AMERICAN Volume 89 = a N aes JOURNAL ; QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY SPECIAL ISSUE The Cultivated Species of the Fern Genus Dryopteris in the United States Barbara Joe Hoshizaki and Kenneth A. Wilson Index to Taxa 99 The American Fern Society Council for 1999 DIANA B. STEIN, Dept. of Biological Sciences, Mount Holyoke College, South Hadley, MA 01075-6418 President BARBARA JOE HOSHIZAKI, 557 N. Westmoreland Ave., Los Angeles, CA oe ice- President W. CARL TAYLOR, 800 W. Wells St., Milwaukee Public Museum, Milwaukee, WI Biel, peas retary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916-1 : 10. Treasurer DAVID B. LELLINGER, 326 West St. NW., Vienna, VA 22180-4151. Membership Secretary JAMES D. MONTGOMERY, Ecology III, R.D. 1, Box 1795, Berwick, PA 18603-9801. Back Issues Curator GEORGE YATSKIEVYCH, Missouri Botanical Garden, PO. Box 299, St. Louis, MO 63166-0299. Journal Editor DAVID B. LELLINGER, U.S. National Herbarium MRC-166, Smithsonian scoteani Washington, DC 20560-0166. Memoir Editor CINDY JOHNSON-GROH, it of Biology, seed Adolphus College, 800 W. College Ave., St. Peter, MN 56082-1498. Bulletin Editor American Fern Journal EDITOR GEORGE YATSKIEVYCH Missouri Botanical Garden, PO. B 99, St. Louis, MO 63166- 0299 ph. (314) 577-9522, e-mail: S aakicich@bkaace mobot.org ASSOCIATE EDITORS GERALD J. GASTONY.......... ee of Spare Indiana agree Bloomington, IN 47405-6801 CHRISTOPHER H. viegees FLER ....Dept. of Botany, University of Kansas, Lawrence, KS 66045-2106 ROBBIN C. MORA en York Botanical Garden, Bronx, NY 10458-5126 JAMES H. PECK Dept. of Biology, University of Arkansas—Little Rock, 2801 S. University Ave., Little Rock, AR 72204 The “American Fern Journal” (ISSN 0002-8444) is an illustrat ed quarterly devoted to the general study of ferns. It is owned by the American Fern Society, and published at oe West St. NW., Vienna, VA 22180-4151. 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FIDDLEHEAD FORUM his editor of the Bulletin of the American Fern Society welcomes contributions from members and bers, including miscellaneous notes, offers to ag or purchase materials, personalia, hocticatferal notes, and reviews of non-technical books on ferns. SPORE EXCHANGE Mr. Roger Boyles, 220 Blakes Dr., Rican g NC 27312-8146, is Director. Spores exchanged and lists of available spores sent on reque GIFTS AND BEQUESTS Gifts and bequests to the Society enable it to expand it i b . Back issues of the Journal and cash or other gifts are src welcomed and are tec diodes fates should be addressed to the Secretary. nteresc American Fern Journal 89(1):1—98 (1999) The Cultivated Species of the Fern Genus Mildred Mathias Botanical Garden, University of California, Los Angeles, Los Angeles, CA 90024 Natural History Museum of Los Angeles County, Los Angeles, CA 90007 Dryopteris in the United States BARBARA JOE HOSHIZAKI KENNETH A. WILSON?! I-SOUR! BOTANICAL WAY 0.4 1999 GARDEN LIBRARY ABSTRACT.—Fifty species of Dryopteris, belonging to three subgenera and ten sections, are known to be in cultivation of the United States. Descriptions, cultural requirements and keys to the sections and the species are provided as an aid to the identification of these species. An addendum lists recently reported species not included in the main treatment. The species treated are: 1) D. 2) D. 3) D. 4) D. 5) D. a 6) D. 7) 22, 8) D. 9) D. 10) D. 11) D. cels 12) D. 137-07, 14) D. 15) 22. 16) D. 24) D. sieboldii cycadina kuratae scottii nis crassirhizoma polylepis pseudo-filix-mas wallichiana a clintoniana cristata ades . sichotensis ota arguta juxtaposita ‘a 25) D. lacer 26) D. marginalis 27) D. mindshelkensis 28) D. stewartii 29) D. sublacera urens 33) D. campyloptera 34) D. carthusiana 35) D. dilatata 36) D. expansa 37) D. intermedia 41) D. erythrosora 50) D. varia The difficulties in understanding Dryopteris, particularly its many species well known to pteridologists. Work continues on the genus, and some new species and hybrids have yet to be delineated and older ones reassessed. The large number of species and the few definitive characters, often a matter of degree and normally based on the dissection of mature fronds, are problems enough without the addition of the inherent variability of the plants. complexes, are ‘ Author for correspondence. Current address: P.O. Box 39512, Los Angeles, CA 90039-0512. 2 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) The fronds tend to vary greatly on the same plant, and the presence of hybrids makes identification particularly troublesome. Cultivated plants compound the problem by the absence of data on their place of origin and their tendency to appear different or underdeveloped under various cultural conditions. As in most ferns, the architecture of the fronds changes as a plant matures. Ju- venile plants tend to have fuller, often broader fronds with closer parts that are less divided. For instance, plants that have bipinnate fronds when mature will, in their juvenile stage, have fronds that are only once pinnate-pinnatifid. The color of the scales also darkens as the plant reaches maturity and in ma- ture plants may be darker at the end of the season than when the fronds are young. Growing conditions, such as humidity, water and nutrient availability, influence the form of the frond. For instance, under suboptimal conditions, the apex of the pinnae can become blunter rather than acuminate; the margins entire rather than crenate or serrate, or mucronate rather than spinulose. With these numerous difficulties in mind, we hope that readers will be patient with our efforts in sorting out the cultivated species of Dryopteris. The primary purpose of this paper is to provide a means of identifying the species of Dryopteris currently in cultivation in the United States. The em- phasis of this work is on species, not infraspecific categories. Though some infraspecific categories (also particularly cultivars) are mentioned, no attempt was made to include all that are known in cultivation. Dryopteris has ca. 225 species and is nearly cosmopolitan. The species grow in wet, shaded forests, open grassy areas, or on rocks and along streams, pri- marily in mountains. The greatest number of species are found in southern, southeastern, and eastern Asia. The genus has been subdivided into 4 subgenera and 17 sections by Fraser- Jenkins (1986). Three of these subgenera and 10 of the sections are represented in cultivation in the United States. We have adopted his taxonomy, but it is beyond the scope of the present study to evaluate this classification. We have prepared a key to the Fraser-Jenkins sections known in cultivation, bearing in mind that many of the characters overlap each other. Fraser-Jenkins (1989) himself wrote that, “Each [section] contains a number of species that vary (in any parameter) so as to make even a general description of the section inap- plicable in many instances though the species form natural assemblages which are separate from one another.” Keys to species are provided in the treament of the sections. Much of the complexity found in the genus is the result of hybridization. Hybrids and apogamous forms are frequent, although some of the species are fertile, sexual, diploids. The basic chromosome number for the genus is X = 41, thus sporophytes of a diploid species has two sets of 41 bivalents, or 82 total chromosomes. Other species are fertile sexual tetraploids with four sets of chromosomes, and a few are fertile sexual hexaploids. However, many of the species reproduce apogamously, rather than sexually. In the species ac- counts, we have taken information on chromosome numbers and sexual vs. apogamous life cycles from various literature accounts (especially Gibby, 1985) HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 3 that we have neither cited nor personally verified, and some cultivated acces- sions may differ from the usual chromosomal condition for a given taxon. Sterile plants of various ploidies are produced when fertile species hybrid- ize, and hybrids between apogamous and fertile species are possible (game- tophytes of apomicts often produce viable sperm cells and can thus act as male parents). We have not detailed sterile hybrids, but growers need to be aware that such hybrids are at times found in cultivation. Sterile plants may be iden- tified by the presence of aborted spores. Because they generally do not repro- duce sexually, hybrids have either been transplanted from the wild or propa- gated by the subdivision of other plants. A concise illustrated treatment of the North American species of Dryopteris was published by Montgomery and Paul- ton (1981). The North American hybrids of Dryopteris were treated by Mont- gomery (1982). These publications illustrate the interrelationships of species and demonstrate the difficulties in deciphering the complexities in the group. For the proper identification of species it is important to have an entire fertile frond from a mature, well developed plant. This should include the entire stipe (petiole) together with its scales and the scales at the very base of the petiole as well as those from the rhizome. The characters important for the identification of the species include: the size and arrangement of the fronds; the relative length and color of the petiole; the density, shape, and color of the scales, hairs or glands; the size, color, texture, and overall shape of the blade and the pattern of its dissection; the shape and pattern of dissection of the pinnae, pinnules, and ultimate segments, particularly of the lowermost pinnae and the lowermost pinnule next to the rachis, and the nature of the margins; the position of the sori on the pinnae; the presence or absence of an indusium: and, when present, the nature of the indusium itself. Of these, the two crucial characteristics are the shape and cutting of the ultimate segments and the color and shape of the stipe-base scales. In general, in this account the description of the dissection of the blade is that exhibited by the lowermost (basal) pinnae; it is common to find that the degree of dissection of the pinnae decreases the closer they are to the tip of the blade. We believe that the illustrations of the species provide the best tools for initial identification. The preliminary deter- mination should be confirmed by checking the specimen against the written description. Even in listing the species, it is not possible to treat each one fully. The search for new garden ferns is an active pursuit, and new species are continually being introduced, making it impossible for us to include all of them. More cultural information may be found in horticultural books. Most Dryop- teris species are terrestrial and adapt to garden soils with a generous amount of humus. Most grow well in moist soil. Even those species known for prefer- ring wet areas will adapt to growing in moist, better drained soil. Most species are more luxuriant with ample humidity. In more arid climates, periodic mist- ing during periods of low humidity will produce more handsome foliage. Some species seem to grow best in acidic soil or basic soil, but most grow in the neutral or slightly acidic range. Partial, but not deep, shade suits most species of Dryopteris. o AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Many temperate deciduous species do not grow well in warmer climates such as southern California, whereas most temperate evergreen species (mostly those in the subgenus Erythrovariae) do well in these climates. The limiting factor is often the temperature tolerances of the species, particularly the min- imum. Where information on temperature is given as the January average, this number represents the temperature mean for that month and is taken from isotherm maps from the area where the fern is native. Please note that this number is an approximation and it does not represent the minimum temper- ature for each species. The minimum temperature tolerances of the more re- cently introduced species are still being gathered, but such information ma be found in the publications of various fern societies and horticultural litera- ture. Remember that even though a plant may survive its minimum tempera- ture in winter, it may be vulnerable when subject to the same or slightly higher temperature in the spring, particularly if new growth has emerged. Data on the tolerance of temperate climate ferns to subtropical and tropical conditions is also incomplete and is affected by many variables. DryopTeris Adans.—Shield fern, buckler fern, wood fern. Plants terrestrial. Rhizome thick, suberect or erect, less commonly creeping, surrounded by close, spirally arranged leaves and old leaf bases; rhizome scales nonclathrate. Leaves usually in a rosette; stipe grooved, scaly, with 3— 7 (9-10) vascular bundles arranged in a C-shaped pattern; blades 1—4 times pinnately compound, bearing scales but lacking needlelike hairs, the pinna midribs grooved. Sori round, dorsal; indusium kidney-shaped, attached at a sinus, rarely absent. This genus of ca. 225 species is cosmopolitan, occurring mostly in temperate forests and montane areas of the tropics. The species are difficult to identify simply because there are so many of them, and there are many similar groups of species. Furthermore, the fronds can vary even on the same plant. Identi- fication requires careful examination of large, mature leaves. About 50 species are in cultivation in the United States, but new species are constantly being added to the trade and older ones are disappearing. The species of Dryopteris pose no special problems in cultivation, except that some of the species native to colder climates do not adapt well to warm- climate gardens and some species thrive only in acidic soils. Most species are easy to propagate from spores or divisions. Offshoots come from the base of erect rhizomes, semi-erect rhizomes or branches of short-creeping rhizomes. Generally, species that are deciduous in cold temperate climates tend to be more evergreen in warmer climates. Fronds that become deciduous wither in place but may or may not promptly lose their green color. The groups used in this treatment are the subgenera and sections of Dryop- teris recognized by Fraser-Jenkins (1986). KEY TO THE SUBGENERA AND SECTIONS OF DRYOPTERIS This key is provided as a rough guide to the sections of the genus. We rec- ognize that many characters of the sections overlap each other and that place- HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 5 ment in a section may be difficult. We have found that the illustrations provide one the most rapid and reliable guides to identification of specimens, followed by a confirmation against the written description. Please consult the introduc- tion for some guidelines on identification. 1, Fronds pinnate with the terminal segment like the lateral ones .... Subgenus 1. Pycnopteris 1. Fronds pinnate or more divided, the apex divided 2. Fronds with very small bullate or bullate-based scales on the underside of the rachis, costae or-costules 23 a ee a ee ey ee (Subgenus 3. Erythrovariae) 3. Basiscopic pinnule on the lowest pinnae shorter than the adjacent pinnules (sometimes equal or slightly longer in D. cystolepidota) ............ Section 3.1. Erythrovariae 3. Basiscopic pinnule of lowest pinnae noticeably longer than the adjacent pinnules .. SRG eg ins a 8 4 ow See es de VG ee sdk ection 3.2. Varia 2. Fronds without bullate or bullate based-scales on the underside of the rachis, costae, or COM Ss ss Hee eee) 6 oa (Subgenus 2. Dryopteris) 4. Fronds once pinnate, the pinnae only We eles amy or lobed only half way or less to the costae, rarely cut to the costae at the pinnae base ....... Section 2.1. Hirtipedes 4. ne fully pinnate-pinnatifid to 3(4)- arora . Fronds mostly 3-pinnate 6. Stipe scales narrow-lanceolate, matt and concolorous; blade triangular-ovate; stipe dark purple-brown at base (D. aemula)............... Section 2.7, Aemulae 6. — otal ovate- a mostly glossy omg usually bicolorous; blade broader, entangular, triangular or aa atti SE gle Section 2.8. Lophodium 5. Pee mostly pinnate-pinnati sed to 2-pinn. . Fronds mostly pinnate-pinnatifid ‘alien ade of some pinnae may be pinnate . . baie cea i ee See eee a esa ails ks) wvera ars ection 2.3. Pandae 7. Fronds nearly 2-pinnate to fully 2-pinnate 8. Fronds thin-coriaceous, dark green, wis above, the seman or pinnules reg- ularly rectangular, the margins not toothed ........ Section 2.2. Fibrillosae 8. Fronds herbaceous, not dark green, ak segments or satis not regularly rectangular, the sides mostly tapering, the margins entire, lobed or toothed 9. Pinnules stalked or with narrow base in basal half of the pinnae, the segments or pinnules entire, lobed or with short acute teeth . . . Section 2.6. Pallidae . Pinnules not stalked nor narrowed at base in basal half of the pinnae; sides and apex of the segments or pinnules with long acute teeth 10. Frond linear-lanceolate to lanceolate (broader in D. eer scales mostly lanceolate to ovate-lanceolate........ Section 2.4. Dryopteris 10. Frond narrowly triangular-lanceolate; scales riangulrlanceoate ere brown, their bases often dark (D. remota) ..... Section 2.5. Rem o Subgenus 1. Pycnopteris (T. Moore) Ching Blade firm-textured, pinnate, the terminal segment resembling the lateral ones. 1. Dryopteris sieboldii (Van Houtte ex Mett.) Kuntze (Rev. Gen. Pl. 2:813. 189 ig. 1 Rhizome erect, more or less stout. Stipe ca. 40 (30-60) cm long, densely scaly at the base, sparsely so above, the scales narrow triangular to ovate tri- angular, subentire to sparsely fimbriate or distantly dentate, dark brown, the rachis sparingly fibrillose-scaly; blade pinnate (young fronds often simple and 6 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) if fi 1 Abb : Fic. 1. Dryopteris sieboldii. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. cordate) with the apex ending in a terminal pinna like the lateral pinnae, Ca. 50 cm long, 37 cm wide, pinna pairs mostly 3 (1-5), pale green beneath, co- riaceous-chartaceous; pinnae broadly linear-lanceolate, 18-30 cm long, 3.5-6 cm wide, base slightly cordate to rounded, sometimes oblique, margins slightly serrate or crenate, sometimes shallowly lobed with the lobes serrate. Sori large, in 2-3 series next to the costa, mostly absent from the marginal and submar- ginal area; indusia large, entire, thin. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 7 Dryopteris sieboldii is native to eastern Asia, where it is common in wooded ravines. This tetraploid, sexual species is distinct in having an apical pinna similar to the lateral ones and with few large lateral pinnae. Although slow growing in cultivation, plants may reach 40 cm or more in height. The spread- ing fronds are coarse and few but tend to form a clump and could be used as an accent in the landscape. Slightly undulating and lobed margins tend to develop in cultivated plants. The plant is hardy to a winter minimum average of ca. 35°F. Semi-deciduous, nearly evergreen in southern California. Subgenus 2. Dryopteris Blade variously dissected, the pinnae gradually reduced to a pinnatifid apex; scales not bullate. Section 2.1. Hirtipedes Fraser-Jenk. Fronds pinnate to pinnate-pinnatifid, if pinnate pinnatifid then usually cut half way or less to the costa (except sometimes more deeply cut on the prox- imal pinnae at their bases). KEY TO SPECIES OF SECTION HIRTIPEDES 3, Indusia ahaa se sos as oe ce os ee 4. D. scottii 1. Indusia present 2. Pinna margins cut mostly less than % way to the costa, the stipe scales dense and shaggy, the sinuses narrow between shallow lobes ..................-2-4:-- 2. D. cycadina 2. Pinna margins cut half way to the costa, the stip ] t pi lyd d shaggy, the sinuses V-shaped between spreading lobes...................... 3. D. kuratae 2. Dryopteris cycadina (Franch. & Sav.) C. Chr. (Index Filicum 260. 1905).— Shaggy wood fern, black wood fern.—Fig. 2. D. hirtipes (Bl.) Kuntze, misapplied D. atrata (Kunze) Ching, misapplied Rhizome erect, stout, infrequently producing offshoots. Stipe stout, to ca. 30—40 cm long, very scaly at the base, less so above, the scales narrowly tri- angular, 10-15 mm long, dark brown to black, margins sparsely slender toothed, apex attenuate, the rachis scales smaller and narrower, some fibril- lose-scaly; blade pinnate, oblong-lanceolate, 50-70 cm long, 20-35 cm wide, pinnae pairs ca. 30, texture thin-leathery; pinnae narrow ovate to long narrow triangular, the base more or less truncate-cordate, sessile or short-petiolate, the margins coarsely serrate to crenate or lobed ca. 4—% way to the costa, the serrations broad and often ending in 1 or 2 small teeth, the proximal pinnae sometimes deeply pinnatifid at their base. Sori 2-6 per segment, absent from the marginal and submarginal area; indusia large, entire, persistent. Basal pin- na pair tending to angle forward and often downward from the adaxial surface of blade; pinnae (proximal) often falcate. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) a DY ) ‘ Fic. 2. Dryopteris cycadina. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Medial pinna [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 9 CUTTY | | (ana ge #0 90-9 * ain Sok: ele, 44s Seg eem oe <¢ ® Fic. 3. Dryopteris hirtipes. A) Frond base [scale=5cm]. B) Stipe scales [scale=5mm]. C) Medial pinna [scale=5mm]. Dryopteris cycadina may reach 30-100 cm in height but is usually rather small. The somewhat leathery, sometimes crisped fronds spread and form a whorl; longer fronds may arch slightly. The species is hardy to a January av- erage of 35°F; fronds do not wither and brown quickly, but remain green for some time and tend to lie prone on the soil during the winter. The edges and tips of the fronds often abort in more arid climates, otherwise the plant is easily cultivated. Dryopteris cycadina is an apogamous triploid fern native to Japan and east- ern China, where it is abundant on wooded hillsides at ca. 1,400—2,700 m. It may circulate in horticulture as D. hirtipes or D. atrata under which names it was previously known in Japan. All three names have been much confused in the botanical literature as well and the entire group is in need of detailed study. As presently interpreted (Fraser-Jenkins 1989), D. hirtipes is a separate species and D. atrata is one of its subspecies (D. hirtipes ssp. atrata (Kunze) Fraser-Jenk.); it is uncertain that any of these plants is cultivated in the U.S. It is uncertain if D. hirtipes (Blume) Kunze from Southeast Asia is actually in cultivation. A plant rarely in cultivation and very tentatively identified as D. hirtipes (Fig. 3) has been circulating under the misapplied names of D. darjeelingensis Fraser-Jenk. and D. stenolepis (Baker) C. Chr. [D. gamblei (C. Hope) C. Chr.]. True D. hirtipes is described as having fronds to 60 cm long, the stipe ca. half the length of the blade, the blade with up to 25 pairs of pinnae, the pinna margins toothed or lobed, and the sori indusiate. The mar- ginal lobes, varying from shallow to more extended, are usually truncate at 10 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) their apices and often bear an obtuse tooth at the distal corner. In contrast, D. cycadina has a shorter stipe, ca. 30 pinnae pairs with the pinnae more closely placed and narrower, and the pinnae margins truncate-serrate. These marginal serrations are oblique-falcate with the tooth at the distal corner acute and in- curved. The plant in cultivation is hardy in the Seattle, Washington, area, but may not be hardy in colder climates. In southern California gardens it is ev- ergreen, although new growth ceases during the winter and old fronds lay prone on the soil. The tips of unfolding fronds tend to abort in this more arid climate. Young plants are eaten by slugs and snail. Better herbarium specimens and more study are needed to resolve the identity of this cultivated plant. Another species similar to D. cycadina is D. commixta Tagawa, an endemic of Japan. It differs by having broader fronds with an herbaceous texture when dry or thicker when fresh, the pinnae stalked, 1-2.5 cm broad, more deeply incised with broader pinnae cut halfway to the costa, usually with 20 or fewer pinnae-pairs, and indusia variable in size, poorly developed. Dryopteris cy- cadina has, in contrast, narrower fronds with a more leathery texture, the pin- nae sessile, 1-2 cm broad, shallowly incised, usually to ca. 30 pinnae-pairs, and the indusia fairly uniform in size. The identification of the currently cul- tivated material is in doubt. The senior author observed that fronds from sub- mature plants that were received from a grower as D. commixta were indistin- guishable from D. cycadina when the plants were mature. Another garden species similar to D. cycadina is D. namegatae (Sa. Kurata) Sa. Kurata from Japan and China. It is thought to be a hybrid, as it appears intermediate between D. cycadina and D. dickinsii (the latter discussed under D. kuratae). Dryopteris namegatae is distinguished from D. cycadina by the veins and their branches being depressed below the adaxial surface of the blade and the presence of shorter basal pinnae. On our cultivated plants the proximal pinnae may bear on their acroscopic side next to the rachis a round- ish to truncate lobe that may be free nearly to the pinnae midrib. The plant is evergreen, as described above in D. hirtipes, and is hardy along the western coast of the U.S., although the frond tips tend to abort in southern California gardens. 3. Dryopteris kuratae Nakaike ex Hoshiz. & K.A. Wilson, sp. nov.—TyPE: Ja- pan, cultivated in Tokyo, originally from Kagoshima Pref., Mt. Takakuma, Osumi Peninsula, 25 July 1959, S. Kurata s.n. (TNS #146476; photo Na- kaike, New FI. Jap. Pterid. 430. 1992).—Fig. 4. Planta D. pycnopteroidi (Christ) C. Chr. similis, sed paleis stipitis et rachidis brunneis usque atro-brunneis, marginibus pinnarum % ad % ad costam lobatis, apicibus loborum plerumque obtusis et obliquis, sinubus inter lobos pler- umque V-formibus, soris plerumque ad apices loborum absentibus et tantum aliquando juxta costam praesentibus differt. Rhizome erect, producing offshoots. Stipe moderately scaly, the stipe and rachis scales mostly brown to blackish brown, narrow triangular to lanceolate, acuminate, irregularly toothed; blade pinnate, oblanceolate, ca. 50 cm long, 15 HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 11 \Olh {7S Fic. 4. Dryopteris kuratae. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Medial pinna [scale=5mm]. 12 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) cm wide, narrowed at base, pinnae linear-triangular, margins mostly lobed 4— ¥% way to the costa, the apex of the lobe obtuse, oblique, bearing 1-3 teeth on the distal side, the sinuses between the lobes mostly V-shaped, the lobes spreading. Sori generally absent from the apical area of the lobe and only occasionally present next to the costa; indusia small. Dryopteris kuratae apparently is an apogamous diploid fern (Kurata and Nakaike, 1985) native to eastern Asia. Fraser-Jenkins (pers. com.) raises the possibility that D. kuratae may be the same as D. hangchowensis Ching, but this has not been resolved. Until recently, D. kuratae was called D. pycnop- teroides by Japanese botanists, but this is a larger species with fronds up to 110 cm long, the sori more costal, and the rachis more densely scaly with only brown or paler scales. True D. pycnopteroides is from western China, and although reported to be cultivated, the plants examined thus far are D. kuratae. Dryopteris pycnopteroides and D. kuratae have been confused with D. dick- insii (Franch. & Sav.) C. Chr., which lacks sori next to the costa. Although D. dickinsii is cultivated in Australia, it has not been found in U.S. cultivation. This complex is in need of more study. Garden plants reach ca. 60 cm in height with many spreading fronds. This species is hardy to a January average of 30°F and slowly becomes deciduous in southern California, where it grows well. It is easily cultivated. 4. Dryopteris scottii (Bedd.) Ching (Bull. Dept. Biol. Sun Yatsen Univ 6:3. 1933).—Fig. 5. Rhizome erect, more or less stout, producing offshoots. Stipes 25-45 cm long, scales dense at the base, narrow-triangular, black, above the stipe base narrower, shorter and more scattered; blade pinnate, ovate, 25-35 cm long by 15-30 cm wide, apex amply foliaceous, base broad; pinnae narrow-triangular, acuminate, subsessile, base mostly oblique, round or to truncate, 6—11 free pairs, dark green, firm herbaceous, margins crenate-lobed to distally serrate, the lobes or serrations oblique at their apex and usually with 1 or 2 short sharp teeth. Sori 2-6 per segment, submarginal; indusia absent. Dryopteris scottii is a tetraploid, sexual species (Fraser-Jenkins 1989) ranging from India to eastern Asia and Malaysia, where it is common in wet ground in dense forest at ca. 900-2,000 m. The absence of indusia is the best distin- guishing character of D. scottii. The broad ovate blade with relatively few broad pinnae with roundish lobes, lightly scaly rachis, and herbaceous texture are also helpful distinguishing features. Dryopteris scottii grows well outdoors in the Seattle area and probably is hardy to a January average of 50°F. It is semi-deciduous and seems to grow best in humid sites. It is eaten by slugs and snails. Section 2.2. Fibrillosae Ching Like Section Hirtipedes except the pinna lobes cut nearly to the costa, leav- ing them connected by a narrow wing of tissue, fronds more or less linear- HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 13 VANE (al) Fic. 5. Dryopteris scottii. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. 14 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) lanceolate, the pinnae segments or lobes are quite regular, rectangular, their sides parallel and untoothed, their apices truncate or rounded-truncate and bearing short teeth. KEY TO SPECIES OF SECTION FIBRILLOSAE — Blade oblong triangular, the basal pinnae ca. equal to or longer than those above. ......... 66 bgp ee 6 cals wes ance + os as MON, OM cM ge ye cete og 7. D. lepidopoda . Blade ovate to lanceolate or oblanceolate, or elliptic, the basal pinnae shorter than those above 2. Veins of segments simple except for the basiscopic segments next to the rachis ........ ee ee ay Oe Saree oe eee eee 8. D. polylepis 2. Veins of segments forked 3. Black blotch or spot present at base of pinna midrib on abaxial side 4. Basal basiscopic pinnule next to the rachis with sides entire to subentire (rarely elon- gate and to shallowly pinnatifid in ssp. borreri); the black blotch at base of pinna midrib on the abaxial side not fading in dried fronds; scales on abaxial side of pinna midrib mostly tan (concolorous), commonly cultivated species with many cultivars ee ee ee 5. D. affinis 4. Basal basiscopic pinnule next to the rachis slightly elongate, lobed to shallowly pin- natifid in mature plants, black blotch at base of pinna midrib on abaxial side fading away on dried fronds, scales on the abaxial side of the pinna midrib mostly dark brown to black with pale margins, infrequently cultivated species .. 9. D. pseudo-filix-mas 3. Black blotch or spot absent from base of abaxial side of pinna costae 5. Blade lustrous, firm; the segment apex typically truncate with larger teeth at the distal corner, the veins on the abaxial surface of segments conspicuous ........... Apes eee wd ie ene Oe, Se ebm cae Oe 10. D. wallichiana 5. Blade not lustrous, firm-herbaceous, the segment apex typically rounded with the teeth on both sides similar, the veins on the abaxial surface not particularly conspic- me uous. 6. Sori on distal pinnae of frond; basal basiscopic pinnule (segment) next to rachis adnate; lateral margins of segment weakly crenate, abaxial side of pinna midrib with many long tapered triangular scales and hair-like scales, scales shaggy, pale BE Pe oe ck ois es eee ee oe... 6. D. crassirhizoma 6. Sori extending to middle of frond or lower; basal basiscopic pinnule next to rachis sessile; lateral margins of segment notched-serrate, abaxial side of pinna midrib with mostly deltoid to triangular scales, many with an abruptly acuminate apex, scales dark brown to black often with pale margins ................... een ee Wl ey wee diets oes 67 es ek ae 17. D. filix-mas (see section Dryopteris) 5. Dryopteris affinis (Lowe) Fraser-Jenk. (Fern Gaz. 12:56. 1979).—Yellow gold- en-scaled male-fern, scaly male-fern, common golden-scaled male-fern.— 1g. 6. D. abbreviata (DC.) Newman ex Manton, non (Schrad.) Kuntze D. pseudomas (Woll.) Holub & Pouzar Rhizome erect, producing offshoots. Stipe ¥,—Y, the blade length, stipe and rachis densely scaly, scales mostly ovate-lanceolate, usually gold-brown often darker at the base; blade pinnate-pinnatifid or to 2-pinnate at the base next to the rachis, mostly lanceolate, to ca. 100 cm long, 30 cm wide, dark bluish green, new growth yellow-green, leathery; pinna costae on underside next to the rachis usually with a blackish blotch, the costal scales lanceolate; pinnules often lobed or slightly auriculate at the base, lowest basiscopic pinnule next HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 15 . 6. Dryopteris affinis. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Pinnules from Pee pinna [scale=5mm]. 16 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) to the rachis adnate to the costa on distal side or winged to the next pinnule; segments parallel sided and subentire, margins slightly recurved, segment apex usually obtuse to subtruncate and bearing wide-based but acute to obtuse pointed teeth. Sori medial; indusia thick and when young with margins tucked under sorus. Dryopteris affinis is native to Europe to the Caspian Sea and northwest Af- rica. This species is best recognized by the dark blotch on the underside of the costa next to the rachis, although it is absent on a plant reportedly from Madeira, and may fade in dried specimens of ssp. borreri (Newman) Fraser- Jenk. It may be faintly present on sun-exposed, old, leathery fronds of D. filix- mas, and is present on fresh fronds of D. pseudo-filix-mas. Should fronds of D. filix-mas have faint blotches, they may be separated by its slightly tapered segments with sharper teeth extending down the sides of the segment and the stipe and rachis scales that are pale, thin, membranous and variable in width. In southern California, D. filix-mas also tends to become deciduous much ear- lier than does D. affinis. Dryopteris pseudo-filix-mas may be separated from D. affinis by the characters in the key. Dryopteris affinis is a very difficult species complex and the delineation of its many variants is quite controversial. Some botanists maintain an informal approach and designate the variants as morphotypes, whereas others take a more formal approach and designate them as subspecies. Even with experi- ence, most variants are difficult to separate. For more details on the subspecies or morphotypes, see Jermy and Camus (1991), Stace (1989), Fraser-Jenkins (1982, 1996), Hutchinson and Thomas (1996), Dyer (1996), Jermy (1996), and Piggott (1997). Studies on D. affinis subspecies (or morphotypes) are incomplete and em- phasize mostly British natives. The native origin of cultivated plants is usually unknown and some may be subspecies different from the British natives. Therefore the following treatment of the cultivated plants should be regarded as tentative. The following subspecies and hybrids have been found in U.S. cultivation and most grow well in southern California: 5.1 Dryopteris affinis ssp. affinis—Western scaly male fern. Description and distribution the same as for the species. Stipe scales dense, deep gold to dark gold; blade glossy, thin-leathery, base more or less tapering; segment apex rounded with obtuse to slightly acute teeth; indusia thick, well tucked under the sporangia, not shriveling but lifting slightly at maturity. At- tractive for its densely scaly stipe, glossy dark bluish-green colored fronds. Particularly noticeable is the yellow-green color of new growth. Diploid, apog- amous. It is hardy to a January average of 25°F, deciduous to semi-deciduous, the old fronds often lasting until spring, easily cultivated.The plant circulating among gardeners as D. affinis from Madeira is distinct by the absence of a dark spot at the base of the pinna costa on the abaxial side, and the presence of very minute glands on the indusial margin and on the tissue protruding from HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 17 the upper surface of the indusium near the center. These observations were made on plants growing in southern California. It is uncertain what the taxo- nomic status of this plant is. It is a sturdy grower and tends to produce a stout rhizome bearing fronds in a well defined fascicular pattern. The stipes are noticeably short and thick. The spores were reportedly collected by Clive Jer- my in Madeira, Spain, and were originally grown and distributed by Judith Jones. 5.2. Dryopteris affinis ssp. borreri (Newman) Fraser-Jenk. (Willdenowia 10: 110. 1980).—Borrer’s scaly male fern, common scaly male fern. Stipe scales moderately dense, pale straw to mid brown with dark bases; blade slightly glossy, base truncate; segment apex squarely truncate to more pointed with sharp acute teeth frequently longer at the corners (resembling cat’s ears) rarely with the lowest basiscopic pinnule next to the rachis elongate and pinnatifid; indusia thin, with partial flat rim, lifting into a disc, then into a cone at maturity. Same range as ssp. affinis. Triploid (2n=123), apogamous, culture as for the ssp. affinis, fronds dying back progressively through the winter. 5.3. Dryopteris affinis ssp. cambrensis Fraser-Jenk. in L.N. Derrick, Jermy & A.M. Paul (Sommerfeltia 6:xi. 1987).—Narrow scaly male fern. Stipe scales, dense, ginger to reddish gold; blade slightly glossy, narrowly elliptic to oblanceolate, base tapering, segment apex rounded-truncate to more pointed with slightly obtuse to acute teeth; indusia thick, with margins just enclosing the sporangia, shriveling and lifting to form a cone upon maturity. Range same as for the species except absent in parts of central and S. Europe. Triploid (2n = 123), apogamous, culture as for ssp. affinis, fronds dying back rapidly after first frost. 5.4. Dryopteris xcomplexa Fraser-Jenk. in L.N. Derrick, Jermy & A.M. Paul (Sommerfeltia 6:xi. 1987). Stipe scales moderately dense, brown; blade lanceolate to ovate-lanceolate, matte, base slightly narrowed, truncate, pinnae outline somewhat uneven, seg- ment margins shallowly crenate-lobed or toothed, segment apex round-trun- cate; indusia shriveling and lifting to form a distorted cone when mature, spores mostly aborted. A hybrid of D. affinis x filix-mas. Range uncertain, probably where both parents exist. Tetraploid (2n=164), apogamous. Widely sold in the trade as D. filix-mas undulata robusta (sometimes as D. affinis x filix-mas ‘Robust’ or D. undulata). Culture as for ssp. affinis, semi-deciduous. Vigorous growing, producing many fronds with pinnae often overlapping slightly to give a full foliaceous appearance. In addition, the following cultivars have been found in the U.S. trade: 18 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Dryopteris affinis ‘Congesta Cristata’. Fronds dwarf to 23 cm, congested and crested. Dryopteris affinis ‘Crispa’.—Frond dwarf and broad, to 20 cm long and 14 cm wide, crisped and congested, the segments held in different planes or some- what twisted giving an irregular outline; from ssp. affinis. Plants by this name in the current trade are normal sized with segments twisted. Dryopteris affinis ‘Crispa Gracilis’—Dwarf, congested, upright leathery fern with the pinnae apices curved and hook-like. It has similar culture require- ments to ssp. affinis, from which it originated. Probably the same as the plant sold in the Dutch trade as D. ‘Crispa Congesta’ or D. ‘Congesta Crispa’. Dryopteris affinis ‘Cristata’ (‘Cristata The King’).—Fronds to 120 cm, arching blade apex and pinnae each ending in a tassel; from ssp. affinis. Dryopteris affinis ‘Cristata Angustata’.—Like cv. Cristata except narrower and smaller, to ca. 45 cm long by 5 cm wide; from ssp. affinis. Current trade ma- terial by this name reaches 80 cm by 7 cm. Dryopteris affinis ‘Grandiceps’.—Frond apex wih a heavy terminal crest. Dryopteris affinis ‘Polydactyla’.—A group of crested forms with flat tassels on the pinnae tips and 2 large crests on the blade apex. Dryopteris affinis ‘Revolvens’.—Tips of pinnae recurved, fronds to 100 cm.; from ssp. borreri. Dryopteris affinis ‘Stableri’.—Erect to slightly arching, narrow fronds, to 1 m. Reported to be a hybrid between D. affinis var. affinis ‘Pinderi’ (an abnormal narrow form of the species) and D. filix-mas (Fraser-Jenkins, 1996)). Dryopteris affinis ‘Stableri Crisped’.—Very upright narrow fronds of medium height, margins crisped. 6. Dryopteris crassirhizoma Nakai (Cat. Sem. Hort. Bot. Univ. Imp. Tokyo 32. 1920).—Fig. 7. Rhizome erect, stout, may produce offshoots. Fronds to ca. 150 cm long, 30 cm wide. Stipe short, 7,—Y, the length of the blade, fasiculate, densely covered with mid-brown to reddish brown scales, the larger scales thin, glossy, firm membranous, usually reddish brown, linear, lanceolate to ovate, attenuate, to 30 mm long or more; blade mostly oblanceolate, gradually narrowed below, deeply pinnate-pinnatifid next to the rachis in the proximal part of the frond; pinnae linear-lanceolate, basal pair usually more triangular but equilateral, underside of the costa next to the rachis with costal scales tan, triangular, variable, larger ones mostly attenuate, kinky; segments (pinnules) typically closely placed, narrowly oblong, weakly falcate, their apices mostly blunt- rounded, dentately toothed or untoothed, segment side margins mostly weakly crenate, lowest basiscopic segment (pinnule) next to the rachis not free and its side margins subentire. Sori medial, borne on distal part of frond. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 19 6 } a dps Wd,\sy3 Kies i ee ua a 7. Dryopteris crassirhizoma. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Pinnules ie 1cm], D) Medial pinna above, distal pinna below [scale=1cm]. 20 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Dryopteris crassirhizoma is a sexual diploid species native to northeastern Asia, where it is abundant. In the past, it has been confused with D. filix-mas, which differs by having teeth along the side margins of the segments and other characters in the key. This species differs from D. affinis by the characters in the key, and D. crassirhizoma has very thick, glossy stipe base scales, segments long, apex mostly round and untoothed, fronds oblanceolate and green rather than bluish-green in color. When D. pseudo-filix-mas lacks the pinnatifid or innate basiscopic pinnule next to the rachis on the lowest pinnae, it may be distinguished from D. crassirhizoma by its truncate to acute pointed more rectangular pinnules with side lobes and acute teeth. Dryopteris crassirhizoma is semi-deciduous in cool winters and probably deciduous in cold winters. It is easy to grow and is hardy to a January average of 30°F. 7. Dryopteris lepidopoda Hayata (Icon. Pl. Formosan. 4:161, fig. 101. 1914).— Fj ig. 8. D. nigra Ching (Bull. Fan Mem. Inst. Biol. 8:430. 1938). Rhizome erect. Stipes usually % the blade length or equal to it, the stipe and rachis scales narrow long-triangular, brownish to black, margins ciliate, cilia often more numerous and longer towards the scale base; young fronds pink; blade pinnate-pinnatifid to 2-pinnate next to the rachis, oblong-triangular, 25— 40 cm long, 10-20 cm wide, slightly lustrous, pinnae mostly falcate often de- flexed at the frond base, acuminate; lowest pinnae ca. as long as the middle pinnae; segments (or pinnules) oblong, many, close, spreading, apex rounded, bearing mostly small narrow-triangular acute teeth, the basal segments con- stricted at their base; pinna costa scales like the rachis scales but smaller and mixed with brown fibrils. Indusia thick, mostly persistent. Dryopteris lepidopoda is an apogamous diploid fern that is native and com- mon in China from the Himalayan region to Taiwan at an elevation of 1,200— 1,550 m. Good distinguishing characters of D. lepidopoda are blades that are usually as wide at the base as in the middle, relatively long stipe with many narrow, black, ciliate scales, and many oblong, lustrous segments that are rounded and sharply toothed at the apex. Sometimes young plants of D. wallichiana and D. lepidopoda are confused. Those of D. lepidopoda are markedly pink when young, whereas those of D. wallichiana are yellowish. Very small somewhat stellate scales on the frond surfaces of D. lepidopoda also distinguish it from D. wallichiana which has lanceolate scales. In both species, these scales are sparsely distributed and shed early. In Seattle gardens, Dryopteris lepidopoda usually grows to 60 cm tall, is evergreen, and probably hardy to a January average of ca. 45°F or lower. 8. Dryopteris polylepis (Franch. & Sav.) C. Chr. (Index Filicum 285. 1905).— Fig. 9. Rhizome erect. Stipe ca. ¥, the blade length, stipe scaly, the scales narrow triangular to ovate, blackish, the margins stiff, ciliate to fimbriate to just below HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION Me Ms NC ( J | we ai B Fic. 8. Dryopteris lepidopoda. