a hairy appear-
ance. The stipe is hollow when old.

The club consists of a very thin wall "enclosing a gelatinous
interior {fig's. <?, j). The subhymenial layer is composed of
very fine, septate, closely-woven, much branched hyphae. On
the inside of this layer the hyphae float out in single branching
strands which show abundant clamp-connections {fig- /). The
clamp-connections appear to agree with those described by
Brefeld (9) in Coprinus, rather than with those which Harper
(10) observed in Hypochnus. For in the latter stages walls are
formed at both ends of the clamp cell rather than only at the
end from which the clamp-cell originated. In some cases the
branch which forms the clamp is of considerable length, form-
ing a large loop (Jig. 5). It was also observed that in many
cases the ends of approaching hyphae are fused, forming a simi-
lar figure to that of a large clamp (fig. 9, a). The hymenial
layer shows differentiation into three kinds of cells. The
" paraphyses " which are probably immature basidia, are small
and closely packed (fig. 4). Large flask-shaped cells pro-
longed into blunt spine-like processes project outward. Some
of them do not reach the surface of the cap while others project
beyond (figs, i, 10). They are of the same shape as the hair-
like cells of the stem and seem to serve the usual protective pur-
pose of the cystidia of the lower hymenomycetes. They are
probably also special excretory cells as is indicated in the abun-
dance of calcium oxalate crystals excreted and in the sunken
posjtionof many. The outer ends of these cells are covered by
an irregular cap of crystals (figs. 6, 8). A third distinct type
of cells in the hymenium are the basidia. They are somewhat
thicker than the " paraphyses " and the sterigmata protrude
beyond the surface. Peck and also Farlow described for this
species two-spored basidia. This material shows, however,
that the latter carry from two to four spores. Numerous

326 MINNESOTA BOTANICAL STUDIES.

examples were noted of basidia bearing either three or four
sterigmata. The basidia are barrel-shaped and considerably
larger in diameter than the hyphae from which they arise. The
sterigmata are borne at the summit in the usual manner for
hymenomycetes. The spores are oval in shape and about
1.6 x 2.5 n in size.

Physalacria inflata apparently enjoys a wide distribution but
seems never to occur in great abundance. It has been col-
lected in S. Car. (i), the White Mountains (8), New York (n),
and in Minnesota. As far as I know these are the only recorded
occurrences.

SYSTEMATIC POSITION.

Physalacria has been assigned to the Clavariaceas. Among
the genera of this family Baumanviella (P. Hennings) seems to
be most nearly related, though this genus contains single-spored
basidia. Glceocephala (Massee) differs considerably from Physa-
lacria, not only in one-spored basidia but also in the localiza-
tion of the hymenium on the lower surface of the more or less
flattened club and in the prominence of its cystidia-like hairs on
the upper surface. Pistillaria and Typhula, on the other hand,
approach in form and texture the true Clavaria types. In
certain aspects Physalacria also shows interesting resemblances
to the Trembling fungi, e. g., to such forms &sDittola amongst
the Dacromycetineae. In Physalacria a somewhat gelatinous
region is found below the subhymenium, though this gelatinous
region is not extensively developed. The forked basidium of
Ditiola removes it far from Physalacria, yet the partial gela-
tinization of the club of the latter is one of but a few such
instances among the Clavariacece.

OTHER SPECIES.

Physalacria langloisii E. & E. (4) is a very small species
found in Alabama in 1888. It grows on rotten wood and is less
than i mm. in height, and is distinguished from P. injlata (S)
Peck, by the smaller size and urn-shaped cystidia.

P. orinocencis Pat. (5) is also a small species about 3 to 4
mm. high. Found in northern South America. It grows in
clusters, has an inflated club which appears (from figures in
Nat. Pflanzenfamilian Fungi, i 1 ** : 131. 1898) to be more
or less conical in form and its hymenium covers the whole
surface of the club.

POLLEY : OBSERVATIONS ON PHYSALACKIA INFLATA. 327

P. stillboidea (Cke.) Sacc. (7) was described by Cooke (6)
(under Pistillina stillboidea Cooke) from New Zealand material
on Panax leaves. It is also very small, about 3 mm. in height,
with hollow, globose, depressed club. Cooke (6) also , states
that Pistillina -paradoxa B. & C. (Crmula -paradoxa B. & C.)
must also be referred to the same genus as Pistillina stilboidea
Cooke. Among others he cites as exsiccatas Thiimen Myc.
Univ. No. 208, and Ellis N. A. Fungi No. 23 on living leaves
of ^tiercus. These two specimens have been examined and
prove to be Cronartium asclepiadeum quercinium B. & C.
They agree with Ellis and Everhardt N. A. Fungi No. 1881,
.second series.