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) 22 cy ei iy! Bi Wi Fic. 9. Dryopteris polylepis. A) Frond [scale= 5cm]. B) Stipe scales [scale=5mm]. C) Pinnules from medial pinna showing unbranched veins [scale=1cm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 23 the scale apex, rachis scales many, like the stipe scales but shorter and paler at the base; blade narrowly elliptic to oblanceolate, to ca. 54 cm long, 18 cm wide, pinnate-pinnatifid in distal part to pinnate-pinnatisect in the proximal part, apex acuminate, pinna elongate linear triangular, sessile, proximal pinnae gradually shortened to 4—5 cm, costa scales like the rachis scales and with small pale brown ovate-fimbriate scales; segments narrow, mostly oblong-fal- cate, many, close, the margins subentire or serrate, the apex rounded to round- truncate, the veins per segment 6-7 pairs, unforked except for those in the basiscopic segments next to the rachis, there the veins often forked. Sori large, marginal, 3-5 per segment, sori in distal % of the blade; indusia round with shallow sinus, gray-brown at center, thick. Dryopteris polylepis is a sexual diploid species from northeastern Asia. The unforked veins, except for those in the basiscopic segments next to the rachis, readily identify this species from other cultivated material of this section, as do the narrower, longish pinnules (or segments). It is hardy to a January average of 35°F; deciduous in southern California. 9. Dryopteris pseudo-filix-mas (Fée) Rothm. (Candollea 10:96. 1945).—Fig. 10. Rhizome erect, stout, producing offshoots. Fronds fasciculate, erect; stipe short, dense scaly at least at base, the scales membranous, mixed, the larger ones triangular to ovate-lanceolate or ovate, to ca. 15 mm, but usually less, brown, many darker at the base and center; blade linear-triangular to oblan- ceolate, to 2 pinnate except 2 pinnate-pinnatifid (to 3 pinnate) if the lowest basiscopic pinnule next to the rachis is developed, 40-80 cm long, 14—25 cm wide; pinnae narrow triangular, mostly sessile, proximal pinnae often shorter and broader triangular, the basal ones usually inequilateral, broader on basis- copic side, underside of costa next to rachis with black blotch at least on fresh material; larger costal scales mostly triangular, base and center darker, often abruptly tapered to a long filamentous apex, margins sparsely fimbriate ciliate; the lowest basiscopic pinnule next to the rachis pinnately lobed to pinnatifid (or pinnate), usually with at least 1 more or less rectangular, lobe cut % way to midrib of pinnule, often elongate, sessile, less often slightly adnate on distal side, pinnules (or segments) oblong, side margins more or less parallel, sub- entire, larger ones shallowly serrate, the apex rounded or more or less truncate and toothed, apices of basal segments acute. Sori medial, mostly in distal part of frond; indusia round reniform or some broader and with a wider sinus, opaque, young indusia with margins tucked under sorus lifting and shriveling upon ripening. Dryopteris pseudo-filix-mas is an apogamous triploid species native to high elevation cloud forests in Mexico and Guatemala. The best distinguishing char- acter of D. pseudo-filix-mas is the often elongate basal basiscopic pinnule next to the rachis that is pinnately lobed to pinnatifid (or pinnate?) with the lobes rectangular. However, fronds from younger plants (even 2-3 years old) often do not develop this distinct basiscopic pinnule or do so only weakly, thus making it difficult to separate D. pseudo-filix-mas from D. affinis, especially 24 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) é * eye? yore 5S. Bod ay Fic. 10. Dryopteris pseudo-filix-mas. A) Frond [scale=5cm]. B) : : Stipe scales [scale=5mm]. C) Low- est pinnae from different fronds showing the margins of the b asal pinnules [scale=1cm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 25 ssp. borreri. Dryopteris filix-mas has teeth extending from the pinnule (or seg- ment) apex down the slightly tapered sides, whereas D. pseudo-filix-mas has more parallel-sided segments with subentire to weakly serrate margins. John Mickel of the New York Botanical Garden introduced D. pseudo-filix-mas into cultivation in the U.S. from Mexico, whereas in Europe it was introduced by Christopher Fraser-Jenkins. Dryopteris pseudo-filix-mas is hardy to a January average of ca. 30°F. This fern is easy to grow and may produce ample offshoots in late summer. It is semi-deciduous in New York and Seattle, but evergreen in Los Angeles. 10. Dryopteris wallichiana (Spreng.) Hyl. (Bot. Not. 1953:352. 1953).—Fig. 11. D. paleacea (D. Don) Hand.-Mazz D. parallelogramma (Kunze) Alston Rhizome erect, stout, may produce offshoots. Fronds fasciculate, erect; stipes 8-25 cm long, % or less the frond length, densely covered with scales, the scales narrow triangular to lanceolate, to 25 mm or more long, 3 mm or more wide, black or very dark brown [in cultivated plants and plants from Asia; mid to pale brown in tropical American and Hawaiian plants], apex ending in a long filament; rachis densely scaly with same type of scales except smaller; blade pinnate-pinnatifid except weakly 2-pinnate in proximal part next to ra- chis, long-ovate to lanceolate, 50-100 cm long, 18-28 cm wide, lustrous dark green above, lighter below and veins conspicuous; pinnae linear-triangular, sessile, base of costa on underside faintly dark or not; segments rectangular, apex truncate or rounded-truncate, margins toothed, subentire to weakly cre- nate-serrate to serrate, often slightly reflexed. Sori medial; indusia round re- niform, entire, convex at maturity, dark brown when dried. Dryopteris wallichiana apparently exists at various ploidies (most reports are of apogamous diploids, but triploid, tetraploid and other counts have been reported). It is distributed in tropical regions from Mexico to South America, Africa, Himalayan region, China, Japan, and Hawaii. It is found in terrestrial habitats in cloud forests at high elevations. This species is set apart by the very narrow black or very dark brown scales (mid brown in tropical American plants) on the stipe and rachis, the evenly placed rectangular, lustrous seg- ments, and the conspicuous veins on the somewhat lustrous underside of the segments. The segment margins tend to be reflexed. Dryopteris wallichiana can be a large fern. It is hardy to a January average of 40°F and is semi-deciduous in cool winters. When not receiving sufficient coolness and humidity, the frond and pinnae tips tend to abort. Section 2.3. Pandae Fraser-Jenk. Fronds 1—2-pinnate, lanceolate to narrow-lanceolate; stipe with scattered, usually pale lanceolate or ovate-lanceolate scales; blade pale-green, somewhat succulent-herbaceous; pinna lobes or pinnules usually with wide, obtuse or rounded apices. 26 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) ; pay “ a i \ Mee nd Ai le ree iH Ted; i re: Bed Ap Fic. 11. Dryopteris wallichiana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Pinnules from medial pinna [scale=5mm] HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 27 KEY TO SPECIES OF SECTION PANDAE 1. Pinnae shallowly lobed, cut ca. % or less deep to pinnae midrib 1. Pinnae deeply pinnately divided, cut % or more deep to pinnae 2. Fertile pinnae noticeably narrower than the sterile ones and feiiiad to - gp % of the Mahe sn 5 i es ces oes ee a D. ludoviciana 2. Fertile and sterile pinnae of approximately the same width 3. Blade ovate-lanceolate, tapering gradually from below the middle to the tip; sory pare ovate-lanceolate; scales dark brown or pale brown with dark center .... . ne EAs, . Blade oblong-lanceolate, tapering in the distal 4; basal pinnae deltate to park rare scales pale brown, with or without a dark brown center 4. Basal pinnae deltate (as long as or only slightly longer than wide); pinnae of the fertile frond twisted at right angles to the blade surface as in an open Venetian blin ; lise 15. D. tokyoensis ww PAs tu. ame eee e.g ee a aie 8 age 8 10 FER 8 Pe) sate 4. Basal pinnae elongate-deltate (ca. 2 times longer than wide); pinnae oo et ais. riage not say Peeisted .. 2. ese a re, ee. . D. clintoniana 11. Dryopteris celsa (W. Palmer) Knowlt., W. Palmer & Pollard (Proc. Biol. Soc. Wash. 13:202. 1900).—Log fern.—Fig. 12 Rhizome short to moderately creeping, branched. Fronds 90-120 cm long, 20-30 cm wide, erect; fertile and sterile fronds and pinnae alike; stipe %—% the length of the frond, the scales dark brown or pale brown, usually with a dark center; blade pinnate-pinnatifid, ovate-lanceolate tapering gradually from below the middle to the tip; basal pinnae ovate-lanceolate, with their first few basal pinnules the same length as or shorter than the adjacent ones. Sori me- dial; indusia without glands. Dryopteris celsa is an uncommon sexual tetraploid species native to the swamps and wet woods of the eastern United States. It is believed to have originated from a cross of D. goldiana X ludoviciana followed by a doubling of the chromosomes. Dryopteris celsa differs from D. clintoniana in the blade ovate-lanceolate, the basal pinnae ovate-lanceolate, and the scales dark brown or pale brown with a dark center stripe. This robust fern is deciduous or semi-deciduous in southern California and is easily cultivated in moist soil. In the wild, it grows in areas where the average January temperature reaches 25°F. 12. Dryopteris clintoniana (D.C. Eaton) Dowell (Proc. Staten Island Assoc. Arts 1:64. 1906).—Clinton’s wood fern.—Fig. 13 Rhizome short creeping, branched. Fronds 40—120 cm long, 15-20 cm wide, fertile fronds longer than the sterile fronds; stipe % the length of the frond or more, the stipe scales pale brown, at times with a dark brown center; blade herbaceous, pinnate-pinnatifid, broad oblong-lanceolate, tapering to the tip in the distal %4, basal pinnae elongate-deltate, ca. 2 times longer than wide, widest at the base, the basiscopic segments longer than the acroscopic ones. Sori me- dial; indusia without glands. Dryopteris clintoniana is a hexaploid sexual species native to northeastern North America, where it grows in moist woods and swamps. This species is believed to have originated from the cross of D. cristata x goldiana followed Fic. 12. Dryopteris celsa. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION Fic. 13. Dryopteris clintoniana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. 30 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) by a subsequent chromosome doubling. Somewhat similar to D. cristata in appearance, it differs in having basal pinnae that are distinctly longer than wide and that, in fertile fronds, are not strongly twisted as in an open Venetian blind. Dryopteris clintoniana is easily cultivated in moist soil in shady gardens. The fertile fronds are deciduous; the sterile fronds, semi-deciduous. In the wild, it grows in areas where the average January temperature reaches 20°F. 13. Dryopteris cristata (L.) A. Gray (Manual, ed. 1, 631. 1848).—Crested wood fern.—Fig. 14. Rhizome short creeping to erect, may produce offshoots. Fronds 30-75 cm long, 7-12 cm wide, the fertile fronds longer and more erect than the sterile fronds, which are %—% shorter than the fertile ones; stipe 4—% the length of the frond, stipe scales light brown; blade herbaceous, pinnate-pinnatifid, nar- rowly oblong-lanceolate, tapering to the tip in the distal %, without glands, fertile and sterile pinnae not markedly different, the fertile pinnae usually twisted at right angle to the blade surface, as in an open Venetian blind; pinnae oblong-triangular widest at the base, basal pinnae not much longer than wide. Sori medial; indusia without glands. Dryopteris cristata is a sexual tetraploid species from northern and eastern North America and Europe, where it grows in bogs, swamps, and wet woods. This species is believed to have originated from a cross between D. ludoviciana and an as yet unknown species followed by a doubling of the chromosomes. The Venetian blind orientation of the pinnae and the deltate basal pinnae help distinguish this species. Dryopteris cristata is easily cultivated in moist soil, particularly favoring wetter areas, where it grows to its maximum size; fertile fronds deciduous, sterile fronds semi-evergreen. In the wild it grows in areas where the average January temperature reaches 0°F, but surprisingly grows well in southern Cal- ifornia, where it barely becomes deciduous. 14. Dryopteris ludoviciana (Kunze) Small (Ferns S.E. States 281. 1938).— Southern wood fern.—Fig. 15. Rhizome short-creeping to erect, branching to produce offshoots. Fronds erect, 60-120 cm long, 15-30 cm wide; stipe % the length of the frond, stipe scales pale brown; blade pinnate-pinnatifid, elliptic-lanceolate, dark-green, herbaceous, semi-evergreen, without glands; fertile pinnae restricted to distal % of the blade and much narrower than the sterile ones, basal pinnae trian- gular, smaller than those above them. Sori medial, indusia without glands. Dryopteris ludoviciana is a sexual diploid species native to the southeastern U.S., where it grows in swamps and wet woods. It is easily recognized by its pinnate-pinnatifid blade in which the pinnate fertile pinnae are distinctly nar- rower than the sterile ones and are restricted to the distal half of the blade. Dryopteris ludoviciana is easily cultivated in moist, rich garden soil. In the HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION ALG f Madd ot Fic. 14. Dryopteris cristata. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Fic. 15. Dryopteris ludoviciana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 33 wild, it grows in areas where the average January temperature reaches 50°F. It is nearly evergreen in southern California. 15. pets opie ters oo (Matsum. & Makino) C. Chr. (Index Filicum 298. 1905).—F Rhizome erect or ascending at times branching and producing offshoots. Fronds erect forming a crown, 35-90 cm long, 11-20 cm wide, fertile and sterile fronds similar, deciduous; stipe ¥,-Y, the length of the frond, the stipe scales light brown; blade pinnate, oblanceolate gradually narrowing to the base; pinnae lobed, mostly cut % or less deep, the basal lobes enlarged to resemble auricles; fertile pinnae narrower than the sterile pinnae and borne on the distal % of the blade. Sori medial; indusia without glands Dryopteris tokyoensis is a sexual diploid from eastern Asia. This popular fern has narrow, erect fronds that form a whorled, crown-like cluster. The shallow lobing of the pinnae distinguishes this species as do the gradual ta- pering of the blade at the base and the auricle-like lobes at the base of the pinnae. D. tokyoensis is very easily cultivated in moist acidic soil and is deciduous. It is, however, well liked by slugs which can rapidly destroy small plants. Plants do not do well in southern California, possibly because of the absence of acid soil or a need for winter chilling. It appears to be related to D. ludov- iciana and may be a partial variant of it (Fraser-Jenkins, pers. com.), although it has been treated as a distinct species. In the wild, it grows in areas where the average January temperature reaches 15°F. Section 2.4. Dryopteris Fronds linear-lanceolate to lanceolate (oblong-triangular in D. goldiana), 2- pinnate, the pinnules widely attached to the costae except at the bases of the proximal pinnae. Pinnules slightly, but not markedly, parallel-sided, usually somewhat tapering to their apices from ca. % their length, toothed at the sides and markedly so at the apices, usually with long acute teeth. Blade matte, herbaceous, the scales of stipe and rachis mostly lanceolate or ovate-lanceo- late KEY TO SECTION DRYOPTERIS 3 peri with the basal pinnae shorter than those above «orenll tetas, fonds less Than 30 cm long «2. ses gees ee ess 18. D. fragrans “ Medium to larger ferns, fronds more than 30 c ng 3. Fronds spreading, blade variable, mostly cat: tip of segment with acute teeth more or less pointing toward the segment apex; indusia thin and white when young, flat or slightly convex 4, Pinnae widest at their base; indusia convex, white when young, brown me older, margins entire; pinnule toothed around apex, teeth not in pairs... . 17. D. filix-mas AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) a ‘ | { { We ate { 4 di 2 \ y Bea — > Fic. 16. Dryopteris tokyoensis. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 35 4. Pinnae widest at their middle; indusia flat, white at all stages, margins lacerate; pin- nule double toothed all arouiid =. : .. . .. . 1. i ee eee 16. D. caucasica 3. Fronds erect, blade lanceolate, more narrowed at base, tip of segment with obtuse teeth pointing away from the segment apex, indusia thick and green when young, wrapping around the sori 5. Fronds pale gray-green, to 50 cm long, stipe ba ] tte, pinnule teeth spreading out in a fan-like fashion around the apex .................... 20. D. oreades 5. Fronds yellowish-green, to 122 cm long, stipe base scales glossy, pinnule teeth not STOOGES eck sk a ee es 21. D. sichotensis 16. Dryopteris caucasica (A. Braun) Fraser-Jenk. & Corley (Brit. Fern Gaz. 10: 221-231. 1972).—Fig. 17. Rhizome usually ascendent at apex, horizontal below, stout, forming off- shoots. Fronds to 105 cm, erect, spreading, fasciculate; stipe %—% length of blade, the scales sparse, pale brown, narrowly triangular to ovate-lanceolate, up to ca. 2 cm long, toothed towards the attenuate apex; blade to ca. 80 cm long, 35 cm wide, mostly ovate-lanceolate to elliptic, flat, herbaceous, 2-pin- nate; pinnae to 20 cm long, 5 cm wide, lanceolate or frequently narrow long triangular with attenuate apex, pinnatisect to pinnate, the pinnules (or seg- ments) with margins entire to mostly lobed, the lobes with very acute, distinct teeth usually arranged in pairs. Indusia very thin, membranous, white, greatly overlapping the sporangia shortly before maturity, edges lacerate, rapidly shrivelling. Dryopteris caucasica is a diploid sexual species native to forests in the al- pine regions of the Middle East (200-850 m elev.). It was introduced into U.S. cultivation very recently, probably from horticultural sources in Britain origi- nating from Fraser-Jenkins’ collections. Dryopteris caucasica is one of the par- ent species of D. filix-mas and is best distinguished from it by the generally paler color of the lamina, the doubly-toothed margins of the segments, and the indusia, which are white at all stages (until shrivelling) and have lacerate mar- gins. The distinct acuteness of the usually paired teeth at the apex of the pinnules (or segments) and their lobes, and the flatness, thinness and lacerate margin of the indusia are important characters of this fern. In case of doubt, its very dark spores distinguish it from D. filix-mas. Wild plants apparently do not form side crowns as readily as those in cultivation. Dryopteris caucasica is easily cultivated, and is deciduous at first frost. It is hardy to at least 30°F and perhaps to 20° F, but doesn’t tolerate summer drought as well as D. filix-mas. 17. Dryopteris filix-mas (L.) Schott (Gen. Fil., plate 9. 1834)—Common male fern.—Fig. 18. Lastrea filix-mas (L.) C. Pres] Rhizomes erect, stout, producing offshoots. Frond 35-150 cm long, 5-30 cm wide; stipe “4—% the length of the blade, sparsely to moderately scaly, the scales mixed, larger ones mostly narrow ovate to broad ovate, to ca. 14 mm long, 6 mm wide, margins erose, sparsely and irregularly fimbriate and toothed, mem- AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) ata C { : ! ‘ yea ' yes Wee i x ni ant ep? ete N PAAR we mS Yn vet \ 1 t Ut" . i IA Tc fg A: ’ 2 no Alt | gti, wr aee : ! \ Wy AC \ j STH f 4:47 i! 4% STi} ' : ntl Arter” et ee SUNS ; B ' “rd Fic. 17. Dryopteris caucasica. A) Frond [scale=5cm] after Fraser-Jenkins & Corley (1972). B) Stipe scales [scale=5mm]. C) Pinnules (segments) from medial pinna [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 37 Pitas peer et Arde AH ene ¢ vette. \ rr ; vA ey eye IG. 18. Dryopteris filix-mas. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Pinnules (segments) from medial pinna [scale=5mm] D) Other frond variations [scale=5cm]. branous, pale brown, rachis scales same as the stipe scales except smaller, narrower and more triangular; blades pinnate-pinnatifid to 2-pinnate at the base next to the rachis, oblong to obovate-lanceolate, usually narrowed to a truncate base, more or less herbaceous, green, slightly paler beneath; pinnae 38 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) short stalked; pinnules (or segments) oblong to slightly tapered, the margins crenate-serrate to serrate, the serrations sometimes toothed, the segment apex rounded with more or less acute, sharp teeth. Sori large, 4-6 per segment; indusia when young to mature, white, round, thin, spreading. Dryopteris filix-mas is a sexual tetraploid species native to Europe, western Asia, the far-western Himalayas, and North America, where it forms large clumps in lowland or alpine forests or in open places on rocks. The species has escaped and become naturalized in New Zealand. Dryopteris filix-mas originated from a cross between D. oreades and D. cau- casica and is intermediate in its morphology between these two species (Fra- ser-Jenkins, 1976, 1989; Fraser-Jenkins and Corley, 1972). It has long been con- fused with D. oreades and D. affinis and its subspecies, which have been called the ”D. filix-mas group” as a group of three species, with D. caucasica sub- sequently being added. The complex was sorted out first by Wollaston (1915) and finally by Manton (1950). Dryopteris filix-mas lacks the dark blotch on the costa where it joins to the rachis (Jermy and Camus 1991), although rarely some plants have faint dark streaks or spots, especially on old leathery fronds exposed to the sun. The segments which are usually slightly tapered and toothed along the sides, the wider, pale membranous stipe scales, the more deciduous habit, the mid-green matte fronds and the less inflexed indusium help to distinguish D. filix-mas from D. affinis which has parallel-sided, more truncate segments usually with entire side margins, narrow golden-brown stipe scales, glossy dark green fronds, and a less deciduous habit. Dryopteris filix- mas hybridizes with D. affinis. A commonly sold hybrid is D. Xcomplexa Fraser-Jenk., which is discussed under D. affinis. This species is part of European folklore. The rhizomes were formed into amulets, called St. John’s hand, and worn as protection from evil spirits. Also, the rhizomes of D. filix-mas have been used as a medication for intestinal worms. Dryopteris filix-mas has spreading to arching fronds that grow to 100 cm long. It is hardy to a January average of 20°F, and is easy to culture in shade. It is deciduous, with fronds that become prone in the autumn then wither during the winter. It is tolerant of somewhat drier sites than other Dryopteris species. Many cultivars are known, but have been greatly confused as to their names. It is probable that many of the named cultivars actually belong to D. affinis. Present-day plants may not correspond to the description found in earlier lit- erature due to confused labeling in gardens. A number of cultivar names not nee here circulate in the U.S. trade. The more common ones in the U.S. trade are: Dryopteris filix-mas ‘Barnesii’.—Barnes’ male fern.—Growth upright, narrow fronds, ca. 130 cm long, 10 cm wide, the pinnae short, wide, the pinnules narrowed at the base, oval, deeply lobed, the lobes often serrate or toothed, oo double toothed. Matches earlier material by same name (Druery, HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 39 Dryopteris filix-mas ‘Crispa Cristata’.—Like ‘Cristata’ of current trade, except the pinnules or segments crisped. Dryopteris filix-mas ‘Cristata’—Crested male fern—A group of cultivars with apices of the blade and pinnae ending in a small to medium sized tassel with- out long finger-like divisions. The current trade plant sold as ‘Cristata’ has a narrow elliptic blade and compact medium size tassels, sometimes the tassel at the blade apex quite large. Dryopteris filix-mas ‘Cristata Martindale’—Wide elliptic blade, blade and pinnae with small crest, pinnae strongly falcate. Dryopteris filix-mas ‘Decomposita’.—Large, broad, foliaceous frond, almost 2- pinnate, 60-80 cm long, fine textured, the pinnules failing to develop properly at the sides so thickened and incised with irregular teeth. Dryopteris filix-mas ‘Grandiceps’.—Large crested male-fern.—Fronds slightly arching, rachis branching some distance from the fronds apex to form very large crests, pinnae narrow and trimly crested, vigorous grower. Dryopteris filix-mas ‘Linearis’.—Pinnules or segments very narrow to nearly filiform. Originated from spore of ‘Decomposita’. Dryopteris filix-mas ‘Linearis Congesta’—Small plant with modestly nar- rowed pinnules or segments, the pinnae short and close together. Dryopteris filix-mas ‘Linearis Cristata’.—Like ‘Cristata’ of current trade, but the pinnules or segments greatly narrowed. Dryopteris filix-mas ‘Linearis Polydactyla’—Slender crested male fern.— Blade broadly elliptic, divisions of the tassels on the blade and pinnae long and finger-like, the segments of the pinnae linear to nearly filiform or depau- perate. Like ‘Linearis’ but with forked pinna apex. Dryopteris filix-mas ‘Polydactyla’.—With tassels on the tips of the pinnae and blade, the divisions of the tassel long and finger-like. Dryopteris filix-mas ‘Ramo-cristata’.—Like ‘Cristata’ of current trade, but stipe branched. Dryopteris filix-mas ‘Undulata Robusta’.—See discussion of D. ote = 5 5 Ss ie] : . 5 n = p st > 3 5 Qu. i?) 3 Oo = e = . is] is) 3 nm c. a] fae) wn i?) 525) Ee oO an oo pf 5 9 Lar J = 5. & i+) Qu 5 he 2) is) 5 - © bar ° sar | ee i<°) n oO (species difficult to separate é. Stipe scales tan with a dark central stripe; fronds erect or slightly arching; North 36. D. expansa reading; North ee ee a ee eee . campyloptera 32. Dryopteris amurensis Christ in Christ & H. Lév. (Bull. Acad. Int. Géogr. Bot. 19:35. 1909).—Fig. 33. Rhizome short-creeping. Fronds 35-60 cm long, stipe longer than the blade, the stipe scales ovate, uniformly light brown or often with slightly darker cen- ter; blade deltoid pentagonal, evergreen, membranous, glabrous above and bearing small scales on veins beneath, 3-pinnate pinnatifid at the base, 2-pin- 62 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) FiG. 33. Dryopteris amurensis. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. natifid above, the basal pinnae % to almost as long as the blade, basal basiscop- ic pinnule of the basal pinna longer than the adjacent one and 4 times longer than the basal acroscopic pinnule; ultimate segment oblong-ovate, pinnately incised or lobed, softly spine tipped. opteris amurensis is native to northeastern Asia where it grows in co- niferous forests. It is a sexual diploid species that may be distinguished by its HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 63 membranous triangular blade which is about as wide as or wider than long and bears small scales along the veins on the abaxial blade surface. This species is easily cultivated but requires moist soil. It is reported to have been able to withstand harsh winter weather (Rush, 1984). In the wild it grows in areas where the average January temperature reaches ca. 10°F. 33. Dryopteris campyloptera Clarkson (Amer. Fern J. 20:118. 1930).—Moun- tain wood fern, eastern spreading wood fern.—Fig. 34. D. spinulosa (O. F. Miill.) Watt var. americana (Fisch.) Fernald Rhizome erect or ascending. Fronds 60-90 cm long, stipe % the length of the blade or more, the stipe scales light brown and usually with a dark brown base; blade without glands, broadly ovate-triangular to pentangular, 3-pinnate- pinnatifid at the base, 2-pinnate-pinnatifid above, pinnae lanceolate-oblong, basiscopic pinnule of basal pinnae longer than the adjacent one and 2—4 times longer than the acroscopic one; ultimate segments oblong to oblong-ovate, pin- nately incised or lobed, finely spine tipped. Sori medial; indusia without glands or rarely with a few glands. Dryopteris campyloptera is native to northeastern North America. It is a sexual tetraploid species that originated from a cross of D. expansa x inter- media. This species is frustratingly similar in appearance to one of its parents, D. expansa. Dryopteris campyloptera displays fronds with a less erect habit and less delicate texture, and less broad and oval than those of D. expansa. In nature, the two species do not overlap in their distribution, except in eastern Quebec and in the Maritime Provinces of Canada. In regions where they do overlap, it is difficult to distinguish D. campyloptera from its parental species. This is also particularly true with cultivated plants when the native source of the plant is unknown. The mountain wood fern is easily cultivated in shady areas or in partial sun in well-drained, moist soil. The fronds are deciduous. In the wild, it grows in areas where the average January temperature reaches 0°F. It is difficult to grow in southern California. 34. Dryopteris carthusiana (Villars) H.P. Fuchs (Bull. Soc. Bot. France 105: 339. 1959).—Spinulose wood fern, toothed wood fern, narrow buckler fern.—Figs. 35, 36. D. spinulosa (O.F. Mill.) Watt Thelypteris spinulosa (O.F. Mill.) Nieuwl. Aspidium spinulosum (O.F. Mill.) Sw. Rhizome ascending to erect. Fronds 45-75 cm long; stipe 4~ the length of the blade, the stipe scales ovate, uniformly tan; blade light-green or yellowish- green, without glands, narrowly triangular-lanceolate to ovate-triangular, 2-(3-) pinnate-pinnatifid proximally, 2-pinnate-pinnatifid distally; ages — triangular, the basiscopic pinnules of the basal pinnae longer than t ' a — ones and 2 times or less longer than the basal acroscopic pinnule; ultimate 64 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Fic. 34. Dryopteris campyloptera. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. segments oblong-ovate, pinnately incised or lobed, finely spine tipped, the margins flat. Sori medial; indusia without glands. Dryopteris carthusiana is found growing in wet woods, stream banks, and swampy areas. It is circumpolar, occurring in North America, Europe, and Asia. It is a sexual tetraploid species that is thought to have originated from a cross between D. intermedia and an as yet unidentified species. Dryopteris HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION Po ACER) J mY Fic. 35. Dryopteris carthusiana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. 66 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) carthusiana has triangular-lanceolate blades, and the basal pinnae have basi- scopic pinnules that are longer than the adjacent basal pinnules. The stipe scales are uniformly light brown. This species is easily cultivated in temperate, moist gardens. It is deciduous and not a vigorous grower in southern California. In the wild, it grows in areas where the average January temperature reaches —5°F. A foliose form is currently available in the trade incorrectly identified as Dryopteris stewartii (Fig. 36). 35. Dryopteris dilatata (Hoffm.) A. Gray (Manual, ed. 1, 631. 1848).—Broad wood fern, broad buckler fern.—Fig. 37. D. austriaca (Jacq.) Schinz & Thell., misapplied Rhizome erect or ascending, producing offshoots. Fronds 30-150 cm long, stipe %—% the length of the blade, the stipe scales mostly ovate-lanceolate, light brown and with a dark central stripe; blade dark green or bluish-green, without glands, triangular-ovate, 3-pinnate proximally, 2-pinnate-pinnatifid distally, pinnae lanceolate-oblong or triangular; the basiscopic pinnules of the basal pinnae longer than the acroscopic pinnules, ultimate segments oblong-ovate, pinnately incised or lobed, finely spine tipped, the margins turning under. Sori medial; indusia without glands or sometimes glandular. Dryopteris dilatata is a common, widespread European and western Asia species in damp woods. It is a sexual tetraploid species and is believed to be derived from a cross between D. expansa and probably D. azorica (Christ) Alston (Gibby and Walker, 1977; Fraser-Jenkins, 1982). The morphological characters of D. dilatata are intermediate between those of the supposed par- ents. It differs from D. expansa in having a less dissected frond and more rectangular ultimate segments that have margins tending to curl under, and in having darker scales and a darker green frond. Dryopteris dilatata of Europe and D. campyloptera (D. expansa X intermedia) of North America have been shown to be genomically similar and appear to have originated from a cross between related species, but probably resulted from independent crosses (Gib- by, 1977). Dryopteris dilatata is phytochemically different from D. campylop- tera, and recent authors have regarded them as distinct (Fraser-Jenkins, 1982). Dryopteris dilatata has pinnules that tend to curl under and stipe scales with a central stripe. This species is semi-evergreen, a robust grower in temperate climates, but in southern California it grows less vigorously. It does best in acidic soils. In the wild, it grows in areas where the average January temperature reaches ca. 25°F. The following cultivars are reported in the U.S. trade by Mickel (1994): Dryopteris dilatata ‘Crispa Whiteside’.—Crisped foliage and lighter frond col- or than species. Dryopteris dilatata ‘Cristata’—Frond and pinna tips crested. Dryopteris dilatata ‘Grandiceps’.—Frond crested forming a dense tassel and crested pinnae. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 67 jade © deg , SaPhee, Fic. 36, Dryopteris carthusiana (foliose form). A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Basal pinna [scale=5mm]. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Fic. 37. Dryopteris dilatata. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 69 Dryopteris dilatata ‘Jimmy Dyce’.—Fronds stiff, erect, blue-green in color. Dryopteris dilatata ‘Lepidota Cristata’—Fronds finely dissected and crested, 12-18 inches long; stipe and rachis with reddish brown scales. Dryopteris dilatata ‘Recurved Form’.—Margins of the segments curving down- 36. Dryopteris expansa (C. Presl) Fraser-Jenk. & Jermy (Brit. Fern Gaz. 11:338. 1977).—Arching wood fern, northern spreading wood fern, northwestern spreading wood fern, northern buckler fern.—Fig. 38. D. dilatata (Hoffm.) A. Gray, in part D. assimilis S. Walker D. austriaca (Jacq.) Woyn. Rhizome erect or ascending, producing offshoots. Fronds 30-90 cm long; stipe %—% as long as the blade, brown at base, pale green above (rarely darker), the stipe scales light brown, occasionally with a dark brown center; blade without glands, herbaceous, broadly triangular to triangular-ovate, 3-pinnate- pinnatifid proximally, 2-pinnate-pinnatifid distally, pinnae lanceolate-oblong, broad at base, basiscopic pinnules of the basal pinnae longer than the adjacent ones and 2-3 times longer than the acroscopic ones, ultimate segments ovate- oblong, pinnately incised or lobed, finely spine tipped. Sori medial; indusia without glands. Dryopteris expansa is native to northern Asia, North America, and Europe. It is a sexual diploid species and represents one of the parents of D. campy- loptera, from which it is difficult to distinguish morphologically. Dryopteris expansa differs subtly from D. campyloptera in having fronds that are nearly upright and with more broadly triangular, thin, delicate blades and more point- ed, falcate segments on proximal pinnae. To differentiate this species with certainty from D. campyloptera, the chromosomes need to be examined. Drop- teris dilatata has been shown to be a taxon distinct from D. expansa (Fraser- Jenkins and Jermy, 1977). This species is easily cultivated in temperate climates. In southern Califor- nia, it tends to produce new growth late in the spring and is deciduous, but not vigorous. It does not tolerate drying in summer. In the wild, it grows in areas where the average January temperature reaches ca. —5°F. 37. Dryopteris intermedia (Muhl. ex Willd.) A. Gray (Manual, ed. 1, 630. 1848).—Evergreen wood fern, glandular wood fern, fancy fern.—Fig. 39. D. spinulosa (O.F. Miill.) Watt var. intermedia (Muhl. ex Willd.) Underw. Rhizome ascending. Fronds 35-70 cm long, stipes usually YY the length of the blade, the stipe scales tan; blade oblong-lanceolate, 3-pinnate-pinnatifid proximally, 2-pinnate-pinnatifid distally, bearing minute stalked re aia cially at the bases of the pinnae and on the rachis; pinnae with ih or margins tapering only towards the tip, basal pinnae with the basa . pinnule usually shorter than the adjacent pinnule, ultimate segments ovate- 70 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) fecgity BAS: \ < ~ A Fic. 38. Dryopteris expansa. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. oblong, pinnately incised or lobed, finely spine tipped. Sori medial: indusia with minute, stalked, glandular hairs. Dryopteris intermedia is native to northeastern North America, where it grows in moist woods and swamp margins. It is a sexual diploid species and HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION = By eX hy, Sa Ads >> FN Ye Fic. 39. Dryopteris intermedia. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. has contributed to the formation of both D. campyloptera and D. carthusiana as one of their parent species. It is characterized by having lacy blades with short basiscopic pinnules on the proximal pinnae and particularly also by the presence of small stalked glands, resembling pin-heads, on the rachis, pinnae 72 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) axis, and indusium. It is genomically similar to D. azorica (Christ) Alston, the probable ancestor of D. dilatata. This species is easily cultivated in temperate, moist climates; it is not a vigorous grower in southern California. Its fronds are evergreen. In the wild, it grows in areas where the average January temperature reaches 10°F. Subgenus 3: Erythrovariae (H. It6) Fraser-Jenk. Blade variously dissected, the pinnae gradually reduced to a pinnatifid apex; abaxial surface of the leaf axis with bullate or bullate-based scales. Section 3.1: Erythrovariae Fronds herbaceous or somewhat leathery, not markedly stiff nor coriaceous, the pinnules without caudate apices, lobes normally rounded, the bullate scales well developed. KEY TO SECTION ERYTHROVARIA 1. Fronds pinnate, the pinnae crenate-serrate 1. Fronds more divided, 2- to 3-pinnate 2. Fronds 3-pinnate at least at the base, the basiscopic pinnules next to the rachis on the lowest pinnae variable in length, often longer than adjacent pinnules 3. Indusia absent, the blade herbaceous ............ ieee cs 43. D. gymnosora 3. Indusia present, the blade thin leathery 4. Rhizome creeping; spinulose teeth of segments mostly turning up from the plane of the fond | oo a ee 39. D. cystolepidota 4. Rhizome ascending to erect; spinulose teeth of segments usually poorly developed, nd D. BE Ser ne 40. D. decipiens tN ies) Lox j o a =| ed >| cs =} j=} 2°] = ® o rol ao) S p> - . io] a j=} = je) Cc. j=] te > og (= yy ro ‘ok Ww re) (=) = a ba v= soit =| j=} a ® =} i) Pal - onal io) oh i ® tS my a S. nm ° =] Be mm ® 5. Not with this combination of characters 6. Pinnules or segments short rectangular-oblong, broadly adnate except sessile next to the rachis on proximal pinnae; indusia to 3 mm in diameter; rarely cultivated species “ble Saas olla e bars eae se ai a A ne ee ee 42. D. fuscipes 6. Not with this combination of characters; indusia 2 mm or less in diameter 7. Pinnae typically distant; stipe and rachis pink-purplish; sori typically submarginal; very rarely cultivated species ......0. . oe. Oe 45. D. purpurella 7. Pinnae adjacent or overlapping; stipe and rachis not pink-purplish (or rarely so); commonly cultivated species SE Nea Me iS alma | 41. D. erythrosora - Blade mostly triangular, gradually tapering to the apex, erect and slightly arch- ing, the pinnules or segments narrow triangular or oblong, their apices mostly [o-] HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 73 rounded; bullate scales fewer on rachis; sori medial (in the wild, larger older plants with the blade more divided, to 3-pinnate or nearly so, and the proximal pinnae conspicuously stalked; this seldom seen in U.S. garden plants) ..... . Ma rireis fre eee. Re ee eee 44. D. hondoensis 38. Dryopteris championii (Benth.) C. Chr. ex Ching (Sinensia 3:327. 1933).— ig. 40. D. pseudo-erythrosora Kodama Rhizome erect, stout. Stipes to ca. 25 cm, clustered, densely covered with shiny, spreading, reddish brown scales, the scales to 1 cm long, lanceolate to ovate-lanceolate, crisped, margins membranous and erose-fimbriate; blade 2- pinnate-pinnatifid, ovate-triangular, to ca. 50 cm long, 25 cm wide; pinnae pinnatisect to pinnate-pinnatifid, falcate, apex pinnatifid acuminate, the scales on abaxial side of costa dense, some with flat bases, others partly or more or less fully bullate; smaller pinnules oblong-ovate to oblong-lanceolate, sessile, margins more or less crenate, apex obtuse, larger pinnules serrate-incised with auricles on both sides, basal basiscopic pinnule next to the rachis usually reduced, veins on abaxial surface with minute tan hairs (to 0.3 mm). Sori medial to often submarginal; indusia round-reniform. Dryopteris championii is an apogamous triploid species from eastern Asia, where it is common on hillsides and open areas with light shade. It is distinct by its dense covering of shiny, reddish brown stipe and rachis scales and, on larger specimens, very regular (neat), dark green, glossy, leathery, fronds. It is similar to Dryopteris erythrosora in general appearance but with larger, less toothed basal pinnules that are eared on both sides. Dropteris championii is a medium to large, evergreen fern and is hardy to a January average above 30°F. 39. Dryopteris cystolepidota (Miq.) C. Chr. (Index Filicum 260. 1905).— ig. 41. D. erythrosora (Eaton) Kuntze var cystolepidota (Miq.) Nakai D. erythrosora var. dilatata (Koidz.) Sugim. Dryopteris nipponensis Koidz. Rhizome medium short-creeping, branched. Stipes to ca. 30 cm long, irreg- ularly clustered, the scales mixed, moderately dense at stipe base, sparser above, dull brown to black, larger scales narrow triangular to lanceolate, ca. 10 mm long or more; blade oblong-triangular to broadly triangular, ca. 50 cm long, 28 cm wide, to barely 3-pinnate in the basal area, the blade apex usually abruptly acuminate, lowest pinnae noticeably the longest, the texture thin leathery, slightly glossy, the costae often conspicuously covered with dark bul- late scales, the scales when young and fresh often tan to pinkish at their bases and blackish brown distally; pinnules lanceolate-oblong to long narrow tri- angular, sessile or adnate, the larger ones eared on one or both sides, the mar- gins lobato-incised to serrate-incised, the teeth tipped with 1 (2) small spines, the spines often incurved and on live plants often turned upwards from the adaxial frond surface, the larger pinnules pinnatisect to pinnate, the basiscopic 74 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) WeIeriy BeoeeaG: badd dah Fic. 40. Dryopteris championii. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Costal scales [scale=1mm]. D) Pinnule from medial pinna [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION C Pic. 4° Dryopteris cystolepidota. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Basal pinna (Note: Basiscopic pinnule not longest as is usual) [scale=1cm]. 76 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) pinnule next to the rachis on the lowest pinnae shorter, equal to, or longer than the adjacent pinnule. Sori small, medial; indusia round-reniform, pinkish red in center at maturity. Dryopteris cystolepidota is an apogamous triploid species that is native to Japan and Korea. The moderately short-creeping rhizome, broad, slightly glossy fronds with an abruptly narrowed tip, the long pinnules, and the often upward-pointing spinulose teeth help in identifying this fern. Although the lengths of the basal basiscopic pinnules next to the rachis vary, they are not as long as those in section Variae. The slightly glossy spreading fronds with the new red growth are particularly attractive. Although the rhizome is mod- erately creeping, the growth is restrained. This species is of medium size, evergreen, of easy culture, and hardy to January averages slightly below 30°F. 40. Dryopteris decipiens (Hook.) Kuntze (Rev. Gen. Pl. 2:812. 1891).— ig. 42. Rhizome ascending to erect, forming offshoots. Stipes 10-35 cm long, clus- tered, the scales denser at the base, sparser above, very narrowly triangular, to ca. 12 mm long; blade narrowly triangular, to ca. 32 cm long, 18 cm wide, 1- pinnate, the proximal pinnae sometimes with a roundish, nearly free lobe next to the rachis, apex pinnatifid; pinnae linear-lanceolate, truncate to cordate, acuminate, falcate, leathery, margins entire or shallowly crenate to serrate at the apex, rachis and costa with bullate scales. Sori generally closer to the midrib than the margin, often more abundant and scattered at the pinnae base; indusia round-reniform. Dryopteris decipiens is an apogamous triploid species native to eastern Asia, where it grows among rocks or in shaded areas, usually at higher elevations. It is distinct from other cultivated 1-pinnate Dryopteris by its pinnatifid acu- minate apex, falcate pinnae, and bullate scales. It is a small to medium fern, hardy to a January average of above 30°F, semi- deciduous in warm climates, and deciduous elsewhere. 41. Dryopteris erythrosora (Eaton) Kuntze (Rev. Gen. Pl. 2:812. 1891).—Autumn fern.—Fig. 43. Rhizome erect-ascending to prostrate, stout, branching to form a few adja- cent crown. Stipes 30-60 cm long, irregularly clustered, the larger scales most- ly very narrow, stiffish, somewhat glossy, blackish brown to black, ca. 10 mm long; blade 30-70 cm long, 15-35 cm wide, typically broadly ovate to oblong, to 2-pinnate; pinnae 8-20 pairs, pinnatisect to pinnate, their apices pinnatifid, acuminate, the bullate scales of the costa fairly persistent, often dense and dark; pinnules narrow-oblong to linear-lanceolate, acute to rounded, the mar- gins subentire, serrate, crenate-serrate or incised-serrate, the teeth mucronate or spinescent and sometimes incurved, the basiscopic pinnule next to the ra- chis on the lowest pinnae usually reduced. Sori often closer to the midrib than HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 77 (AMY A) Fic. 42. Dryopteris decipiens. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Basal pinna [scale=1cm]. medial; indusia round reniform, quite evenly placed, at maturity red; for green- ish white indusia, see f. viridosora. Dryopteris erythrosora is an apogamous triploid species from eastern Asia, where is grows in woods on low mountains and hills. It is a very common and very variable species. Because of its variability, this species is often con- fused with others. The 2-pinnate, broad frond, the short basiscopic pinnule next to the rachis on the proximal pinnae, the many non-opposite pinnae, and the frequently incurved spinulose tipped teeth, often dark-tipped bullate scales, help somewhat to distinguish it from most U.S. cultivated Dryopteris. In the southern California trade, it is much confused with the similar appear- ing D. hondoensis (which see). Plants currently circulating in the Pacific Northwest as D. bissetiana and D. purpurella are D. erythrosora variants, these variants often have more triangular fronds and deeply serrate lobed ee than typical D. erythrosora. True D. bissetiana and true D. purpurella (whic see) are very rare in U.S. and appear distinctly different. 78 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Sy cat if Fic. 43. Dryopteris erythrosora. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Lowest pinna [scale=1cm]. D) Pinnule from medial pinna [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 79 The range of variability of Dryopteris erythrosora in U.S. gardens includes differences in height of the plant, the arching or spreading habit of the fronds, the fullness and shape of the blade and its divisions, particularly the degree of lobing in the pinnules, the intensity of the green blade color, the length of the stipe, thinness or brittleness of the blade tissue, color of the indusia, and also other features. Dryopteris erythrosora has medium to large fronds and is hardy to a January average somewhat above 30°F. It is a robust grower and easily cultured. The new growth is often a pinkish or reddish bronze, which is more pronounced on some plants than on others. This species is valued for its shiny evergreen foliage. Plants with red or maroon-red indusia provide an added interest. The following are found in cultivation. These were originally published as formae, but some authors believe that they are best regarded as cultivars: Dryopteris erythrosora f. prolifica (Maxim. ex Franch. & Sav.) H. It6 in Takai & Honda (Nov. Fl. Jap. 4:41. 1939).—Blade deltoid, often with buds; pinnules strongly contracted, linear shaped, apex sharply pointed. The original plant was found among wild plants in Japan. Small-medium plant of easy culture. Dryopteris erythrosora f. viridosora (Nakai ex H. It6) H. It6 (Bot. Mag. (Tokyo) 50:68-69. 1936 [also see H. It6 in Nakai and Honda, Nov. Fl. Jap. 4:41. 1939]).—Indusia when mature whitish green. Native to Japan. The form with white indusia belongs to f. viridosora. Judith Jones of Fancy Fronds Nursery, Seattle, Washington (pers. comm.) reports that spores from D. erythrosora with the normal greenish white indusia produce plants with red indusia and less frequently with very white indusia. Martin Rickard of England (pers. comm.) also reports that sowings of spores from plants with greenish white indusia yield some plants with red indusia. 42. Dryopteris fuscipes C. Chr. (Index Filicum, Suppl. 2, 14. 1917).—Fig. 44. Rhizome ascending to erect, slow to form offshoots. Stipes 20-30 cm long, clustered, the scales brown or more normally reddish brown, narrowly tri- angular, mostly confined to the stipe base, less persistent above and into the rachis; blade triangular, 20-40 cm long, 15-25 cm wide, to 2-pinnate; pinnae pinnatisect to pinnate, lanceolate to narrow triangular, apex pinnatifid, acu- minate to attenuate, costae with bullate scales; pinnules mostly oblong, fal- cate triangular at base of proximal pinnae on large fronds, less than 2.5 cm long, sessile or adnate, apex rounded to truncate, undersurface with fuga- cious, dark, hairlike scales, margins entire to crenate-serrate, the basiscopic pinnule next to the rachis on the lowest pinnae usually reduced. Sori close to midvein; indusia round-reniform, relatively large, to 3 mm in diameter, ' whitish tan. Dryopteris fuscipes is an apogamous triploid species from East Asia. This species is a medium-large fern with few fronds, giving a sparsely foliated as- pect. Its narrow pinnae seem to be farther apart from each other than those found on other cultivated species, and, along with the many broad truncate 80 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) MAAN) Mer arance Fic. 44. Dryopteris fuscipes. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. pinnules, help in the recognition of the species. Some of the introduced plants were grown from spores collected in China by the senior author. It is hardy to a January average above 30°F, and is evergreen, although the fronds tend to lie prone in the winter in southern California. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 81 43. Dryopteris gymnosora (Makino) C. Chr. (Index Filicum 269. 1905).— Nephrodium gymnosorum Makino Rhizome short creeping. Stipes 30-50 cm long, slender, sparsely scaly, the basal stipe scales narrowly triangular, 6-8 mm, nearly entire, brown to black- ish brown; blade narrowly to broadly ovate, sometimes nearly deltoid, 24—45 cm long, 18-30 cm wide, herbaceous, somewhat narrowed in distal part, acu- minate; pinnae broadly lanceolate or narrow long-triangular, 3-6 cm wide, nearly sessile, apex caudate-acuminate, proximal pinnae with basiscopic side larger, and the pinnule next to the rachis often the longest; pinnules mostly broadly lanceolate, ca. 1.5 cm long, 5-22 mm wide, sessile, pinnately lobed to parted, toothed, apex rounded to subacute. Sori medial; indusia absent. Dryopteris gymnosora is an apogamous triploid species native to eastern Asia. This species is occasionally listed in specialty catalogs in the U.S., and we have not been able to obtain U.S. material for verification. The herbaceous blade, the often long basiscopic pinnule next to the rachis on the proximal pinnae, and the absence of indusia are the easiest identifying features. 44. Dryopteris hondoensis Koidz. (Acta Phytotax. Geobot. 1:31. 1932).— Rhizome ascending to sometimes short-creeping, branching to form incon- spicuous crowns slightly distant from one another. Stipes in clusters, the scales light brown to blackish brown, larger scales narrowly triangular to ca. 10 mm long, dull, margins entire or sometimes with occasional fimbriations; blade triangular (frond 50-70 cm long), 2-pinnate to 3-pinnate or nearly so on larger fronds of old plants; bullate scales of the costa mostly light to medium brown, usually falling, frequently absent; pinnae pinnatifid to pinnate, apex pinnatifid to an acute or acuminate apex (proximal pinnae conspicuously stalked on more divided fronds on native plants); pinnules oblong, the margins incised- lobed or serrate, toothed, the apical teeth mostly acute or short spinescent, the basiscopic pinnule next to the rachis on the lowest pinnae usually reduced. Sori medial; indusia round-reniform, grayish white (for red indusia see f. rub- isora). Dryopteris hondoensis is an apogamous triploid species from Japan. When young, this species looks very much like D. erythrosora and is often sold as such in the trade. At maturity, Dryopteris erythrosora is a larger, less compact plant, and its fronds tend to be more oblong and arching rather than more triangular and spreading with little arching as in D. hondoensis. Dryopteris erythrosora has a very stout crown that may divide to form more crowns ad- jacent to each other, whereas crowns of D. hondoensis are smaller and more distant. Stipe scales of D. erythrosora are darker, appearing somewhat stiff and glossy with smooth margins, but those of D. hondoensis are a lighter appearing softer, dull with slightly irregular margins sometimes bearing a few fimbriations. Other less consistent differences are: the pinnules or segments of D. erythrosora are narrower and more pointed, whereas those of D. hondoensis 82 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) C Fic. 45. Dryopteris gymnosora. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Lowest pinna [scale=1cm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 83 Fic. 46. Dryopteris hondoensis. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Basal pinna [scale=1cm]. D) Pinnule from medial pinna [scale=5mm]}. 84 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) are broader and rounder at the apex; the indusia are generally closer to the pinnule midrib, neatly aligned and closely placed, in D. erythrosora, whereas they are more medial, and not so closely placed in D. hondoensis; the darker more persistent bullate scales and more brittle fronds of D. erythrosora differ some from the lighter colored, less persistent bullate scales (said to be absent on some wild forms), and less brittle fronds of D. hondoensis. Japanese bota- nists place much emphasis on the conspicuously stalked proximal pinnae found only on well developed fronds of older plants. This stalk is not con- spicuous on U.S. cultivated plants. Dryopteris hondoensis is a medium size fern hardy to a January average above 30°F. This species is more or less evergreen, with new growth often reddish, and it is easily cultivated. Plants with red sori are known as follows: Dryopteris hondoensis f. rubisora Kurata (J. Geobot. 13(2):42. 1964).—Differs in the red rather than grayish white color of the indusia. 45. Dryopteris purpurella Tagawa (Acta Phytotax. Geobot. 1:307. 1932).— D. erythrosora (Eaton) Kuntze var. purpurescens H. It6é D. indusiata (Makino) Makino & Yamam. var. purpurescens (H. It6) Kurata Rhizome very short-creeping to ascending. Stipes 20-30 cm long, purplish, the scales narrow triangular, cordate at base and then often narrowing gradu- ally or abruptly to an attenuate gland tipped apex, pale margined or not, brown and black or all greenish, to 7.5 mm long; blade ca. 30-45 cm long, 20-35 cm wide, triangular to broad-ovate, mostly 2-pinnate, abruptly narrowing to a pointed apex, rachis purplish; pinnae lanceolate to narrow triangular, pinnat- isect to pinnate, acute to acuminate, attached at right angles or nearly so to the rachis, often somewhat distant, costa with tannish bullate scales: pinnules oblong to elongate elliptic, the teeth somewhat narrow triangular-mucronate; the pinnule next to the rachis on the lowest pinnae only slightly reduced, sometimes pinnatifid. Sori typically submarginal; indusia round-reniform, whitish tan and faintly pink at center, slightly convex. Dryopteris purpurella has been reported as having both triploid and tetra- ploid races, both apogamous (Hirabayashi, 1974). It is native to Japan. It is an attractive, medium-sized fern that grows with restraint and has an interesting color in the new foliage. Only a few plants exist in U.S. and have recently been introduced from Yakushima, Japan, by the senior author, The purplish stipe and rachis, the 2-pinnate flatter fronds with ca. 6 pinnate pinnae well spaced apart, the proximal pinnae at or nearly at a right angle to the rachis, and the submarginal sori help to discriminate D. purpurella. According to several Japanese botanists, some of the U.S. cultivated material sold as D. purpurella is not that species, although the stipe and rachis may initially be pinkish. In this U.S. material, the larger fronds are basally 2 pin- nate-pinnatifid to almost 3 pinnate, with ca. 10 pinnae (contrast with descrip- tion above) that are broader and closer together, and the basiscopic pinnule HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 85 Sf Ne Yo’ ai ws WC / v my = aan Ayevae <#) : Fic. 47. Dryopteris purpurella. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Basal pinna [scale=1cm]. D) Pinnule from medial pinna [scale=5mm]. 86 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) next to the rachis is shorter than in typical D. purpurella. Young fronds of such plants do look somewhat like D. purpurella, but older fronds are too foliaceous and the pinnae are markedly crowded. They have been identified as D. erythrosora, the variability of which is discussed above under that spe- cies’ treatment. Dryopteris purpurella is hardy to a January average of above 40°F, is more or less evergreen, and has new growth that is purplish pink to bronze. Section 3.2: Variae Fraser-Jenk. Fronds markedly stiff, coriaceous, the pinnules often with caudate apices and pointed lobes, the basiscopic pinnule of the basal pinnae noticeably longer than adjacent pinnules, the stipe scales all narrow and the costa and costules with slightly bullate-based scales (vs. more bullate scales of Section Erythro- variae). KEY AND DESCRIPTION TO SECTION VARIAE 1. Blade usually pentagonal with conspicuously elongate, basal basiscopic pinnules, segment teeth short-aristate, young rachis not mottled with dark and light colored scales ee eo aipowce wa ce ace Naaera noni Ue tit ois, Oe a oc 47. D. formosana 1. Blade triangular or weakly pentagonal, basal basiscopic pinnule not conspicuously elongate, segment teeth absent or rarely aristate, young rachis mottled with light and dark colored scales 2. Blade gray-green, matt or dull, thickish, somewhat stiff 3. Frond to ca. 90 cm long, blade thick, stiff and rough textured, typically abruptly narrowed before tapering to the apex, scales on stipe and rachis ascending ....... 50. D. varia 3. Frond to ca. 40 cm long, blade firm, gradually tapered to the apex, scales on stipe and rachis spreading of recuitving |... 666. eS 49. D. saxifraga 2. Blade dark green, slightly glossy, thin leathery, flexible 4. Basal stipe scales with pale margins ..................05.s 48. D. sacrosancta 4. Basal stipe scales without pale margins ...................... 46. D. bissetiana 46. Dryopteris bissetiana (Baker) C. Chr. (Index Filicum 245. 1905).—Beaded wood fern.—Fig. 48. D. varia (L.) Kuntze var. setosa (Thunb.) Ohwi Rhizome short-creeping to ascending or erect, sometimes forming offshoots. Stipes to ca. 55 cm long, the larger stipe scales mostly black with or without a faint narrow pale margin, very narrow triangular, base more or less cordate and pale, the medium to smaller scales stouter, often tan, their margins often fimbriate or sometimes irregularly dentate-erose; blade mostly 2 pinnate, to 3- pinnate at the base in larger older fronds, to ca. 55 cm long, 32 cm wide, glossy, dark green, texture firm, medium leathery, the pinnules narrow triangular, fal- cate, the margin incised-serrate to crenate-serrate, slightly reflexed. Sori sub- marginal to medial, many ultimate segments or lobes each bearing 1 sorus; indusia at maturity greenish, round-reniform. Dryopteris bissetiana is an apogamous triploid species native to eastern Asia. The somewhat blunt lobes with recurved margins and slightly embossed ad- axial surface gives pinnules of this species a bead-like look, hence the common HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 87 Fic. 48. Dryopteris bissetiana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. name, beaded wood fern. Some plants currently circulating as D. bissetiana in the trade are actually a variation of D. erythrosora (recognized by the _ rather than long basiscopic pinnules of D. bissetiana). Other presently - - vated plants were grown from spores that were collected in a fruit orchar 88 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) near Guilin, China, by the senior author. The longer stipes give the plant a more open look than others of this group. Dryopteris hikonensis (H. It6) Kurata (D. pacifica (Nakaike) Tagawa), although presently rarely cultivated, may be confused with the closely related D. biessetiana. The blades of D. hikonensis abruptly narrow near the apex, and its segments are minutely dentate, whereas blades of D. bissetiana are gradually narrowed toward the apex and its seg- ments are nearly entire. Dryopteris bissetiana is a medium-sized, evergreen fern with dark green fo- liage and a bead-like texture to the frond. It is hardy to a January average of ca. 25—30°F. 47. Dryopteris formosana (Christ) C. Chr. (Index Filicum 266. 1905).— ig. 49 Rhizome erect-ascending to short creeping, branching to form a few crowns. Stipes to ca. 30 cm long, the basal stipe scales black-brown, a pale margin inconsistently present, narrowly triangular, the larger ones to 15 mm long, base slightly cordate to truncate, distal stipe scales narrowly lanceolate, black, slightly irregularly dentate, the costa scales long lanceolate or bullate, the bul- late scales often with the narrowed part dark; blade broadly pentagonal, to 3 pinnate, the basal pinnae with the basiscopic side larger, the pinnule next to the rachis often noticeably longer than adjacent pinnules, the larger pinnules often slightly auriculate, more so on the distal side, the margins of the pinnules and segments mostly serrate-aristate, the segments oblong to narrowly ovate, rounded to acute, often with two teeth at the apex. Sori medial; indusia round- reniform. Dryopteris formosana is an apogamous triploid species from Japan and Tai- wan. The fairly consistent, broad, pentagonal frond is a helpful identifying feature for this species, along with the relatively long basiscopic pinnule next to the rachis. These and other features indicate that this species is not closely related to the other three species treated in this section. This species is a medium-sized, nearly evergreen fern with erect-arching growth and broad, finely divided fronds. It is hardy to January averages of 35°F or lower. Dryopteris formosana is of restrained growth and is easily cultivated. 48. Dryopteris sacrosancta Koidz. (Bot. Mag. (Tokyo) 38:108. 1924).—Fig. 50. D. varia (L.) Kuntze var. sacrosancta (Koidz.) Ohwi Rhizome ascending to erect, occasionally producing offshoots. Stipes to Ca. 40 cm long, the larger basal stipe scales very narrow triangular, ca. 10 mm long, shiny black with a very narrow pale margin, margins more or less den- tate, the stipe scales above weakly cordate, lighter colored at the base, clath- rate, the cells oriented in swirls; blade narrowly triangular, to ca. 50 cm long, 3-pinnate, somewhat glossy, green, the margins quite flat, the pinnae tending to overlap, the larger segments subentire to weakly and irregularly serrate to serrate-lobed. Sori submarginal; indusia reniform. Dryopteris sacrosancta is an apogamous triploid species from eastern Asia. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 89 Ai id Fic. 49. Dryopteris formosana. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. C) Pinnule from medial pinna [scale=5mm]. The larger, flatter blade and margins help to distinguish it from D. bissetiana, which has a smaller blade with the divisions positioned in slightly different planes and the margins reflexed. The adaxial blade surface of D. bissetiana is often slightly embossed on the surface, and the plant has a more open habit AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) iN f lJ Fic. 50. Dryopteris sacrosancta. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 91 Zo: \ ; Vy, ; ee Vy tri j — | ee 18 L | 4 Fic. 51. Dryopteris saxifraga. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. mainly, due to the longer, more erect stipes. Plants in the trade have been grown from spores taken from a plant collected in 1982 at Toho University, Chiba, Japan, by the senior author. This species is hardy to a January average of 30°F. It is evergreen and slower- growing than D. bissetiana. 49. Dryopteris saxifraga H. It6 (Bot. Mag. (Tokyo) 50:125. 1936).—Fig. 51. Polystichum varium (L.) C. Presl, misapplied Rhizome ascending to erect, branching to form offshoots. Fronds 35-40 cm long, 10-15 cm broad; stipes ca. half the length of the blade; stipe scales black- brown, to 2 cm long, larger scales long, narrowly triangular with a narrow pale margin, long-tapering to a hair-tip, base pale, cordate, scales on distal part of stipe becoming paler and broader at their base; blade ovate-triangular, 2-pin- nate-pinnatifid, smaller pinnules with margins slightly turned under, sinuate, the rachis scales mixed, larger ones ovate-triangular, their bases hardly to slightly pouched, the apices long tapered, hair-like, dark, the smaller scales short-ovate, bullate, light tan, with hair-like apices, the costa scales half the size of rachis scales, roundish ovate, very bullate. Sori in 2 rows on pinnules, medial or closer to midrib; indusium ca. 1 mm in diameter. 92 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) Dryopteris saxifraga is a sexual diploid fern native to Japan, Korea, and Manchuria, where it grows among rocks in high mountains. It is similar to D. bissetiana in often being slightly embossed on the adaxial surface of the blade and with recurved margins, but differs in being a smaller, stouter plant with shorter stipes with browner scales, pinnae closely placed rather than distant, dull rather than glossy, and medium green rather than dark green in color. This species is easily cultured and is evergreen in southern California, and is hardy to a January average of ca. 10°F. 50. Dryopteris varia (L.) Kuntze (Rev. Gen. Pl. 2:814. 1891).—Fig. 52. Polystichum varium (L.) C. Pres Rhizome ascending to very short creeping. Stipes to ca. 45 cm long, the larger stipe scales dark brown-black, often paler toward the base, cordate; blade more or less erect, stiffish, mostly oblong triangular, to ca. 45 cm long, 43 cm wide, 2-pinnate-pinnatifid to 3-pinnate at the base, proximal pinnae more developed on the basiscopic side (on older plants the blades texture thick and hard, somewhat abruptly narrowed and then tapering to the apex and dull gray-green in summer), the segment margins slightly turned under, weakly serrate, teeth mostly acute, rarely aristate, the rachis scales linear-triangular, the costa scales linear lanceolate, many dilated at the base, their base flat to very slightly convex. Sori medial to mostly submarginal; indusia round reni- form, ca. 1.5 mm in diameter, margins with sparsely distributed minute hairs. Dryopteris varia has been reported to consist of both diploid and triploid, apogamous races (Hirabayashi 1974; Fraser-Jenkins; 1989). It is native to north- eastern India (rare) and eastern Asia. In southern California the foliage is bron- zish when emerging and somewhat gray-green when old and stiff. The overall thicker stiff texture and triangular to oblong pentagonal frond are useful for overall identification. The abrupt narrowing of the apex before tapering to the blade tip is not always strikingly apparent on all fronds. Of more definitive help are the flatter bullate scales which may have a noticeable long apex, and the thick, rougher texture of the frond. This species was known in cultivation in southern California in 1970s. It was grown from spores collected in Japan. This species is hardy to a January average of ca. 35°F. It is more or less evergreen, has sparse fronds, the emerging ones yellow to rusty bronze. It is of slow growth but not difficult to cultivate. ADDENDA The following taxa of Dryopteris have not been treated in the preceding text. These have recently been reported by commercial growers and hobbyists, but have not been confirmed or are very recent introductions. Should the reader desire more details beyond those we have given here, we refer you to the description of the Sections and then also to appropriate floras. Most of the species from the Sino-Himalayan area were introduced into cultivation by Christopher Fraser-Jenkins and are described in Fraser-Jenkins (1989). Japa- HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION & aerate) Fic. 52. Dryopteris varia. A) Frond [scale=5cm]. B) Stipe scales [scale=5mm]. 94 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) nese fern species are described in English, without illustrations, in Iwatsuki et. al (1995). There are several recent illustrated publications on Japanese ferns, and although the text in these is in Japanese, the scientific names are usually given in Latin. We are most familiar with the eight-volume work of Kurata and Nakaike that has names and indices in Latin. Volume I (1979) and Volume IV (1985) contain the Dryopteris species, hybrids in Dryopteris are included in Volume VII (1994), and Volume 8 (1997) has additional distribu- tion maps and supplementary information for taxa covered in earlier volumes. Dryopteris blandfordii (C. Hope) C. Chr. (Index Filicum 254. 1905).—Plants listed by this name were not available for verification. The name represents a large fern with fronds to ca. 90 cm tall. Mid-size plants look like D. filix-mas, but are more lobed on proximal pinnules, whereas larger plants may look like D. stewartii, but with darker stipe scales. A native of western Himalaya, Tibet, and China, the species is in section Remotae. Dryopteris xboottii (Tuck.) Underw. (Native Ferns, edition 4, 117. 1893).— Plants listed by this name were not available for verification. The name rep- resents a hybrid between D. intermedia (section Lophidium) and D. cristata (section Pandae). The hybrid is native to the northeastern U.S. Dryopteris buschiana Fomin (Flora Sibiriae et Orientalis Extremi 5:52. 1930).—Plants listed under this name were not available for study and the name is of uncertain application. “Dryopteris claytoniana,” Hort—Plants listed by this name were not available for study, and valid publication of this epithet in the genus Dryopteris could not be verified. The name is probably a misspelling of Dryopteris clintoniana (which see). “Dryopteris coreano ssp. montana,” Hort.—Plants listed by this name were not available for study. The name is probably a misspelling for Dryopteris coreano-montana, a synonym of D. sichotensis (which see). Dryopteris fructuosa (Christ) C. Chr. (Index Filicum 267. 1905).—Plants listed by this name were not available for verification. The name represents a variable fern with markedly leathery, glossy, dark green fronds to ca. 100 cm long, 2- pinnate to 2-pinnate-pinnatifid at the base. This native of the Sino-Himalayan region belongs to the section Pallidae. Dryopteris guanchica Gibby & Jermy (Bot. J. Linn. Soc. 74:258. 1977).—Plants listed by this name were not available for verification. The name represents a plant with concolorous rhizome scales and proximal pinnae with the basal acroscopic pinnules usually shorter than the next. This native of the Canary Islands belongs to the section Lophidium. Dryopteris koidzumiana Tagawa (Acta Phytotax. Geobot. 2:190. 1993).—This name refers to an evergreen fern similar to D. erythrosora with few scales, narrower pinnules with dentate margins, incurved teeth, and larger sori. The attractive red new fronds last longer than in D. erythrosora. Plants were intro- HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 95 duced from Yakushima, Japan, by the senior author and spores were distrib- uted to growers. This native of Japan belongs to section Erythrovariae. “Dryopteris megalodus,” Hort.—Plants listed by this name were not available for study and the name, which could not be verified as validly published, is of uncertain application. Dryopteris munchii A.R. Sm. (Proc. Calif. Acad. Sci., ser. 4, 40:218. 1975).— Plants listed by this name were not available for verification. The name rep- resents a fern with triangular fronds to ca. 100 cm long, nearly 3-pinnate; the stipe and rhizome scales are tan with dark heavy streaks. It is native to Chia- pas, Mexico, and belongs to the section Cinnamomeae Fraser-Jenk. (not treated or keyed above), which is characterized by having fronds 1- to nearly 3-pin- nate, lanceolate to narrowly triangular, thinly herbaceous, brittle, and the pin- nules markedly obliquely sloping. Dryopteris namegatae (Sa. Kurata) Sa. Kurata (J. Geobot. 17:89. 1969).—Plants listed by this name were not available for verification. The name represents a fern native to Japan and China that may be a hybrid between D. cycadina (D. atrata) and D. dickinsii (see treatment of D. kuratae), both belonging to section Hirtipedes. Dryopteris odontoloma (Bedd.) C. Chr. (Acta Horti Gothob. 1:59. 1924).— Plants listed by this name were not available for verification. The name rep- resents a fern with fronds to ca. 65 cm long, elongate triangular-lanceolate, 2- pinnate, with the pinnae markedly cordate at their base and small submarginal sori (to 1 mm diameter). The species is in section Pallidae. Dryopteris pacifica (Nakai) Tagawa (Coloured Illustrations of the Japanese Pteridophyta, 100, 211; plate 36, figure. 204. 1959).—Dryopteris varia (L.) Kuntze var. hikonensis (H. It6) Sa. Kurata.—This species is currently being grown from spores by commercial and amateur growers. It is an evergreen, medium-size fern with dark green, glossy, triangular fronds to 3-pinnate and is classified in section Variae. The stipe scales are usually all blackish brown or black, the segment margins are flat, and the pinnae generally do not overlap. The indusium margin is sometimes ciliate. It is similar to D. bissetiana, but the latter has reflexed segment margins. Dryopteris pacifica maybe confused with D. sacrosancta, but the latter has stipe scales with pale margins, lighter green fronds that are dull or hardly shiny, pinnae that tend to overlap, and indusia with entire margins. Dryopteris pacifica is evergreen and reportedly hardy in USDA zone 6 (average annual minimum temperature 0 to —10°F). It tends to be a more compact growing plant than D. sacrosancta. Some of the plants from which spores have been distributed in the U.S. were collected in Yakushima, Japan, by the senior au- thor. The species is native to Japan, Korea and China. In Japan, it grows in areas with a minimum January average temperature of 30°F or warmer. Dryopteris paleacea (T. Moore) Hand.-Mazz. (Verh. K. K. Zool.-Bot. Ges. Wien 96 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) 58:100. 1908).—Plants listed under this name were not available for verifica- tion. The name is a synonym for D. wallichiana (section Fibrillosae) and plants in the trade may represent that species (which see). Dryopteris sordidipes Tagawa (Acta Phytotax. Geobot. 3:29. 1934).—This is an evergreen ca. 50-90 cm tall and reminiscent of D. dilatata except firmer and more coarsely cut. About 20 years ago, the name appeared in a Chicago catalog. This listing was not verified. The senior author made a recent introduction from Yakushima, Japan, and spores have been distributed to growers. The spe- cies is native to Japan and Taiwan and belongs to the section Variae. Dryopteris yigongensis Ching in C.Y. Wu (Fl. Xizangica 1:253. 1983).—Plants listed by this name were not available for verification. The name represents a fern with glossy fronds to ca. 50 cm long, narrow triangular-lanceolate, to 2- pinnate at the base and with long stipes bearing black, glossy scales. A native of the Sino-Himalayan area, it belongs to the section Fibrillosae. Dryopteris villarii (Bell.) Woyn. ex Schinz & Thell. (Vierteljahrsschr. Naturf. Ges. Ziirich 60:339. 1915).—Plants listed by this name were not available for verification. Cultivated materials may represent D. mindshelkensis (which see), a tetraploid that has D. villarii as one putative parent. The species belongs to the section Pallidae. ACKNOWLEDGMENTS We are immensely grateful for the help of so many people during this project. Translators, — ieee eins growers, and scientists were most generous with their help. Japanese bot- anists, Norio Sahashi, Toshiyuki Jereeow nine Heri Hirsbayashi have been of invaluable assis- tance Pease a number of years in t d by providing us with much of the literature otherwise not available to us. In the summer of 1993, we had the privilege of joining many other Japanese botanists in the field in southern Japan, where we were made familiar with many of the native fern species, including a large number of Dryopteris. We are indebted to these botanists, particularly Mituyasu Hasebe, Noriaki Murakami, and Shigeo Ma- suyama. We are especially grateful to Keisuke Yasuda for his patience and good humor in pointing out to us the Dryopteris species and in later sending us specimens of species that we had not been able to see in the field. Wu Su-gong of China was able to confirm the identity of a particularly troublesome species. Rose Murphy and Christopher Page of England kindly shared with us their expertise on the British species. We would also like to thank Alan R. Smith, University California at Berkeley, for allowing us the use of the herbarium facilities. The herbarium of the New a Botanical Garden kindly pro- vided us with the loan of some American and European species. We acknow ledge, also with gratitude, the use of the facilities of the herbaria of the Galvani) of California, Los Angeles, California State University, Northridge, the Natural History Museum of Los Angeles County, and of the Missouri Botanical Garden. We relied heavily on the publications and the personal counsel of Christopher Fraser-Jenkins. He has always been available to identify specimens that puzzled us. He has also reviewed our manuscript and suggested revisions and corrections; we are greatly indebted to his efforts. Any errors in this publication, however, are entirely our ow We wish to express our gratitude to Raymond Tom 2 California, who translated the Chinese literature, and the late Nami Hoshizaki and Kaoru Wood of California, and Noriaki Murakami of Tokyo, who helped us with translations from the Japanese. Figure 22 was adapted and reprinted HOSHIZAKI & WILSON: DRYOPTERIS IN CULTIVATION 97 with kind phere from Kurata and Nakaike’s 1979 first volume of their Illustrations of Pteri- dophytes of Jap There were seciatlalisae from many growers that made this work possible. Carl Starker of Washington. We are particularly appreciative of Judith Jones of Fancy Fronds, Washington, for providing both live plants and herbarium material throughout this study. Naud Burnett of Casa tei Texas, kindly provided assistance in publication. gratefully acknowledge the assistance of Robbin Moran of the New York Botanical Garden, and retina Halley, Bob Halley, and Phyllis Bates of the San Diego Fern Society, who reviewed this paper and provided us with corrections and many helpful editorial suggestions. LITERATURE CITED DrRueERY, C. T. 1910. British ferns and their varieties. Routledge and Sons Ltd., London. Dyer, A. 1996. What should we do about Dryopteris affinis? Pteridologist 3:25—-27. gg C. R. 1976. Dryopteris caucasica, and the cytology of its hybrids. Fern Gaz. 11: 263-267. ae Dryopteris in Spain, Portugal, and Macaronesia. Bol. Soc. Brot. 55: 175-335. . 1986. A classification " boi — Dryopteris (Pteridophyta: Dry ). Bull. British Mus. (Nat. Hist.), Bot. 14 19 monograph ot preieieet a Dryopteridaceae) in the Indian subcon- Hnane Bull Brit. Mus. (Nat. Hist.), Bot. 1 reaffirmation of the oe sno! of Dryopteris affinis (Dryopteridaceae: Peele. Fern Gaz. 15:77-81. FRASER-JENKINS, C. R., and H. V. CorLEY. 1972. Dryopteris caucasica—an ancestral diploid in the male fern aggregate. Brit. Fern Gaz. 10:221-231. Pants -JENKINS, C. R., and M. Gipsy. 1980. Two new hybrids in the Dryopteris villarsii aggregate (Pteridophyta, a and the origin of D. submontana. Candollea 35:305—310. eager oy C. R., and A. C. Jermy. 1977. Nomenclatural notes on Dryopteris: 2. Fern Gaz. 11: 8-340. ae Me 1977. The origin of Dryopteris campyloptera. Canad. J. Bot. 55:1419-142 . 1985. Cytological observations on Indian subcontinent and Chinese ° Dropters and Polys- tichutn ar aide hi Dryopteridaceae). Bull. Brit. Mus. (Nat. Hist.), Bot. 14:1-42. Gipsy, M., and S. WALKER. 1977. Further pdneac nig studies and a reappraisal a the diploid siibaekey in the ipcapieets carthusiana complex. Fern Gaz. 11: 315-32 HIRABAYASHI, H. 1974. AS i Seay studies on Dryopteris of Japan. Biedioto Co., Tokyo. ome, G., and B. A. THomas. 1996. Welsh ferns, edition 7. National Museum and Galleries Wales, Cardiff bewniate: K., T. YAMAZAKI, D. BOUFFORD, and H. Onsa. 1995. Flora of Japan. Volume I. Pteri- dophyta and Gymnospermae. Kodansha Ltd., Tokyo JERMY, C. 1996. What should we do about Dryopteris affinis? A response by Clive Jermy. Pteri- dologist 3:27-28. JERMy, C., and J. CAMus. 1991. The illustrated field guide to ferns and allied plants of the British Isles. Natural History Museum Publications, London KuraTA, S., and T. NAKAIKE, eds. 1979. Phawaions of pteridophytes of Japan, volume I. University of Tokyo Press, Tokyo . 1985. Mlustrations of pteridophytes of Japan, volume IV. University of Tokyo Press, Tokyo. ieee Illustrations of pteridophytes of Japan, volume VII. University of Tokyo Press, Tokyo. a fiber ot of pteridophytes of Japan, volume VII. University of Tokyo Press, Tokyo. eee © ae Problems of cytology and evolution in the Pteridophyta. Cambridge University Press, baa MickEL, J. T. 1994. Ferns for American gardens. MacMillan, New York. 98 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 1 (1999) MONTGOMERY, 4 D. 1982. Dryopteris in North America, part II. The hybrids. Fiddlehead Forum 9(4):23-3 MONTGOMERY, h D., and E. M. PAULTON. 1981. Dryopteris in North America. Fiddlehead Forum 8(4):25-3 NAKAIKE, T. seh New flora of Japan, Pteridophyta, revised and enlarged. Shibundo Co., Tokyo. PiccotT, A. 1997. Morphotypes of the “Dryopteris affinis’’ co mplex i in Britain and Ireland. Affinis 199 WOLLASTON, G. B. 1915. Three species of Lastrea Filixunds: Brit. Fern Gaz. 3:20-24. INDEX TO VOLUME 89, NUMBER 1 99 Index to Taxa (Taxa treated as accepted names have page numbers in boldface; taxa treated as synonyms have page numbers in italics; taxa treated as part of discussions have page numbers in the normal font.) Aspidium Dryopteris (continued) — 63 eiasae det 73, 76 (Fig. 41) Dry darjeelingensis, tanta 14, 41, - sce, "16 rh be aemula, 58, 61 (Fig. 32) dickins affinis, 14, 17, 18, 23, 38, 39, 44 (Fig. 6) panies "6, < eg 96 (Fig. 37) ‘Congesta Cristata,’ 18 ‘Crispa Whiteside,’ 66 ‘Congesta Crispa,” 18 “Cristata,’ “Crispa Congest, ois ‘Grandiceps,’ 66 ‘Crispa,’ 18 ‘Jimmy Dyce,’ 69 ‘Crispa. Gracilis,’ 18 ‘Lepidota Cristata,’ 69 Cristata,’ 18 ‘Recurved Form,’ 69 ‘Cristata Angustata,’ 18 erythrosora, 73, 16, 77, 79, 81, 84, 86, 88, “Grandiceps,’ 1 ; 43) ‘Pinderi,’ 18 i sane 77, 79 ‘Polydactyla,’ 18 f. viridos ‘Revolvens,’ 18 var. eens. 73 ‘Stableri,’ 18 var. dilatata, 73 ‘Stableri Crisped,’ 18 var. purpuresce ssp. affinis, 16, 17, 18, 46 expansa, 46, 63, ee 69 (Fig 38) ssp. seta os 17,18, 25 filix-ans. 16-18, 35, 30; 29, “42, 94 (Fig. 18) . cam si ‘Barnesii,’ 38 amurensis, a, a Ge 33) ‘Crispa Cristata,’ 39 arguta ‘Cristata, assimilis, ‘Cristata Martindale,’ 39 al f ‘Decomposita,’ austriaca, 66, 69 "Grandiceps, 39 azorica, 66, 72 ‘Linearis, bissetiana, 77, 86, 87, 88, 91, 95 ‘Linearis Congesta,’ 39 ig. 48) ‘Linearis Cristata,’ 39 blandfordii, 94 ‘Linearis Polydactyla,’ 39 x boottii, 94 ‘Polydactyla,’ 39 buschiana, 9: : o-cristata,’ 39 campyloptera, 63, 66, 69, 71 (Fig. 34) ‘Robust,” 17 carthusiana, 63, 64, 66, 71 (Figs. 35, 36) ‘Undulata a 39 caucasica, 35, (Fig. 17) formosana, 88 (Fig. 49) celsa, 27 (Fig. 12) fragrans, 39, 41 Fig. 19) championii, (Fig. 40) var. remotiuscula, 41 claytoniana fructuosa, 94 clinoniana, 27, 30, 41, 94 (Fig. 13) fuscipes, 79 (Fig. 44) ommixta, 10 gamblei, poemaniee 17, 38, 39 goeringiana coreano goldiana, 27, ee (Fig. 20) ssp. montana, 94 guanchica, 94 coreano-montana, 44, 45, 94 gymnosora, 81 (Fig. 45) crassirhizoma, .: 20 (Fig. 7) hangchowensis, 12 var. setosa hikoensis, 88 cristata, 27, 30, ee 94 (Fig. 14) hirtipes, ie 9, a (Fig. 3) ssp. a cycadina, 7, 9, 10, 95 (Fig. 2) 100 AMERICAN FERN JOURNAL: VOLUME 89, NUMBER 1 (1999) Dryopteris (continued) Dryopteris (continued) hondoensis, 77, 81, 84 (Fig. 46) ~— Hirtipedes, 7, 12 rubisora, 84 Section Lophodium, 58, 61, 94 indusiain Sie Pallidae, 46, a 94, 95, 96 Section ee losemedin: 63, 4. e 8, 70, 94 (Fig. 39) Section Rem ig juxtaposita, 48, 51 (Fig. 25) Section Cetus 6, 86, 95, aaa 9 sichotensis, 44, 94 (Fi hes e sp. nov., 10, 12, 95 (Fig. 4) sieboldii, 5, 7 (Fig. hy ce, 51 (Fig. 26) sordidipes, 96 idopoda, 20 (Fig. 8) spinulosa, 63 Ri 27, 30 (Fig. 15) var. americana, 63 mar _— 51 (Fig. 27) var. intermedia, 69 megal stenolepis, 9 redial 54, 55, 96 (Fig. 28) stewartii, 55, 66, 94 (Fig. 29) munchii, Subgenus Dryopteris, 7 namegatae, 10, 95 Subgenus Erythrovariae, 4, 72 nigra, 20 Subgenus Pycnopteris, 5 — 73 sublacera, age ey (Fig. 30) toloma submonta 55 isi 38, 41, 44 (Fig. 21) yoens 3 yi 16) “Crispata,’ 44 undulata, ‘Cristata,’ 44 sor 3 aed 31) ‘Incisa Crispa,’ 44 ‘Cris : pacifica, 88, 95 pide ra perma crispata, 58 p var pata, ssp. pallida, 55 varia, 92 (Fig. 52) arallelogra var, sacrosancta, 88 ses 20, 23 so 9) var, hikoensis, 95 pseudo-erythrosora, 73 var. setosa, 86 wove Hisaiak ¢ 16, 20, 23 (Fig. 10) villarii, 55, pseudomas, 14 ssp. submontana, 54 purpurella, 77, " . a 47) wallichiana, 20, 25, 96 (Fig. 11) pycnopteroides, yigongensis, 96 mota, 46 (Fig. fy strea sacrosancta, 88, 95 (Fig. 50) filix-mas, saxifraga, 91, 92 (Fig. 51) propinqua, 4] scottii, 12 (Fig. 5) rigida, 55 ction Aemulae, 58 — gymnosorum, 8] Pscm varium, 91, 92 Thely elypte Section Fibrillosae, 12, 96 plana 63 3 ii 1753 0 863 INFORMATION FOR AUTHORS Authors are encouraged to submit manuscripts pertinent to pteridology for pub- lication in the American Fern Journal. 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Montgomery, Ecology III, Inc., R.D. 1, Box 1795, Berwick, PA 18603. VISIT THE AMERICAN FERN SOCIETY’S WORLD WIDE WEB HOMEPAGE: http://www. visuallink.net/fern AMERICAN Volume 89 FERN Number 2 April-June 1999 JOURNAL QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Some Commercial Uses of Pteridophytes in Central America Barry A. Thomas Genet Composition of Diphasiastrum complanatum in Western Hungary: a Case Study Agnes Major and Péter Odor Spore — and Sterilization Sa Germination and Early seas Development of Platycerium bifurc rjana Camloh Isoetes in Alaska and the Aleutians Donald M. Britton, Daniel F. Brunton, and Stephen S. Talbot Spore Germination and Early Gametophyte Development in Stromatopteris ean P. Whittier and Jean-Christophe Pintaud Effects of Temp Spore Germination in Some Fern Species from Semideciduous cities Forest Marli A, Ranal Studies on Cryptogramma crispa Spore Germination milia Pangua, Lorena Garcia-Alvarez, and Santiago Pajarén SEM Studies on Vessels in Ferns. 13. Nephrolepis Edward L. Schneider and Sherwin Carlquist Review: The Feciil and Allied Plants of New England Ferns of the Tropics 101 106 124 133 142 149 171 The American Fern Society Council for 1999 DIANA B. STEIN, Dept. of Biological Sciences, Mount Holyoke College, South Hadley, MA 01075-6418 President BARBARA JOE HOSHIZAKI, 557 N. Westmoreland Ave., Los Angeles, CA pirouette Vice-President W. CARL TAYLOR, 800 W. Wells St., Milwaukee Public Museum, Milwaukee, WI 53233- Bs Aah ecretary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN sea tom Treasurer DAVID B. LELLINGER, 326 West St. NW., Vienna, VA 22180-4151. Membership Secretary JAMES D. 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BOTANICAL American Fern Journal 89(2):101—105 (1999) g 1999 JUL2 Some Commercial Uses of Pteridophytes caw“ in Central America Barry A. THOMAS Geography Department, University of Wales Lampeter, Lampeter, Ceredigion SA48 7ED, United Kingdom ABSTRACT.—Leatherleaf fern is grown for the floral trade in at least three countries in Central America. Polypodium leucotomos is also cultivated in Honduras for production of DIFUR for the treatment of skin complaints such as psoriasis. Wild-collection of tree ferns for their fibre appears widespread and a limited number of other pteridophytes are wild-collected for medicinal use. In late 1995, I was fortunate to be awarded a Winston Churchill Travelling Fellowship which enabled me to visit Central America to talk to many people about the relationship between plant conservation, agriculture, and horticul- ture in this botanically rich area. So, while travelling through the eight diverse countries, I was able to spend some studying the commercial production of two species of ferns and the sale and local use of wild-collected pteridophytes. Although I anticipated seeing the leatherleaf fern, Rumohra adiantiformis (Forst.) Ching, I was surprised to see the extent to which it was cultivated in Costa Rica, Guatemala, and Honduras. In these three countries, this fern is locally cultivated over extensive areas, where whole hillsides can be covered with sheeting to shade vast numbers of plants (Figs. 1-3). The areas covered in this way were certainly very much larger than any I have seen in Florida. I saw no leatherleaf fern cultivation in Belize, El Salvador, Nicaragua, and Panama, and all enquiries gave negative replies. However, this cannot be taken to be definitive, because I was also led to believe by several Guatemalan ag- ricultural botanists that no Rumohra was being grown in Guatemala. Even a professional guide that accompanied me had no idea what was being grown under the rather obvious and extensive shading. Stamp (1992) has outlined the propagation techniques of this species used in Florida, but conditions in Central America appear rather better. Land is not a problem and irrigation is only very infrequently necessary. Indeed, drainage seems to be more of a problem, because the rows of plants are separated by shallow ditches. Certainly, cold damage is not a potential problem, as it can be in Florida. Every site I visited had people working on the crop, either weed- ing, harvesting, sorting, or packing the fronds for transportation and ultimate export (Fig. 4). Although I have as yet no detailed export information on des- tinations or quantities, I did discuss them with some producers. I was led to believe that the annual crop was steadily increasing and that it was mainly bound for Europe or North America. In Guatemala City airport, I did watch boxes of fern fronds being loaded into KLM airliners, presumably bound for Amsterdam and then for sale on the international market. This is almost cer- 102 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) in Honduras. 3) Rows of Ruhmora adiantiformis in cultivation near Zamorana, Honduras. 