The genus Physalacria as at present known therefore, com-
prises four species, two North American, one South American,
and one Australian.

FIGURE ON PLATE XLIX.
Figure 16 Photograph of plant, natural size.

DESCRIPTION OF PLATE LIII.

Figures 2 and 3 are magnified 5.5 all the remaining figures art
magnified 455 times.

Figure i. Detail of section vertical to surface of club.

Figure 2. Diagram of vertical median section through whole plant;
«, hymenium ; b, closely packed hyphae ; <;, interior gelatinous portion ;
d, layer of flask-shaped cells; £, interior portion shown in fig. 7; _/",
hollow central portion of old stem.

Figure 3. Same as fig. 2 except that it is taken at right angles to
flat surface. Lettering same as fig. 2.

Figure 4. Small portion of hymenium showing basidia with spores
and also sterile cells between.

Figure 5. Clamp connections of loose inner hyphae. Stages in
formation.

Figure 6. Detail of flask-shaped cells on surface of stipe.

Figure 7. Cross section of inner portion of stipe, 0, intercellular
space; b, large cells; c, smaller cells.

Figure 8. Cystidial cell of hymenium.

Figure 9. Peculiar hyphal septations ; <z, anastomosing of two ap-
proaching hyphal ends.

Figure 10. Cross section of outer portion stipe: a, flask-shaped
cells ; b, hyphae beneath the surface.

Figure IT. Longitudinal section of portion shown in fig. 10.

Figure 12. Detail of large cells in stipe.

328 MINNESOTA BOTANICAL STUDIES.

BIBLIOGRAPHY.

1. Schweinitz. Syn. Carol. No. 1120. 1822.

2. Fries. El. Fung. I : 234. 1828.

3. Cooke. M. C. Mycographia 344. 1879.

4. Ellis and Everhardt. Journ. Myc. 4: 73. 1888.

5. Patouillard. Bull. Soc. Myc. 41 t XIII., fig. 2.

6. Cooke. Grev. 19: p. 2. 1890.

7. Saccardo. Sylloge Fungorum, 9: 2^6. 1891.

8. Farlow, W. A. Appalachia, 3 : 232-251. 1884.

9. Brefeld. Botanische Untersuchungen aus der Gesammtgebiet,

etc., 3. 1877.

10. Harper, R. A. Botanical Gazette, 33: 1-25. 1902.

11. Peck. Bull. Torr. Bot. Club, 9: 2. 1882.

12. Hennings. Engler and Prantl, Natiirlichen Pflanzenfamilian, i-

i2: 130. 1898.

VOL. III. MINNESOTA BOTANICAL STUDIES.

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XXIX. SYMBIOSIS IN THE GENUS LOLIUM.

E. M. FREEMAN.

The existence of fungus hyphse in Lolium tcmulentum, L.
•perenne and L. italicum furnishes interesting relationships be-
tween host and parasite. Nothing has as yet been done with
the two latter species but the life-cycle of the Lolium temulen-
tum fungus is perhaps completely known. There is however
no reason for supposing that they differ at all from L. temulen-
tum in their life-histories. It cannot be affirmed without reser-
vation that the entire life-history of L. temulentum is understood,
but it can be affirmed that the yearly life-cycle is known, and
that the parasite can live on indefinitely, infecting generation
after generation of Lolium plants without spore intervention.

The facts of the life-history are briefly as follows : certain
grains of L. temulenlum, varying in number in ordinary com-
mercial mixtures of the grains, from 85 per cent, to 98 per cent,
of the total number, show a layer of hyphae in the hyaline
layer just exterior to the aleurone cells. Occasionally these
hyphae penetrate into the endosperm, between the aleurone and
starch cells, but never enter them. The fungus does not appar-
ently thrive in the endosperm. On the convex side of the grain
the hyphae extend almost or quite to the tip of the scutellum but
never enter the scutellum from this point. Along the grain
groove the hyphae are missing except at the very base where an
infection patch exists, from which hyphae can be found penetrat-
ing into the base of the scutellum of the embryo and from here
to the growing point of the plumule, where a considerable patch
of mycelium is developed and remains dormant until the grain
germinates. On germination the hyphae keep pace in their
growth with that of the growing point and can be found here
throughout the remaining life of the plant. The hyphae develop
in all of the branches and also in the leaf-bases. The appear-
ance in the latter is explained in the similar chemotactic proper-
ties of the basal leaf-meristem to those of the stem growing-
point. In the young ovaries the hyphae permeate the nucellus
and here develop luxuriantly. They are pushed back by the

329

330 MINNESOTA BOTANICAL STUDIES.

elongation of the embryo-sac and at the time of fecundation of
the egg the hyphae along the funicular region have ceased
growth, thus causing the isolation of a patch of hyphae between
the point of the attachment of the ovule and the micropyle.
This later becomes an infection-layer, for from this patch arise
those hyphae which penetrate the embryo. The hyphae do not
enter the embryo until the latter shows the rudiments of the
scutellum and growing point. As soon as the latter is estab-
lished hyphae grow from the infection-layer directly to the
growing point and by the time the grain is mature have built
up a patch of mycelium. Thus the mycelium passes from gen-
eration to generation of the host by direct infection and is appar-
ently able to dispense with spore formation.

Since these facts were established I have attempted in various
ways, by altering the conditions of growth of the fungus to in-
duce it to form spores but all efforts have as yet failed. The
most obviously probable way of succeeding in this attempt
seemed to be the following. If the growth of the young embryo
were prevented the fungus might establish an extraordinarily
vigorous mycelium and in the absence of the embryo might
revert to the spore- forming habit, for it would seem probable that
the ancestral form might have formed its spores or a sclerotium
in the ovary. In one respect expectations were realized, for a
vigorous mycelium in the nucellus was produced. In fact the
hyphae were so densely compacted as to constitute a sclerotium-
like body ; but this was very small. Whether or not the fungus
inhibited the production of the normal amount of nucellar tissue
was not to be determined, but could now be ascertained since
plants with and plants without the fungus can be raised at will.
However the fungus refused to form spores. Two explanations
might be offered for this failure : (i) the habit of mycelial in-
fection may be so well established that ability to form spores
has been lost entirely ; (2) the fungus may be some ergot-form-
ing parasite or one which forms spores in some other organ
of the host plant. Analogy with other forms however indicate
the greater probability of the first proposition.

As the development of some parasitic fungi seems to be
favored by weakness in the host plants, L. temulentum plants
with the fungus were partially starved by retention in dark
chambers for certain periods. It was hoped that the consequent
etiolation would favor the parasite at the expense of the host.

Freeman: SYMBIOSIS IN THE GENUS LOLIUM. 331

This experiment also failed and a consideration of the nature
of the fungus might lead one to predict this result. Very
closely adapted parasites thrive best on healthy plants, so that
during the vegetative period conditions favorable to the host
would favor the parasite also. Whatever the true nature of the
Lolium fungus or its ancestors may be, there is no doubt that
it is now a very highly specialized parasite. Consequently the
weakening of the host did not favor a tendency toward spore
formation, in the predominance of parasite over host. These
experiments therefore only strengthen the supposition that the
spore forming power of the fungus has disappeared entirely.
Further cultural investigations have confirmed the result of
previous anatomical research on the life-history of the fungus.
Twelve plants of L. temulentum with the fungus were planted
in pots of three each at the Cambridge Botanical Gardens in
Cambridge, Eng., and six were left in the open while six were
placed under glass bell jars during the flowering season. This
precaution was taken to guard against the possibility of the
transferrence of fungus spores with the pollen. It should be
mentioned here however that no fungus hyphag have been found
in any part of the stamens except at the base as in other leaves.
Twelve plants without the fungus were placed under similar
conditions. The entire crops of these plants were sent to me
in the fall of the same year. The number of grains received
in each case was as follows :

From plants with the fungus in the open 3?59^

From plants without the fungus in the open 222

From plants with the fungus covered at flowering 1*071

From plants without the fungus covered at flowering 824

Out of these 100 of each were examined to determine the
presence of the fungus. In every case 100 per cent, came
true to the parent plant. Of another lot of with-fungus plants
100 grains were examined and again 100 per cent, were found
infected. This should establish beyond a reasonable doubt
both the accuracy of this method of infection and the existence
of two- races of L. temulentum one with and the other without
a fungus and further supports my previous researches on the
modus operandi of infection.

It is a very noticeable fact that the crops of these Cambridge
plants show a considerable difference in the number of grains
produced by the two races and that the plants with the fungus

332 MINNESOTA BOTANICAL STUDIES.

seem to be the more vigorous of the two. From the seeds of
this crop I planted this last summer an approximately equal
number of with- and without-fungus grains under conditions as
nearly alike as possible. The number of grains obtained was
not determined but the experiment was undertaken merely to
observe the growing plants.