4) Sort- ing and drying the fronds just outside the growing area. 5) Bottled and boxed calaguala from the factory at Tegucigalpa, Honduras. tainly the reason that producers in Florida have experienced a decline in wholesale prices with production costs sometimes exceeding income (Smith et al., 1988). Although it was fascinating to come around a corner or over a hill and see yet more extensive leatherleaf fern cultivation, it was even more exciting to encounter commercial production of a plant locally described as Polypodium leucatomos Poir. (the name is generally regarded as a synonym of Phlebodium aureum (L.) J. Sm., but the locally native species are P. decumanum (Willd.) . Sm. and P. pseudaureum (Cav.) Lell.). Rhizomes of this fern are sold in markets, some pharmacies, and even on the pavements in Guatemala and Hon- duras as a cure for skin complaints, urinary and liver disorders, gastritis, rheu- matism, and arthritis. But in Honduras, ten acres of this fern are commercially grown under sheeting about 200 km north of the capital city, Tegucigalpa. An estimated 2.5 million plants are grown on strips separated by ditches prepared for flooding when the plants are sprayed with water in the summer. Harvesting THOMAS: COMMERCIAL USES OF PTERIDOPHYTES 103 OQ Fics. 6-9. Commercial utilization of pteridophytes in Mexico and Honduras. 6) A “swan” made from tree fern in a garden center in Xico, Veracruz, Mexico. 7) Wild-collected orchids in tree fern pots for sale at a roadside in Guatemala. 8) A wild-collected cycad in a tree fern urn in a garden center in Xico, Veracruz, Mexico. 9) Packaged Equisetum bogotensis hanging for sale in the central market at Tegucigalpa, Honduras. is every three or four months, giving an estimated yield of between one and two million fronds. They are dried at about 50°C, ground up, and taken to the factory at Tegucigalpa for warm water extraction and cleaning with ion ex- change charcoal. The final extract is concentrated at 40°C under vacuum. The concentrated liquid is marketed locally in Central America under the local name for the fern, “calaguala” (Fig. 5). However, it is also sold in capsule form as a prescribed drug in Spain, France, and the U.S.A., where it is marketed under the brand name DIFUR. This drug is claimed to have curative effects for skin complaints such as psoriasis (Azami, 1989; Gonzalez S. et al., 1994). I was horrified to see the extent to which tree fern stems are being used as plant pots in several of the countries (Fig. 6). They were all clearly taken from wild-collected plants, for nowhere did I see any evidence of their commercial growth. The environmental effects of such indiscriminate collecting of tree 104 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) ferns is compounded by their use as containers for other wild-collected plants. In Guatemala, I saw the roadside sale of wild-collected orchids in tree fern pots (Fig. 7) and in the so-called nursery I was shown piles of tree fern stems ready for pot production. In a garden center in Veracruz, Mexico, I found more wild-collected orchids in tree fern pots, but even worse were the large, clearly wild-collected cycads in carved pots of much larger tree ferns (Fig. 8). There is a noticeable difference between the countries in the extent to which plants and plant products are used medicinally. In Guatemala and Honduras, it was comparatively easy to find people selling plants and herbal medicines, but this no doubt reflects the much larger percentages of indigenous peoples still living in these countries. In Belize and Costa Rica, there seems to be a resurgence of interest, but this is often initiated by town dwellers or even foreign nationals living there. Nicaragua still has a largely and artificially ur- banized population, because of the many years of civil war, and much of the economy of Panama is influenced or even controlled by the Panama Canal. In both Guatemala and Honduras, I saw the horsetail, Equisetum bogotense Kunth (Fig. 9), the clubmoss, Selaginella pallescens (C. Presl) Spring, and the ferns, Phlebodium spp., on sale in markets and sometimes even spread out on pavements by roadside hawkers. The large horsetail, Equisteum myriochaetum Schltdl. & Cham., locally called “cola de caballo” or “‘barba de jolote,” is re- putedly good for kidney infections, colic, inflammations, and rheumatism, and as a vaginal wash. Equisetum bogotense is also said to be good for these dis- orders. Selaginella is reputedly good for kidneys, coughs, gripe, and intestinal discomfort. According to the literature, both the local S. pallescens, called locally “doradilla” or “rosa de Jerico,” and S. lepidophylla (Hook. & Grev.) Spring, imported from Mexico, are available. Local books on medicinal plants list a number of other pteridophytes (House et al., 1995; Chinchilla, 1995), although I saw none of these being sold in a recognizable form. They are: Adiantum concinnum Humb. & Bonpl. ex Willd. (culantrillo or pata de sante), A. andicola Liebm., A. capillus-veneris L., A. tenerum Sw., and Asplenium scolopendrium (hoja de ciervo or lagua de ciervo [this temperate species does not occur in Central america and the species used is possibly A. serratum L.]), for fever and bronchitis; Elaphoglossum latifolium (Sw.) J. Sm. (ciervo), for coughing, menstrual pains, and as a blood regulator; Lygodium venustum Sw. and Pteridium aquilinum (L.) Kuhn, for inflammation, muscular pains, and rheumatism. Acrosticum aureum L. rhizome is also said in Belize to be a cure for ‘“‘madness.”’ ADDENDUM I have no idea of the effects that last year’s devastating floods have had on the cultivation of ferns in Honduras. ACKNOWLEDGMENTS I would like to thank Efrain Medina Campo (University of San Carlos, Guatemala City), Manfred Petres (Ministry of Natural Resources, San Jose, Costa Rica), Jorge Mendoza (Helechos Interna- THOMAS: COMMERCIAL USES OF PTERIDOPHYTES 105 tional, E] Picacha, Honduras), and Andrew Vovides (Instituto de Ecologia, Xalapa, Veracruz, Méx- ico) for their valuable help while I was in their respective countries. Finally I must acknowledge with grateful thanks the financial support from the Winston Churchill Memorial Trust, because without this my visit to Central America would have been impossible. LITERATURE CITED AMaAZI, M. 1989. Vitiligo repigmentation with anapsos Polypodium leucatomos. Int. J. Dermatol, 28:479. mags S. V. 1995. Plantas ee ee het edition. Grafa Print S.A., San Jose, Costa Rica. GONZ , P. C. Joust, and M. A, PATHAK. 1994. Polypodium leucotomos extract as an anti- Oudaat agent in the therapy of skin dipories: J. Invest. Dermatol., 102:651. Hous, P. R., C. TORRES, S. LAGOS-WITLE, T. MEJIA, L. OcHoA, and M. Rivas. 1995. Plantas medi- cinalis communes de Honduras. Lopez S. de R. L., Tegucigalpa, Hondur SMITH, S. A., T. G. TAYLOR, and L. L. LOADHOLTZ. 1988. Cost of Sandaisitoli for tisdale ferns in Florida: Staff Paper 342. University of Florida Institute of Food and Agricultural Sciences, Gainesville, FL. Stamps, R. H. 1992. Commercial leatherleaf fern culture. Pp. 243-249 in J. M. Ide, A. C. Jermy, and A. M. Paul. Fern horticulture: past present and future perspectives. Intercept, Andover, Great Britain. American Fern Journal 89(2):106—123 (1999) Genet Composition of Diphasiastrum complanatum in Western Hungary: a Case Study AGNES Major Department of Genetics, Eétvés Lorand University, Muzeum krt. 4/a, 1088 Budapest, Hungary PETER ODOR Department of Plant Taxonomy and Ecology, Eétvés Lorand University, Ludovika tér 2, 1083 Budapest, Hungary ABSTRACT.—The study includes an investigation of the genetic composition of a marginal ground pine Dipplicisisatesien complanatum) population consisting of patches of different size. The genetic analysis was performed on the basis of 15 isozyme loci. The proportion of polymorphic loci was o>) clonal diversity index was D=0.898. Wright’s fixation indices over polymorphic loci were —0.07 (0.063) and —0.026 (0.091) at the ramet and genet-level, respectively. The observed and expected ramet-level and genet-level average heterozygosities were not significantly different in spite of the fact that at some loci genotype numbers significantly differed from Hardy-Weinberg proportions. of intragametophytic selfing. Results clearly showed that in addition to clonal growth, sexual reproduction played a substantial role in the establishment and maintenance of the study popu- lation at the boundary of the species’ distribution, and indicated the importance of microsite conditions. Populations of colonizing and clonal plants are usually genetically depau- perate because of bottlenecks and selfing during colonization, and different forms of vegetative reproduction that overwhelm sexual recruitment (Darling- ton, 1958; Stebbins, 1950). During clone establishment, successfully competing genets occupy all available microhabitats, so populations frequently are as- sumed to originate from only a small number of founder genets. In addition, homosporous ferns and other pteridophytes are apt to undergo intragameto- phytic selfing, further decreasing the genetic variability within the population while genetic divergence among populations can increase. In contrast, gene flow into established populations, recombination, plus secondary (repeated) seedling recruitment can result in increased genotypic variability within pop- ulations. Ellstrand and Roose (1987) compiled a summary of 27 studies that revealed a tendency of 20 clonal plant species to form multiclonal populations of in- termediate diversity and evenness, and most clones proved to be limited to few populations. Eriksson (1993) summarized the state of the art of genet dy- namics in clonal plants, and called attention to the fact that the whole life cycle and genet structure (independent or connected ramets and growth form) must be taken into account to obtain a clear picture of dispersal, recruitment, and the genetic composition and structure of clonal plant populations. More- MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 107 over, a particular species can produce different patterns under different con- ditions, the genet and ramet dynamics can alter in different habitats (Kull, 1995), and the genetic structure of a population partly depends on the history of the site (Eriksson and Bremer, 1993). Therefore, in spite of their extremely long lifespan and considerable size of individual clones, clonal organisms can maintain relatively high genotypic variation that has large differences between populations (Kemperman and Barnes, 1976; Jonsson, 1995; Jonsson et al., 1996). Usually, the role of generative reproduction increases with small scale disturbances (Eriksson, 1989; 1993), and on more stressed sites, where the intragenet competition is lower (Wu et al., 1975). Concerning the Lycopodiaceae, Levin and Crepet (1973) in their earliest study investigated the genetic variation of Lycopodium lucidulum Michx. (Hu- perzia lucidula (Michx.) Trevis.) in 16 New England populations. Individual populations showed little to moderate genetic variation; the proportion of polymorphic loci varied from 5—28%. A low level of variability within the populations frequently was coupled with uniform heterozygosity for the same alleles at some loci, and with regional differentiation. Among the 242 individ- uals of 16 populations, they found only 19 multilocus genotypes; the clonal diversity ranged 0.14-0.77. This observed low diversity was partly explained by the phylogenetically relic state of the species and the disproportionate bal- ance between asexual and sexual reproduction. The clonal growth characteristics of the European clubmoss Lycopodium annotinum L. were investigated thoroughly in Swedish Lapland (Callaghan et al., 1986a, b; Svensson and Callaghan, 1988a, b; Carlsson et al., 1990). They found that the most important factors in the maintenance of the populations were an opportunistic guerilla growth-form, and long survival of the genets. In his comparative study, Oinonen (1967; 1968) found that the guerilla growth- form was not so obviously expressed in L. complanatum (Diphasiastrum com- planatum (L.) Holub), because the annual growth of the horizontal branches proved to be less intensive than that of L. annotinum and L. clavatum L. (Ly- copodiaceae); moreover, the distribution of the vertical branches was more closely packed. More recently, several papers have been published on the mating system and genetic structure of other clonal fern species, or have analysed intraga- metophytic selfing in clonal ferns and in those with subterranean gameto- phytes (McCauley et al., 1985; Soltis and Soltis, 1986; Holsinger, 1987; Soltis and Soltis, 1987; Soltis and Soltis, 1988a; b; c; Soltis et al., 1988a; b; Wolf et al., 1987; 1988). Soltis and Soltis (1990) published fixation indices ranging from —0.590 to 0.672 for lycopods (Lycopodium, Huperzia), genetic diploidy (Soltis and Soltis, 1988a), and variable intragametophytic selfing among pop- ulations. For Diphasiastrum complanatum and D. digitatum (A. Braun) Holub, no intragametophytic selfing was reported in spite of their subterranean ga- metophyte development. The aim of this study was to investigate the importance of the sexual repro- duction, genetic diversity, clonal growth, and intragametophytic selfing within and between the patches of D. complanatum in a population. Diphasiastrum 108 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) complanatum is a very rare, protected species in the Hungarian flora, because it reaches the southern boundary of its distribution in the Carpathian basin. It was an important consideration that the sampling should not damage the rel- atively small population. The simplest method to detect the genet structure of the population was the application of isozyme analysis. The number of genets in the patches can be revealed by this method, and it is possible to estimate their size, to compare the genetical and topographical distances among them, and to give elementary characterization of the genetic structure of the popu- lation. MATERIALS AND METHODS STUDY AREA, PLANT MATERIAL, SAMPLING.—This study was carried out at the western boundary of the Carpathian basin in Hungary near Szentgotthard (““Vendvidék”; latitude 46°53’N, longitude 16°15’E). The bedrock of the area is alluvial drift-boulder, clay, and adobe. The study area is located at 300 m above sea level. The average annual precipitation is approximately 800 mm and the mean annual temperature is 9.1°C. The vegetation is zonal mixed coniferous- deciduous forest (Genisto nervatae-Pinetum [Pécs, 1965]) and extrazonal, probably artificial spruce forests. The soil is illuviated brown forest soil with slight podsolization. In most parts of Hungary the zonal vegetation at this al- titude is Turkey oak wood (Quercetum petraeae-cerris). The appearance of the mixed forest at this low altitude is explained by the vicinity of the Alps and occurrence of nutrient poor acidic soil, as well as by the history of the vege- tation (Odor, 1996). The sampling was carried out in seven patches of D. complanatum in May 1996, for the apices of horizontal branches, and in June 1996, for the young tips of the vertical modules. Voucher specimens are on deposit at the Genetics Department of Eétvés Lorénd University. The patches were very different in size and shape (Fig. 1). The sampling points were allocated along the boundary of the patches except G, N, and P. Patch II was sampled more intensively, as patches II and III were denser, whereas the others were looser. Some parts of patch V seemed to be physically isolated. Patch VI was situated in a 20-30 year old planted spruce wood in which shrub, herb, and moss layers were less developed than the vegetation of the other patches, which belonged to similar old stands of a deciduous-coniferous mixed forest with minor difference among them (Odor, 1996). ISOZYME ANALYsIS.—1) Sample preparation: For the separation of GOT (glu- tamate-oxaloacetate transaminase, E. C. 2.6.1.1), EST (colorimetric esterase, E. C, 3.1.1.1), LAP (leucine aminopeptidase, E. C. 3.4.11.1), PER (peroxidase, E. C. 1.11.1.7), PGI (phosphoglucose isomerase, E. C. 5.3.1.9), and ACP (acidic phosphatase, E. C. 3.1.3.2), 200 mg of the actively growing horizontal branches were ground in a chilled mortar in 1 ml of the following extraction buffer: 0.1 M K-phosphate, pH=7.5, 0.029 M Na-tetraborate, 0.017 M K-metabisulfite, 0.2 M L-ascorbic acid, 0.016 M dithiothreitol, 4% soluble PVP, with 2 g sucrose MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 109 a v Hl. uZ u U - U U U ieee | U M V U r 5m R O al N s L M 5m 5m Vi A B A 5m Vil. 6 E C D 5m IM. I i. a : H 409 om \! ‘ 5m H Fic. 1. Bird’s-eye view of the location of the patches (bottom left), and the location of the sampled ramets in patches I-VII. and 10 pl 2-mercaptoethanol as additives per 10 ml grinding buffer. For the extraction of MDH (malate dehydrogenase, E. C. 1.1.1.37), IDH (isocitrate a hydrogenase, E. C. 1.1.1.42), and SKDH fahikimata dehydrogenase, E. 1.1.1.25), 200 mg of the branches were homogenized in a chilled mortar in - ml of the following grinding buffer: 0.04 M Tris-HCl, pH=7.8, 0.01 M MgCL, 0.001 M EDTA, 0.005 M dithiothreitol, 0.013 M K-metabisulfite, 4% soluble 110 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) PVP, with 2 g sucrose, and 10 pl 2-mercaptoethanol per 10 ml buffer (modified after Soltis et al., 1983). The slurry was transferred to Eppendorf tubes and centrifuged at 6000g for 10 minutes. Aliquots of the supernatant were stored at —80°C until use. Twenty pl of this extract were absorbed onto filter paper wicks that were inserted into the horizontal starch gels. 2) Electrophoresis: Horizontal discontinuous electrophoresis was performed on 11% (ACP, EST, GOT, LAP, PER, PGI) or 11.5% (MDH, IDH, SKDH) starch gels (Connaught starch, 26X12%X0.8 cm) with buffer systems as follows. The ACP, EST, GOT, LAP, PER, and PGI isozymes were separated with Arulsekar and Parfitt’s (1986) “A” system at 130 V constant voltage. The MDH and SKDH isozymes were separated according to the first system of Soltis et al. (1983) at 100 mA constant current until the bromophenol blue marker migrated 7 cm. The IDH isozymes were assayed by both systems. 3) Staining schedules: All the staining procedures were performed on 2mm thick starch slabs according to Soltis et al. (1983), with some modifications. For EST, 0.1 M Tris-malic acid, pH=6.4, buffer was used; for LAP isozymes 0.1 M Tris-0.1 M maleic acid, pH=4.5 (titrated with NaOH); for GOT an ad- ditional 10 mg bovine serum albumin per 50 ml staining buffer were applied; for PER O-dianisidine substrate was included instead of 3-amino-9-ethylcar- bazole. The IDH, SKDH, and PGI systems were stained with the agar-overlaying method in 0.1 M Tris-HCl, pH=8.0, buffer solution with 0.5% final concentra- tion of agar, at 37°C; LAP and PER usually showed acceptable staining at room temperature. Data ANALYSES.—The clonal growth form of D. complanatum may bias the analysis of genetic variation. Therefore, the number of distinct multilocus ge- notypes was conservatively regarded as the number of genetically distinct in- dividuals (genets). Sampled sporophytes were initially regarded as ramets without knowing anything about their relationships. In Table 1, the locus (zone) of the most anodally migrating isozymes is labelled as “1”, and alleles within a zone are designated in the same way. The proportion of polymorphic loci (P) was calculated in the case of isozymes with clear separation and in- terpretable patterns. The allele frequencies at all loci were calculated for ra- met-level and genet-level, and the effective number of alleles (A,) was deter- mined. The latter was calculated as A, = 1/2p,,? for the kth locus, where p,, is the frequency of the ith allele of the kth locus, and averaged over loci as A., and A,, on ramet- and genet-level, respectively. The clonal diversity in the population was characterized with the modified Simpson diversity index (Pie- lou, 1969): D = 1 — & {[n,(n, —1)]/[N(N —1)]}, where n, is the number of the ramets of the ith multilocus genotype and N is the number of the sampled ramets. Furthermore, observed (H,,) and expected genetic diversity (H,, = 1— p,.2) was averaged over all polymorphic loci for both ramet- and genet- level (H,,, H,, and H,<, H,<). Deviation from Hardy-Weinberg equilibrium at each variable locus was tested by chi-square test. Wright’s (1965) fixation in- dices were calculated for each polymorphic locus, and averaged as F, and F, at the ramet-level and genet-level, respectively. MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM Lit TABLE 1. Genotype of genets at the polymorphic loci and the number of ramets in a genet. Genotype Genet Ramet MDH-1 MDH-4 SKDH EST-1 EST-4 PGI-2 PER A 2 12 11 11 22 Ze 33 22 B 1 12 11 11 22 22 22 1] Cc 1 12 11 11 12 22 22 12 D 2 12 1] 23 22 22 Za 12 E 1 12 11 ZS 12 22 23 12 r 1 11 bs 11 22 12 AD. 12 G 1 12 11 11 12 22 23 12 H 2 12 1] 13 12 22 23 12 I 1 12 11 22 12 12 22 12 J 1 12 12 Ze 22 12 23 ee K 2 11 1a | 11 22 12 23 us L 1 22 11 23 22 22 2 11 M 1 22 11 23 22 12 22 12 N ] 12 11 11 22 12 22 12 O 1 22 1 25 22 11 22 11 r 1 22 11 23 12 11 12 12 Q 1 11 11 33 22 11 33 {2 R 2 12 12 23 22 11 23 12 S 2 11 1] 23 22 11 22 11 U 12 12 11 33 22 11 ee 22 Vv 1 12 11 22 22 11 23 22 For further data analyses multivariate methods were used on the basis of the genet multilocus genotype distribution (Table 1). For the ordination of the genets, principal coordinates analysis (PCoA, [Gower, 1966]) was carried out using the chord-distance function of Orléci (1975) and Euclidean-distance function of Podani (1980). All calculations were performed by the SYN-TAX program package of Podani (1993). Geographical evenness of the genetic composition of the genets was inves- tigated by spatial autocorrelation analysis (Sokal and Oden, 1978a, b). Warten- berg’s SAAP version 4.3 (1989) Spatial Autocorrelation Analysis Program was used to calculate Moran’s I coefficient and its statistical significance. Moran’s I is a special type of the Pearson product-moment correlation coefficient that describes the deviation of the neighboring points from the mean of all obser- vations. Values of I significantly larger than the expected value [E(I) = —(n—1)-*] show that the points (individuals) in that distance class are more similar than would be expected by chance, and values significantly lower than E(I) show that individuals are less similar. In both cases some factors are af- fecting the distribution and not chance alone. All univariate autocorrelograms and the average autocorrelogram were analysed for different sets of distance classes (different numbers of distance classes of equal size or with equal num- bers of point pairs). 412 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) RESULTS VARIABILITY OF ISOZYME SYSTEMS.—ACP and LAP enzyme activity could be detected only at a single monomorphic band. GOT isozymes showed a mono- morphic zone with strong activity and occasionally a second, faintly stained, more cathodal zone which was unscorable. EST isozymes migrated into both the cathodal and anodal regions. The bulk of activity appeared in the anodal part where several zones could be distin- guished. The separation and activity of isozymes were interpretable and scor- able in the fastest zone (EST-1), and in the fourth region (EST-4). There ap- peared two putatively allelic isozyme forms in both zones. The three-banded pattern of the putative heterozygotes reflects a dimeric esterase coded by locus EST-4. The inheritance of these latter electromorphs should be tested by di- rected crosses. PGI isozymes were resolved in two zones probably coded by two loci. The activity in the faster region (PGI-1) was monomorphic but frequently faint and hardly visible. In the slower region (PGI-2), there occurred three allelic forms, with three-banded patterns in putative heterozygotes, indicating a dimeric en- zyme (Fig. 2a). IDH isozymes separated by the method of Soltis et al. (1983) appeared in two zones of the anodal part of the gel, and may most likely be coded by two loci. In the more cathodal region there occurred a monomorphic single band with faint intensity (IDH-2). The more anodal region (IDH-1) showed a three- banded pattern (Fig. 2b). We could observe four (a—d) phenotypes in this re- gion with slight differences in the position of the bands. There may be alter- nate interpretations for this region. On one hand, there may be a polymorphic locus with four alleles. The minute differences in the mobility of the bands seem to contradict this interpretation. On the other hand, this could be a du- plicated locus with homozygosity at the individual loci and an interlocus het- erozygote isozyme band. This possibility is indicated by the fact that among the sampled 38 ramets there was not a single individual with the characteristic one-banded homozygote phenotype. According to a third interpretation, the pattern may be produced by a monomorphic enzyme that suffered special modification/degradation processes. We decided on separating these isozymes with the “A” system of Arulsekar and Parfitt (1986) to check the stability of the patterns. In this case, the phenotypes were quite different: there appeared a monomorphic active isozyme followed by two faint ghost bands (Fig 2c). Therefore, IDH-1 was conservatively interpreted as a monomorphic locus pro- ducing multiple banded phenotype originating from secondary modifications or degradation. MDH isozymes separated in the anodal part of the gel in four different zones. Zones “1” and “4” were variable, probably as independent loci (mdh-1 and mdh-4) with two alleles and three-banded heterozygote patterns (Fig. 2d), whereas in zones “2” and “3” monomorphic activity was detected. These latter may be two independent loci (mdh-2 and mdh-3). PER isozymes showed very complex and different patterns in the vertical MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 113 ON oP @' RR ck BS Fic, 2. Patterns of some isozymes of Diphasiastrum complanatum. a) PGI-2 isozymes of genets Q, D, C, F, I, B, M, N, P, and S. b) IDH-1 isozymes separated with system 1 of Soltis et al. (1983); letters a-d show the variants detected only by this system. c) IDH-1 isozymes separated with Arulsekar and Parfitt’s (1986) system A; all the ramets produced the same pattern; light bands of SOD eee dismutase, E. C. 1.15.1.1) isozymes are visible more anodal from the IDH-1 activity. d) MDH isozymes of genets N, O, P, Q, and R. e) PER isozymes of young ge vertical branches of genets A, B, and C. See Table 1 for detailed genotyping at the individual loc green branches and the underground horizontally growing branches. In the underground branches, both the cathodal and the anodal isozymes showed high activity, but in the green vertical branches the activity of the cathodal isozymes decreased, and the resolution was unsatisfactory. The most inten- sively stained anodal region could be interpreted as a putative locus with two allelic forms (Fig. 2e). SKDH activity appeared in a single zone of the anodal part, near the origin. The pattern was interpretable as a locus with three allelic electromorphs. 114 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) BLE 2. F indices of polymorphic loci. (Standard deviations of means are in parentheses). Significant difference between expected and observed genotype numbers. * P < 0.05; ** P < 0.01; *** P < 0.001. Locus Ramet-level: Genet-level: EST-1 —0.102 =0.167 EST-4 —0.102 —O.107 MDH-|1 “UAE —0.238 MDH-4 —0.040 PAP) PER 0.106 —0.145 PGI-2 =0:375" 0.103 0.456*** 0.356*** Means (S.D.) FR = —0.076 (0.063) F, = —0.026 (0.091) Out of the investigated 15 loci 7 proved to be polymorphic giving P = 0.46 as the proportion of polymorphic loci. The effective number of alleles ond standard deviation) was A,, = 1.33 (0.14) and A, = 1.38 (0.15) for ramet- and genet-level, respectively. Because there was no ramet with a homozygous genotype at all loci, no intragametophytic selfing was detected. Among the 38 investigated ramets there were 21 different multilocus genotypes (Table 1, Fig. 1) that could be considered as distinct genets. Simpson’s clonal diversity index was D = 0.898, indicating a high level of clonal diversity: 66.6% (14) of the 21 genets was present as unique ramets in the samples, a further 28.6% (6) was present only in 2 ramets. The genet structure of the individual patches showed substantial variability. The smallest patch I contained a single genet (S), although its ra- mets (24 and 25) showed a minute difference in their IDH-1 pattern, probably by modification or degradation (Fig. 2b). Other smaller patches consisted of different genets: three ramets of patch IV and VI belonged to 2 genets each; four ramets in patch VII were from three genets, five ramets in patch III were from four genets, and the eight ramets of patch V belonged to seven genets. On the contrary, 12 of the 13 ramets in patch II belonged to the same genet, showing the essentially different structure of that patch. Genetic diversity was calculated on the ramet- and genet-level. Ramet-level average observed heterozygosity was H,, = 0.174 (0.066) and the average ex- pected heterozygosity was H,, = 0.165 (0.059), whereas genet-level values were H.,,. = 0.187 (0.062) and H, = 0.184 (0.062). The difference between these mean observed and expected heterozygosities was not significant (t = 0.1, df = 3, P > 0.05). Fixation index (F) for each polymorphic locus was estimated and averaged over loci for both ramet- and genet-level (Table 2). Ramet-level averaged at F, = —0.076 (0.063), genet-level at F, = —0.026 (0.091), with no significant deviation between them (t = 0.17, df = 6, P > 0.05). Slight heterozygote excess (negative F value) was detected at five of seven loci on both ramet-level and genet-level, heterozygote deficiency (posi- tive F value) at two loci, but the sign of deviations was not uniform for MDH- 4 and PER. For SKDH, a highly significant heterozygote deficiency was indi- cated in both cases. The chi-square test of observed versus expected genotype numbers showed significant deviation from Hardy-Weinberg proportions at ra- MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 115 fis 2 > = 18.11% 0.5 0.0 ‘\ A] = 19.48% -0.5 0.0 0.5 fede 1 . 3. Principal coordinates analysis of multilocus genotypes based on their isozyme patterns. ee was performed on the basis of chord-distance function of Orldéci (1975). Symbols of the genets growing in the same patch are bordered. |' and |? mean the Eigen-values as percentages. met-level in the case of MDH-1 (P < 0.01), SKDH (P < 0.001), and PGI-2 (P < 0.05), but at genet-level only in the instance of SKDH (P < 0.001) The result of principal coordinates analysis based on the chord-distance function of Orléci is presented in Fig. 3. Axes I and II together contained 39.4% of the variation. The same ordination was produced with the Euclidean- distance function. Both distance functions ordinated most of the genets as independent of their geographical position. Only genet pairs G-H, L-Q, M-O, and U-V were genetically more related according to their ordination and be- longed to the same patches. Spatial autocorrelation coefficients were Calculated with different sets of dis- tance classes. The values of univariate (one locus) autocorrelation coefficients and the average I indices are shown in Table 3a and 3b for the ten distance classes of equal size and equal numbers of point pairs in ten distance classes options, respectively. In some distance classes for some loci there appeared univariate Moran coefficients that significantly differed from those for inde- pendent spatial structure. EST-4 and SKDH appeared among these isozymes in both analyses. However, the average I indices definitely detected no signif- icant spatial substructure in the population: I=—0.001 (0.002) for the equal size distance classes and I=—0.050 (0.033) for the latter case. The average TABLE 3A. Moran’s I (spatial autocorrelation) coefficients of polymorphic loci in 10 distance classes of equal size. Distance class 9 contained no point pairs. Significant deviations from the expected values are labelled * P < 0.05; ** P < 0.01. The standard deviation of the average I index is in parentheses. Last column shows the significance of the correlograms (C. Pr.) D. class 1 2 3 4 zs" 6 7 8 10 No. of pairs 104 = 24 12 15 41 4 2 3 C.-Pr MDH-|! —0.05 0.01 —0,15 0.02 —0.36 0.05 ~0.51 0.29 0.29 0.710 MDH-4 —0.06 —0 —0.08 0.11 =—0.12 0.00 —0.17 11 11 tr: SKDH —O il 1.20** —0,.24 0.11 —0.02 —0.02 0.46* —0.62* —0.02 0.012* EST-1 —0.06 0.12 0.05 —0.30 =—0.25 0.05 0 —0.53 i EST-4 —0.02 0.40 —0.07 —0.07 —0.21 0.35** 0.40 0.40 0.40 0.036* PGI-2 —0.06 —0.01 —0.1 —0.11 0.02 0.01 0.04 —0.30 0.03 I —0.03 —0.89* —0.08 0.14 0.28 —O21 0.41 —0.38 0.36 0.191 AVERAGE —0.06 0.07 —0.10 0.01 —0.03 —0.07 0.10 —0.02 0.09 —0.001 OIL (6661) Z YAAWOAN 68 ANNIOA “TVNUNO!l NYA NVOMANV TABLE 3B. Moran’s I (spatial autocorrelation) coefficients of polymorphic loci in 10 distance classes of equal size. Significant deviations from the expected values are labelled * P < 0.05; ** P < 0.01; *** P = 0.001. The standard deviation of the average I index is in parentheses. Last column shows the significance of the correlograms (C. Pr.). D. class 1 2 3 4 | 8 9 10 No. of pairs 21 21 21 21 ZA 21 21 21 21 21 C.F MDH-1 0.70** —0.09 —0.01 —0.47* —0.36* —0.16 0.11 —0.40* 0.24 —0.05 0.001 *** MDH-4 —0.0: —0.26 0.13 0.00 —O.11 ~0.11 —0.05 0.00 —0.05 0.00 0.911 KD 0.24 —0.34 0.02 —0.08 —i2 | =—0.27 0.02 0.18 —0.11 0.00 0.658 EST-1 0.03 0.17 —0.27 —0.30 0.07 —0.07 —0.13 —(0).13 0.10 0.03 0.968 EST-4 0.17 0.20 —0.20 —0.13 —0.13 0.27* =0.27 0.00 ~0.53%" 0.13 0.039* PGI-2 ~(),25 —0.09 0.00 0.02 0.01 —0.16 _ 05 0.02 —0.01 PER 0.48* 0.23 —0.93** 0.85** —O37" =0.50** 0.34** 0.14 = G71 —0.16 0.000*** AVERAGE 0.19 —0.09 “18 —0.02 ~O16 —0.14 —0.01 —0.03 —0.06 0.00 —0.050 WALVNVTdNOS WAYLSVISVHdId AO SLANAD ‘MOdO 8 YOLVW Lit AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) 1 VA Bs \ *K MDH-1 Fay Ye? ae ie | athe. » - + M --MDH-4 0,2 + 1 \ 7 . ae yy Oe ee, ee \ eC! Lusi 8 — : ais She ees : ~ — % — EST ee aie ~~ ; — m— EST4 = “ AY. . eu -- y ~~ ay —e— PGi-2 0,2 ‘7 \ — 3 : a YW x in | o +— = PE y 3 Ae ‘ é x \ N AVERAGE 04 \ I ~T 7 Ney : \ ; ~ / O68 , ! T * oe eT \ 08 sil eee Soler r el Ree Seeconeee \ . : pay 0 1 2 3 4 5 6 7 8 9 10 Distance classes IG. 4. Spatial autocorrelation analysis of the genet multilocus genotypes. Moran’s I autocorre- lation coefficients are shown for individual polymorphic loci, and their average is presented in each distance class. a) Correlograms for ten distance classes of equal size (data from Table 3a); distance class 9 contained no point pairs. b) Correlograms for ten distance classes with equal numbers of point pairs in each (Data from Table 3 autocorrelograms show a random series of nonsignificant values for all dis- tance classes (Fig. 4a and 4b). The diagrams obtained by other options were similar. DISCUSSION The observed numbers of isozymes were in accordance with the data for eight lycopods reported by Soltis and Soltis (1988a) for those enzymes in com- mon in the two studies (GOT, LAP, PGI, and SKDH). In addition to these re- MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 119 sults, we managed to visualize ACP, (NADP) IDH, and MDH isozymes of D. complanatum, detected two loci for colorimetric esterase (EST), one of them with a dimeric enzyme, and found a locus of PER. Two loci for IDH and four loci for MDH are the numbers usually detected for diploid seed plants, further supporting the concept of genetic diploidy of lycopods. In addition, with the first electrophoresis buffer system IDH-1 isozymes behaved similarly to that was observed by Hickey for Isoetes (cited by Duff and Evans [1992] as “Hickey, personal communication”). They explained a fixed uniform three-banded pat- tern as the product of a monomorphic locus after posttranslational modifica- tion or degradation. Actually, in the case of D. complanatum the second sep- aration system proved the monomorphic state of the IDH-1 isozymes, but the pattern with the first system was not uniform; there appeared reproducible minute mobility differences between the bands even for two ramets classified as the members of the same clone on the basis of the other isozymes, i.e., ramet 24 and 25 of genet S. These differences can be based on a genetically variable modifier system similar to that suggested by Hickey et al. (1989) for TPI zone III of Isoetes species or can show different stages of the same modification/ degradation process. Because the background of these alterations could not be clarified, we did not take into account this heterogeneity when evaluating di- versity. On the other hand, it would be worth investigating the genetic back- ground of this modification process. Its presence or absence might even have phylogenetic aspects like the TPI modification proteases of some Isoetes spe- cies. The high number of unique multilocus genotypes in the D. complanatum population studied parallels the results of Ellstrand and Roose (1987). In the review of the genotypic diversity of clonal plants they indicate that these spe- cies are prone to having multiclonal populations of intermediate variability and unique genotypic composition. They found that out of 238 populations only 17 consisted of a single genotype; that is, 93% of the surveyed popula- tions were multiclonal. The 17 uniclonal populations belonged to as few as two uniclonal species. At the same time, Ellstrand and Roose (1987) demon- strated that the number and frequency of the genotypes detected were depen- dent upon the number of characters included in the study, so the estimation of genetic diversity must have been biased. In spite of the fact that in our case the number of characters investigated was not large (15 isozyme loci), we hope that because the sample size exceeded Holsinger’s (1987) recommendation of 25 and we detected a relatively high ratio of polymorphic loci, the estimation of the genet number and diversity was not overestimated, and we reliably registered the lack of intragametophytic selfing. Including this study, three observations have been reported about intragametophytic selfing in D. com- planatum, and none of them revealed any. Soltis and Soltis (1990) found no intragametophytic selfing in two populations examining 17 isozyme loci. The analyzed population showed considerable genetic variability, with a 0.898 clonal diversity value, 1.81 (0.519) ramet/genet ratio, and 46.7% poly- morphic loci, comparable with the average value (39.9%) reported by Hamrick and Godt (1989) for long-lived herbaceous perennials. The observed relatively 120 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) high P value and the genetic diversity measures of the present study may characterize the genetic composition of the species, as it is a widespread club- moss. In their review of genetic differentiation in homosporous ferns, Soltis and Soltis (1990) reported average F indices for different lycopods from —0.141 to 0.016 on the basis of 14 population analyses. The —0.076 ramet- level and —0.026 genet-level values observed in this study fit in with their series. The lack of intragametophytic selfing among the 21 genets, the slight tendency to heterozygote excess, and the fluctuation of the F values among loci (on the ramet-level —0.477 to 0.456 and on the genet-level —0.238 to 0.356, with significant heterozygote deficiency at some loci or excess of het- erozygotes at another locus) not only clearly show the importance of sexual reproduction but call attention to the fact that microsite conditions may have had strong effects on the population. In addition, the data might reflect founder effect. Thus, high clonal diversity and the structure of this small marginal population may be the consequence of the history of the population and the site. The relatively large and probably old patch II was practically formed by one clone, only a single ramet belonged to a distinct but genetically close genet that differed in its SKDH genotype. Other patches, like patch V, which had a similar size and was about 100 m from patch II, showed high clonal variation: 7 genets per 8 ramets. This pattern shows that under microsite conditions sexual reproduction must have played an important role. Principal coordinates analysis visually demonstrates that the genetic distance of the genets is inde- pendent of their position in the population. As distant genet pairs as M-S (nearly 460 m) and B-N (nearly 200 m) are genetically closely related accord- ing to the ordinaton, whereas genet pairs A—B and J-K in small patches (di- ameter about 6 m) are independently ordinated. Only four genet pairs listed in the Results were genetically related and belonged to common patches. Spa- tial autocorrelation analysis, supporting the results of the principal coordinates analysis, did not reveal any trend in the distribution of genetical characters. Irregular significant difference from the value expected for random structure might have occurred because of the fact that the above mentioned genet pairs were related and might be the consequence of the small number of data. The type of recruitment, which is of primary importance for the substruc- turing of the population, was not recognizable because the development of gametophytes and young sporophytes could not be observed. Although the vegetation of the patches seemed to be superficially similar, it could have been more heterogeneous from the “plant’s point of view” resulting in different patterns in the patches. Moreover, the coenological and ecological conditions are not favorable at the boundary of the area of D. complanatum (Odor, 1996, 1997). The presence of the strongly competitive plants of the deciduous forest could have increased the clonal variation. Bruce and Beitel (1979) published their observations on a gametophyte com- munity of six lycopods growing in a 30 year old artificial jack pine (Pinus bank- siana Lamb.) plantation. One of the most important characteristics was the clus- tering of gametophytes for all species but one. The most spectacular case was L. lucidulum: all of the observed 125 gametophytes grew in a single square 45 MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANATUM 121 cm on a side. This clustering is also a common feature of pteridophytes with surperficial gametophytes, and clearly indicates the strong controlling effects of the microenvironment. Spore dispersal presumably is not a limiting factor of initialization because of abundant spore production. Meusel and Hemmerling’s (1968) and Oinonen’s (1968) observations support these findings. They reported that even different clubmoss species could frequently be found in mixed patch- es, and often not only along the area boundary, but even in the Alps, although their ecological claims were different. As a further consequence, the presence and proximity of gametophytes originating from different sporophytes can in- crease the chance of outcrossing, which can produce a genetically variable pop- ulation, maintain genetic diversity, and explain the lack of intragametophytic selfing even at species with subterranean gametophytes. The presence of the endophytic symbiont fungi is crucial to the survival and development of the prothalli (Freeberg and Wetmore, 1957; Freeberg, 1962), so theoretically it must be a limiting factor of initial colonization and recruit- ment, and may cause the patchy appearance of the gametophytes. It may also be supposed that the presence of older sporophytes or prothalli can increase the concentration of fungi in the local soil so that it has a positive effect on the recruitment of new genets sustaining diversity. According to Oinonen’s (1967) observations, “Young plants of ground pine start to develop at a normal rate about 4-6 years after they have penetrated the soil surface, and manifestation occurres about ... 8-20 years after the spores were sown.” Therefore, during the early period of the population es- tablishment or under frequent local disturbances and recolonization, genetic diversity can be substantial, but later the number of genets will decrease as a consequence of competition. Despite of the supposed facilitating effect on re- peated recruitment the genet dynamics of established populations cannot be intensive because of the slow development of D. complanatum. New genets from repeated recruitment are likely to appear around the edges of the areas occupied by established old clones. Considering these results, the patches of the analysed population can represent different colonization states. The unique small genet V of patch II may be a genetically related genet that is a “loser” in a long competition, or the case of a repeated recruitment inserted near the verge of the spot among the ramets of genet U. In the studied D. complanatum population, the patches may show a younger successional state of colonization or a permanently stressed conditon, except patch II, in which colonization must have been earlier. According to these findings the genetic diversity may be substantial even in the case of a small marginal D. complanatum population. In the early phase, the genetic composition mainly depends on sexual reproduction, then local conditions, history, and age influence the genetic structure of the population. Clonal growth plays more important role in established populations. ACKNOWLEDGMENTS We thank Gabor Vida for his helpful review of an earlier version of the manuscript and Randall Small for his detailed review and useful comments. This work was supported by the Pro Reno- vanda Cultura Hungariae Foundation. 122 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) LITERATURE CITED ARULSEKAR, S., and D. E. PARFITT. 1986. Isozyme analysis procedures for stone fruits, almond, grape, walnut, pistachio and fig. Hort. Science 21:928-933 Bruce, J. G., and J. M. BEITEL. 1979. A community of Lycopodium gametophytes in Michigan. Amer. Fern J. 69:33-41 CALLAGHAN, T. V., A. HEADLEY, J. A. LEE, B. M. SVENSSON, L. LIXIAN, and D. K. LINDLEY. 1986b. Modular growth and function in the vascular cryptogam Lycopodium annotinum. Proc. Roy. Soc. Lond. B Es 195-206. CALLAGHAN, T. V., B. M. SVENSSON, eis A. HEADLEY. 1986a. The modular growth of Lycopodium annotinum. Fern. Gaz. 13:65-7 rapt B. A., I. S. 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Budapest 19:1 JONsson, B. O., I. S. tor. ae N. CRONBERG, 1996. Clonal diversity and allozyme variation in population of the arctic sedge, Carex bigelowii (Cyperaceae). J. Ecol. 84:449-459. KEMPERMAN, J. A., and B. V. BARNES. 1976. Clone size in American aspens. Canad, J. Bot. 54:2603— 2607. KULL, T. 1995. Genet and ramet dynamics of Cypripedium calceolus in different habitats. Nové- nyféldr. Tansz., Eétvés pac eter ce Budapest 19:95-104. ee D. A., and W. L. CREPET. 1973. Genetic variation in Lycopodium lucidulum: a phylogenetic relic. Evolution 27: ae McCaul ey, D. E., D. P. WHITTIER, and L. M. REILLY. 1985. Inbreeding and the rate of self-fertiliza- tion in a grape a sam i dissectum. Amer. J. Bot. 72: 1978-1981 MAJOR & ODOR: GENETS OF DIPHASIASTRUM COMPLANA TUM 123 E. Téth and R. Horvath, eds. Proceedings of the “Research, Conservation, and Management” ape Aggtelek National Park Directorate, Jésvafé, Hungary. OINONEN, E. 1 Sporal regeneration of ground pine (Lycopodium complanatum L.) in southern Pala | in ee light of dimensions and the 5 fi of its clones. Acta Forest. Fenn. 83: 1968. The size of Lycopodium clavatum L. and L. annotinum L. stands as compared to that of L. complanatum L. and Pteridium cepilinum (L.) < apa the age of the tree stand, and the dates of fire on the site. Acta Forest. Fenn. 87:1— Or. ocI, L. 1975. Multivariate analysis in vegetation research. W. Junk oan Haag. PIELOU, E. C. 1969. An introduction to mathematical ecology. Wiley Interscience: New York. Pocs, T. 1965. A magyarorszégi tilevelti erd6k cénolégiai és ékolégiai viszonyai [The coenological d ecological relationship of coniferous forests in Hungary]. Ph.D. Thesis, Esterhazy Karoly Teacher Training college, Eger, Hungary. [in Hungarian] pap J. 1980. SYN-TAX: Szdmitégépes programcsomag ékolégiai, cénol6giai és perppoane 3 ant lyozdsok végrehajtasdra [SYN-TAX: Computer program package for cluster of, phytosociology and taxonomy]. Névenyrendsz. Novényféldr. Tansz., Eétvés Lorand Tu- igh ds he Budapest 6:1—158. [In Hungarian] SYN-TAX version 5.0, user’s guide. Scientia Publishing, Budapest SOKAL, R. ~ and N. L. ODEN. 1978a. Spatial autocorrelation in biology. 1. Methodology. Biol. J. Linne. Soc. 10:199-228. . 1978b. Spatial autocorrelation in biology. 2. Some a iin implications, and four ap- plications of evolutionary and ecological interest. Biol. J. Linn. Soc. 10:229-249. SoLtis, D. E., and P. S. SoLtis. 1986. Electrophoretic evidence for inbreeding in the fern Botry- chium virginianum (Ophioglossaceae). Amer. J. Bot. 73:588-592. . 1987. Breeding system of the fern Dryopteris expansa: evidence for mixed mating. Amer. J. Bot. 74:504—-509. ma. LORRE. tas lycopods with high chromosome numbers ancient polyploids? Amer. J. Bot. 75:238—-24 . 1988b. cuales variation and population structure in a fern Blechnum spicant (Blech- nacetis) from Western North America. Amer. J. Bot. 75:37—4 . 1988c. Estimated rates of intragametophytic selfing in cell Amer. J. Bot. 75:248—256. . 1990. Polyploidy, breeding systems, and genetic differentiation in homosporous pterydo- nhiex Pp. 241-258 in D. E. Soltis and P. S. Soltis, eds. Isozymes in plant biology. Chapman and SoLtis, D. E., C. H. HAUFLER, D. C. Darrow, and G. J. GASTONY. 1983. Starch gel a of ferns: a compilation of grinding buffers, and staining schedules. Amer. Fern J. 73:9-27. Sotis, D. E., P. S. SoLtis, and K. E. HOLSINGER. 1988a. Estimates : ioe gs selfing and a eas BP eis gene flow i in homosporous ferns. Am 1765- Sottis, D. E., P. S. Sots, and R. D. Noyes. 1988b. An th Gr tate of intraga- gee ae in Equisetum arvense. Amer. J. Bot. 75:231-237. paiement G. L. 1950. Variation and evolution in pants, Columbia University hinge aed tom SVENSSON, B. M., and T. V. CALLAGHAN. oe Apica p lati ation dynamics in Lycopodium annotinum. Oikos 51:331-342. hee Small-scale vegetation ee related to — ong of Lycopodium annotinum and v. s of a micro-environment. Vegetatio 76:16 Warren D. 1989. SAAP version 3, a spatial eeelcioe analysis program. Exeter Soft- East sethiket, NY WoLr, P GC. C. H. HAUFLER, and E. SHEFFIELD. 1987. sor ins names evidence for genetic diploidy in bracken fern (Pteridium aquilinum). Science 236: 949. . 1988. Electrophoretic variation and mating system 2 the clonal weed Pteridium aquilinum (L.) Kuhn (bracken). Evolution 42:1350—-1354 WRIGHT, S. 1965. The interpretation of population structure by F-statistics with special regard to systems of mating. Evolution 19:395—420. Wu, L., A. D. BRADSHAW, and D. A. THURMAN. 1975. The potential for evolution of heavy metal tolerance in plants. Heredity 34:165-187. American Fern Journal 89(2):124—132 (1999) Spore Age and Sterilization Affects Germination and Early Gametophyte Development of Platycerium bifurcatum MARJANA CAMLOH National Institute of Biology, Vetna pot. 111, 1000 Ljubljana, Slovenia ABsTRACT.—The effects of spore age and sterilization on spore germination and early gametophyte development were investigated in the fern, Platycerium bifurcatum. The highest germination per- The viability of spores varies enormously among ferns, ranging from a few days to a few years (reviewed by Miller, 1968). In recent years it has become clear that genetic and certain physiological attributes of spores have to be considered to explain spore longevity (Raghavan, 1989). Storage conditions also are very important for spore viability. Fully hydrated spores showed com- plete ability to germinate after storage for two years at room temperature (Lind- say et al., 1992). Another possibility for extending the viability of spores is storage at —70°C. This was successfully used for ripe strobili of Equisetum hyemale L. (Whittier, 1996). Despite these discoveries, pteridologists generally store fern spores under dry, cold, and dark conditions (Miller and Wagner, (Raghavan, 1989). These experiments concentrated on the effect of spore age on germination, but further gametophyte development has received much less attention. The study of Smith and Robinson (1975) provides valuable information concerning Polypodium vulgare L. spores. In their experiments, spores stored for 0-7 years were used, but at intervals of one year only. In another study, spores of Pteris vittata L. were stored for 10-100 days and tested at intervals of 20 days (Beri and Bir, 1993). Although different aspects of spore germination and gameto- phyte development in Platycerium have been investigated in detail (Nagmani and Raghavan, 1983; Thentz and Moncousin, 1984; Camloh and Gogala, 1992; Camloh, 1993; Camloh et al., 1996), to the best of our knowledge spore age CAMLOH: GAMETOPHYTE DEVELOPMENT IN PLATYCERIUM 125 effect on developmental processes has never been studied. In the present study, experiments were conducted to determine the effect of spore age (2-14 month old spores were used) and sterilization on germination and particularly on early gametophyte development of the fern P bifurcatum. MATERIALS AND METHODS Spores of the fern Platycerium bifurcatum (Cav.) C. Chr. were kindly pro- vided by Dr. B. J. Hoshizaki, Univ. of California, Los Angeles. They were col- lected in September 1991 from mature leaves of a single plant and stored in the dark at 5°C. Spores stored for 2 to 14 months were used in the experiments. They were isolated from sporangial and other debris according to Camloh (1993). The culture method has been described in detail elsewhere (Camloh et al., 1996). Briefly, spores of P. bifurcatum sterilized with 70% (v/v) ethanol and 10% (v/v) solution of commercial bleach (4% NaOCl) were sown on the surface of 5 ml modified Knop’s solution (Miller and Greany, 1974). Some experiments were also performed with unsterilized spores. The pH of the me- dium was adjusted to 5.7-5.8 before autoclaving. The media were placed in tubes 24 mm in diameter and covered with plastic caps. Cultures were main- tained at 23+2°C, under a 16h photoperiod at 36-50 pmol-m ?-s~', provided by cool white lamps (Osram L 65W/20S). Spores were grown in these condi- tions for 3 to 10 days. The percentage of germinating spores was evaluated at 3 and 6 days after sowing. The criterion for germination was the breakage of the exine and pro- trusion of the rhizoid. Samples of at least 100 spores per replicate were ex- amined. The length of the primary rhizoid was measured at three, four, six, and eight days after sowing, and the gametophyte cell number was determined at six, eight, and ten days after sowing. At the end of the experiment, rhizoids were counted. All measurements were made on gametophytes cleaned and stained with acetocarmine-chloral hydrate according to Edwards and Miller (1972) in a microscope fitted with an ocular-micrometer. For the determination of all parameters, except where otherwise specified, at least 25-35 samples were examined per replicate. There were two or three replicates per set of spores of different age. Mean values and standard error (SE), which are rep- resented in the figures as vertical bars, were calculated from the data. The 22 Chi-squared test (x?) and Student’s t-test were used for evaluating levels of statistical significance (P) between the data obtained with two months old ster- ilized spores and those obtained with other spore samples. RESULTS The effects of spore age and sterilization on germination are shown in Fig. 1. The percentage of germination was determined at three and six days after sowing. Maximum germination of sterilized spores occurred after two to three months of storage, but we must point out the considerable variations in ger- mination obtained in experiments with three months old spores compared to 126 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) 100 > A UNSTERILIZED SPORES 4 3 DAYS ME 6 DAYS Z * 80 — 3 = 60- 2 bee! y+] = £ * = 40 as ote 7 ® * Be Shs ete x 20 - 0 EE 2 2A 3 4 8 14 14A Spore age (months) Fic. 1. Effects of spore age and sterilization on germination of P. bifurcatum spores. Vertical bars indicate SE. Chi-squared test was used for evaluating the level of statistical significance; *P <0.05 FUP <0,01, 4777: <6,001; ’ others. In older spores, substantially decreased germination was observed. Be- cause the criterion for germination was the protrusion of the rhizoid through the exine, this result indicates that spore age affects the process of rhizoid initiation. However, the highest germination percentage was obtained with un- sterilized spores. An additional and unexpected result was noted: in contrast to sterilized spores, the germination percentage of unsterilized spores did not decrease with increased storage time (Fig. 1). Although three days after sowing older spores germinated slightly better than younger ones, after six days the germination responses of unsterilized spores of both ages (2 and 14 months) were almost identical. This result suggests that during the sterilization pro- cedure certain changes in spores occurred that resulted in altered germination. The effects of storage time and sterilization on rhizoid length are shown in Fig. 2. It is evident that as the age of sterilized spores increases, there is a decrease in rhizoid length. This was observed as early as three days after spore sowing. The same age effect also was obtained at four, six, and eight days after sowing. Only when eight month old spores were cultured for eight days was CAMLOH: GAMETOPHYTE DEVELOPMENT IN PLATYCERIUM 127 240 — , : A __UNSTERILIZED SPORES ‘ x [] 3DayYs xX a Be 4 DAYS “ 6 DAYS ol x : (] 8 pays E 460 - RS Ms : = RS RY £ Ry eS Ry x s : 0 me me * : © D Re % me z aa c RI Q % RI § 120- Poke ia ch 2 RS XS i 9 re) RY “ i q %, 2, * 2 = 80- RS i > % rg RS RS KS RS RY RS RS Ry d x) xX] xX Xs * <, %, NX KS Ry Ko x S 40 + x i“ x x R RS xs RS Ry RY Ry eS RS KS Ry Ry ey Ry Ry Ry xy " “ RY > Won x "aS Spore age (months) Fic. 2. Effects of spore age and sterilization on the rhizoid length of P. bifurcatum gametophytes. Vertical bars indicate SE. Student's t-test was used for evaluating the level of statistical signifi- cance; *P <0.05, **P <0.01, ***P <0.001. no further decrease in rhizoid length detected in comparison to four month old spores. Regardless of spore age, the most intensive elongation of the pri- mary rhizoid occurred after between four and six days in culture (Fig. 2). Ster- ilization also affected rhizoid length. Two month old unsterilized spores cul- tured for four or more days have longer rhizoids than sterilized spores. This effect was also observed with fourteen month old spores, although their rhi- zoid length was much smaller in comparison to two month old spores (data not shown). In addition to rhizoid elongation, spore age also affected rhizoid number (Fig. 3). After ten days of culture, the highest number of rhizoids developed on gametophytes from two month old spores. With increasing spore age the number of rhizoids decrease. The sterilization had no effect on the rhizoid number of two month old spores (Fig. 3). Similarly, in an experiment with fourteen month old spores, no effect of sterilization on rhizoid number was detected (data not shown). Increasing spore age also leads to a decrease in cell number (Fig. 4). At day 128 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) 3 i A - UNSTERILIZED SPORES 15> Rhizoid number 1,0 - 2 2A 3 a 8 Spore age (months) Fic. 3. Effects of spore age and sterilization on the rhizoid number of P. bifurcatum gametophytes 10 days after spore sowing. Vertical bars indicate SE. Student's t-test was used for evaluating the level of statistical significance; *P <0.05, **P <0.01, ***P <0.001. six, spore age already has a significant effect, but after two and four more days the decrease in cell number is even more pronounced. After ten days, game- tophytes grown from two month old spores have nearly twice as many cells as those grown from older spores. The greatest decrease in cell number oc- curred between two and three month old spores. Thereafter, increased storage time did not cause a substantial further decrease in cell number. After six and eight days of culture, the cell number of gametophytes grown from sterilized spores is quite comparable with those grown from unsterilized spores. How- ever, after ten days gametophytes with more cells were grown from sterilized spores. DISCUSSION Spore age and sterilization affect spore germination and early gametophyte development in P. bifurcatum. Sterilized spores gave the highest germination response after two to three months of storage. Three month old spores even CAMLOH: GAMETOPHYTE DEVELOPMENT IN PLATYCERIUM 129 R A UNSTERILIZED SPORES C] é6pays M@ spays N 10 DAYS = * Cell number > | UB Spore age (months) Fic. 4. Effects of spore age and sterilization on the cell number of P. bifurcatum gametophytes. Vertical bars indicate SE. Student's t-test was used for evaluating the level of statistical signifi- eance;: *P.<0.05, **P O.1 ¢ 5.8 +:0.2-a 64+0.1b 331382 GA Sg fats P. pleopeltifolium 912024 10 = O01 ¢ 6.5 2-0. Eb 5.626. a 331314 80.56** P. poly, les Blt 02a $9 = 02b 78+02a 115+ 06c 3:49 150" * Adiantopsis radiata E+ 0.2 62> 01 4 5.9'+:0.1l-a 6.2 +024 S1212 166.21+* Polypodiu tipes 8.2 + 0.02 d 7.4 + 0.04 b 6.6 + 0.03 a 7.8 + 0.05 c 3;7309 545.05** Pteris denticulata 93+ 0.14 To = OG 64+ 0.1a 70) = 0.1b SP a! 130.43** PSt (6661) @ YAGWNN 68 ANNTIOA “TVNYNOL NYAd NVORWV RANAL: EFFECTS OF TEMPERATURE ON SPORE GERMINATION 155 70) Microgramma lindbergii 80) Polypodium polypodioides = - 50 x 60 7 = = 30 = 40 J : ree reel = 20 cI 8 I days after inoculation 1 4 B days after inoculation > = 50 Microgramma squamulosa I = 50 = = 30 = 30 = = E F Ee 10 E 10 = = 4 8 4 8 days after Inoculation 2 days after inoculation 6 100 Polypodium hirsutissimum ci J 100 Polypodium latipes A Libs a 3 °°] rare eget . o aT ? > Kad ----- 21.2 ane as j = = 60] ---253 4 = 60 i. = —~ 40 z 40 3 s 20 20 4 8 12 4 8 days after inoculation 3 days after Ineculation Fd 100 Polypodium pleopeltifolium 80) pteris denticulata sieges | 80 ai I aa x —— en = De a4 ’ ] = -~ Yr sep. | 2° 3 A ; . = i = 20 = 4 8 days after inoculation 8 a 8 days after Inoculation 4 Fics. 1-8. aro of fern spores at four different temperatures. Vertical bars represent the oud errors of the me TABLE 5. Germinability of fern spores (mean + standard error; Sanaa at different temperatures, 12—13 days after inoculation. Temperatures are average temperatures of the experiments | and 2. Means followed by the e letter in each line are not significantly different based on the Tuke ey test (a ). F indicates the values of Snedecor’s distribution (* significative at sm, « ** significative at 1% of probability) and C.V. the coefficient of variation. = 0.05 Temperature (°C) Species 18.4 21,7 25.2 29.4 F C.V. (%) Microgramma lindbergii 40.6 + 82a 56.5 + 64a 52.0 = 2.6 4 17.7. 2340 10.13** 17.10 M. squamulosa 579 +234 415 +324 376 2 158 22.8 +28b 11.08** 8.73 Polypodium hirsutissimum 75.8+63a 90.7 + 6.6a 85.6 + 3.6a 95.7+40a 1 14.47 P. pleopeltifolium 453 +165 Jia + 8.7 ab 84.1+ 5.7 a 63.3 2°77 ab 5.14* L722 polypodioides 458 + 55a 64.3 + 96a 38.7: 5A-a 67-2275 17.33** 21,52 diantopsis radiata 28.4 + 3.2b 426+39a 38.7 + 3.3 ab 51.8 © 2.2 ab 3 10.31 Polypodium latipes 83.9+09a PA fae 84.7+ 16a TIS E27 a 1.26 6.37 Pteris denticulata 54.7 + 10.0 a 60:9 + 5:1a 54.5 42.7 a 67:2 +944 0.66 19.34 9ST (6661) 2 MAGINON 68 AWN'TOA ‘TVNYNOl NYA NVOMANV RANAL: EFFECTS OF TEMPERATURE ON SPORE GERMINATION 157 Although October is considered a wet month, in 1984 and 1985 it was atypi- cally dry (Ranal, 1995a). Thus, under natural conditions the germination pro- cess for these species may be most effective in November and December (av- erage temperature of 23.3 and 24.0°C, respectively, for the period 1970-1985), when spores are abundant in the environment, the water supply is suitable and constant, and the temperature adequate for most of the day. After these months, the young gametophyte will have sufficient time to develop a plate structure before the next dry season. The results of this study suggest that for some species, especially the gen- eralists, other factors are probably more limiting in their establishment than habitat temperatures, at least in the range of temperatures studied. For exam- ple, the humidity of the environment may be the limiting factor for Pteris denticulata. The low endurance of hydric stress by gametophytes and sporo- phytes of this species support this idea (Ranal, 1991a, 1995a). Moreover, prob- ably the limiting factors for the subsequent phases of development may be different in relation to the factors which are important to germination and can interfere in the establishment process of each species in the different micro- habitats. Although there is no quantitative data about gametophyte development in seasonally dry tropical areas, Barreiro Rico included, the results obtained in this study indicate that in these environments the most important limiting factor to the establisment of ferns is water, not temperature. According to Kor- nds (1985), water deficiency is the key factor limiting the occurrence of pte- ridophytes in seasonally dry tropical areas, acting on their adaptations in re- lation to specific habitat, life-forms, phenological patterns, and reproductive biology. ACKNOWLEDGMENTS This study was supported by the Conselho Nacional de Desenvolvimento Cientifico e Tecno- légico (CNPq) as part of a M.S. dissertation. 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Effects of oe and temperature on the ger- mination of fe ei of pees delgadii. Rev. Brasil. Bot. 7:53-56. MILLER, J. H. 1968. Fern gametophytes as experimental material. ca Rev. (Lancaster) 34:361—440. PaGE, C. N. 1979. Experimental aspects of fern ecology. Pp. 551-589 in A. F. Dyer, ed. The exper- imental biology of ferns. Academic Press, London See bee B., and R. RIBA ee s sciaatdis de explores de Cyatheaceae bajo diversas tem- per Wh por 14:281- reliceneviae? v 1980. Cytology, ohsililoas: and biochemistry of germination of fern spores. Int. Rev. Cytol. 6 samal 1989. Developmental biology of fern gametophytes. Cambridge University Press, Cam- bridge, Great Britain. RANAL, M. A. 1983. 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Temperature and photocontrol of Onoclea sensibilis spore germination. Pl. Physiol. 62:116-119 TRYON, R. M., and A. F. TRYON. 1982. Ferns and allied plants, with special reference to Tropical America. Springer-Verlag, New York. Warne, T. R., and R. M. LLoyp. 1980. The role of tion and gametophyte devel in habitat selection: ene yids responses | in certain temperate and tropical ferns. Bull. Torrey Bot. Club 107:5 American Fern Journal 89(2):159-170 (1999) Studies on Cryptogramma crispa Spore Germination EMILIA PANGUA, LORENA Garcia-ALVAREZ, and SANTIAGO PAJARON Departamento de Biologia Vegetal I, Facultad de Biologia, Universidad Complutense, E-28040 Madrid, Spain ABSTRACT.—In order to study the germination capacity of Cryptogramma crispa, spores were cul- tured on sterilized Petri dishes with nutritive medium solidified with agar. Germination was checked at 10, 15, 20 and 25°C, and, as in most of homosporous ferns, the germination optimum was at temperatures above 20°C. Two light intensities were used, 10 and 40 pEm~?s~', to reproduce of the spores. After these processes, the spores are able to germinate and reach similar germination rates, although the frozen spores delay the beginning of germination and show a decreased ger- mination rate. The results of these experiments point toward the possibility that the spores of C. crispa are dispersed at the end of the growing season and go through a dormancy until next spring. In the Iberian Peninsula (Spain and Portugal), Cryptogramma crispa R. Br. grows on ecologically very particular and well defined habitats. It occupies especially siliceous stone fields, such as granite, gneiss, sandstones, quartzites, or slates in high mountain zones, usually above the timber-line. In the Iberian Peninsula, its optimum is about 2000 m. The plants grow preferentially in rock fissures or cracks and in hollows between rock blocks. In these habitats, which are mostly over 2000 m elevation, the growing season may be very short, scarcely two months in extreme conditions, and usually no more than four months (Rivas-Martinez, 1987). The considerations of the distribution of pteridophytes suggest the need for more detailed investigations on the life-cycles of species to determine the im- portance of specific variations in the life-cycle in limiting the distributions of plants. Variations in the distributions of species might be accounted for by random processes, such as dispersal, or in a more deterministic manner by subtle and specific variations in life cycle characteristics (Woodward, 1987). In ferns, it is important to study the factors that can affect the development of the gametophyte that would lead to the establishment of the sporophytic gen- eration. Probably due to the short period to complete their development, C. crispa sporophytes reach the end of the summer with practically all the spores still retained (Peck et al., 1990; pers. obs.), which are released at about the same time that the leaves shed. Thus, the spores are released at the end of the grow- ing season. It is difficult to reproduce wild conditions in short-term laboratory experi- ments. But some climate changes, especially temperature ones, that may influ- ence spore germination can be tested in the laboratory, provided that in nature conditions may be operating over a longer period. Thus, a few experiments 160 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) TABLE |. Localities of and acronyms for populations of Cryptogramma crispa used in the present study. Except for population SCO, the localities are all located in Spain. Acronym Locality HU Prov. Huesca, Torla, Barranco de la Pazosa, slates, 2000 m, Herrero s.n. on 2 Oct 1995. LNE Prov. Soria, Sierra de Urbién, Laguna Negra, 1900 m, Pajarén & Pangua s.n. on 26 Sep 1996. MON Prov. Zaragoza, Sierra del Moncayo, near Ermita de la Virgen, 1620 m, Prada & Pan- gua s.n. on 4 Oct 1996. PEN Prov. Madrid, Sierra de Guadarrama, Peijialara, granite stone field, 2100 m, Pajarén & Pangua s.n. on 10 Oct. 1996. SCO SCOTLAND, Central Region, near Aberfoyle, slate stone field, 550 m, Jermy, Pajaron, Pangua & Lindsay s.n. on 28 Aug 1995. si Prov. Soria, Sierra de Freguela, near Puerto de Santa Inés, siliceous stone field, 1799 m, Pajarén & Pangua s.n. on 25 Sep 1995. were designed to check how temperature and light intensity affect spore ger- mination, how low storage temperatures can change spore viability and wheth- er spores are able to undergo a dormancy until spring, and if more or less sharp temperature changes can affect spore germination rate. These experi- ments are described below. MATERIALS AND METHODS We have used spores from six populations at different localities scattered in the distribution area of this species in the Iberian Peninsula. We also included a population from Scotland, which originated from a similar habitat but at lower elevation. The localities of the populations studied are listed in Table 1. Climatic data at the nearest meteorological stations were obtained from Elias Castillo & Ruiz Beltran (1977). The climatic conditions in Scotland were sim- ilar to those at the Iberian populations because of increased latitude. All spores were stored at a room temperature of 20°C. To study germination, the spores obtained from several plants in each pop- ulation were sown on sterilized 6 cm diameter Petri dishes with nutritive me- dium solidified with agar (Dyer, 1979). For each of the following culture con- ditions and each locality two dishes were sown. In all cases, the germination rates were calculated by counting 50 spores from each plate and expressed as the mean value of the two dishes. Germination was scored as spores in which the first rhizoid had emerged. In the four experiments we used white light and a 12 hour light/dark photoperiod. Temperature and light intensity conditions varied in the four experiments as described below. In experiment 1, spores from all the populations were studied. In experiments 2-4, we used only spores from the SCO, STI, and HU populations. EXPERIMENT 1.—Plates were cultivated at 30yEm~2s~' and at 10, 15, 20, and 25°C. The lowest temperature was chosen because spring months are still cold, and mean temperatures of about 10°C in May are common at these localities (Elias Castillo and Ruiz Beltran, 1977). This experiment was developed to PANGUA ET AL.: CRYPTOGRAMMA CRISPA SPORE GERMINATION 161 check the effects of temperature on the germination of spores, and to check if the spores could initiate germination at low temperatures, similar to the usual mean temperatures ocurring at the begining of spring. EXPERIMENT 2.—The temperature was maintained at 12°C for 31 days and then raised to 18°C, with samples cultivated at 10,Em~s~' and at 40nEm~?s~'. Sharp changes in temperature are not uncommon in montane climates. We chose this jump from 12° to 18°C because these are the usual mean tempera- tures in June and July respectively at these localities (Elias Castillo and Ruiz Beltran, 1977). We were attempting to check if this kind of change could ac- celerate the germination process. The different light intensities were used to check if there is an influence of this parameter on germination of the spores. Considering the habitat of these plants, it is possible for the spores to fall either on the rock surface or the soil, where they are exposed to direct sun-light, or in rock fissures and under rock blocks, where illumination is lower. EXPERIMENT 3.—After sowing, half of the plates were kept in darkness for 15 days in a refrigerator at 4°C and the other half received 15 days of darkness in a freezer at —18°C. After this period, all plates were moved to a growth-cham- ber and cultured at 21°C and 30 pEm~’s~'. If spores do not germinate in au- tumn, they should be subjected to low winter temperatures, although most of them should be protected by snow cover at these altitudes. Under snow cover, the temperatures rarely fall below freezing, and this cover also protects against wind action and desiccation as there is almost no evaporation (Geissler, 1982). In case no snow cover protection occurs, the spores would be exposed to freez- ing temperatures. EXPERIMENT 4.—In this experiment conditions of experiments 2 and 3 were combined, but at low light intensity only. The cultures were maintained at 10 yEm-~2s~' and 12°C for 14 days, then the temperature was raised to 18°C. Before the experiment, the plates were kept at chilling and at freezing temperatures as in Experiment 3 for 15 days. We wanted to check the effect of low temper- atures on the viability of the spores as in Experiment 3, but at a lower light intensity, and to check the effect of a sharp change in temperature as in ex- periment 2, but with spores previously kept in cold. A statistical analysis was carried out to test if the variations observed rep- resent real differences between treatments or merely chance differences. First, we searched the best fitting regression model, and afterwards we compared the regression lines within, ever, and among experiments when necessary. To test the goodness of fit of the model, the r-squared statistic was calculated, and also the F-ratio and p-value obtained from the analysis of variance of the model were used. A further analysis of variance for variables was carried out, and F- ratio and p-values for intercepts and slopes were obtained as well. All analyses were carried out with STATGRAPHICS Plus 3.0. RESULTS EXPERIMENT 1.—The results of germination experiments at a range of temper- atures from 10 to 25°C are presented in Fig. 1. In all samples, the spores cul- Germination % 00 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) ee oa nm aM ee hr hh tr fe] ce) ok, ie) NSN ao © ae | a 2 | = | Pe Low 13 15 17 21 24 28 31 37 39 days | 10°C 415°C 20°C 2590 PANGUA ET AL.: CRYPTOGRAMMA CRISPA SPORE GERMINATION 163 tivated at 20° and 25°C began germination four days after sowing, and after two weeks practically all viable spores had germinated. At 15°C, germination was delayed about a week, and the delay was greater in 10°C cultures, in which germination started 15-20 days after sowing. Population HU showed a differ- ent behavior (Fig. 1c), as only spores cultivated at 20° and 25°C germinated and the germination rate was less than 10%. Higher germination rates varying from 84—-100% were reached in cultures at 20° and 25°C, except in SCO, which only reached 50% (Fig. 1b). In cultures at 10° and 15°C, PEN and MON (Fig. 1a, d) had relatively high germination rates at the end of the experiment, similar to the rates obtained at higher tem- peratures. Nevertheless, in SCO and STI (Fig. 1b, f) a small decrease of spore viability was observed at 10° and 15°C and a greater decrease was seen in LNE (Fig. 1e). Statistical analysis (Table 2) shows no significant differences among the slopes, but highly significant (p<0.01) differences among intercepts, compar- ing the results of each population at each culture temperature. EXPERIMENT 2—Germination rates from SCO, STI, and HU kept at the same temperature but at two different light intensities were similar in both cases. The germination rates varied from 85-95%, except in HU. However, at a higher light intensity (Fig. 2a), germination started 22 days after sowing, whereas at 10 pEm~?s~', it began four days earlier (Fig. 2b). After day 31, at which tem- perature increased from 12 to 18°C, spore germination rates increased abruptly in all samples. No significant differences are shown in the statistical analysis between both light intensities, except for population STI (Table 2). However, there are highly significant differences when compared with the results of experiment 1. EXPERIMENT 3.—Half of the replicates were kept in darkness at 4°C for 15 days (Fig. 3a) and the other half were kept in darkness at —18°C for 15 days (Fig. 3b), before transfer to a growth chamber at 21°C and 40 »Em *s"'. The spores kept at 4°C began germinating 5 days after transfer to the growth chamber, and in a few days reached their highest germination rates. The spores kept at — 18°C began germinating 12 days after transfer to the growth chamber. The highest germination rates were reached a few days after transfer to the growth chamber. In all samples, the germination rates were clearly lower in samples exposed to freezing temperatures, especially in SCO, in which germination decreased from 80% to 10%. In both treatments, STI had the highest germination rates, followed by SCO, with the lowest germination rates in HU. Comparing these results with the ones obtained in experiment 1, it is apparent that SCO lost viability when kept at 4°C, and much more so if spores were kept at —18°C, relative to spores fone Fic. 1. Germination rates of all six populations at 10, 15, 20, and 25°C (experiment 1), during 39 days. 164 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) TABLE 2. Results of the statistical analyses of each experiment and of comparison between experiments. Columns 2-4: r° statistic indicating the percentage of variability explained by the model as fitted; F-ratio and p-value are results of the analysis of variance of the model. Columns 5-8: F-ratio and p-value for intercepts and slopes are results of the further analysis of variance for variables. Intercepts Slopes Population r F Pp F ) r Pp Experiment | EN 78,09 38,30 0,0000 11,961 0,0000 0,35 0,7901 SCO 84,27 30,61 0,0000 21,66 0,0000 1,51 0,2259 MON 83,80 29,56 0,0000 16,48 0,0000 0,11 0,9543 LNE 99 41,89 0,0000 44,68 0,0000 2351 0,0911 HU 84,21 30,47 0,0000 23515 0,0000 24,70 : STI 80,30 23,29 0,0000 15,62 0,0000 0,40 0,7514 Experiment 2 SCO 85,40 54,61 0,0000 0,37 0,5470 1,64 0,2111 STI 91,26 97,42 0,0000 7,86 0,009 1 4,26 0,0483 HU 93,47 133,63 0,0000 0,17 0,6842 1,07 0,3103 Experiment 2 vs Experiment | 66 44,56 0,0000 7,32 0,0003 8,22 0,0002 STI 84,88 38,49 0,0000 27,84 0,0000 11,38 0,0000 HU 94,32 114,02 0,0000 10,58 0,0000 51,08 0,0000 Experiment 3 96,68 252,76 0,0000 535,99 0,0000 90,29 0,0000 STI 87,26 59,37 0,0000 23,38 0,0000 0,85 0,3639 HU 95,63 189,53 0,0000 323,91 0,0000 104,35 0,0000 Experiment 4 SCO 85,17 49,78 0,0000 67,09 0,0000 33,46 0,0000 STI 85,24 50,07 0,0000 23,14 0,0001 4,55 0,0426 HU 83,91 42,95 0,0000 36,28 0,0000 37,92 0,0000 Experiment 3 vs Experiment 4 (— 18°C) 78,88 32,38 0,0000 0,62 0,4372 0.92 0,3473 STI 85,00 49,10 0,0000 15,46 0,0006 0,72 0,4030 HU 41,17 6,06 0,0028 0,85 0,3646 0,92 0,3466 Experiment 3 vs Experiment 4 (4°C) 82,28 40,23 ,0000 8,31 0,0078 147 0,2892 STI 83,14 42,74 0,0000 6,25 0,0190 0,18 0,6750 HU 85,12 49,58 0,0000 26,29 0,0000 0,19 0,6669 maintained at room temperature. The same phenomenon occurred in STI, but at a lower proportion. These results show statistically significant differences among intercepts and among slopes, except for population STI, in which differences between the slopes of both treatments were not significant (Table 2). EXPERIMENT 4.—As in experiment 3, after sowing the plates were maintained at 4° and —18°C. Afterward, they were cultivated for 14 days at 12°C followed by cultivation at 18°C, always at 10 pEm~?s-', Results are shown in Fig. 4. PANGUA ET AL.: CRYPTOGRAMMA CRISPA SPORE GERMINATION ermination % 100 2 . &ECO «STi HU -o BA i O 18 20 22 24 27 29 31 34 36 38 41 43 45 48 50 56 days 5 germination % ie =SCO «STI +HU PE CNG ERS SaaS ota A 60 |— PO a ee a OMe 20 22 24 27 D9 31 34 36 38 41 43 45 48 50 56 days Fic. 2. Germination rates obtained in experiment 2. Temperature was 12°C until day 31, then 18°C, Light intensities: a) 40.Em~*s~'; b) 102Em~?s~". Spores kept at 4°C germinated about 9 days earlier than the ones kept at —18°C. As in experiment 3, the germination rates of the latter treatment were lower, especially in population SCO, which decreased its germination from 80% to 10%. It is noteworthy that when the temperature was changed, the germination rates increased abruptly in the samples kept at chilling temperatures, whereas 166 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) germination % 80 jas 4) 60 a 4SCO «STI +HU 20 3 $57 10 12 14 17 19 21 24 26 28 31 33 39 days germination % 100 =SCO «STI +HU i (0S 8 7 10 12 14 17 10 21 24 6 oe a1 gs days Fic. 3. Germination rates obtained in experiment 3. Temperature was 21°C and light intensity 30.Em~*s~'. a) Spores pretreated at 4°C. b) Spores pretreated at —18°C. in the samples kept at freezing temperatures this increase showed up five days later. Comparing the results of experiments 3 and 4, it is apparent that similar overall germination percentages were reached at all combinations of temper- ature and light intensity. These germination rates were different in the spores pretreated at 4°C than in the ones kept at —18°C. PANGUA ET AL.: CRYPTOGRAMMA CRISPA SPORE GERMINATION : germination % 10 2 =SCO «STI HU pions 80 pie —— x eva | on jones § a 40 | 20 a sae ie mage pt eo 7 10 12 14 17 19 21 24 26 28 31 days Or ens F germination % 33 39 ie €SCO «STl *+HU 60 20 |— Of -# # #% ip 14 17 19 21 24 26 28 31 33 39 Fic. 4. Germination rates obtained in experiment 4. Light intensity was 10,Em *s'. Temperature was 12°C until day 12, then 18°C. a) Spores kept at 4°C. b) Spores kept at —18°C. As in experiment 3, statistical analysis showed significative differences among intercepts and among slopes of all three populations, although for STI it was only significant at the 95% level (p<0.05). When comparing results from experiments 3 and 4, no statistically significant differences appear among slopes in experiments with spores kept at —18°C, nor in experiments with 168 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) spores kept at 4°C. Intercepts showed statistically significant differences among experiments with spores kept at 4°C, and only in population STI in experi- ments with spores kept at —18°C. DISCUSSION The most favorable temperature for germination, as in most homosporous ferns, is about 20°C (Dyer, 1979; Raghavan, 1989). At 10° or 15°C, the spores germinate as well, although germination is delayed. The significant differences among intercepts corroborate that this delay is due to differences in culture conditions. The small differences in germination rates may be due to chance events. In the wild, 20°C can be reached easily at the altitude where C. crispa grows, at least during the summer. Above the timberline, intensive insolation produces a favorable microclimate for low-growing plants (Geissler, 1982). However, even if the temperatures remain more or less cold, spores of C. crispa could begin germinating, although they would do so later. The absence of C. crispa at lower elevations, where temperatures around 20°C are common over a long period, might be explained by competition events that are outside the scope of the present study. Germination rate varies for each population, as has been noted in some As- plenium species (Pangua et al., 1994; Prada et al., 1995). The populations PEN, STI, and MON showed the highest viability at all of the studied temperatures, whereas the population HU, for unknown reasons, had spores with very low germination. The increase in temperature in experiment 2 corresponded to an abrupt in- crease in germination rate in all cases, independent of the light intensity. In nature, especially in Mediterranean montane climates, abrupt changes in tem- perature are common at the begining of the summer (Rivas-Martinez, 1987). Spores can respond to these changes by reaching high rates of germination in a shorter time. Although temperatures may fluctuate diurnally and there are other factors of the microhabitat that can influence germination, a general warming, with related mean temperature increase, at the begining of the sum- mer, would result in accelerated germination. Different light intensity treatments do not show significant differences in germination rates, except in population STI. Nevertheless, there is a clear trend for earlier germination at a lower light intensity. This perhaps gives some ad- vantage to spores that disperse into rock crevices or under rock blocks. The cold temperatures that spores endure during the winter can affect their viability in different ways. Spores kept at 4°C, a temperature probably common in hollows between rock blocks with snow cover that protects them from freez- ing, show germination rates similar to spores kept at room temperature (20°C). Spores kept at —18°C (experiments 3, 4), a temperature easily reached in these habitats where there is no snow cover protection during winter, delay their germination and lose viability at a higher or lower percentage (depending on the population) although some germination still occurs. Of course, in nature at high elevations, freezing temperatures may be acting over a longer period PANGUA ET AL.: CRYPTOGRAMMA CRISPA SPORE GERMINATION 169 than those in our experiments. This would presumably mean that spores in exposed sites would lose viability to a greater or lesser degree. But in rock boulder areas, the common habitat of C. crispa, there are a lot of safe sites that would keep spores warmer under the snow cover. Hill (1971) observed in Thelypteris Stee Schott, Woodwardia virginica (L.) Sm., and Adiantum pedatum L. that spores stored both at 6°C and freezing temperatures retained most of their viability. He concluded that it seemed that spores are able to retain their viability during winter, although evidence suggested that the usual overwintering stage is the gametophyte. We have not observed gametophytes in the wild in any of the studied populations, either during the spring or summer; this agrees with the observations of Peck et al. (1990) for Cryptogram- ma stelleri (S.G. Gmel.) Prantl. On the other hand, Dyer and Lyndsay (1992, 1996) detected C. crispa spores in British soil spore banks that retain their germination capacity. Our results suggest that spores released by C. crispa at the end of the summer may delay their germination until the following spring. Spores that fall in cracks or other protected sites with a higher winter temperature (Young, 1985) and lower light intensity probably would germinate more quickly with faster gametophyte development. ACKNOWLEDGMENTS We are grateful to Dr. Carmen Prada for discussions and criticisms on an earlier version of ae paper, as well as to two anonymous reviewers for their comments and suggestions. Also to Felix Pérez-Garcfa and Jose Maria Iriondo, both from E.U.LT. Agricolas, for allowing us to use a growth chambers in their Departament. This research was supported by D.G.C.Y.T., project number PB94-0317 LITERATURE CITED — A. F. 1979. The culture of fern gametophytes for experimental investigation. Pp. 253-305 n A, F. Dyer, ed. The _ biology of ferns. Academic Press. Lon en. A. F,, and S. Linpsay. 1992. Soil spore banks of temperate ferns. Amer. Fern “ 82:89-122. . 1996. Soil spore banks, a new resource for conservation. Pp. 153-160 in J. M. Camus, M. cibby, and R. J. Johns, eds. Pteridology in Perspective. Royal Botanic Gardens, Kew, a ELIAS Carls F,, and L. Ruiz BELTRAN. 1977. Agroclimatologia de Espana. Ministerio de Agri- cultura. Madrid sag pa P. 1982. rg pet Communities. Pp. 167-189 in A. J. E. Smith, Bryophyte ecology. Chap- an and Hall. London. HILL, R H. 1971 ae habitat requirements for spore ace and prothallial growth of three ferns in southeastern Michigan. Amer. Fern J. 61:171—182. PANGUA, E., S. LinDsAy, and A. DYER. 1994. Spore germination ae gametophyte development in three species of Asplenium. sai Bot. (Oxford) 73:587-593. PECK, J. H., C. J. PECK, and D. R. FARRAR. 1990. Influences of life mB attributes on formation of local and oe fern isis Amer. Fern J. 80:126—142. Prapa, C., E. PANGUA, S. PAJARON, A. HERRERO, A. ESCUDERO, ny A. Rusio. 1995. A comparative study of enseniek morphology, gametangial ontogeny, and sex expression in the Asplen- ium adiantum-nigrum complex (Aspleniaceae, Pteridophyta). Ann. Bot. Fenn. 32:107-115. 170 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) RAGHAVAN, V. 1989. Development of fern gametophytes. Cambridge University Press, Cambridge, Great Brita RNY Fees Oy a 1987. Memoria del mapa de series de vegetacién de Espana. I.C.0.N.A., Ma- drid. YOUNG, J. E. 1985. Some effects of temperature on germination and egos paeeet in Asplen- ium ruta-muraria and A. trichomanes. Proc. Roy. Soc. Edinburgh 86B Woopwakrbp, F. I. 1987. Climate and plant distribution. Cambridge oe iene Cambridge, Great Britain. American Fern Journal 89(2):171-177 (1999) SEM Studies on Vessels in Ferns. 13. Nephrolepis EDWARD L. SCHNEIDER and SHERWIN CARLQUIST Santa Barbara Botanic Garden, 1212 Mission Canyon Road, Santa Barbara, CA 93105 ABSTRACT.