Both races seem to thrive well but the with-fungus plants were
without doubt on the average more vigorous. All observers
who have worked with L. temulentum agree that the fungus
exercises no noticeably injurious effect upon the host. The
above comparisons indicate that not only is this true but that we
have here experimental evidence of a case of true symbiosis —
a symbiosis differing in many respects from mycorrhizal symbi-
osis and the symbiosis of the lichens. This condition is not so
remarkable when we consider the well-known cases of those
grass smuts where the presence of the fungus is not betrayed
until spore formation. In fact from the analogy of stimulation
of many rusts as well as smuts in hypertrofication, etc., stimu-
lation of the Lolium plant would almost be expected. The
later destructive action in spore formation is here unnecessary
on account of the new device for infection. One may therefore
consider that this symbiosis has arisen through a previous con-
dition of destructive parasitism.

The nature of the fungus still remains an open question. 1
have previously enumerated the objections to the assignment of
this fungus to the ergot-forming parasites and it certainly has
little or no resemblances to the Uredineae. Nor has it any simi-
larity to the Hyphomycetes and Pyrenomycetes of molded
grains. The Ustilagineae seem to furnish the closest affinities.
The growth of the hyphae in the growing point and the infec-
tion of the nucellus are quite similar to certain smuts. The
hyphae seem to be intercellular and in no case was I able to
demonstrate the penetration of the cell walls either by the ordi-
nary hyphae or haustoria. In this respect and in the abundance
of septations in the nucellar hyphag the fungus seems to differ
from smuts. In regard to the latter point the fungus seems at
this stage to have departed from the normal ustilagine methods
and the septations may perhaps be regarded as relics of those
septations preceding the chlamydospore formation. It is well
known that oat smut infects seedlings. And it would not be
unnatural to expect a still earlier infection. But if this were

Freeman: SYMBIOSIS IN THE GENUS LOLIUM. 333

pushed further forward it must necessarily take place in the
intraseminal life of the host. Granting this to be the case, one
would expect to find the infection finally taking place as soon
as the stem growing point is established, for not until then are
the chemotactic substances present to attract the hyphge. And
this is what actually occurs.

An analogous driving forward of the period of infection has
probably taken place among those parasites attacking the host
plant in the extraseminal seedling stages. It seems not improba-
ble that the oat smut may have formerly been able to infect any
immature tissues of the host as the corn smut does. The ovary
is a particularly favorable organ both on account of nutrition
and distribution facilities and hence, the preference for the
ovary as a spore-bearing region for the parasite. The estab-
lishment of an attraction by the growing point of the stem and
the localization of the sporiferous region in the ovary would
naturally be followed by an earlier infection since the presence
of the fungus in the unbranched stem multiplies the results of
the infection when the stools are mature. The proximity of
spores to the grains in sowing becomes obviously advantageous
and hence the infection would be pushed forward to the earliest
stages of the extraseminal development of the host, as in the
oat smut. If now, one presupposes a chemotactic attraction of
the growing point for the hyphae of a smut and also at the same
time a failure of the fungus to destroy the endosperm and young
embryo during spore formation, then nucellar hyphae crowded
back by the endosperm and embryo, might easily effect direct
infection of the growing point of the young embryo. This
period would then be shoved forward to the formation of the
earliest rudiments of the growing point. It seems probable that
the fungus is therefore an ustilagine which has brought forward
its infection period to the intraseminal stages of the host and
more particularly to the first appearance of the growing point,
and has lost the power of spore formation.

Apparently infection of a without-fungus plant of L. temulen-
turn is^impossible as is also the eliminating of the fungus from
the with-fungus plants, barring the possibility of failure to in-
fect all ovaries in an inflorescence, which case seems highly
improbable. There exists therefore not only two races of L.
temutentum, but also of L. -perenne and of L. linicola and
probably of other species of Lolium, in each case one with, and
the other without, the fungus symbiont.

334

MINNESOTA BOTANICAL STUDIES.

The question now naturally arises whether the symbiosis was
established previous to or subsequent upon the origin of these
species. In other words has the symbiosis arisen once or three
times ? The simpler explanation seems at first to be that the
symbiosis was previous. This would however presuppose the
development in each of the three species of two parallel lines
one with and one without the fungus. On the other hand a
subsequent establishment of the symbiosis would require like
results for the same fungus or for several closely related fungi
attacking three species of Lolium. A fuller knowledge of the
inter-relations between these species as well as of the remaining
species of Lolium is necessary before an intelligent opinion on
this point can be framed.

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Minnesota botanical
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Call Number:

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221037