—Vessels are present in roots, rhizomes, and stolons of N. exaltata; vessel elements are vessel surfaces; this characteristic has been observed in other ferns from habitats in which marked fluctuation of water availability occurs. As shown in other papers of this series as well as the present paper, adaptation to ecological conditions is more important than phylogenetic position in explaining the presence and degree of specialization of vessel elements in ferns. Nephrolepis exaltata (L.) Schott is a fern that has been described as epi- phytic or epipetric (Lellinger, 1985); our specimen was collected in crevices of recent lava flows in the Hawaiian Islands. The species extends from central Florida into tropical Central and South America and tropical portions of the Pacific and the Old World (Hillebrand, 1888; Lellinger, 1985; Tryon and Tryon, 1982). Thus, N. exaltata is a tropical and subtropical species that occupies exposed and pioneering habitats within its range, although it is also found in shady and moist localities, as in the Hawaiian Islands (Neal, 1965). The pio- neering characteristics of N. exaltata and its ability to withstand full sun and periodic drought are features of interest with respect to morphology of trache- ary elements. In at least some ferns of dry localities, vessel elements with more specialized perforation plates occur, as in Pteridium (Carlquist and Schneider, 1997a), Astrolepis (Carlquist and Schneider, 1997b), Woodsia scopulina D.C. Eaton (Schneider and Carlquist, 1998a), and Woodsia ilvensis (L.) R. Br. (Carl- quist and Schneider, 1998a), whereas in ferns of moist localities, vessels may be present, but perforation plates are like lateral wall pitting except for absence of pit membranes (e.g., Osmundaceae and Schizaeaceae; Carlquist and Schnei- der, 1998b). The habit of Nephrolepis offers distinctive organs that invite study with respect to potential diversity in morphology of tracheary elements. In addition to presence of relatively thick rhizomes, Nephrolepis exaltata plants bear rel- atively slender stolons. Tubers are formed on the slender stolons of N. cordi- folia (L.) C. Presl. Because flow rates might be expected to be slower in the tubers because they function in storage of water and probably other substances, one might not expect perforation plates adapted to promoting rapid conduc- tion rates in the tubers. Our studies have concentrated on ferns from habitats that show pronounced fluctuation of temperature and water availability, such as Polystichum from areas that freeze in winter and Phlebodium, a tropical epiphyte (Schneider 172 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) and Carlquist, 1997). Nevertheless, our choices have also been made with the aim of surveying a diversity of ferns with respect to systematic position. Ne- phrolepis belongs to a family we have not studied previously, Davalliaceae, although recently it has been placed in a monotypic family, Nephrolepidaceae (see Tryon and Tryon, 1982, who place it in Davalliaceae but with some res- ervations). MATERIALS AND METHODS Roots, rhizomes, and stolons of N. exaltata were collected on a lava flow at the 92.5 mile marker on Highway 11, near Kipahoehoe Natural Area Reserve, on the island of Hawaii, Hawaii, September, 1997. Tubers of N. cordifolia (L.) Pres] were obtained from plants cultivated at the Ganna Walska Lotusland Foundation, Santa Barbara, California. Portions were preserved in 50% aque- ous ethanol. Macerations of vascular tissue from roots, rhizomes, stolons, and tubers were prepared using Jeffrey's Fluid and stored in 50% aqueous ethanol. Macerations were spread onto the surfaces of aluminum stubs, air dried, sputter-coated, and examined with a Bausch and Lomb Nanolab scanning electron microscope (SEM). Our earlier studies (e.g., Carlquist and Schneider 1997a) showed that macerations were as reliable as sections in preserving pit membranes. Pit mem- brane removal due to processing is evident in the form of tearing of membranes and membrane remnants at the edges of pits, and the pattern of complete absence of pit membranes on areas corresponding to perforation plates (i.e., end walls of tracheary elements) in other vascular plants confirms this inter- pretation. RESULTS In tracheary elements of roots (Figs. 1-4), facets that lack pit membranes and are therefore perforation plates are common. These perforation plates resemble lateral wall pitting in all respects but pit membrane presence (a lateral wall is shown at left in Fig. 1 and at left in Fig. 3). In Fig. 2, a portion of a tracheary element in which three of the four facets are perforation plates is shown (two of these are on the back side of the cell); the narrow cell facet, right, contains pit membranes. Likewise, several adjacent facets that are perforation plates are present in the wide vessel at center in F ig. 3. Pit membrane remnants can be seen in some perforations at ends of perforation plates. In Fig. 4, the perfora- tion at left contains threadlike remnants of the pit membrane; the two perfo- rations, center, have small portions of weblike pit membranes; and the pit at right contains a striate pit membrane that is nearly intact. In rhizomes, long perforation plates with numerous bars, much like lateral wall pitting except for absence of pit membranes, are present (Figs. 5, 6). The vessel element shown in Fig. 5 shows a cell tip that bears a perforation plate, only a small portion of which is shown. The cell facets of the vessel element shown in Fig. 6 are probably mostly lateral walls (we were not able to delineate SCHNEIDER & CARLQUIST: VESSELS IN NEPHROLEPIS oo ee ie * *. Say “e VOee wp Ves. Si bist me a 2, 43 SA lhe tran = & € ~“_ Da: Fics. 1-4. Portions of tracheary elements of Nephrolepis exaltata from root macerations. 1) Ad- jacent tracheary elements showing (extreme left), lateral wall pitting in metaxylem element; per- foration plate in metaxylem element; and, at right, protoxylem elements. 2) Tracheary element in which facing wall and two walls on the back side are perforation plates; facet at right bears lateral wall pitting. 3). Adjacent tracheary elements showing (extreme left) lateral wall pitting, and (cen- tral element) several facets bearing perforation plates. 4) Portion of perforation plate showing strandlike pit membrane remnants (left), weblike remnants (center two perforations), and nearly intact pit membrane (right). Scale bars in all figs. = 5 ~m AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) WAHL Hier AWN \{{ SEU - nM Rl) and cetolon 6) ana stoion Fics. 5 mac pits, these may be artifacts, and all of the facets shown appear to bear lateral wall pitting. Scale bars in all figs. = 5 SCHNEIDER & CARLQUIST: VESSELS IN NEPHROLEPIS 175 end walls in this particular cell, however). Because debris behind the cell may clearly be seen through the cell, pit membranes are absent on most of the facets (at least some pit membranes are present on the facet at right). Thus, lateral wall perforation plates are present. Tracheary elements of a stolon (Figs. 7, 8) show a range of expressions. End- wall perforation plates are present on some vessel elements (Fig. 7). The very wide tracheary element in Fig. 8 has numerous facets. All of these facets have lateral wall pitting; some pits have slitlike gaps in their membranes, which we interpret as probable artifacts. Tubers of N. cordifolia (Fig. 9) are borne on slender stolons a short distance below the substrate surface. Our preparations of these stolons consisted mostly of parenchyma cells in which we observed very little starch. Portions of a few tracheary elements (Figs. 10-12) were present in these preparations, however. The cell tip in Fig. 10 does not possess well-defined perforation plates. Three facets from this tip, enlarged in Fig. 11, have striate pit membranes. The walls in Fig. 11 also have some gaps in pit membranes. Some of these gaps may be artifacts, in our opinion. In Fig. 12, facets from the lateral wall of a tracheary element are illustrated. These facets bear striate pit membranes but also some membrane-free pits. Some of these we interpret as perforations. DISCUSSION AND CONCLUSIONS Vessel elements are clearly present in the roots, rhizomes, and stolons of N. exaltata. Tracheary elements of the tubers of N. cordifolia are not so easily identified as vessels, although some perforations were identified; tracheids as well as vessels may be present in the tubers. The presence in tubers of tra- cheary elements with poorly developed perforation plates correlates with the probable water storage function of the tubers. Tracheary elements adapted to rapid conductive rates are less likely to be found in tracheary elements in storage organs than in organs in which more rapid conductive rates probably occur (Schneider and Carlquist, 1998b). Nephrolepis exaltata occurs in a variety of habitats, but some of these (in- cluding the locality at which the rhizomes, stolons, and roots were collected) have marked fluctuation in water availability. Lellinger (1985) characterized many habitats of N. exaltata and N. cordifolia as epipetric or epiphytic. Sim- ilarly rhizomatous ferns of such habitats, notably Phlebodium (Schneider and Carlquist, 1997), have vessel elements in which perforation plates are clearly present. Like Phlebodium, the perforation plates of Nephrolepis do not differ from lateral walls except in absence of pit membranes. Such vessel elements contrast with vessel elements of Woodsia species that grow in habitats with marked fluctuation in temperatures and water availability, such as W. scopu- lina (Schneider and Carlquist, 1998a) and W. ilvensis (Carlquist and Schneider, 1998a). In the Woodsia vessel elements, perforation plates have few bars and wide perforations, in contrast with lateral walls, in which the pits are narrower and shorter than perforations. Perforation plates on lateral walls as well as end walls are present in vessel 176 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 2 (1999) Fics. 9-12. Tuber (9) of Nephrolepis cordifolia and tracheary elements (10-12) from maceration of a tuber of N. cordifolia. 9) Habit of tuber, attached to stolon. 10) Tip of tracheary element, showing several facets. 11) Enlarged portion of the tracheary element shown in Fig. 10; most pits contain pit membranes, and perforation plates are either absent or poorly developed. 12) Central portion of tracheary elements, with pit membranes absent in some pits but present in most. Fig. 9, X1; scale bars in Figs. 10-12 = 5 : SCHNEIDER & CARLQUIST: VESSELS IN NEPHROLEPIS 177 elements of N. exaltata. Perforation plates on lateral walls have been reported in a number of ferns, such as Pteridium (Carlquist and Schneider, 1997a) and Phlebodium (Schneider and Carlquist, 1997). Lateral perforation plates occur in ferns of seasonally dry habitats in which numerous vessel elements in con- tact with each other occur in the vascular strands. Nephrolepis appears among more highly derived ferns in cladograms, whether based on macromorphology (Smith, 1995) or molecular evidence (Pry- er et al., 1995). The vessels of Nephrolepis are not as specialized as those of Pteridium, Astrolepis, or Woodsia (which occupy positions of moderate to high specialization in the references just cited). Degree of specialization in perfo- ration plates does not correlate with phylogenetic position as much as it de- pends on ecological and physiological factors. LITERATURE CITED CARLQUIST, S., and E. L. SCHNEIDER. 1997a. SEM studies on vessels in ferns. 2. Pteridium. Amer. J. Bot. 84:581-587. . 1997b. SEM studies on vessels in ferns. 4. Astrolepis. Amer. Fern J. 87:43-50. . 1998a. SEM studies on vessels in ferns. 6. Woodsia ilvensis, with comments on vessel origin in ferns. Flora 193:179-185. . 1998b. SEM saat on vessels in ferns. 10. Selected Osmundaceae and Schizaeaceae. Int. J. Pl. Sci. 159:788-797. HILLEBRAND, W. ier8 erie of the Hawaiian Islands. Williams & Norgate, London LELLINGER, D. B. 1985. A field manual of the ferns se fern-allies of the United States and Canada. aeigren. insttion Press, Washington, D' NEAL, M. C. 1 In gardens of Hawaii. Bishop spc Press, Honolulu, HI. PRYER, K. M., ee SMITH, and J. E. Skoc, 1995. Phylogenetic ralationships of extant ferns based n evidence from Giarphology and rbcL sequences. Amer. Fern J. 85:205—282. SCHNEIDER, E. L., and S. CARLQUIST. 1997. SEM studies on vessels in ferns. 3. Phlebodium and j 349. . 1998a. SEM studies on vessels in ferns. 5. Woodsia scopulina. Amer. Fern J. 88:17—23 . 1998b. Origin and nature of vessels in monocotyledons. 4. Araceae subfamily Philoden: droidees. ‘ Torrey Bot. Soc. 125:253-260. SMITH, A. R. 1995. Non-molecular phylogenetic hypotheses for ferns. Amer. Fern J. 85:104—122. TRYON, R. M, ic A. F. TRYON. 1982. Ferns and allied plants, with special reference to tropical America. Springer Verlag, New York. American Fern Journal 89(2):178—-179 (1999) REVIEWS The Ferns and Allied Plants of New England, by Alice F. Tryon and Robbin C. Moran. 1997. Center for Biological Conservation, Massachusetts Audubon Society, 208 South Great Road, Lincoln, MA 01773. xv, 325 pp. Hardcover (ISBN 0-932691-23-4) $49.95 plus $3.00 shipping/handling. This marvelous book, the second in the Massachusetts Audubon’s Natural History of New England series, is a most useful guide to identification of the nearly 100 ferns and fern allies of New England’s six states: Maine, Vermont, New Hampshire, Massachusetts, Connecticutt, and Rhode Island. It was care- fully prepared and tightly edited. The reader is first provided a phylogenetic list of families, genera, and species and a key to genera. Then the body of the book is organized into generic treatments, with the ferns first and then the fern allies, arranged phylogenetically. For each genus there is a generic description and key to species, with black frond sillouhettes that explain or confirm the language of the key. Species descriptions include scientific binomials, syn- onomy, common name, morphological characteristics, habitat, New England range, world range, chromosome number, remarks regarding habitats, habit, hybrids, floristics, economic use, and derivation of scientific name. Locality dot maps are provided for New England collections; tone maps for world range. The black and white photographs by W. H. Hodge and Robert L. Coffin are coffe-table art-book quality. A most distinctive feature of this work is that the text is supplemented with 142 scanning electron photomicrographs of spores showing clearly their shapes and surface features. These show how ferns of remarkably similar frond morphology may have remarkably different spores, making identifications by spores often easier than by fronds alone. A brief section on New England climate and geology is an aid for interpretive naturalists. The section on good choices for the garden presents ferns by size categories. References are listed for further study of horticultural uses of ferns. Access to any part of the book is made easy with a glossary, reference list of technical literature, and index to scientific and common names. Synonomy is sufficient to relate the taxonomy employed here to other floristic works. The authors are to be congratulated for summarizing New England pteri- dophyte floristics with an economy of words, wonderful photos, maps, and silhouettes, and very usable keys. The care and attention they took in the preparation and editing of the book contributed to the simple elegance of the final product. All this and six state fern floras, all for about $8.00 a State!— JAMES H. Peck, Department of Biology, University of Arkansas at Little Rock, 2801 S. University Ave., Little Rock, AR 72204. REVIEWS 179 Ferns of the Tropics, by Wee Yeow Chin. 1998. Timber Press, 133 S. W. Second Ave., Suite 450, Portland, OR 97204. 190 pp. Hardcover (ISBN 0- 88192-458-X) $34.95 plus shipping and handling. Dr. Wee, recently retired from National University of Singapore, has long been involved in nature conservation and nature education. This work contin- ues his efforts to provide popular books on the flora of Southeast Asia. The first half of the book is an in-depth (85 pp.) introduction to ferns and fern allies: general morphology, life cycles, economic uses, propagation, cultiva- tion, habitats, folklore, and superstitions. The second half (105 pp.) is the spe- cies section that presents a carefully selected cross-section of fern diversity in the Southeast Asian tropics. Seventy-seven species are arranged alphabetically by genus, with brief descriptions of morphlogical characteristics, habitat, habit, and cultivation requirements. The work is supplemented with a glossary, bib- liography, index to common and scientific names, and most importantly to new fern fanciers, fern society addresses, e-mail addresses, and websites. The text is lean and simply written. It supplements the numerous gorgeous color photos that will make any reader a fern fanatic. There is a good mix of close- ups, habitat and habit shots, and stunning full page photos that emphasize the incredible form diversity of ferns in the tropics. This slim volume packs a lot of the tropics into a small space, and it will captivate any reader in temperate regions.—JAMES H. Peck, Department of Biology, antici of Arkansas at Lit- tle Rock, 2801 S. University Ave., Little Rock, AR 7 DTT INFORMATION FOR AUTHORS Authors are encouraged to submit manuscripts pertinent to pteridology for pub- lication in the American Fern Journal. Manuscripts should be sent to the Editor. Acceptance of papers for publication depends on merit as judged by two or more referees. 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VISIT THE AMERICAN FERN SOCIETY’S WORLD WIDE WEB HOMEPAGE: http://www.visuallink.net/fern AMERICAN ate FERN — JOURNAL a QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Two New Fern Species from Southern Mexico 181 Francisco G. Lorea-Hernandez and Alan R. Smith Rush Quillwort (Isoetes junciformis, sp. nov.), a New Pteridophyte from Southern 187 Georgia Daniel F. Brunton and Donald M. Britton Some Observations on the Reproductive Anatomy of Isoetes andicola Eric Karrfalt 198 Ontogeny of the Sporangia of Sphaeropteris cooperi Kenneth A. Wilson 204 Shorter Notes Salvinia minima in Arkansas 215 Two Additional Stations for the Southern Woodfern Hybrid, Dryopteris x australis in 216 Maryland 3-C-(6''’-O-Acetyl-B-cellobiosyl) Apigenin, a New Flavonoid from Pteris vittata 217 The American Fern Society Council for 1999 DIANA B. STEIN, Dept. of Biological Sciences, Mount Holyoke College, South Hadley, MA 01075-6418 President BARBARA JOE HOSHIZAKI, 557 N. Westmoreland Ave., Los Angeles, CA 90004-2210. Vice-President W. CARL TAYLOR, 800 W. Wells St., Milwaukee Public Museum, Milwaukee, WI 53233-1478. Secretary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916-1110. Treasurer DAVID B. LELLINGER, 326 West St. NW., Vienna, VA 22180-4151. he gp ig Secretary JAMES D. MONTGOMERY, Ecology HI, R-.D. 1, Box 1795, Berwick, PA pis 2 9801 ‘ack Issues Curator GEORGE YATSKIEVYCH, Missouri Botanical Garden, PO. Box 299, St. eho ‘MO 63166-0299. Journal Editor DAVID B. LELLINGER, U.S. National Herbarium MRC-166, Smithsonian Institution, Washington, DC 20560-0166. Memoir Editor parent JOHNSON-GROH, Dept. of Biology, Gustavus Adolphus College, 00 W. College Ave., St. Peter, MN 56082-1498 Bulletin Editor American Fern Journal EDITOR GEORGE YATSKIEVYCH Missouri Botanical Garden, P.O. Box 299, St. Louis, MO 63166-0299 ph. (314) 577-9522, e-mail: gyatskievych @lehmann.mobot.org ASSOCIATE EDITORS GERALD J. GASTONY.......... port : ies ereaes raeg id “ce IN 47405-6801 CHRISTOPHER H. HAUFLER .. ersity oO s, Lawrence, KS 66045-2106 ROBBIN C. MORAN aaa sonteres aca: Bronx, ‘NY 10458-5126 JAMES H. PECK A ‘ agi University of Arkansas—Little Rock, 1 S. University Ave., Little Rock, AR 72204 e “American ie Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the general study of ferns. It is owned by the American — Society, and aver at shoes West St. NW., Vienna, VA 37180-4151. Periodicals postage paid at Vienna, VA, and addition Claims for missing issues, made 6 months aan to 12 months cae after the date of i a and orders for back issues should be addressed to Dr. James D. Montgomery, Ecology III, R.D Berwick, PA 18603-9801. Changes of address, dues, and applications for membership should be sent to the Membership cpm Gene quiries rorya ferns should be addressed to the Secretary. Sohieciolies $20.00 gross, $19.50 net if paid through an agency oni fee $0.50); sent free to members of the ppg Fern Society (annual dues, $15.00 + $5.00 mailing surcharge beyond U.S.A.; life membership, $300.00 + $140.00 mailing surcharge eae U.S.A. Back volumes are available for most years as printed issues or on microfiche. Please contact the Back Issues Curator for prices and pvallabiliey POSTMASTER: Send address changes to AMERICAN FERN JOURNAL, 326 West St. NW., Vienna, VA 22180-4151. FIDDLEHEAD FORUM The editor . foe Bulletin of the American Fern Society welcomes contributions from members and non-members, including patient notes, offers to Apron or purchase materials, personalia, horticultural geek and reviews of non-technical books on fern SPORE EXCHANGE Ms. Donna McGraw, 8207 Colonial Oaks Ln., Spring, TX 77379 (e- — a com), is Director. Spores exchanged and lists of available spores sent on reques GIFTS AND BEQUESTS Gitts and bequests to the Society enable it to expand its services to members and to others interested in ferns. Back issues of the Journal and cash or other gifts are always welcomed and are tax-deductible. Inquiries should be addressed to the Secretary. MISSouR} BOTANICAL American Fern Journal 89(3):181—186 (1999) SEP 2 3 1999 Two New Fern Species from Southern MéXicw«r FRANCISCO G. LOREA-HERNANDEZ Instituo de Ecologia, A.C., Apdo. Postal 63, 91000 Xalapa, Veracruz, Mexico ALAN R. SMITH University Herbarium, 1001 Valley Life Sciences, #2465, University of California, Berkeley, CA 94720-2465 ABSTRACT.—Diplazium errans and Polystichum schizophyllum, endemic to the state of Guerrero, Mexico, are described and illustrated. The former is distinct by its long lanceolate once-pinnate leaves with numerous pinna-pairs and the presence of rhizogenous buds on the rachis toward the blade tip. The latter is distinguished by its tripinnate blades, persistent light-tan indusia, and by marginate, most often black-tipped scales on the stipe and rachis. The relationships of these spe- cies within the corresponding genera and especially to their Mexican congeners is discussed. UMEN.—Se describen e ilustran Diplazium errans y Polystichum schizophyllum, especies en- démicas del Estado de Guerrero. La primera se distingue por sus largas hojas lanceoladas una vez claro y la presencia con frecuencia de escamas marginadas con puntas negras en el pecfolo y raquis. Se comentan ademas las relaciones de estas especies con otras de los géneros correspon- dientes, en particular con sus congéneres mexicanos In the early 1980s, the state of Guerrero in southern Mexico was targeted as one of the regions for which floristic knowledge was poor. For the last fifteen years, different Mexican research institutions have collected extensively in the state in order to amass collections sufficient to publish a proper floristic account. Part of the outcome of these years of field work has been the discovery of several new plant species. Here we name two of the previously undescribed ferns. Ferns in Mexico are fairly well known, although relatively few fern floras have been published (Matuda, 1956; Knobloch and Correll, 1962; Smith, 1981; Mickel and Beitel, 1988; Mickel, 1992). The number of pteridophyte species in Mexico is expected to be around 1000 (Riba and Pérez-Garcia, 1994; Mickel and Smith, in prep.). In pteridophyte diversity, the state of Guerrero, with 373 species (Lorea and Velazquez, 1998), is fourth among the Mexican states, be- hind Oaxaca, Chiapas, and Veracruz. With the description of the following new species, Guerrero now has three endemic pteridophytes, including Selaginella rzedowskii Lorea-Hern. (Lorea, 1983). Diplazium errans Lorea-Hern. & A.R. Sm., sp. nov. (Fig. 1).—TyPe: Mexico, Guerrero, Mpio. Petatlan, 10 km NNE of El] Mameyal, dirt road Papanoa— Corrales, 1000 m, 3 Mar 1985, M. G. Campos 1531 (XAL; isotypes NY, UC) Diplazio werckleano H. Christ affinis, a qua imprimis differt laminis lon- gioribus, pinnis plus quam 20 paribus per frondem (vs. 5-10 paribus in D. werckleano), rhachidibus distaliter 1-2 gemmis rhizophoris praeditis. AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) 182 ogg LF Wes E LE Ss AE SO MS SS WAS gs ) WA g SS =» WS ( () ; M \ ) | y Ng NZ > Ve izes Diphziom Mls. E Saavedra LOREA-HERNANDEZ & SMITH: TWO NEW MEXICAN FERNS 183 Rhizomes ascending to erect; rhizome scales dark brown, lustrous, 0.5 X 0.5—1 mm, lanceolate, entire; fronds clumped, stipes 34-40 cm, Y,—¥, the frond length, pale gray-green or pale yellow-green, adaxially grooved, glabrous ex- cept for some scales at base, these dark to light brown and rather dull; blades 52-66 X 17-22 cm, lanceolate, 1-pinnate, free pinnae (20—) 24 (—28) pairs, apices pinnatifid; rachises grooved, pinna rachis groove open to main rachis groove, lacking hairs but minutely papillate (papillae 0.1 mm long), pale yel- lowish green, one or two buds developing in pinna axils adaxially on distal fourth of rachis; pinnae ascending, slightly falcate, bases inequilateral, cuneate basiscopically, slightly auricled acroscopically, margins shallowly lobed to un- dulate and with lobes and undulations faintly serrulate, largest pinnae (usually third pair) 11-15 X 1.7—2.4 cm, lanceolate, short-stalked (5-7 mm), apices caudate-acuminate, smallest pinnae 0.7-1.3 X 0.4—0.7 cm, elliptic or rhombic, sessile, apices acute or obtuse; costae and blades glabrous abaxially; veins free, branched 4—6 times (2-3 pairs); sori along 1-3 (—4) acroscopic and also along 1-3 basiscopic veins of a vein group, double sori uncommon, when present along the 1-3 most proximal veins of a vein group; indusia 2.5-9.5 x 0.2-0.4 mm, entire; spores ca. 64 per sporangium, 44-50 < 26-32 ym (including the perine), perine 4—6 pm thick. The closest affinity of this species is with D. werckleanum H. Christ, which agrees in the presence of once pinnate blades, slightly lobed pinnae, venation, and soriation. However, D. werckleanum has at most 10 (usually 5-10) free pinna pairs per frond and lacks buds on the rachis, whereas D. errans has more than 20 free pinna-pairs per frond and bears buds on the main rachis. Diplazium werckleanum is known from southern Mexico (including uncom- monly in Guerrero) to Panama and Colombia. Diplazium werckleanum belongs to a group of species that have usually been distinguished by the degree of lamina dissection and the inequilateral pinnae. It is especially close to D. cris- tatum (Desr.) Alston, which has more deeply lobed pinnae. Other species in this group, for example D. drepanolobium A.R. Sm. and D. lonchophyllum Kunze, have free basal acroscopic pinnules and are morphologically less close- ly related to D. errans. None of the species of the D. werckleanum group has rhizogenous buds on the rachis. Other gemmiferous Mexican diplaziums include D. altissimum (Jenm.) C. Chr. (bipinnate- pinnatifid blades; syn. = D. entecnum Mickel & Beitel), D. neglectum (H. Karst.) C. Chr. (once pinnate blade with equilateral pinnae), D. obscurum H. Christ (once pinnate blade with a terminal conform pinna), D. plantaginifolium (L.) Urb. (simple blade), D. ternatum Liebm. (blade ternate, with two lateral pinnae and a conform apical one), D. urticifolium H. Christ (once pinnate blade with equilateral pinnae), and D. vera-pax (Donn. Sm.) Hieron. (once pinnate blade with pinnatifid apex). Of these, the most similar to D. errans is D. vera-pax, which occurs in Veracruz and Chiapas. This species oe Fic. 1. Diplazium errans. a) rhizome and stipe base; b) stipe base scales; c) blade; d) proximal pinna; e) sorus and indusium, side view. 184 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) was synonymized by Adams (1995) under D. riedelianum (Bong. ex Kuhn) Kuhn ex C. Chr., a Brazilian species that we regard as distinct. Diplazium vera- pax has only 1-3 free pinna pairs, and is perhaps the hybrid between D. plan- taginifolium, with simple blades, and D. werckleanum. It seems unlikely that D. errans is a hybrid, as no other once-pinnate Diplazium with inequilateral pinna bases, more than 20 pinna pairs, glabrous blades, and gemmiferous buds is known to occur in Mexico or Mesoamerica. Moreover, the spores of D. errans are well-formed (kidney-shaped and perispore with a loose reticulate wing); this argues against a hybrid origin or hybrid status for D. errans. Currently, Diplazium errans is known only from the type collection; how- ever, the species was not rare in the area, according to observations by the collector. However, pine-oak forests at ca. 1000 m on the western slopes of the Sierra Madre, where D. errans grows, are not common in Guerrero, having been mostly cut for farming and logging operations. Polystichum schizophyllum Lorea-Hern. & A.R. Sm., sp. nov. (Fig. 2).—TypeE: Mexico, Guerrero, Mpio. Malinaltepec, 4 km S of Paraje Montero, 2000 m, 7 May 1989, F. Lorea 4574 (XAL; isotype UC). Differt a P. hartwegii (Klotzsch) Hieron. laminis 3-pinnatis, 2-10 (—16) seg- ments discretis per pinnulam, segmentis basi constrictis, stipitum paleis in- distincte bicoloribus, in medio fuscis, ad apicem interdum denigratis, ad mar- ginem fulvis, indusiis pallide fulvis, ca. 1 mm diam., persistentibus. Rhizomes erect, massive; fronds clumped, 1-1.4 m, stipes 30-55 cm, stra- mineous to dark red-brown, sparsely to densely scaly, scales of two types, some somewhat bicolorous with a central, shining, dark brown to blackish band (the distal part sometimes blackish) and wide, translucent, light brown to brown margins, structurally marginate toward the base, 6-20 X 0.8—4.5 mm, lanceolate to ovate-lanceolate, entire to erose or fibrillose toward the tip, more abundant toward the rachis base, not persistent, others light brown, concolor- ous, dull, 2-9 x 0.1-1.5 mm, capillary to lanceolate, entire or denticulate to long ciliate, more abundant toward the rachis tip; blades 70-92 x 28-42 cm, deltate, tripinnate, free pinnae 32-36 pairs, strongly ascending toward the blade apex, gradually reduced in size to a pinnatifid tip, largest pinnae the third or fourth pair, lowest pinnae 13-22 x 2.5-8 cm, lanceolate-deltate, apices pinnatifid, caudate, free pinnules 20-25 pairs in largest pinnae, larger pinnules (on largest pinnae) each with 2-10 (-16) free segments, these constricted to the midrib, penultimate blade segments inequilateral at base, basiscopically excavated, acroscopically the lobes larger and more spreading or slightly au- riculate, margins denticulate-spinulose; rachises and higher order axes grooved, sparsely to conspicuously scaly, scales lanceolate to capillary, light brown, dull, mostly entire or remotely denticulate to short-ciliate, laminar tis- sue glabrous adaxially, with sparse hairlike scales abaxially; sori 1-6 per ul- timate segment; indusia (0.6—) 1 (-1.7) mm in diameter, circular or nearly so, entire, light tan. LOREA-HERNANDEZ & SMITH: TWO NEW MEXICAN FERNS 185 %Y & * Y 1 Ng Sy, = YSU ws She), SY SSS = URS SY SSSA PEN TS a a ae ey SC oe: REA \ gris Ron Ye op ow se A AES oe yak Ae 3 ON = , SANS Eee er II ESE Fs PASS 1 Dre T RST Es ele AS SOS, St S05 pws S's Bite Os Oss DS OSIR IN ones EAS NDS One SN SON Ras ea rs x % : a eS “t Mt Pe = Aiea sat KARSN e/ S e x “RN Nie FE aN t = * id Brel rae es + Ace By OZ, et wv S27 ANA we ‘ AT AR : RABICS AS 3¢ aes 2 i le Pa A SASF VU ACN ANN Mess % RAS y See 2s UNAS Ee Nee . ASG ees ¥ SS SY ee’ ZN WENA, RUERES ic BS Rese Vig ge tg Sly) rap LEZ SEN SP Shs ETA ERR RAS 1a Se pee ~ eep ee y¥ wv ay Sy ae SS GL SHALES fy, RP IS WES . Lx Pe a=W ENG Ne EES CED LS . ~ Ney: oe Bi op fps Oy fet Ash ue gine «el ; My Fic. 2. Polystichum schizophyllum. a) general view of leaf (basal pairs of pinnae not shown); b) stipe scales; c) proximal pinnule, abaxial view. 186 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) ARATYPES.—MEXICO. Guerrero. Mpio. Malinaltepec, ca. 5 km N of Paraje Montero, 2210 m, F. Lorea 4538 (XAL, UC). Aside from P. schizophyllum, there is only one other Mexican species of Polystichum that is fully tripinnate: P speciosissimum (A. Braun ex Kunze) Copel. Nevertheless, these species seem not to be closely related, as suggested in the latter species by the copious concolorous scales along stipes and ra- chises, the several to many (to 15) pairs of reduced proximal pinnae per frond, the beadlike segments with strongly revoluted margins, and the presence of exindusiate sori. The presence of black-tinged scales mixed with fibrillose scales along the stipes and rachises in P. schizophyllum suggests a relationship to P. distans E. Fourn. However, the latter species has bipinnate fronds, more clearly bicolor- ous scales with a shining black center and narrow light brown margins, and generally smaller indusia. It is likely that P schizophyllum is most closely related to P. hartwegii (Klotzsch) Hieron., which also differs in its bipinnate fronds, but with a tendency, in some specimens, to have a nearly free acro- scopic lobe on the pinnules. Polystichum hartwegii is widespread and variable in many Mexican states, and extends through Mesoamerica and even into northern South America. Polystichum schizophyllum is known from a small area in the southern half of the Sierra Madre in Guerrero with mixed oak forest. The species is expected to thrive at least in several places where patches of oak forest occur in the mountains of southern Guerrero. ACKNOWLEDGMENTS We thank E. Saavedra for rendering the meticulous drawings that illustrate these new species. LITERATURE CITED ApaMs, C. D. 1995. Diplazium Sw. Pp. 228-246 in G. Davidse, M. Sousa S., and S. Knapp, general eds. Flora Mesoamericana. Vol. 1. Psilotaceae a Salviniaceae (volume eds., R. C. Moran and R. Riba.). ae Nacional Auténoma de México, Instituto de Biologia, Mexico City. KNOBLOCH, I. W., D. S. CoRRELL. 1962. Ferns and fern allies of Chihuahua. Texas Research undation ee r, Texas LorEA, F. 1983. sae iar reedowskil, una nueva especie de selaginela heteréfila del estado de Guerrero. Bol. Soc. Bot. México 44:2 LorEA, F., and E, sepsis 1998. Peridot, lista de los taxa y su distribucién en la entidad. Estudios agape en Guerrero MatTupaA, E. 1956. Los helechos del Valle ri México y alrededores. Anales Inst. Biol. Univ. Nac. México ae MICKEL, J. 1992. Pteridophytes. Pp. 120-431 in R. McVaugh, ed. Flora Novo-Galiciana, Vol. 17 Gymnosperms and pteridophytes (gen. ed., W. R. Anderson). University of Michigan Har. barium, Ann Arbor MICKEL, J., and J. BEITEL. 1988. Pteridophyte flora of Oaxaca, Mexico. Mem. New York Bot. Gard. 1-5 Risa, R. and B. Poergiinieaee 1994. Perspectivas en el estudio de las pteridofitas. Bol. Soc. Bot. México 55:129-13 SMITH, A., R. 1981. Pacidnayeis Pp. 1-370 in D. E. Breedlove, ed. Flora of Chiapas. Part. 2. California Academy of Sciences, San Francisco. American Fern Journal 89(3):187—197 (1999) Rush Quillwort (Isoetes junciformis, sp. nov.), a New Pteridophyte from Southern Georgia DANIEL F. BRUNTON 216 Lincoln Heights Road, Ottawa, Ontario K2B 8A8, Canada DONALD M. BRITTON Department of Molecular Biology and Genetics, University of Guelph, Guelph, ntario N1G 2W1 Canada ABSTRACT.—A previously undescribed pteridophyte, Isoetes junciformis, is reported from Tift County, Georgia. This new tetraploid appears to be a rare endemic of the upper Coastal Plain region. It is suspected to be an allopolyploid, possibly arising from the hybrid between I. flaccida and I. melanopoda. Quillworts (Isoetaceae) have traditionally been considered rare pterido- phytes in the Coastal Plain region of southern Georgia. Snyder and Bruce (1986) reported only the diploids, Isoetes engelmannii A. Braun and Isoetes flaccida Shuttlew. ex A. Braun, from this area. They also noted a number of suspected sterile hybrids, viz., those from southern Georgia listed in Boom (1982). All of those specimens of suspected hybrids, however, have been re- vised to one or another species, as have the southern Georgia specimens of I. engelmannii listed in Snyder and Bruce (1986). Despite these reductions, the total number of quillwort taxa in southern Georgia has increased dramatically in recent years. Luebke (1992) described two new hexaploid species, Isoetes georgiana Lueb- ke and I. boomii Luebke, as endemics of the upper coastal plain. The former subsequently has been found to be locally common in several southern Georgia watersheds (Brunton and Britton, 1996b). The recently described tetraploid, I. hyemalis D.F. Brunt. (Brunton et al., 1994), has now been recorded from three counties in southwestern Georgia, from a turn-of-the-century collection (Brun- ton and Britton, 1996a) and through contemporary field work (J. R. Allison, pers. comm.). Another newly described tetraploid, Isoetes appalachiana D.F. Brunt. & D.M. Britton (Brunton and Britton, 1997), was also found in two counties in southern Georgia during field investigations by R. Carter and J. R. Allison (pers. comm.). Finally, collections from 1949 have been seen recently that confirm the occurrence of Isoetes melanopoda Gay & Dur. from Miller County in southwestern Georgia (Big Drain below Babcock Pond, Thorne & Muenscher 9114 [GA, PH]). The identities of other Isoetes populations in southern Georgia have yet to be settled, indicating that the discovery of other taxa, including previously undescribed species, is possible. In this paper we report an addition to the list of Isoetes in southern Georgia. The taxon described below is from a population first discovered in 1970 by 188 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) W. R. Faircloth and apparently represents a rare, previously unknown coastal plain endemic. MATERIALS AND METHODS As part of ongoing systematic studies of Isoetes in North America, approx- imately 1,500 herbarium specimens of Isoetes have been studied from the southeastern United States deposited at CAN, DFB (D. F. Brunton personal herbarium), DUKE, FLAS, FSU, GA, MICH, NCSC, NCU, NYS, OAG, PH, PSU, UNA, UNCC, USCH, USF, VDB, VPI, and VSC, as well as selected specimens from GH, MO, NY, and US. Specimens that could not be attributed to estab- lished taxa were detected during these herbarium studies and, where possible, site investigations of the populations of origin were undertaken. Scanning elec- tron microscope (SEM) photographs of selected megaspore and microspore samples were taken using the standard methods of Britton and Brunton, (1989, 1992). Microspores were measured in Euparol, as described by Britton (1991). Megaspore widths (to the outer edges of spore ornamentation) were measured at a magnification of 40x or 50 on SEM stubs or in sporewells (Brunton, 1990) using a binocular stereo microscope equipped with an ocular micro- meter. Chromosome counts were obtained from living material of Brunton & Mc- Intosh 11,818 and Brunton & McIntosh 1 3,525 from Chula, Tift Co., GA. Plants were grown in distilled water in a growth cabinet. The developing root tips were excised and pre-treated in aqueous paradichlorobenzene (PDB) at room temperature for four hours. Then, they were washed in distilled water, fixed in acetic alcohol (3:1 absolute ethyl alcohol to glacial acetic acid) for 30 min- utes or more, hydrolysed in Warmke’s solution (1:1 concentrated HCl to ab- solute ethyl alcohol) for 7-10 minutes at room temperature, and stained in leucobasic fuchsin (Feulgen) for two hours. The meristems were squashed un- der a cover glass in 45% acetocarmine stain and examined under a light mi- croscope. RESULTS We could not attribute a collection identified as Isoetes flaccida from Chula, Tift County, Georgia (W.R. Faircloth 6690), to any known Isoetes taxa. Cyto- logical investigation of living plants obtained from the site indicated that these plants are tetraploid (2n = 44), a chromosome number previously undetected in Isoetes in Georgia. The population also proved to be morphologically dis- tinct from the two newly described tetraploids, I. appalachiana and I. hye- malis, which have been discovered recently in this region. The following de- scribes the morphological characteristics of the unnamed tetraploid from Chu- la, Georgia. Gross MorPHOLOGY.—The Chula tetraploid is a large quillwort, with + erect, BRUNTON & BRITTON: A NEW ISOETES FROM GEORGIA 189 Fic. 1. Isoetes junciformis plant (arrow) among graminoids (Tift County, GA, 4 May 1994) dull, pale green to grayish green leaves reaching lengths of 35-40 cm (Fig. 1). Although many quillworts bear a superficial resemblance to newly developing graminoid plants, the Chula tetraploid appears remarkably like the young Jun- cus or sedges with which it associates (Fig. 2). Its pale-colored leaves are white to hyaline at the base. Many young plants have a distinctly pinkish purple wash through the pale basal 2-3 cm of their leaves. The oval, heavily short-brown-streaked sporangia are each topped by a rel- atively large, narrowly-triangular, blunt-tipped and delicate ligule. Fresh, in- tact ligules (mostly on immature leaves) reach 35-40% the length of the spor- angium. The velum extends from the base of the ligule across ca. 40% of the sporangium (Fig. 3). The rounded or two-lobed corm supports a dense mass of round, hollow, relatively straight, grayish brown roots that branch dichotomously near the ends. The roots become flattened and a darker dusky brown color upon drying. MEGASPORE SIZE AND MorPHOLOGY.—Well-formed, intact megaspores average ca. 460 ym in diameter. Megaspores have a glazed, porcelain-like surface. The proximal hemisphere is densely covered with low, irregular protuberances and short, broad mounds (Fig. 4a). In lateral view, a broad band of subdued, ob- scure mounds usually can been seen bordering the distal side of the equatorial ridge (Fig. 4b). The distal hemisphere of well-formed megaspores is promi- nently ornamented with a broken-reticulate pattern of low, broad, intercon- necting ridges (Fig. 4c). The megaspores of some plants of the Chula tetraploid are variable in size 190 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) Fic. 2. Young Isoetes junciformis (arrow) with an immature rush (Juncus sp.) (Tift County, GA, 19 Mar 1998). and ornamentation. Some of the smaller spores (430-440 .m) are misshapened and with a dense, broken-reticulate ornamentation pattern. The variable, some- what polymorphic megaspore ornamentation observed on these specimens re- sembles the condition observed in primary sterile hybrids. Despite careful searching of the site and its vicinity in 1994 and 1998, however, no other taxa have been discovered, although Isoetes georgiana was reported at ‘‘many dif- ferent areas of swamp forest” in the adjacent floodplain of Whiddons Mill Creek (Musselman & Allison 96-207 [ODU]). This is in contrast to finding at least one and usually both of the putative parents growing with al] confirmed hybrid populations in North America (Britton 1991; Britton and Brunton, 1996; Musselman et al. 1997). Stronger evidence yet against a hybrid origin of the Chula population is provided by the determination of tetraploid chromosome counts even from plants with some megaspores of variable size and polymorphic ornamentation (e.g., Brunton & McIntosh 13,525) (see also Origins, below). Accordingly, be- cause all plants in the Chula population are tetraploids whether they produce either uniform or some polymorphic megaspores, we believe that megaspore variation in this case is most likely the result of developmental polymorphism. It is likely environmentally induced in the Chula tetraploid, as it appears to be in many populations of other southeastern quillworts that also develop in ecologically stressful situations, Examples of such taxa include the ephemeral BRUNTON & BRITTON: A NEW ISOETES FROM GEORGIA 191 Im_ -leaf margin (hyaline) li = ligule (35 - 45% of sporanium length) la -labium (sharp-poi ve fe -fenestra (window over sporangium) Sp -sporangium (short-brown-streaked) 0 5 10mm Fic. 3. Basal adaxial side of megasporophyll of Isoetes junciformis (tracing from Brunton & Crins 11,848 [DFB)). wetland species, I. melanospora Engelm., I. piedmontana (N. Pfeiff.) C.F. Reed, I. virginica N. Pfeiff., and I. melanopoda. MICROSPORE SIZE AND MORPHOLOGY.—The oval microspores average approxi- mately 30 wm in length (Fig. 4d). The microspores are strongly spinulose, being densely covered with fine-tipped but relatively broad-based spines on all surfaces. SITE EcoLoGy.—The Chula tetraploid is found in a low, seasonally flooded swale at the base of a northwest-facing sandy slope (Fig. 5). The plants are found most commonly as scattered individuals in areas of the swale that re- main most deeply flooded for the longest period. They are usually growing somewhat in isolation of associated graminoid vegetation. The quillworts grow with their corms at the bottom of a 3-5 cm deep layer of silty-clay (pH +6.0) with their roots extending beneath that into coarse sand. No plants were found in those portions of the swale where the substrate lacks a substantial silt or clay component. The site was submerged by 15-30 cm of quietly flowing water during the height of spring floods in March 1998 (pers. obs.). In recent years (1994-1997), the site was virtually dry by early May. It occupies a narrow intermediate zone between the adjacent upland area and the outer edge of a mature, deciduous floodplain swamp forest dominated (at its edge) by red maple (Acer rubrum L.) and sweet gum (Liquidamber styraciflua L.). The original character of the upland vegetation is unknown as the site was logged about 1990, then planted with loblolly pine (Pinus taeda L.) seedlings. Plants become more difficult to distinguish from associated graminoid veg- etation as the swale dries out; the site is typically dominated by graminoid vegetation by May or June (W. R. F aircloth, pers. comm.; pers. obs.). The few small plants detectable in August 1994 among the relatively dense graminoid vegetation possibly represent new growth responding to periodic mid-summer 192 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) Fic. 4. Isoetes junciformis spores (Brunton & Crins 11,848 [OAC)). a) proximal view of megaspore; b) lateral view of megaspore; c) distal view of megaspore; d) microspores. flooding. Although pine saplings in the adjacent plantation have dramatically increased in height over this period, it is not apparent that any significant change in the amount of light reaching the quillwort site has occurred. DISTRIBUTION AND STATUS.—More than fifty plants were observed along a 30- 40 m length of the quillwort swale in March and May 1994. Approximately 75-100 plants were observed here during the height of the March 1998 flood- ing. No other populations of the Chula tetraploid have been confirmed. Speci- mens from near Leary, Calhoun County, in southwestern Georgia (Kral s.n., 11 May 1977, VDB 158307, VDB 158308) appear to have comparable morpholog- ical characteristics to the Chula tetraploid, including megaspore and micro- BRUNTON & BRITTON: A NEW ISOETES FROM GEORGIA 193 . 5. Isoetes junciformis site in seasonally flooded swale between riparian swamp forest (right) a cleared (formerly forested) upland area (left) (Tift County, GA, 19 Mar 1994 spore size and ornamentation, but chromosome counts from plants of the Cal- houn County population have not been made. DISCUSSION Sufficient morphological and cytological evidence has been gathered from live and preserved material to indicate that the Chula tetraploid represents a previously undescribed species. Accordingly, the following binomial is pro- posed: Isoetes junciformis D.F. Brunt. & D.M. Britton, sp. nov.—TyPE: U.S.A. Georgia, Tift County, 7 km WSW of Chula, Whiddons Creek near Little River, 3 May 1994 D.F. Brunton & WJ. Crins 11,848 (OAC; isotypes MICH, MIL, DFB) Herba erecta, inucea; folia glauca, velum tegens +40% sporangii, maculis bruneis maculati; megasporae +460 ym ornatae iugis humilibus atque latis, inter se concurrentibus; microsporae ovales, echinatae, +30 y»m. Chromoso- matum numerus 2n = In the following summary description of Isoetes junciformis, features partic- ularly helpful for its identification are in boldface. Robust (25-40 cm tall), amphibious, perennial herbs from a 1.5-2.5 cm wide, rounded to two-lobed corm with numerous round, hollow, gray-brown, mostly unbranched roots; 194 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) leaves stiffly erect to somewhat reflexed, pale, grayish lime-green, white to hyaline, basal 2-3 cm lightly washed with pinkish-purple (at least when young); sporangia oval, ca. 7.5 < 4.0 mm, hyaline to white surface densely short-brown-streaked; velum covering 38.9% (SD 6.62%, N = 23) of sporan- gium; ligule delicate, narrowly triangular, obtuse, 35-40% the length of the sporangium; megaspores 458.9 ym (SD 39.64 pm, N = 50) in diameter when well-formed; some variable in size and apparently developmentally polymor- phic, with ornamentation of low, broadly rounded ridges in a ragged, broken- reticulate pattern and conspicuous broad to narrow band of subdued, obscure ornamentation bordering the distal side of the equatorial ridge; microspores oval, densely spinulose with broad-based, fine-tipped spines on all surfaces; 29.9 wm (SD 1.26 wm, N = 20) long; Cytology: 2n = 44. The epithet reflects the rush-like appearance of well-developed plants. PARATYPES.—U.S.A. Georgia. Tift County, WSW of Chula, Whiddon’s Mill Creek near its junction with Little River, W.R. Faircloth 6690 (GA, VSC); D.F. Brunton & K.L. McIntosh 11,818 (DFB, OAC); D.F. Brunton & K.L. McIntosh 12,051 (DFB); D.F. Brunton & K.L. McIntosh 13,525 (DFB, OAC). SIMILAR SPECIES.—The diploid Isoetes melanopoda appears to be morpholog- ically the most similar species to tetraploid I. junciformis, particularly with regard to their grayish green, stiffly-erect to reflexed leaves, often with a pink- ish purple wash in the pale basal section. Indeed, I. junciformis looks like a robust I. melanopoda with atypically bold megaspore ornamentation and an exceptionally large velum. Isoetes melanopoda, however, has smaller mega- spores (404 um, N = 60), with a substantially more obscure megaspore orna- menation (moderately to densely covered in small, low tubercles or short, ver- miform crests and mounds) and a velum coverage rarely exceeding 15%. The other swampland diploid in south Georgia, I. flaccida, also has smaller (391 ym, N = 45), typically more obscurely ornamented megaspores (sparely to densely low tuberculate or with short, irregular, vermiform ridges), dark green leaves, and a sprawling, flaccid stature. It also is characterized by an extensive (80-100%) velum coverage of the sporangium. Of the possible tetraploids, Isoetes hyemalis can be discriminated from I. junciformis by its more tuberculate megaspore ornamentation, shorter (15— 20%) velum coverage, and dull olive-green to dark green, strongly reflexed leaves (Brunton et al., 1994). Tetraploid I. appalachiana also exhibits a shorter (20-25%) velum coverage and has dull olive-green to dark-green, strongly re- flexed leaves, as well as a high-walled, strongly reticulate megaspore orna- mentation pattern (Brunton and Britton, 1997). Isoetes louisianensis Thieret, the tetraploid endemic of coastal plain swamp forests in southern Louisiana and adjacent Mississippi (Lark, 1996), exhibits an even more congested, high- walled and reticulate megaspore ornamentation patttern with a distinctive equatorial band of short spines. It has a relatively large velum coverage (+30%) approaching that of I. junciformis, but also has substantionally larger mega- spores (+ 530 pm). OriGINs.—Most if not all North American sexual polyploid Isoetes species are believed to represent allopolyploids, as has been demonstrated for I. riparia BRUNTON & BRITTON: A NEW ISOETES FROM GEORGIA 195 (Taylor et al., 1985; Taylor and Hoot, 1997) and I. appalachiana (W.C. Taylor, pers. comm). Polyploid sterile primary hybrids are also known, such as hexa- ploids Isoetes Xfairbrothersii J. Montgom. & W.C. Taylor (Montgomery and Taylor, 1994) and I. Xhickeyi W.C. Taylor & Luebke (Taylor and Luebke 1988). An allopolyploid origin for I. junciformis would most likely result from the doubling of the hybrid between two diploids (2x x 2x = sterile 2x hybrid; doubled = fertile 4x species). Of the known southeastern diploids, the mor- phologically most similar and likely progenitors for I. junciformis would be I. flaccida, I. melanopoda, or I. engelmannii. None of these taxa is presently known to occur within ca. 80 km of the Tift County site. A combination of the wide (80—100%) velum character of J. flaccida with the narrow (10-15%) velum of I. melanopoda, as well as their similarly low tuberculate to vermiform megaspore ornamentation patterns, would likely re- sult in a plant demonstrating a similar morphological appearance to that of I. junciformis. An I. engelmannii X I. melanopoda combination is not a candi- date as this hybrid represents the origin of I. louisianensis (Taylor and Hoot, 1998). A hybrid combination involving Isoetes engelmannii, in any event, would be expected to demonstrate a more evenly reticulate megaspore orna- mentation (Brunton and Britton, 1996c). Development of I. junciformis from the doubling of the as-yet undiscovered primary diploid hybrid, I. flaccida x I. melanopoda, therefore seems plausible. An alternative explanation for the development of I. junciformis could be the formation of a fertile population from the sterile hybrid between a hexa- ploid and a diploid species (6x < 2x = sterile 4x; selection for fertility over time = fertile 4x species). An I. georgiana X I. melanopoda hybrid, for in- stance, would presumably have many of the characteristics expected of the sterile progenitor of I. junciformis. Evidence for the utilization of this evolu- tionary pathway, however, has not been established for any North American quillwort (cf., Taylor et al., 1993). Regardless of its origins, I. junciformis constitutes an addition to the growing number of endemic vascular plants known from this small area of the Georgia Coastal Plain. These include such wetland/riparian species as Rhynchospora solitaria Harper (Sorrie, 1998), and I. georgiana and I. boomii (Luebke, 1992; Brunton and Britton, 1996b). As noted above under the Megaspore Size and Morphology section, available evidence indicates that I. junciformis is not a primary sterile hybrid. In addi- tion to uniformly tetraploid chromosome counts being obtained from the Chula population, a wide range of plant sizes is evident within that population, in- dicating that on-going, in situ reproduction is taking place. With no other Js- oetes taxa being found within the Chula population, this constitutes strong evidence that sexual reproduction is occurring there. FURTHER RESEARCH.—The determination of the morphological characteristics and taxonomic significance of this population has been complicated by the rarity of living and preserved material. Some expressions of the variation ob- served in spore morphology, for example, are represented by only one or two 196 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) known specimens. Morphological and cytological examination of living ma- terial of I. flaccida and I. melanopoda elsewhere in southern Georgia and southeastern Alabama could be important in clarifying the status and nature of I. junciformis. Molecular studies of the known and suspected I. junciformis populations and of adjacent southern Georgia Isoetes populations will likely be necessary to provide a clearer insight into the origins of this apparently rare coastal plain endemic. ACKNOWLEDGMENTS We wish to acknowledge the assistance and co-operation of the curators of the various herbaria from which material was borrowed. We are also grateful to Professor Victor Matthews, University of Guelph, for the Latin translation of the species diagnosis. Wayne R. Faircloth of Valdosta State University provided valuable information i - the location and circumstances of his orig- inal discovery of Isoetes a iformis. Jame pre Georgia Department of Natural Resources, Richard Carter, Valdosta State Deine: and Lytton J. Musselman, Old Dominion University, provided important information concerning in Isoetes populations in southern Geor- and Agri-food Canada, Ottawa, provided valuable assistance by arranging for some of the loan material employed in this investigation. LITERATURE CITED Boom, B. 1982. erepees of Isoetes in the southeastern United States. Castanea 47:38-59 BRITTON, sy ay 1991. A hybrid Isoetes, I. Xharveyi, in northeastern North America. Gonad. J. Bot. 69:634-640. BRITTON, oi M., and D. F. BRUNTON. 1989. A new Isoetes hybrid (Isoetes echinospora X riparia) for Canada. Canad. J. Bot. 67:2995-3002 - 1992. Isoetes xjeffreyi, hyb. nov.; a new Isoetes (Isoetes macrospora x riparia) from Que- hee. Canada. Canad. J. Bot. 70:447-452. ——.. 1996. Isoetes ee a new triploid hybrid from western Canada and Alaska. Canad. J. Bot. 7 —59. BRUNTON, D. F. 1990. se ee for the protection of spore samples from Isoetes (Isoetaceae) voucher specimens. Taxon 39:226—228. BRUNTON, D. F., and D. M. BRITTON. 1996a. Noteworthy collections: Alabama and Georgia. Castanea 61:398-399. . 1996b. The status, distribution and identification of Georgia quillwort (Isoetes georgiana: Isoetacene) Amer. Fern J. 86:105—113. 1996c. Taxonomy and distribution of Isoetes valida (Isoetaceae). Amer. Fern J. 86:16—25. . 1997. Appalachian quillwort, wae appalachiana, i nov.; Isoetaceae), a new pterido- phyte from fg eastern United States. Rhodora 99:118— — D. F., D. M. BriTTon, an ae c TAYLOR. 1994, pad hyemalis, sp. nov. (Isoetaceae); a ew quillwort from the southeastern United States. Castanea 59:12—21 Cae J. 1996. Louisiana quillwort (Isoetes louisianensis Thieret) recovery lun. U.S. Fish & Wild- life eRe Jackson, Mississippi. LueBKE, N. T. 1992. Three new species of Isoétes for the southeastern United States. Amer. Fern 2s : 3-2 MonTcomery, J. D., and W. C. TAYLOR. 1994. Confirmation of a hybrid Isoétes from New Jersey. Amer. Fern J. a 115-120. MUSSELMAN, L, J., R. D. BRAY, and D. A. KNEPPER. 1997. Isoetes xcarltaylorii (Isoetes acadiensis x Isoetes engelmannii), a new interspecific quillwort hybrid from the Chesapeake Bay. Can- ad, J. Bot. 75:301-309. BRUNTON & BRITTON: A NEW ISOETES FROM GEORGIA 197 SNYDER, L. H., AND J. G. BRUCE. 1986. ae guide to the ferns and other pteridophytes of Georgia. University pr Georgia Press, At SORRIE, B. A. 1998. Npreabiks collections Georgia. Castanea 63:496—5 Tay.or, W. C., AND S. B. Hoor. 1997. Evolutionary relationships of mages species based on ITS bpp out Amer. J. Bot. af ‘ee. 163. [ABSTRACT 998. Origin of the Louisiana quillwort, Isoetes louridiemnensis. Amer. J. Bot. 85 (supple- a 101-102. [Abstract]. TAYLOR, W. C., and N. T. LUEBKE. 1988. Isoetes pare’ a naturally occurring hybrid between I. echinospra and I. macrospora. Amer. Fern J. 78:6—13. TAYLOR, W. C., N. T. LUEBKE, D. M. BRITTON, R. J. Matin and D. F. BRUNTON. 1993. Isoetaceae Reichenbach. Pp. 64-75 in Flora of North America Editorial Committee, eds. Flora of North America, ret ee Mexico, volume 2: Pteridophytes and Gymnosperms. Oxford University Press, New TAYLOR, W. C., N. T. 8 UEBKE, and M. B. SMITH. 1985. Speciation and hybridization in North Amer- ican Quillworts. Proc. Roy. Soc. Edinburgh 86B:259-263. American Fern Journal 89(3):198—203 (1999) Some Observations on the Reproductive Anatomy of Isoetes andicola ERIC KARRFALT 10261 Fry Rd., McKean, PA 16426 and contained about twice h DNA as of those in adjacent gametophytic cells. Such embryos, which were not associated with archegonia, are interpreted to have arisen via some form of apog- amy. Plants that are now referable to Isoetes andicola (Amstutz) L.D. G6mez were originally described as two separate species, Stylites andicola E. Amstutz and S. gemmifera Rauh. Although Gémez (1980) had maintained the latter taxon as a variety of I. andicola, this distinction is dubious (Karrfalt and Hunter, 1980; Karrfalt, 1984), and although Gémez (1980) did not provide any new evidence to support the combination of Stylites with Isoetes, Karrfalt (1984) showed that the elongate, monopolar stem characterizing the genus Stylites is secondarily derived during the ontogeny of each young Stylites plant and that these plants begin life with bipolar corms that are morphologically indistin- guishable from those of any other species of Isoetes. As an adjunct to this study of stem development, plants of J. andicola maintained in cultivation yielded a large number of gametophytes that subsequently produced correspondingly large numbers of new sporophytes. These gametophytes and sporophytes pro- vided an unexpected opportunity to study some aspects of the reproductive anatomy of this taxon. These anatomical observations seem to suggest that apomixis exists in this species. METHODS AND MATERIALS The material used in this study was collected and maintained as reported previously (Karrfalt and Hunter, 1980; Karrfalt, 1984). The observations re- ported here were made over a period of eight months beginning when the megaspore walls first began to open and continued until the food reserves of the gametophytes (as seen in sectioned material) was essentially exhausted. Because the original intent of growing the gametophytes was simply to obtain additional sporophytes, both megaspores and microspores were sown together. The possibility of conducting the present study only became apparent after KARRFALT: REPRODUCTIVE ANATOMY OF ISOETES ANDICOLA 199 the spores had germinated. Inferences about the developmental fate of the archegonia and hence their involvement or not in sexual reproduction were possible because the serial sections were all complete and the series of sections uninterrupted. All cells of all archegonia could be accounted for readily. The material used in this study was processed by standard paraffin methods. A total of 378 archegonia in 62 gametophytes was studied in serial sections. The megagametophytes mostly were sectioned at right angles to their exposed surfaces so as to cut the maximum number of archegonia in longitudinal sec- tion. This orientation was obtained readily by embedding the megagameto- phytes under a dissecting microscope so that a plane tangential to the center of the exposed surface of the gametophyte was perpendicular to the bottom of the embedding vessel. Sections were then cut parallel to the lower surface of the paraffin block. In a few cases, sections were cut parallel to the exposed surface of the gametophyte so as to view the archegonia in serial cross sections. The megagametophytes were sectioned at 6 ym, except those used for micro- spectrophotometry, which were sectioned at 20 pm. The microgametophytes were processed within the dead microsporangia and sectioned at 4 pm. The microspectrophotometry was done by the two wavelength method of Ornstein (1952) using light wavelengths of 550 nm and 495 nm. The swimming spermatozoids were photographed under phase contrast mi- croscopy. RESULTS AND DISCUSSION When the megaspore suture first opens, the exposed surface of the mega- gametophyte does not protrude at all beyond the spore wall. With further growth, however, the megagametophyte enlarges considerably and eventually reaches a volume of which only half or less can be contained within the spore wall. Subsequent to the opening of the suture, new archegonia are produced continually throughout the life of the gametophyte. The largest number of ar- chegonia on a single gametophyte was 23. A detailed study of gametophyte development was not attempted, but the process seems to be the same as in other species of the genus. There is an initial stage of free-nuclear divisions followed by a gradual cellularization beginning under the trilete scar portion of the megaspore wall. Eventually, cellularization is essentially complete, al- though occasional binucleate cells may be found. After sporelings began to emerge from the gametophytes in the cultures, a search was begun to identify earlier stages of sporophyte development. Sam- ples consisting of ten megagametophytes were selected from time to time and processed so as to examine for embryos within a few days after removal from cultures. Each sample was selected to include a more or less full spectrum of of gametophyte sizes. Initially, the only embryos found were in an advanced state of development, with well-defined primordial organs. These were always located deep within the tissue of the gametophyte and not associated with an archegonium (Fig. 1). In every megagametophyte containing an embryo not associated with an archegonium, all of the archegonia were still intact, com- 200 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) te ws ‘te, c ie Fics. 1-11. Isoetes andicola. 1) Section of a megagametophyte containing a sporophyte not as- sociated with an archegonium. 2, 3) Longitudinal sections of mature archegonia; arrows mark spurious neck canals. 4) Cross-section of an archegonial neck through the second tier from the distal end. 5) Cross-section of an archegonial neck through the third tier from the distal end. 6) Longitudinal section of an archegonium lacking the outer two tiers of neck cells and containing vigorously growing embryo; arrows mark mitotic figures. 7) Longitudinal section of a microspo- 8 crogametophytes. 9) Longitudinal section of a mature microgametophyte containing four coiled spermatozoids, remnants of jacket cells, and a prothallial cell (at the righthand end). 10) Sper- matozoid during the slowly swimming stage. 11) Spermatozoid during the rapidly swimming stage. Scale bars: 1, 7 = 100 ym; 2-6, 11 = 25 wm; 8-10 = 10 pm. KARRFALT: REPRODUCTIVE ANATOMY OF ISOETES ANDICOLA 201 plete with an undivided egg cell in the venter. The nuclei of the embryo cells were about twice the diameter of those in the surrounding gametophyte tissue, suggesting that in spite of the lack of participation of an egg cell, the origins of these embryos nonetheless involved an increase in ploidy level. Microspec- trophotometric measurements of gametophytic nuclei and sporophytic nuclei confirmed that this was the case. The mean relative amount of DNA for the gametophytic nuclei (9.75 + 1.11, n = 9) was about half that of the sporophytic nuclei (18.4 + 1.79, n = 11), and that of the maximum measurement for ga- metophytic nucleus (13.1, presumably 2c) was very similar to the minimum measurement from among the sporophytic nuclei (13.7; also presumably 2c, where the quantity of DNA in a haploid nucleus prior to DNA synthesis = c, the quantity of DNA of the completion of DNA synthesis and in a diploid nucleus prior to DNA replication = 2c, and at the completion of DNA synthe- sis = 4c). The mature archegonia had necks consisting of four tiers of neck cells. At first, it appeared that a canal ran through the length of the neck from the venter to the exterior, but closer examination showed that this was not the case. Some archegonia appeared to show a canal in longitudinal section (Figs. 2, 3, arrow), but others showed no “canal” in the outer two tiers of neck cells anywhere in the complete series of relatively thin sections (Fig. 3, left archegonium). Com- parisons of longitudinal sections of serial cross sections (Figs 4, 5) revealed that there were no neck canal cells and no neck canal in the outer two tiers of the neck, but that the ventral canal cell extended through the inner two tiers of neck cells. The superficial appearance of a canal through the outer two tiers of some of the necks in longitudinal sections resulted when the plane of section was such as indicated by the opposed arrows in Fig. 4. The “canal” through the outer two tiers in these cases was either simply a face view of the wall between the longitudinal files of neck cells on opposite sides of the neck (Fig. 3, arrow) or some combination of that wall and the lumen of one of the cells sharing that wall (Fig. 2, arrow). Rauh and Falk (1959) refer to an un- specified number of neck canal cells passing throught the outer two tiers of the archegonial neck, but their micrograph of such an archegonium (Rauh and Falk, 1959, Abb. 34 III) is so similar to Fig. 2 that I suspect their identification of neck cells may have been mistaken. Of the 62 megagametophytes that were studied in serial sections, 3 pos- sessed a few archegonia that lacked the outermost tier of neck cells. These archegonia were obviously moribund, with most of their remaining cells ap- parently suberized. In the entire study, only one archegonium was found that was missing the outer two tiers of neck cells, i.e., its neck canal was open to the exterior. The venter of this archegonium contained a vigorous embryo (Fig. 6, note three mitotic figures at arrows). Attempts to manually break off the outer two tiers of neck cells to facilitate fertilization resulted in no embryo formation. Near the end of this study, embryos began to occur within arche- gonia fairly frequently. Twenty such embryos were found, but nine of these appeared to be in poor condition or were dead. One intact archegonium con- 202 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) tained a living embryo with each of its four cells in late anaphase and hence presumably in good health. Whether or not gametic union ever occurs in this plant, apparently normal male gametophytes and spermatozoids are produced. Although some micro- sporangia contain small abortive megaspores in addition to microsproes (Fig. 7) and all microsporangia examined contained substantial numbers of abortive microspores with collapsed protoplasts (Fig. 8, to the right and left of center), numerous mature and immature male gametophytes were seen as well (Fig. 8, center). The mature male gametophytes contain four large coiled spermato- zoids surrounded by the remnants of jacket cells and a single prothallial cell (Fig. 9), as in other species of Isoetes. The spermatozoids swim vigorously and in this respect at least appear perfectly normal. A wet mount of some contents of a microsporangium would usually include some swimming spermatozoids. If not, slight pressure on the cover slip with a dissecting neeedle would cause one or more mature antheridia to dehisce. Upon emergence, each spermato- zoid uncoils slightly (Fig. 10) and begins to rotate and swim slowly. The nu- merous flagella then begin to beat more rapidly and the spermatozoids uncoil further (Fig. 11). They progress so rapidly through the water that it can be difficult to keep them in view. The rapid swimming persists for about one minute, after which the spermatozoids slow down and recoil to essentially their initial shape. The embryos that occur deep within the gametophytic tissue and are not associated with archegonia are apparently apogamous, The anatomical obser- vations definitely preclude the participation of either egg or spermatozoid in the formation of these deep-seated embryos. The present observations do not reveal specifically how these embryos are initiated, but the microspectropho- tometric results show unequivocally that the ploidy level of these embyos is double that of the surrounding gametophytic tissue. Possibly the deep-seated embryos are initiated by endoreduplication in single vegetative gametophytic cells or possibly the nuclei of binucleate cells fuse. Occasional binucleate cells have been seen in I. taiwanensis De Vol (Huang and Chiang, 1986) and are not unique to I. andicola. In I. andicola, however, they may have acquired a novel function, that of embryo formation. possibility cannot be excluded, however, that under field conditions or in ma- terial from other localities the archegonia might develop differently. KARRFALT: REPRODUCTIVE ANATOMY OF ISOETES ANDICOLA 203 The usual persistence of the outer two tiers of neck cells and the absence of a neck canal through this part of the neck apparently ensures that if fertiliza- tion ever occurs it is a rare event. In any case, the embryos that are not asso- ciated with archegonia and those contained within archegonia with intact un- opened necks are most likely apomictic. Regardless of their origin, the spo- rophytes produced by megagametophytes probably play a relatively minor role in the maintenance of populations of this species. Gémez (1980) found that vegetatively produced plants of J. storkii T.C. Palmer are much more likely to grow into adult plants and in less time than sexually produced plants, because the latter are very much smaller and more fragile than the former. The abun- dant robust gemmae produced by I. andicola would likewise seem to have a competetive advantage over the minute sporophytes produced by gameto- phytes in the crowded conditions (Karrfalt and Hunter, 1980) in which these plants grow. ACKNOWLEDGMENTS I thank Dr. Marion Himes for the use of the microspectrophotometer and especially for her generous assistance concerning its operation. LITERATURE CITED BrERHORST, D. W. 1968. On the Stromatopteridaceae (fam. nov.) and the Psilotaceae. Phytomor- phology 18:232—268. Gomez, L. D. 1980. ie reproduction in a Central American Isoetes (Isoetaceae), its mor- phological, systematic, and taxonomical significance. Brenesia 18:1—-14. HUANG, S.-F., and S.-H. T. CHIANG. 1986. The development of the female gametophyte in Isoetes SniWenisiial De Vol. Taiwania 31:15-32. KARRFALT, E. 1984. The origin and early development of the root-producing meristem of Isoetes andicola L.D. Gémez. Bot. Gaz. (Crawfordsville) 145:372—377. KARRFALT, E., and M. HUNTER. 1980. Notes on the natural history of Stylites gemmifera. Amer. Fern J. 70:69—72. esp htinay L. 1952. The distributional error in microspectrophotometry. Lab. Invest. 1:250—262. U and H. FALK. 1959. Stylites E. Amstutz, eine neue Isoetacee aus den Hochanden Perus. L alien Morphologie, anatomie, und ee i eae Sitzungs- ber. Heidelberger Akad. Wiss. Math. Naturwiss. Kl. 1 American Fern Journal 89(3):204—214 (1999) Ontogeny of the Sporangia of Sphaeropteris cooperi KENNETH A. WILSON Rancho Santa Ana Botanic Garden, 1500 North College Avenue, Claremont, California 91711 initials or “segments.” Segment 0, located at the level of the surface receptacular cells, does not become subdivided and does not contribute further to the structure of the mature sporangium. Segments I, II, Ill and IV each become subdivided through a series of divisions to produce the mature sporangia. The four-rowed sporangial stalks are formed from Segment I and part of Seg- ment II, and the capsules develop from a part of Segment II and Segments III and IV. The annulus develops in Segments II and IV. The developmental pattern of the sporangia of Sphaeropteris cooperi is compared to that of the sporangia of higher leptsoporangiate ferns. The most familiar and most frequently illustrated leptosporangia are those of the higher leptosporangiate ferns. The development of the sporangia of the higher leptosporangiate ferns was described in a series of papers (Wil- son,1958a, b, 1960) and is now well understood. In the sporangia of the ad- vanced leptosporangiate ferns, as illustrated by species in the Polypodiaceae, Grammitidaceae, and Vittariaceae, it was shown that the stalk and the capsule of the leptosporangium develops from a single epidermal primordial cell that becomes divided into five initials or “segments,” rather than from the activity of an apical cell. Each one of these “segments” in turn divides, through a series of divisions to produce the mature sporangium. Segment 0 contributes only to the formation of the stalk; Segment I to a portion of the stalk and part of the proximal face of the capsule; Segment II to the stomial region, the stalk, and to the proximal and distal faces of the capsule; and Segments III and IV to the rest of the annulus and to both the proximal and distal faces of the capsule. Although the stalk may be one-, two- or three-rowed at its base, the capsule is always subtended by a three-rowed stalk. The one-rowed stalk results di- rectly from the horizontal orientation of the first division of the sporangial initial, whereas the two- and three-rowed stalks depend on the orientation of both the first division and also the division that produces Segment I. A review of the history of our knowledge of the nature of the leptosporan- gium and its development was published in the introduction to the study of the ontogeny of the sporangia of Phlebodium aureum (L.) J. Sm. (Wilson, 1958a). Recent descriptions of sporangial development continue to reproduce the erroneous pattern apparently originated by Campbell (1905) that the spo- rangial initial produces a three-sided apical cell that cuts off several basal cells to form the stalk until a transverse division stops its activity by cutting off the cap cell. Other accounts are unclear, incomplete and often incorrect. (see Gif- ford and Foster, 1989; Bold et.al., 1987; Holttum et. al., 1970). No detailed ontogenetic studies have been published since the appearance of the paper on WILSON: SPORANGIAL ONTOGENY IN SPHAEROPTERIS 205 the sporangium of Anarthropteris lanceolata (Hook. f.) Pic. Serm. (as A. dic- tyopteris (Mett.) Copel.) (Wilson, 1960). As pointed out in a study of mature sporangia of species of the Polypodi- aceae, Grammitidaceae, and Vittariaceae (Wilson, 1959), the cell arrangement in the sporangia reflect the ontogeny of these structures and, with but few exceptions, there is no reason to doubt that the development of the capsules follows the pattern of those of Phlebodium (Wilson, 1958a), Xiphopteris and Pyrrosia (Wilson, 1958b), and Anarthropteris (Wilson, 1960). Edwards (1996) expanded the examination of the structure of mature sporangia by initiating a survey of the cellular structure of the capsules of more than 110 species in 20 families. Three of these species were illustrated in his published abstract. Sporangia with four-rowed stalks, however, are known in several fern genera including Dipteris, Cheiropleuria, and members of the Cyatheaceae. Wilson (1959) pointed out that it was not possible to homologize the sporangial faces of Dipteris and Cheiropleuria with those of the higher leptosporangiate ferns. The known developmental patterns cannot give rise to a four-rowed stalk. Bower (1915) wrote that in Cheiropleuria sporangia with four-rowed stalks, the segmentation of the young sporangium, ‘‘Appears to show a regular cleav- age of the segments in two opposite rows,” and the ‘‘Subdivision of the two rows of segments of the stalk by walls in the plane of the drawings has given rise to the four rows of cells of the stalk, as seen in later stages.” This suggests a distinctly different developmental pattern in these sporangia than is known. The only studies of the development of sporangia with four-rowed stalks are those of Bower (1915, 1923, 1926). Holttum and Sen (1961), in their paper ‘Morphology and classification of the tree ferns” did not make a detailed ex- amination of the sporangia, but based their comments mostly on Bower’s pub- lications. For a clear understanding of the structure of the sporangia with four- rowed stalks their ontogeny needs to be studied in detail. Because it is readily available in cultivation in southern California, Sphaer- opteris cooperi (F. Muell.) R.M. Tryon [Cyathea cooperi (F. Muell.) Domin] was chosen for study to serve as a model for the pattern of development of spo- rangia with four-rowed stalks. MATERIALS AND METHODS The material used in this study was collected from plants in cultivation in Los Angeles, California. A specimen of this fern has been deposited in the herbarium of Rancho Santa Ana Botanic Garden (Wilson 2067, RSA). Slides are deposited at Rancho Santa Ana Botanic Garden. Fertile pinnae of Sphaeropteris cooperi in early and increasingly mature stages of development were preserved in formalin-aceto alcohol (FAA). Sori were processed by three different methods: 1) Fertile pinnules were infiltrated with the tertiary butyl alcohol series, embedded in paraffin, and sectioned at 10um. The sections were then stained in the Sharman (1943) series. 2) Fertile pinnules were cleared in 5% NaOH, bleached in 50% chlorine bleach, and stained in 3% tannic acid in 50% alcohol and 3% ferric chloride in 50% 206 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) alcohol (the alcoholic stains were used to prevent maceration). After dehydra- tion in alcohol, the sori were dissected from the pinnule lamina and placed on a slide in Diaphane. The sori were then teased to separate the sporangia and a coverslip was mounted. This technique resulted in an enormous amount of damage, but methodical searches of the slides and a large number of dis- sections revealed undamaged sporangia. This procedure of clearing and stain- ing the sporangia allows both sides of each developing sporangium to be stud- ied. 3) Young cleared fertile pinnules stained in tannic acid and ferric chloride were imbedded in paraffin, sectioned at 201m, and mounted on slides. These preparations were studied to confirm the early division patterns observed in the other preparations. Mature sporangia were studied mounted in Crystal/Mount (Biomedia Corp., Foster City, California) on temporary slides. This process reduced the dehis- cence of the capsules and at the same time prevented movement of the spo- rangia while being examined. All illustrations were made with the aid of a Leitz drawing tube on a Leitz microscope. Both sides of each cleared sporangium were drawn and each spo- rangial “Segment” is shaded to facilitate comprehension of the cell lineages in the developing sporangium. The shading patterns used conform with those used in earlier sporangial ontogenetic studies in order to allow for easier com- parisons. RESULTS Sporangia of Sphaeropteris cooperi begin their development by the swelling of a single superficial cell of the receptacle, which soon becomes segmented by a diagonal wall that extends from approximately the surface of the recep- tacle, or a little above it, toward the base of the cell (Fig. 1, arrow). This first- formed wall segments the sporangial initial into a terminal cell, the “mother initial,” and a basal cell, Segment 0. Segment 0 is found at the level of the receptacular superficial cells and takes no further part in the formation of the sporangium. Sporangia teased from the receptacle rarely show any evidence of the presence of Segment 0 and, therefore this segment can be identified clearly only in sectioned preparations (Figs. 1-5). The cell called the mother initial divides to produce three segments that contribute to the formation of the stalk and capsule. Segment I is formed by a wall approximately perpendicular to that producing Segment 0 and intersect- ing this wall so as to divide the basal portion of the initial into two roughly equal halves (Fig. 2. arrow). The formation of Segment I is followed by a di- vision of the mother initial that produces a wall roughly perpendicular to that of Segment I and parallel to the first-formed wall, giving rise to Segment II (Fig. 3, arrow). At this stage, the mother initial is 2-sided and wedge-shaped at the base. Segment III forms from a division more or less at a right angle to that of Segment II, directly above Segment I and parallel to it (Figs. 4, arrow; 14a, b). Although this series of divisions suggests the activity of an apical cell, the divisions of the mother initial always produce Segments I, II, and II, and WILSON: SPORANGIAL ONTOGENY IN SPHAEROPTERIS 207 is i. A) xy) iy ~ on bas iy nul \ Sy a \W) iat eS Ay lig _ — Fics. 1-28. Sporangial ontogeny in Sphaeropteris cooperi. 1-10) Internal segmentation. All fig- ures drawn from sectioned material. 1) Formation of Segment 0. 2) Formation of Segment I. 3) ficial segmentation, earlier stages. All figures from cleared, stained whole mounts. Both sides of each sporangial primordium are illustrated and are designated “a” and “‘b.” Arrows point to newly formed walls or cells. 0 = Segment 0. Scale bar = 10 pm. 208 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 3 (1999) then it divides to produce a transverse wall which cuts off the cap cell of the sporangium, Segment IV. The mother initial thereby becomes completely en- closed by its daughter cells (Figs. 5, arrow; 15a, b, arrows). Segments I, II, and III become subdivided by a series of intercalary divisions. The first divisions of Segments I, II, and III are usually by horizontal walls (Figs. 14a, b; 15a, b), although in rare instances the first wall to subdivide the segment is a vertical one (see Segment III, Fig. 14b). These early divisions usually take place before the mother initial gives rise to Segment IV. Segment I contributes only to the formation of the portion of the stalk di- rectly below Segment III, and Segment II also contributes to the stalk formation and to the wall of the capsule as well. Thus, the sporangial stalk is formed by both Segment I and the proximal portion of Segment II. Segment I and Segment II first become subdivided by horizontal walls (Figs. 14a, b; 15a, b). Soon after, as the young sporangia enlarge, anticlinal, longitudinal divisions in each of the cells function in bisecting each segment and give rise to a sporangial stalk, which at its base is composed of four rows of cells (Figs. 13a, 14b, 17a, b). In the more distal region of the stalk, directly below the capsule, additional sub- divisions associated with capsule formation become evident (Fig. 13b). The transverse division that produces Segment IV encloses the mother ini- tial, which is three-sided and cymbiform. The enclosed mother initial now divides in the same manner and sequence as it did in producing Segment II, III and IV, so that it becomes enclosed by three cells that separate it from the cells of the sporangial wall (Figs. 6-8). These innermost daughter cells give rise to the tapetum by means of periclinal and anticlinal divisions (Figs. 8, 9). A cross section through the capsule at this stage reveals that the enclosed cell appears elliptical (Fig. 10), whereas in longitudinal section it appears trian- gular (Figs. 8, 9). No attempt was made to follow the divisions of the tapetal cells or of the central cell. Additional horizontal and vertical divisions accompany the enlargement of the developing sporangium. In the distal portion of Segment II these give rise to a row of cells on each side of the segment contiguous to Segment III that contribute to the formation of the annulus (Figs. 20b, 21b, 22b, 23b). In addi- tion, in Segment II, two cells, roughly occupying the mid-region of the seg- ment, are formed that link the lateral vertical rows of the annulus in Segment II and complete the U-shaped line of cells that produce the lower arc of the annulus (Fig. 23b arrows). A series of divisions in Segment IV gives rise to the cells that complete the ring. The first subdivision of Segment IV is by a wall parallel to the one that produces Segment I (Fig. 16a, b). Divisions in each daughter cell perpendicular to this first wall complete the delimitation of the annular region in Segment IV (Fig. 18a). Additional divisions in the cells next to Segment III produce a row of cells that are contiguous to the distal portion of Segment III (Fig. 19 a,b). These cells become continuous with the annular cells of Segment II (Figs. 20b, 21b). The U-shaped row of cells in Segment II together with the row of cells in Segment IV produce the uninterrupted ring of cells that is the annulus (Figs. 27a, 28b). The annulus, therefore, always borders Segment III along its WILSON: SPORANGIAL ONTOGENY IN SPHAEROPTERIS 209 Si Ne: | Ui, | ee i ‘ ‘ WK i ul TH \G - ay nan thi 4 pe I ill, : ay? CLUL/) Md Wp i OM) G : iN, if) | F oy —S Trial 1 —e— Trial! _ —+— Triat2 20 —*— Trial2 5 —= Trial3 — Trial3 5 404 = 2 = =} as x C) S B 204 a 2 = ps é 10 0 0.0 r T r r ——— 0 105 104 103 102 10! 0 105 104 103 102, 1¢1 Concentration Concentration Fic. 1. Response of Onoclea sensibilis gametophytes from the por GA, site to varied doses (i.e F concentrations) of antheridiogen A,;, as characterized by mean number of antheridia Nfs gametophyte (left) and gametophyte surface area, estimated as tlt x Waldth (right). developed archegonia in the vicinity of their meristematic notches. The ten- dency for these hermaphrodites to appear seemed to be greater in 10-* CAF cultures than in 10-? CAF. Similar observations were reported by Naf (1979). To determine the suitability of various criteria that could characterize and quantify antheridiogen response, initial experiments using a single accession (Gray, GA) evaluated three observable attributes of response by O. sensibilis gametophytes over A;; concentrations of 10-'-10-° CAF. These attributes were: 1) mean number of antheridia per gametophyte, 2) mean surface area per ga- metophyte, and 3) percentage of male gametophytes. Antheridiogen response characterized as mean number of antheridia per ga- metophyte is illustrated in Fig. 1A. Number of antheridia per gametophyte was very close to 0 in both the control and at 10°° CAF. Maximum response was observed at concentrations of CAF 10~° or higher. Two of the three replicates showed very similar response levels across CAF concentrations 10° '—10~°, sug- gesting that at this concentration mean number of antheridia exhibits a satu- rated response. However, the first replicate showed a substantially higher re- sponse at 10-2 than at either 10~° or 10-*. All three replicates showed an in- termediate response at 10°-* CAF. In replicate 3, the mean number of antheridia per gametophyte was substantially lower than in the first two trials. This is believed to be due to differences in the spore sowing density. The second criterion, gametophyte size, was evaluated because it is known to be affected by antheridiogen extracts (Naf, 1979) and to influence the num- ber of antheridia per gametophyte, as demonstrated in Cystopteris (Haufler and Ranker, 1985). To determine the most effective means to estimate gametophyte surface area, gametophyte outlines were traced onto paper using a camera- lucida. These were cut out, their length and width measured, and their surface area determined using a leaf area meter. Area was regressed against the follow- 226 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) TABLE 2. Regression analyses of the relationship between gametophyte surface area and four estimators. Square of Square of Statistic Length Width length width Length X width Ue 0.78 0.70 0.78 0.64 0.93 F 65.55 43.56 68.48 34.01 Pare he P <0.001 <0.001 <0.001 <0.001 <0.001 N 20 20 20 20 20 ing independent variables: 1) length, 2) width, 3) square of length, 4) square of width, and 5) length x width. The product of two dimensions (length x width) provided a substantially better approximation of area than any of the other variables (Table 2), and this product was therefore used. Variation in gametophyte surface area (as estimated by the product of length and width) in response to CAF concentration is illustrated in Fig. 1B. The area response was not as consistently predicted by CAF concentration as was either mean number of antheridia or percent male (see next paragraph), and furthermore was in- consistent across experiments. However, a tendency for area to decrease with increased CAF concentration was clearly indicated, consistent with previous reports (Naf, 1979). Antheridiogen response characterized as percent male is illustrated for the Gray, GA, population in Fig. 2A. Response at CAF concentration 10-° was at or near zero and indistinguishable from the control. Response at CAF concen- trations 10°', 10-7, and 10°* was at or near 100% males (96.5—100 %), indi- cating a saturated response for this criterion. At 10-* CAF an intermediate response was observed (68.9—79.3% males), indicating that the threshold for substantial response lies between 10-° and 10-* CAF. This concentration seems highly comparable to Naf’s (1956) determination of 3.2 X 10-5 as a response threshold concentration for A,, exudate. Values for percent male at each con- centration were highly consistent across the three replicates, much more so than for either number of antheridia or area. The relationship between gametophyte surface area and antheridium num- ber was explored. To determine whether the number of antheridia per game- tophyte increased with area, regression analysis was performed within each treatment (Table 3). Number of antheridia was positively associated with ga- metophyte surface area for all three replicates at 10-? CAF, for all at 10-2, two of the three at 10-* and two at 10~*. In all other treatments, there was no significant relationship between area and antheridial number. Thus, significant regressions tended to be observed in cultures where most or all gametophytes possessed antheridia, indicating that for male gametophytes, number of an- theridia increased with size of the gametophyte. However, the strength of this association was not great, the highest value of r? being 0.722. Nor was the — . 2, Response of Onoclea sensibilis gametophytes from six populations to varied doses of antheridiogen A,,;, as characterized by percentage of male gametophytes. STEVENS & WERTH: ANTHERIDIOGEN RESPONSE IN ONOCLEA 227 Percent Male Percent Male Percent Male Gray GA 105 104 103 102 101 Concentration Richmond VT =—@— Triel 1 106 «$105 104 103 102 10! Concentration Nashville TN ag Teh i —ir Trial 2 1e6 105 «104 1¢° 10? §61¢1 Concentration Eagle WI 100 5 80 + 2 S «0-4 = —e— Triall = —_— Trish 2 r-F) 40 44 5 -0 20 o 106 «105 104 103 102 19! Concentration Berwick PA 100 5 80 - Ans 2? e § 40 4 fot —@e— Triall F —a— Trisl2 20 - 0- 0 106 «6105 «6104 «103 «6102 101 Concentration Vienna VA 100 + 80 - i Biv z § 40 + = —@— Triall -¥ —t— Trial? 20- 0-4 0 176 «6105 «6104 «6103 ) «6102 101 Concentration 228 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) TABLE 3. Relationships between number of antheridia (X Anth.) and surface area (X Area) of gameto- phytes for each of the three replicates (Rep.) at each concentration of CAF (Treatment). The abbreviation r refers to the regression coefficient, df to degrees of freedom, and P to resultant probability. Asterisks (*) next to r? values indicate a significant relationship at the P < 0.05 level. Treatment Rep. X Anth. X Area df r P 10-1 1 40.50 0.72 1,28 0.704* 0.000 2 STL 0.65 1,28 0.3597 0.000 3 19.90 0.60 1,28 0.722* 0.000 10-2 1 49.23 O77 1,28 0.632* 0.000 2 35.67 0.58 1,28 0.409* 0.000 3 PEPE 0.80 1,28 0.657* 10-3 1 34.67 1.14 1,28 0.725 Z 35.96 0.83 1,28 O.713* 3 19.07 0.91 1,28 0.538* 0.000 10-4 1 16.77 1.68 1,28 0.138* 0 Z 15.40 1.73 1,28 0.158* 0.029 2 9.87 1.05 1,28 0.00' 0.619 10-5 1 0.03 1.92 1,28 0.051 0.231 2 0.60 1.11 1,28 0.049 0.238 2 0.00 L235 1,28 0.000 NA Control 1 0.00 123 1,28 0. 2 0.10 £53 1,28 0.101 0.086 3 0.00 LST 1,28 0.000 NA number of antheridia per surface area unit of gametophyte even approximately constant across replicates, as can be seen by comparison between Figs. 1A and Clearly, percent male gametophytes not only provided a more consistent response criterion, but also could be more rapidly scored than number of an- theridia. Therefore, subsequent experiments were evaluated using only the percent male criterion. COMPARISON OF POPULATIONS.—Five additional populations were compared to each other and to the Gray, GA, population for antheridiogen response using the percent male criterion (Fig. 2). To allow for the possibility that some pop- ulations might exhibit greater sensitivity, the range of A,, concentration was extended to include 10-° CAF. The level of response was similar in all popu- lations. Response approached saturation at 10-? CAF, with means across rep- licates for populations varying from 96.6—-100% male (the lowest value for an individual replicate was 85%). In most populations, response at 10-* and 10-° CAF was similar to that of the control, i.e., few to no males. The exception was the Eagle, WI, population, in which substantial numbers of males were observed in some replicates at both 10-* (21% males) and 10-* (16% males); control cultures for this population contained no males. All cultures exhibited intermediate levels of response at 10-* CAF. Mean response values at this dose differed substantially among populations, ranging from 18% (Nashville, TN) to 78% (Gray, GA) Relative to the amount of variation among populations, there was a substan- STEVENS & WERTH: ANTHERIDIOGEN RESPONSE IN ONOCLEA 229 tial amount of variation in response at 10-* between replicates for some pop- ulations. Variation in response among replicates was greatest in the Richmond, VT, population (26-84% males at 10-* CAF) and least in the Gray, GA, pop- ulation (68.9—79.3% males at 10-* CAF). DISCUSSION Herein we have quantified three components of response by O. sensibilis to varied concentration of antheridiogen A,,;. Percent males and mean number of antheridia per gametophyte increased and gametophyte surface area decreased with increasing doses of A,;. Of these, percent male was the most predictable, and provides a consistent and convenient means to characterize antheridiogen response. Using this criterion, threshold and saturation doses of A,; concen- tration were found to differ by approximately two orders of magnitude (ca. 10-5 and 10° CAF, respectively), with the intermediate concentration (10~+ CAF) eliciting an intermediate response. This response profile is by and large consistent among populations sampled across the species range. However, there is evidence of at least some variation among populations at the inter- mediate dose (10-* CAF). Notably, the Nashville, TN, population exhibited a substantially lower response (mean of 18% males) at 10-* CAF than all other populations, which exhibited mean response levels in excess of 50% males at 10-* CAF. There were also some populations in which males appeared at con- centrations below 10~*, notably the Eagle, ia mapa which exhibited substantial numbers of males at 10-° and 10 The fundamentally similar results among dics could reflect a lack of genetic variation determining antheridiogen-response phenotype in O. sensi- bilis. Alternatively, substantial genetic variation for response may exist, but be evenly distributed among populations such that population means appear equivalent. Phenotypic differences among genetically different individuals would be small if antheridiogen response variation is polygenically deter- mined. It is uncertain whether the differences in response by individual ga- metophytes exhibited at 10-* CAF are a result of genetic differences among gametophytes or instead represent stochastic response variations among ga- metophytes that are genetically uniform with respect to determinants of A,; response. The lower response value at 10-* CAF in the Nashville, TN, popu- lation and the occasional response of some gametophytes (e.g., in the Eagle, WI, population) at extremely low doses (10-° and 10-° CAF) suggests that at least some genetic variation may exist. However, the heritability of antheridi- ogen response is completely unknown in O. sensibilis or in any other poly- podiaceous (sensu lato) fern. In Ceratopteris richardii (Parkeriaceae), mutation- induced lack of antheridiogen sensitivity was shown to have a simple (two gene) mode of inheritance (Scott and Hickok, 1987; Warne et al., 1988). How- ever, it remains possible that this species includes more subtly differentiated, continuously varying phenotypes that are determined polygenically. As with most studies on antheridiogen response, the present experiments were carried out under highly artificial conditions that constrain their rele- 230 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) vance to the natural life history characteristics of O. sensibilis. Unnatural con- ditions included use of agar solidifed media instead of soil, heat sterilization of media and antheridiogen via autoclaving at temperatures greatly exceeding those encountered in nature, continuous illumination, and use of antheridi- ogen derived from bracken rather than from Onoclea itself. Use of definable artificial conditions make such experiments more feasible, repeatable, and comparable to the broad literature which has used similar conditions. The validity of this and other similar experiments and relevance to nature could be evaluated by repeating them under more natural conditions, as has been done but rarely (Haufler and Ranker, 1985). Irrespective of whether the present results reflect life-history attributes of O. sensibilis, they do have bearing on the use of this species as a standard assay organism for detecting antheridogen A. The similar response profile observed across a large portion of the species range further validates the reliability and consistency of O. sensibilis for this purpose. The means by which antheridi- ogen response is quantified has varied among studies, having included deter- mining the minimum dose that can produce any response (e.g., Naf, 1956; Naf et al., 1975), percentage of males in cultures (Naf, 1965; Klekowski and Lloyd, 1968; Schedlbauer and Klekowski, 1972; Rubin and Paolillo, 1983; Scott and Hickok, 1987; Nester-Hudson et al., 1997), or number of antheridia per ga- metophyte (Haufler and Ranker, 1985). The response variation among popu- lations at the lower concentrations indicates that inconsistencies may be ex- perienced using the minimal response criterion, unless the same spore source of Onoclea is always used. We found that the most easily scored response attribute, percent male, is also the most reliable for judging A,, concentration. The strength of A,, extracts could be standardized by determining the dilution at which 50% of the gametophytes are male, analogous to the LD50 criterion used in toxicology. Moreover, the discovery of individuals such as the one from Nashville, TN, with lower sensitivity could provide an additional assay point for more precisely standardizing antheridogen concentrations. Such standardization will have importance as research on antheridiogen response continues, as different CAF extractions may vary in their Ap; concentration and diminish over time. ACKNOWLEDGMENTS We are grateful to Paul Wolf for providing bracken spores, to Ralph Brooks, Carol Kuhn, Chris Haufler, John Miller, Carl Taylor, James Montgomery, Dean Whittier, David Barrington, and Cathy Paris for providing collecti f Onoclea sporophylls, and to Lisa Wellborn for help in developing methodology. Helpful comments by an anonymous reviewer resulted in improvement of the man- uscript. This research was supported by a REU supplement to NSF grant BSR-851184 and by DEB- 9220755. LITERATURE CITED Dopp, W. 1950. Ein die Antheridienbildung bei Farnen férdernde Substanz in den Prothallien von Pteridium aquilinum (L.) Kuhn. Ber. Deutsch. Bot. Ges. 63:139-147. STEVENS & WERTH: ANTHERIDIOGEN RESPONSE IN ONOCLEA 231 GasTONY, G. J., and M. C. UNGERER. 1997. Molecular systematics sn a revised taxonomy of the Onocleoid ferns (Dryopteridaceae: Onocleae). Amer. J. Bot. 84:840-8 HAUuFLER, C. H., and G. J. GAstony. 1978. Antheridiogen and the breddina ehateu in the fern genus Bommeria. Canad. J. Bot. 56:1594-1601. HAUFLER, C. H., and T. A. na he 1985. Eade: antheridiogen response and evolutionary mechanisms in a s. Amer. J. 7659-6 KLEKOWSKI, es J. JR. 2 Reproductive pistons of the ptaridophyta. Ill. A study of the Blechna- te]. a. rae 62:361-377 Pp a — J. JR., and R. M. Lioyp. 1968. Reproductive biology of the agian ai 1. General Segre: and a study of Onoclea sensibilis L. J. Linn. Soc., Bot. 60:315—32 siailiien J. H. 1968. Fern gametophytes as experimental material. Bot. Rev. “i ama 34: 361-426. MILLER, J. H., ar P. M. MILLER. 1970. Unusual dark-growth and antheridial differentiaion in some gametophytes of the fern Onoclea sensibilis. Amer. J. Bot. 57:1245—1248. NAr, U. 1956. The demonstration of a factor concerned with the initiation of antheridia in Poly- podiaceous ferns. Growth 20:91-105. 1965. On antheridial metabolism in the fern species Onoclea sensibilis L. Pl. Physiol. 40: 888-890. —. 1979. Antheridiogens and antheridial development. Pp. 435-468 in A. F. Dyer, ed. The mipeaneninl biology of ferns. ap ea Press, London NAr, - K. NAKANISHI, and M. ENpo. 1975. On the physiology and chemistry of fern antheridi- ns. Bot. Rev. (Lancaster) 41: henge reise HUBEON .E., C. LApas, and A. McC.Lurp. 1997. Gametophyte ween age and antheri- me pce activity in Thelypteris ovata var. lindheimeri. Amer. Fern J. 87:131—142 Peck, J. H., C. J. Peck, and D. R. Farrar. 1990. Influences . 8 agar ior on fortiation of local and pring fern populations. aes Fern J. 80: NKER, T. A 7. Experimental systematics and Lue lies of the fern genera Hemio- nitis aia Gril with reference to Bommeria. Ph. D. dissertation, University of Kansas, Lawrence. Ruin, G., and D. J. PAOLILLO. 1983. Sexual development in Onoclea sensibilis on agar and soil media without the aig ioc of antheridiogen. Amer. J. Bot. 70:811-815. SCHEDLBAUER, M. D., and E. J. KLEKOWSKI . 1972 Antheridiogen activity in the fern Ceratopteris thalictroides (L.) Brogn. Bot. J. Linn. Soc. 65:399-413. SCHNELLER, J. J., C. H. HAUFLER, and T. renin . 1990. fe Giicidiontlh and natural gametophyte ae Amer. Fern J. 80: 143 152. Scott, R. J., and L. G. HICKOCK. 1987. rm analysis of antheridiogen sensitivity in Ceratopteris richardii. Amer. J. Bot. 74:1872-18 VOELLER, B. is 1964. Antheridiogens in eae Colloq. Intern. Centre Nat. Rech. Sci. (Paris) 123: 665-6 WARNE, T. iy ae G. Hickok, and R. J. Scott. 1988. Characterization and genetic analysis of an- eridiogen- pee pie mutants in the fern Ceratopteris. Bot. J. Linn. Soc. 96:371-379 Hieecees 3. Antheridiogen response in Phanerophlebia and related ferns. Amer. Fern American Fern Journal 89(4):232—243 (1999) Phylogeny of Aspleniaceae Inferred from rbcL Nucleotide Sequences NorIAKI MURAKAMI? Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake-cho, Kyoto 606-8502, Japan SATORU NOGAMI Department of Plant Sciences, School of Science, University of Tokyo, Hongo, okyo 113-0033, Japan MIkIO WATANABE Department of Biology, Aichi Kyoiku University, Kariya 448-0001, Japan KUNIO IWATSUKI Faculty of Science, Rikkyo University, Tokyo 171-0021, Japan Aspleniaceae are a well-defined family of leptosporangiate ferns, character- ized by an elongate sorus type along the leaf veins with an elongate indusium, -shaped leaf traces in the upper parts of fronds, clathrate rhizome scales, and a basic chromosome number of x = 36. However, intrafamiliar relationships of the Aspleniaceae are poorly known and no widely accepted system of clas- sification for the family has been established (Iwatsuki, 1975; Tryon and Tryon, Boniniella Hayata, and Hymenasplenium Hayata. The status of these genera is too obscure to be widely accepted because the relationship to other members In this study, we determined the rbcL sequences of 25 species of Aspleni- aceae with various leaf and rhizome morphologies and conducted a molecular phylogenetic analysis in order to elucidate intrafamiliar relationships. A mo- ‘ Author for correspondence. MURAKAMI ET AL.: rbcL PHYLOGENY from rbcL NUCLEOTIDE SEQUENCES 233 lecular phylogeny has already been constructed for a portion of the family, the Hymenasplenium group, using both restriction site analysis of chloroplast DNA (Murakami and Schaal, 1994) and rbcL sequencing (Murakami, 1995). However, no molecular study for the whole family has previously been per- formed. Significant nucleotide sequence variation in the rbcL gene was found in the earlier study on Hymenasplenium, and even higher levels were to be expected for Aspleniaceae as a whole. It is widely believed that rbcL is a slowly evolving gene and not suitable for intrafamiliar or intrageneric phylo- genetic analyses, but this is not in the case for the Aspleniaceae. Considerable amounts of sequence variations were observed even within a species complex, such as the H. obliquissimum (Hayata) Sugimoto et Kurata complex (Murakami et al., 1998) and the A. nidus L. complex (Murakami et al., 1999). MATERIALS AND METHODS PLANT MATERIALS.—Fresh leaves of 19 species and 2 varieties of Aspleniaceae were collected, mostly in Japan. Only A. ensiforme and A. nidus were col- lected from Thailand and Laos, respectively. For Hymenasplenium, six rep- resentative Old and New World species that were shown to be distantly related in our earlier study (Murakami, 1995) were selected. In total, 25 species and 2 varieties (listed in Table 1) were used for molecular phylogenetic analyses of Aspleniaceae. Although the number of taxa sampled is too small to cover the entire spectrum of variation in Aspleniaceae, which has more than 700 species, it still covers most of the variation in leaf shape (simple to finely pinnatifid), presence or absence and various position of gemma, and rhizome shape (erect, ascending, to long creeping). The samples also contained repre- sentatives of all five of the segregate genera noted in the introduction. We do not necessarily recommend the adoption of all of these segregate genera; these names are used in this paper merely for convenience. Voucher specimens are deposited at the herbarium of the Faculty of Science, Kyoto University (KYO). For allozyme analyses, A. prolongatum and all Aspleniaceae species that grow together with it on Yaku Island were collected (Table 2). For A. xkenzoi, plant materials were obtained from cultivated stocks whose origins are known. They originated from at least two different localities: Yaku Island and Ohsumi Peninsula, both Kagoshima Prefecture, Japan. Ten individuals from each spe- cies, and five individuals of the hybrid were analyzed. RBCL SEQUENCING.—Total DNAs were isolated from a single plant of each spe- cies using a modified CTAB method (Doyle and Doyle, 1987). When necessary, the DNAs were purified using a Qiagen column (tip 20) according to the vend- er’s instructions. This purification procedure improved PCR amplification. Three fragments overlapping each other and covering most of the rbcL gene were amplified by PCR using two sets of primers developed by Hasebe et al. (1994) and one modified by ourselves. For amplifying the middle fragments of the rbcL gene (307-1016 nucleotide position of the tobacco rbcL gene), we designed a new set of primers, 5’-TATCCCTTAGACCTCTTCGAAGAAGGTTC LE 1. Source of plant materials and voucher information for taxa sequenced for this investigation. All localities are in Japan, unless otherwise noted. oru B Information for six species of Hymenasplenium sequenced for an earlier investigation (Murakami 1995) is also included. Collector abbreviations: SN, Sat ogami; KO, Koichi Oohora; RI, Ryuji Ito; YT, Y. Takahashi; NM, Noriaki Murakami; YH, Yuichi Higuchi. Species Locality Voucher and DNA database accession number Asplenium antiquum Makino (Neottopteris antiqua (Makino) M asam A. cariceksliam (Hance) Baker (Hymenasplenium cardiophyl- lum (Hance) Nakaike; Boniniella ikenoi (Makino) Hayata) . cheilosorum Kunze ex Mett. (aailivietes cheilosorum > (Mett.) Tagawa) ensiforme Wall. ex Hook. et Grev. griffithianum Hook. > > A. hondoense N. saa et Hatanaka (H. hondoense (N. Murak. ke) laetum Sw. a ‘lagen (Sw.) N. Murak.) nidus L. (N. nidus (L.) J. Sm.) normale D. Don (var. normale) normale var. boreale Ohwi ex Kurata normale var. shimurae H. Ito >> >>> DD oligophlebium Baker prologatum Hook. pseudo-wilfordii Tagawa riparium a4 (H. riparium (Liebm.) N. Murak. ritoense ruprechtii = ae (Camptosorus sibiricus Rupr.) sarelii Hook. SC ae eis L. 2k gs scolopendrium (L.) Newm.) tenuicaule Hayat. trichom s Sous Nakai wildfordii Mett. ex Kuhn wrightii Eat. ex. Hook . yoshinagae Makino ee een obliquissimum (Hayata) Sugimoto et Kurata (H. obliquissi- Mt. Nishiyama, Hachijo Is. Cult., Bot. Gard., Univ. of Tokyo China, = cep crema Yunnan prov. Thailand, Doi Inth Yakushima Is., Kagvahinng Pref. Hayama, Kouchi Pref. Funada, Kiho, Mie Pref. Ko. , Shin Choshidani, Miyama, Mie Pref. Yakushima Is., Pref. abari, Mie Pr Funads, Kiho, Mis Pref. Iwayadani, Shia Nara Pref. Costa Rica, Virgen del S Deai, Nchi-katuura, Wik nein Pref. Toyama Pref. Nigishima, Kumano, Mie. Pref. Fukushima, Univ. of Tokyo Shingu, Wakayama Pref. Nigishima, Kumano, Mie. Pref. Tamakiguchi, pap ees Mie Pref. Funada, Kiho, Mie Pref. Choshi-dani, tad Mie Pref. Choshi-dani, Miyma, Mie Pref. SN, RI & YT 34 (ABO13235) NM 59692] (ABO14706) NM and X. Cheng 93-C358 (AB014704) Fukuoka et al. 93-T619 (AB014709) NM J93-001 (ABO14688) NM 596920 (ABO14705) 07) Iwatsuki et al. 93-L585 (ABO14687) SN, RI, YH 15 (ABO14701) SN 22 (AB014702) SN, RI, YT &YH 23 (ABO14703) NM 596902 (U30605) SN 2] (ABO14700) SN & KO 25 os SN 14 (ABO1469 SN & KO 13 (ABO14698) SN & KO 12 (AB014699) SN & KO 29 (ABO014695) SN, RI, YT & YH 3 (AB014690) SN, RI, YT & YH 8 (AB014689) PET (6661) b HAAWON 68 ANNTIOA ‘TVNYNOl Nad NVORANV £2. Voucher information of plant materials for allozyme analyses. All localities are in Japan. Collector abbreviations: NM, Noriaki Murakami; YH, TABL Yuichi Higuchi “ie aage-gawa, Yaku Is. Kagoshima Pref. Species Locality Voucher Asplenium antiquum Makino See antiqua (Makino) Masam.) Suzunoko, Yaku Is. Kagoshima Pref. NM 93-J45 A. nidus L. (Neottopteris nidus (L.) J. Sm aku Is. Kagoshima Pref. NM 93-J46 A. griffithianum Hook. Suzunoko, Yaku Is. Kagoshi e NM 93-J47 A. wrightii Eat. ex Hook ana-agegawa, Yaku Is. — Pref. 1 A, prolongatum Hook. Suzunoko, Yaku Is. Kagoshima A. cataractarum Rosenst. (Hymenasplenium cataractarum (Rosenst.) N. Murak.) arses Yaku Is. pag aS NM 93-J48 A A . wilfordii Mett. ex Kuhn Xkenzoi Kurata Pree Bot. Gard., Univ. of Tokyo M 93-J32 NM 93-J50-J54 SHONANOAS ACLLOFTONN 71991 Woy ANFDO'TAHd 7991 "TV LA INVAVUNWN SEZ 236 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) (TW-NP1, 307 Forward) and 5’-ACTGTTGTAGGTAAACTAGAAGGTGAACG (TW-2PR, 1016 Reverse), which eare more effective than those used by Hasebe et al. (1994), not only for Aspleniaceae but also for a wide range of vascular plants including angiosperms (Chayamarit 1997). The amplified fragments were isolated on agarose gels and purified using GeneCleanlII (BIO 101 Inc.). Purified double-stranded DNA fragments were sequenced directly in both di- rections using an ALF autosequencer and AutoCycle sequence kit (Pharmacia) with FluoroPrime (the labeled primers for sequencing) having the same se- quence that was used for amplification. Assembly and alignments of the sequences were performed using the GE- NETYX computer program (Software Development, Tokyo). For phylogenetic analyses, PAUP version 3.1.1 (Swofford, 1993) was used. Most parsimonious trees were searched by a heuristic procedure with 100 random taxon-additions to find all equally optimal islands. This analysis was conducted with TBR, MULPARS, and unordered options in which all changes were equally weight- ed. In order to evaluate the internal support for monophyletic groups, boot- strap analyses (Felsenstein, 1985) were conducted with 1,000 replicates using heuristic searches. Diplazium esculentum (Retz.) Sw. was used as an outgroup. We also tried to use Athyrium otophorum (Miq.) Koidz., Dryopteris dickinsii (Fr. et Sav.) C.Chr., and Deparia petersenii (Kunze) M.Kato as outgroups. Their validity as outgroups of Aspleniaceae was confirmed by Hasebe et al. (1994, 1995) when these authors conducted a molecular phylogenetic analysis of most fern fam- ilies. ENZYME ELECTROPHORESIS.—AIl procedures for enzyme electrophoretic analy- ses followed Shiraishi (1988). We analyzed seven enzyme systems: aspartate amino transferase (AAT), isocitrate dehydrogenase (IDH), leucine aminopep- tidase (LAP), shikimate dehydrogenase (SKD), 6-phosphoglucose dehydroge- nase (6PG), alcohol dehydrogenase (ADH), and phosphoglucomutase (PGM). RESULTS We determined 1,191 nucleotides of the rbcL sequences of 25 species and 2 varieties of the Aspleniaceae (Table 1). All sequences aligned easily without any insertions or deletions. The percentage difference of the nucleotides be- tween the most distantly related species was about 7-9%. Percent sequence divergence among typical Asplenium species was 4-6%. Thus, rbcL sequence divergence was appropriate for phylogenetic inferences. ara MURAKAMI ET AL.: rbcL PHYLOGENY from rbcL. NUCLEOTIDE SEQUENCES 100 Diplazium (outgroup) Asplenium normale var. shimurae var. boreale A. oligophlebium A. trichomanes A. incisum A. sarelii A. tenuicaule Campntocoriic sibiricus fi wrightii A. ritoense A. wilfordii A. pseudo-wilfordii | A. yoshinagae A. ensiforme 98 100 98 60 A. griffithianum A. prolongatum Neottopteris nidus Phyllitis scolopendrium H. hondoense Boniniella ikenoi H. obliquissimum H. cheilosorum H. laetum H. riparium BeewpOerU np pos < m 238 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Pe Sao S778 AAP Ok sa nd Ao LS tee eh LAP ae ea SK Dacha aa ne 6PG|/ == — Fic. 2. Schematic view of zymograms showing electrophoretic banding profiles for eight species of Asplenium in five different enzyme systems. Species are numbered as follows: 1) A. prolongatum, 2) A. Xkenzoi, 3) A. antiquum, 4) A. wrightii, 5) A. nidus, 6) A. griffithianum, 7) A. cataractarum, and 8) A. wilfordii. Note that A. xkenzoi has additive profiles of those characterizing A. prolongatum and A. antiquum, which is especially evident for dimeric enzymes, such as AAT and IDH. Cladistic analyses of the rbcl, sequences with all characters equally weighted and Diplazium as an outgroup produced 15 equally parsimonious trees of 585 steps with consistency indices of 0.638 and 0.525, with and without uninfor- mative characters, respectively, and a retention index of 0.743. The strict con- hybrid, A. xkenzoi. AAT, IDH, LAP, SKD, and 6PG were well resolved. No polymorphisms were detected in each species as far as we examined, but large variation was found among the species. The patterns of the obtained zymo- grams are shown schematically in Fig. 2. DISCUSSION The rbcL tree obtained in this study was well resolved. We will here discuss the strict consensus tree with bootstrap values (F ig. 1). The molecular tree most- Sera Fic. 3. Gross morphologies of four species of Asplenium. a) A. antiquum. b) A. prolongatum. c) A. Xkenzoi. d) A. wrightii. Scale Bar = 5 cm. Note that A. Zlolo0327 ZlolOoFi2 SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 247 first time away from its usual distribution in southern Chile and Argentina (Ibisch et al., 1996). The pore heat significance of these disjunct popu- lations remains to be exp d. Although the new — records reported here significantly add to our knowledge of Bolivian pteridophytes, they also show how little is still known about this botanically neglected country. Recent in-depth studies of Bolivian orchids (Ibisch, 1998) and bromeliads (Krémer et al., in press) have also re- vealed a large number of new country records and undescribed taxa, and have placed Bolivia among the ten most species-rich countries worldwide for these groups (Ibisch, 1998). Among pteridophytes, we are aware of a large number of undescribed species already represented in herbarium collections, and de- spite greatly intensified collecting activities in the last few years, we expect that a significant number of taxa remain to be found. The number of undes- cribed pteridophyte species from Bolivia probably approaches the number of new records listed below. ADIANTUM AMAZONICUM A. R. SM.—La Paz. Prov. Caranavi, Comunidad apg Unidos, 36 km de Caranavi hacia Sapecho, 15°41'S, 67°30'W, 1250 m, Kessler 11380 (LPB, UC). Previously known from Venezuela and Amazonian Brazil (Smith in Steyermark et al., 1995). DIANTUM CAJENNENSE WILLD. EX KLOTZSCH—Cochabamba. Prov. José Carrasco Torrico, Valle del sat, 17°08’S, 64°50'W, 220 m, Kessler 8826 (LPB, UC). Previously known from Colombia, Vene- uela, Guianas, Trinidad, Ecuador, Peru, and Amazonian Brazil (Lellinger, 1991; Smith in Stey- pein et al., 1995). This species was regarded as a synonym of the polymorphic A. tetraphyllum Humb. & Bonpl. ex Willd. by Tryon and Stolze (1989b), but it seems clearly distinct in Bolivia and elsewhere in South America (Lellinger, 1991). ADIANTUM DEFLECTENS MART.—Santa Cruz. Prov. Nuflo de Chaves, San Ramon, Estancia Castado, linea 105, 16°38’S, 62°27’W, 370 m, Arroyo P. 10 (NY); Prov. Chiquitos, Cerro Muttin, 7 km NE de la pista, 25 km al S de Puerto Suérez, 18°11.3’S, 57°52.7’W, 750 m, Vargas 3326 (NY). Previously known from Mexico, Guatemala to Panama, Colombia, Venezuela, French Guiana, Ecuador, Peru, Brazil, and Paraguay (Tryon and Stolze, 1989; Zimmer in Davidse et al., 1995) ADIANTUM HUMILE KUNzE?—Beni. Prov. Gral. Ballividn, 16 km por el camino maderero, 12 km de cumo-Rurrenabaque, 15°05’S, 67°07’W, 800 m, Kessler 10930 (LPB, UC); Prov. Vaca Diez, 13 11°02'S, 66°15’W, Moraes 215 (LPB, UC). Previously known from Belize, Panama, Colombia, Ven- ezuela, Guianas, Trinidad, Ecuador, and Peru (Jermy in Davidse et al., 1995). 2. ADIANTUM LUNULATUM BURM. F.?—Beni. Prov. Gral. Balliviaén, 16 km por el camino maderero, 12 m de Yucumo a Rurrenabaque, 15°05’S, 67°07’W, 700 m, Kessler 10896 (UC). Previously known from Cuba, ing Guatemala to Panama, Colombia, Venezuela, Old World tropics (Zimmer in Davidse et al., 1995) ADIANTUM RUIZIANUM KLOTZSCH—Cochabamba. Prov. José Carrasco Torrico, 130 km antigua carre- era Cochabamba-—Villa Tunari, 17°07'S, 65°36’W, 2000 m, Kessler 7210 (LPB, UC). Previously known only from Peru (Tryon and Stolze, 1989b). ADIANTUM SUBVOLUBILE METT. EX KUHN, VEL AFF.—Cochabamba. Prov. Ayopaya, 10 km Cocapata— Cotacajes, 16°38'S, 66°41'W, 3000 m, Kessler 9372 (LPB, UC); same general locality, 2900 m, Kes- te (LPB, UC); same general locality, 2700 m, Kessler 9483 (LPB, UC); same general oi 0 m, Kessler 9540 (LPB, UC); Prov. Ayopaya, 4 km S S de Saila Pata, 16°54’S, 66°56’W, 3 pce 12373 (LPB, UC). Previously known from Ecuador and Peru (Tryon and Stolze, hey 248 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) 2060249 |9 ADIANTUM VILLOSISSIMUM METT. EX KUHN—Beni. Prov. Gral. Ballividn, 20 km por el camino madere- ro, 12 km de Yucumo—Rurrenabaque, 15°08’S, 67°07'W, 9 Ecuador (Fay & Fay 3981, MO, QCNE, UC; Fay & Fay 4463, MO, UC; Mexia 7217, UC). Tryon and Stolze (1989b) referred the Peruvian specimens that Tryon (1964) had previously regarded as this species to A. tetraphyllum Humb. & Bonpl. ex Willd., but the two species seem amply distinct to us. The type is from Colombia (Turbo), not Panama, as stated by Tryon (1964). ALSOPHILA SETOSA KAULF., VEL AFF.—La Paz. Prov. Caranavi, Serran{a Bellavista, 41 km de Caranavi hacia Sapecho, 15°41'S, 67°30’W, 1450 m, Kessler 11451 (LPB, UC). Previously known from s. Brazil, ne. Argentina, and Paraguay (Gastony, 1973; Conant, 1983). This species is remarkable for its skeletonized aphlebiae at the base of the petiole. 266\A8C3 ANEMIA ELEGANS (GARDNER) C. PRESL—Santa Cruz. Prov. Velasco, Parque Nacional Noel Kempff M., Campamento Las Gamas, frente al Farellon, 14°49'24"S, 60°23'20"W, 900 m, Arroyo et al. 192 (MO, UC, USZ). Previously known only from Brazil (Mickel, 1962). Z lo Ole \\ ANEMIA VILLOSA HuMB. & BONPL. EX WiLLD.?—Santa Cruz. Prov. Valle Grande, 5 km de Loma Larga a Masicurf, 18°43’S, 63°54’W, 2150 m, Kessler 6318 (LPB, UC). Previously known from Colombia, Venezuela, Guianas, Ecuador, Peru, and Brazil (Mickel, 1962; Smith in Steyermark et al., 1995). Specimens from Bolivia determined as this in various herbaria are mostly other species, particu- larly A. flexuosa (Sav.) Sw. and A. tomentosa (Sav.) Sw. 2 (6609977 ARACHNIODES OCHROPTEROIDES (BAKER) LELLINGER—La Paz. Prov. J. Bautista Saavedra M., Pauji-Yuyo, entre Apolo y Charazani, 15°03’S, 68°29’W, 1350 m, Kessler 10116 (LPB, UC). Previously known ica, Panama, Colombia, Venezuela, Guyana, Surinam, Ecuador, and Peru (Tryon and Stolze, 1991; Smith in Steyermark et al., 1995). 2\o(0070\% — ASPLENIUM DELITESCENS (MAXON) L. D. GOMEz—Beni. Prov. Cral. Ballividn, 12 km por el camino ] Colonia San Pedro, 470 m, Seidel 8253 {LPB UG): Previously known from Cuba, s. Mexico, Gua- temala to Panama, Colombia, Venezuela, Ecuador, Peru, and Amazonian Brazil (Murakami and Moran, 1993). ?(ol060\@ 75 ASPLENIUM EXTENSUM FE, VEL AFF.—Cochabambaa. Prov. Ayopaya, 2 km de Casay Vinto—Choro, 16°52’S, 66°38’W, 3350 m, Kessler 9239 (LPB, UC). Previously known from Colombia and Peru (Tryon and Stolze, 1993). Notable for the short papillate hairs along the rachises adaxially. 2(o4O\0 80 ASPLENIUM INAEQUILATERALE WILLD.—Chuquisaca. Prov. Sud Cinti, entre El Palmar y Rinconada del ufete, 20°51'03”S, 64°19'10’W, 1500-1600 m, Arroyo et al. 869 (USZ, UC). La Paz. Prov. J. Bautista Saavedra M., 18 km de Charazani hacia Apolo, 15°10’S, 68°45’W, 2150 m, Kessler 10558 (LPB, canes, 3 km al NE de Bermejo, 18°06'S, 63°36’W, 1050 m, Kessler 12211 (LPB, UC). Previously known from s. Brazil, Africa, Madagascar, Mascarenes, s. India, and Sri Lanka (Sehnem, 1968). This species has often been known as A. brachyotus Kunze, but plants from Africa and South America appear identical (Moran and Smith, unpublished data). ~ iA | ASPLENIUM PALMERI MAXON—Cochabamba. Prov. Ayopaya, 20 km Cocapata—Cotacajes, 16°46’S, Y LlobOA\S I Zlbolol bl (co SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 249 66°44'W, 2000 m, Kessler 9569 (LPB, UC). La Paz. Prov. Inquisivi, 6 km de Inquisivi a Sita, 16°53’S, 67°08'W, 2500 m, Kessler 5559 (LPB, UC). Previously known from Arizona, Mexico, Belize, Gua- temala, Honduras, Colombia, w. Venezuela, Ecuador, and nw. Argentina (de la Sota, 1977; Stolze et al., 1994; Adams in Davidse et al., 1995). This species is flagelliform and gemmiferous at the apex, and hae sometimes been included within the non-flagellate A. heterochroum Kunze (e.g., by Stolze et al., 1994); we believe the two are distinct species. ASPLENIUM PEARCE] BAKER—La Paz. Prov. Abel Iturralde, Rio San Antonio, 46 km de Ixiamas a Alto Madidi, 13°38'S, 68°26’W, 300 m, Kessler 11119 (LPB, UC). Previously known from Venezuela, Guianas, Ecuador, Peru, and Amazonian Brazil (Tryon and Stolze, 1993; Smith in Steyermark, 1995). ASPLENIUM PURPURASCENS METT. EX KUHN, VEL AFF.—Beni. Prov. Gral. Ballividn, 12 km por el camino maderero, al SW del km 12 Yucumo-Rurrenabaque, 15°04’S, 67°07'W, 450 m, Kessler 10819, 10844 ee: UC). La Paz. Prov. Iturralde, San Buena Ventura, 14°15’S, 67°40’W, 1400 ft., Williams 1080 (NY). Santa Cruz. Prov. Ichilo, Parque Nacional Amboro, steep slopes above and 1 km S of Rio Saguayo, 17°41’S, 63°44’W, 750 m, Nee 36020 (LPB, NY). Previously known only from w. Ecuador (Murakami and Moran, 1993). All four Bolivian specimens differ from those in Ecuador in having the proximal pinnae with segments more prolonged along the basiscopic side and thinner blades. 2lololSQOO AspLenium RADDIANUM GAUDICH.—Cochabamba. Prov. José Carrasco Torrico, 107 km antigua carre- ZLEOS ULE SCOSSH (oY SOCOIE 7 ClbO27 1y 26615123 era Cochabamba-—Villa Tunari, 17°10'S, 65°38’W, 3050 m, Kessler 6669 (LPB, UC); Prov. José Car- rasco Torrico, 141 km antigua carretera bac> ein Tunari, 17°07'S, 65°33’W, 1400 m, Kes- sler 7727 (LPB, UC); Prov. Ayopaya, Pujyani 0 km Cocapata—Cotacajes, 16°38'S, 66°41'W, 2900 m, Kessler 9307, 9316 (LPB, UC); same general oa, 2850 m, Kessler 9441 (LPB); same general locality, 2700 m, Kessler 9507 (LPB, UC). L Prov. J. Bautista Saavedra M., Pauji-Yuyo, entre o y Charazani, 15°02'S, 68°29’W, ae aaa 9807 (LPB, UC); Prov. Caranavi, Comunidad easel Unidos, 36 fa de Caranavi hacia Sapecho, 15°41’S, 67°30’W, 1450 m, Kessler 11687, ; ; one (LPB, UC). Santa Cruz. Prov. Valle Grande, 6 km de Loma Larga a Valle Grande, 18°43’S, 54’W, 2150 m, ered 6330 (LPB, UC). Previously known from Colombia, Venezuela, Peru, and 8. iain (Tryon and Stolze, 1993). BLECHNUM BLECHNOIDES KEYSERL.—Santa Cruz. Prov. Valle Grande, 13 km de Loma Larga a Valle Grande, 18°39’S, 63°55'W, 2300 m, Kessler 6474 (LPB, UC). Previously known only from Chile (Rodriguez in Marticorena and Rodriguez, 1995). BLECHNUM LAEVIGATUM CAV.—Santa Cruz. Prov. Valle Grande, 1 km de Loma Larga a Masicuri, 18°46’S, 63°54'W, 1900 m, Kessler 6199 (LPB, UC). Previously known from se. Brazil, n. Argentina, and Uruguay (Murillo, 1968). BLECHNUM OBTUSIFOLIUM ETTINGSH.—Santa Cruz. Prov. Valle Grande, 2 km de Loma Larga a Valle Grande, 18°45'S, 63°54’W, 2050 m, Kessler 6411 (LPB, UC). Previously known from Peru, se. Brazil, nw. Argentina (Tryon and Stolze, 1993). BOLBITIS NICOTIANIFOLIA (Sw.) ALSTON—Beni. Prov. Gral. ge 12 km por el camino maderero, al SW del cumo-—Rurrenabaque, 15°04’'S, 67°07’W, 450 m, Encalie 10698 (LPB, UC). Cochabamba. Prov. co Cavernas del Repech6n, te Nacional Carrasco, 17°02'S, 65°26'W, 550 m, Kessler 8319 (LPB, UC). La Paz. Prov. J. Bautista Saavedra M., Pauji-Yuyo, entre Apolo y Charazani, 15°02’S, 68°29’W, 1450 m, Kessler 9896 (LPB, UC); Prov. Abel Iturralde, Rfo San An- tonio, 46 km de Ixiamas a Alto Madidi, 13°38’S, 68°26’ W, 300 m, Kessler 11167 (LPB, UC), Pre- viously known from the Antilles, Guatemala to Panama, Colombia, Venezuela, Guianas, Ecuador, and Peru (Hennipman, 1977; Tryon and Stolze, 1991; Hennipman and Moran in Davidse et al., 1995). Cal (K {) Fée—Cochabamba. Prov. José Carrasco Torrico, 94 km a piteitens Cochabamba-Villa Tunari, 17°12'S, 65°41'W, 3500 m, Kessler 6783 (LPB, UC). La Paz Prov. Inquisivi, “Rio Jancha Kaihua,” opposite side of headwaters of Rio Chekha, 1 km NW ZhIND LBZ ZlollOb3 Zlo&(O04Z2Z “4 ‘ 207114 oO 707 250 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Laguna Huara Huarani, Lewis 39967 (LPB, MO, UC). Previously known from Colombia and Ec- uador (Leén in Tryon and Stolze, 1993). CASSEBEERA PINNATA KAULF.—Beni. Itenez, Serranfa San Simon, 14°25’S, 62°03'W, 205 m, Quevedo et al. 986 (MO, UC, USZ). Santa Cruz. Prov. Velasco, Parque Nacional Noel Kempff M., 14°49’S, 60°23’W, 900 m, Killeen et al. 4785 (MO, UC, USZ). Previously known from s. Venezuela and Brazil (Tryon, 1942; Smith in Steyermark et al., 1995). CASSEBEERA TRIPHYLLA (LAM.) KAULF.—Chuquisaca. Prov. Hernando Siles, 42 km de Monteagudo a Padilla, 19°38’S, 64°03’W, 1200 m, Kessler 4916 (AAU, LPB). Previously known from se. Brazil, Uruguay, Paraguay, and n. Argentina (Tryon, 1942; de la Sota, 1977). Usually treated as Doryopteris triphylla (Lam.) H. Christ, but belonging better in Cassebeera, and allied to C. pinnata. CHEILANTHES MICROPTERIS Sw.—Cochabamba. Prov. Cercado de Cochabamba, entre los pefiascos de lo Cerros de San Pedro y S. Miguel, 2575 m, Steinbach 6 (UC); Prov. Esteban Arce, 1 km de La Vifia—Anzaldo, 17°57'S, 65°52'W, 2100 m, Kessler 4586 (AAU, LPB); Prov. Narciso Campero Leyes, 10 km al NW de Novillero a Santiago, 18°19’S, 65°15’W, 2400 m, Kessler 4662 (AAU, LPB). Pre- viously known from Brazil, Uruguay, and nw. Argentina (de la Sota, 1977). CHEILANTHES TWEEDIANA HOOK.—Chuquisaca. Prov. Yamparaez, 5 km de Icla a Uyuni, 19°24'S, 64°48'W, 2450m, Kessler 4744 (AAU, LPB); Prov. Jaime Zudafiez, Presto, 18°55’S, 64°57’W, 1500 m, Moraes & Vargas 1827 (LPB, UC). Cochabamba. Cerro San Pedro, 2400 m, Steinbach 8759 (UC); Prov. Cercado de Cochabamba, Colina de San Pedro, 2575 m, Steinbach 8 (UC). Dpto. unknown. Bolivian Plateau, Bang 1057 (UC). Previously known from Argentina and Paraguay. CNEMIDARIA ULEANA (SAMP.) R.M. TRYON VAR. ULEANA—Cochabamba. Prov. José Carrasco Torrico, 136 km antigua carretera Cochabamba-Villa Tunari, 17°06'S, 65°35'W, 1900 m, Kessler 7289 (LPB, UC); Prov. José Carrasco Torrico, 137 km antigua carretera Cochabamba-—Villa Tunari, 17°06’S, 65°35’W, 1600 m, Kessler 7413 (LPB, UC). Previously known from Peru and s. Brazil (Stolze, 1974; Tryon and Stolze, 1989a), and also, now, Ecuador (Stolze & Stolze 1708, F, UC); var. abitaguensis (Domin) Stolze occurs in Colombia and Ecuador (Stolze, 1974). CTENITIS NIGROVENIA (H. CHRIST) CopEL.—Santa Cruz. Prov. Valle Grande, 12 km de Loma Larga a Masicurf, 18°48’S, 63°49’W, 1000 m, Kessler 6056 (LPB, UC); Prov. Florida, Refugio Los Volcanes, de Bermejo, 18°06’S, 63°36’W, 1050 m, Kessler 12268 (LPB, UC). Previously known from s. Mexico, Guatemala to Panama, Colombia, Venezuela, Trinidad, and Peru (Tryon and Stolze, 1991; Moran in Davidse et al., 1995). CTENITIS PEDICELLATA (H. CHRIST) COPEL., VEL AFF.—Santa Cruz. Prov. Valle Grande, 12 km de Loma Larga a Masicurf, 18°47’S, 63°51’W, 1300 m, Kessler 6071 (LPB, UC). Previously known from se. Brazil (Christensen, 1913; Sehnem, 1979). CTENITIS REFULGENS (METT.) VARESCHI—La Paz. Prov. Sud Yungas, Alto Beni, Sapecho, Colonia Tar- apaca, 15°32'S, 67°21'W, 610 m, Krémer et al. 50 (LPB, UC). Previously known from s. Mexico, Guatemala, Panama, Colombia, Venezuela, Guianas, Ecuador, Peru, and Amazonian Brazil (Tryon and Stolze, 1991; Moran in Davidse et al., 1995). CTENITIS SLOANE! (SPRENG.) C.V. MorTON—Beni. Prov. Moxos, Chimanes forest, 15°10’S, 66°37’W, 260 m, Fay & Fay 2810 (MO, UC); Prov. Yacuma, Campamento Campo Monos, bajando por el Rio Curiraba, 14°38’S, 66°04'W, 195 m, Acebey 42 (LPB, UC). Previously known from Florida, Antilles, s. Mexico to Panama, Colombia, Venezuela, Ecuador, and Peru (Tryon and Stolze, 1991; Moran in Davidse et al., 1995). CYATHEA MICRODONTA (DeEsv.) DoMIN—Beni. Prov. Gral. Ballividn, 12 km por el camino maderero, al SW del km 12 Yucumo-Rurrenabaque, 15°04’S, 67°07'W, 450 m, Kessler 10733 (LPB, UC). Cochabamba. Prov. José Carrasco Torrico, Valle del Sajta, 17°08’S, 64°50’'W, 220 m, Kessler 8823, 8871 (LPB, UC); Prov. José Carrasco Torrico, Projecto Valle del Sacta, 241 km W of Santa Cruz, SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 251 17°12’S, 64°43’W, 290 m, Fay & Fay 2283 (LPB, MO, UC). Previously known from the Greater Antilles, s. Mexico, Guatemala to Panama, Colombia, Venezuela, Guianas, Trinidad, Ecuador, Peru, and Brazil (Barrington, 1978; Tryon and Stolze 1989a). 2(o02\4y DANAEA Noposa (L.) SM.—Beni. Prov. Gral. Ballividn, 12 km por el camino vrei al SW del km 12 Yucumo-—Rurrenabaque, sib 67°07'W, 450 m, Kessler 10737 (LPB, UC); ext ae a 16 km por el camino maderero, 12 km de Yucumo a Rurrenabaque, 15°05’ S, 6 m, Kessler 10901 (LPB, UC). Cochahalabe: Prov. Chapare, Cavernas del Repechén, ‘Pelee + Carrasco, 17°02'S, 65°26’W, 550 m, Kessler 8257 (LPB, UC). La Paz. Prov. Abel Iturralde, Rio San Antonio, 46 km de Ixiamas a Alto Madidi, 13°38’S, 68°26’W, 300 m, Kessler 11148, 11225 rAd anavi hacia Sapecho, 15°39’S, 67°28'W, 1200 m, Kessler 11626 (LPB, UC); Prov. Sud Yungas, Alto Beni, Sapecho, Colonia Tarapaca, 15°32’S, 67°21’W, 610 m, Krémer 45 (LPB, UC). Previously known from the Antilles, s. Mexico, Central America, Colombia, Venezuela, Trinidad, Guianas, Ecuador, Peru, and s. Brazil (Tryon and Stolze, 1989a; Camus and Pérez-Garcia in Davidse et al., 1995). 206294 Dennsragvria spruceI T. MooRE—Cochabamba. Prov. José Carrasco Torrico, 117 km antigua car- etera Cochabamba—Villa Tunari, 17°08’S, 65°38’W, 2350 m, Kessler 7071 (LPB, UC); Prov. José Carrasco Torrico, 130 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°36’W, 2000 m Kessler 7194 (LPB, UC). Previously known from Costa Rica (Hammel 19265, MO), Ecuador, and Peru (Tryon and Stolze, 1989b). 26007¢0Ob Diptazium pipLaziowwEs (KLOTZSCH & H. KARST. EX KLOTZSCH) ALSTON—Cochabamba. Carrasco Torrico, 143 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 1300 m, aaa 7561 (LPB, UC); Prov. José Carrasco Torrico, 137 km antigua carretera lees Tunari, 17°07’S, 65°35’W, 1500 m, Kessler 7764 (LPB, UC); Prov. José Carrasco Torrico, 137 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°35’W, 1500 m, Kessler 7802 (LPB, UC); v. Chapare, 1.5 km al SE de El Palmar en camino a Avispas, 17°04'S, 65°34’W, 910 m, Kessler 1 a7 (LPB, UC). Previously known from Guatemala to Panama, Colombia, Venezuela, Trinidad, and Ecuador (Stolze et al., 1994; Adams in Davidse et al., 1995). 2002793, DIPLAZIUM EXPANSUM WILLD.—Beni. Prov. Ballivian, 25 km from Yucumo on Yucumo—Quiquibe road, in the Pilén Lajas, 15°17'S, 67°04'W, 950 m, Fay & Fay 2755, 2784 (MO, UC). Coidebanbe Prov. Chapare, Incachaca, 2200 m, Steinbach 8918 (UC); Prov. Chapare, 1 km de la Caverna Re- pech6n, hacia Villa Tunari, 17°02’S, 65°27'W, 500 m, Kessler 8370 (LPB, Ue), La Paz. Prov. Car- anavi, Serranfa de sabia 46 km de Caranavi hacia Sapecho, 15°39'S, 67°28’W, 1200 m, Kessler 11619 (LPB, UC). Santa Cruz. Prov. Valle Grande, 12 km de Loma Larga a Masicuri, 18°47'S, 63°51'W, 1250 m, prec 6030 (LPB, UC); Prov. Florida, Refugio Los Volcanes, 3 km al NE de Bermejo, 18°06'S, 63°36’W, 1050 m, Kessler 12247 (LPB, UC). Previously known from s. Mexico to Costa Rica, Colombia, Venezuela, Guianas, Ecuador, Peru, and Brazil (Tryon and Stolze, 1991; Stolze et al., 1994; Adams in Davidse et al., 1995). That this relatively common and widespread species, represented in herbaria by old collections, has not heretofore been recorded from Bolivia is further evidence of the paucity of our knowledge of Bolivian ferns. “e6017°° Diptazium LONCHOPHYLLUM KUNZE—La Paz. Prov. Caranavi, comunidad Cultural Unidos, 36 km de Caranavi hacia Sapecho, 15°41'S, 67°30'W, 1450 m, Kessler 11695 (LPB, UC). Previously known from s. Mexico, Guatemala to Panama, Colombia, and Ecuador (Stolze et al., 1994; Adams in Davidse et al., 1995). 2c\\S0 25 Diptazium MOCCENIANUM (SopiRO) C. CHR., VEL AFF.—Cochabamba. Prov. José Carrasco Torrico, 149 km antigua carretera Cochabamba-Villa Tunari, 17°07'S, 65°34'W, 1000 m, Kessler 7980, pt. (LPB, UC). Previously known from Colombia and Ecuador (Stolze et al., 1994), and also, now, Brazil . (Minas Gerais, Salino 2409, BHCB, UC). 2oelUS¢ | Doryorreris COLLINA (RADDI) J. SM.—Santa Cruz. Prov. Vallegrande, 18 km de Masicuri hacia Bo- 252 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) yuibe, 18°59'S, 63°44’W, 550 m, Kessler 5262 (AAU, LPB); Prov. Nuflo de Chavez, Lomerfo, ca. 63 km S of Concepcién to Las Trancas, then ca. 10 km N on access road, 16°30’S, 61°53.5’W, 500 m, Abbott 16375 (UC, USZ). Previously known from Guyana, Surinam, Brazil, and Paraguay (Tryon, 1942; Kramer, 1978). Z lolol2\b2% BLAPHOGLOSSUM AMAZONICUM ATEHORTUA EX MICKEL—La Paz. San José, 1600 ft, Williams 1027 (NY). Santa Cruz. Prov. Ichilo, 4 km al SW del Campamento Macufiucti, 17°44’S, 63°35’W, Kessler 8670 (LPB, UC). Cited only for Peru in the original description (Mickel in Tryon and Stolze, 1991), but at that time already known from Bolivia (Williams 1027) and Mato Grosso, Brazil (Hatschbach 36206, NY, UC) (J. Mickel, pers. comm.). Zlol\73 \4 ELAPHOGLOSSUM AMBIGUUM (METT. EX H. CHRIST) ALSTON—Cochabamba. Prov. José Carrasco Torrico, 137 km antigua carretera Cochabamba—Villa Tunari, 17°07’S, 65°35’W, 1500 m, Kessler 7755 (LPB, UC); Prov. José Carrasco Torrico, 7 km de Siberia hacia Karahuasi, 17°47'S, 64°41’W, 2200 m, Kessler 9085 (LPB, UC). La Paz. Prov. Caranavi, Serran{fa Bellavista, 41 km de Caranavi hacia Sapecho, 15°41'S, 67°30'W, 1450 m, Kessler 11444 (LPB, NY, UC). Previously known from Panama, Colombia, Venezuela (Mickel in Davidse et al., 1995), and Ecuador (Fay & Fay 3796, MO, UC). 2b I2I62 ELAPHOGLOSSUM AMPHIOXYS MICKEL—Cochabamba. Prov. José Carrasco Torrico, 143 km antigua car- retera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 1300 m, Kessler 7537 (LPB, UC); Prov. José Carrasco Torrico, 149 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 1000 m, Linares 39 km hacia Caranavi, después de la cumbre, 1410 m, Beck 479 (LPB, NY); Prov. Caranavi, Serranfa Bellavista, 41 km de Caranavi hacia Sapecho, 15°41’S, 67°30'W, 1450 m, Kessler 11475 (LPB, NY, UC); Prov. Caranavi, Serrania Bellavista, 44 km de Caranavi hacia Sapecho, 15°40’S, 67°29'W, 1300 m, Kessler 11561 (LPB, NY, UC); two other collections seen. Previously known only from Peru (Mickel in Tryon and Stolze, 1991). Zlob (2\o\ ELAPHOGLOSSUM AMPLUM MICKEL—Chuquisaca. Prov. B. Boeto, 1 km de Nuevo Mundo a Padilla, arretera Cochabamba-Villa Tunari, 17°08’S, 65°38’W, Kessler 7090 (LPB, NY, UC). La Paz. Prov. Murillo, Valle de Zongo, subiendo del final del camino 22 km hacia la cumbre, 3200 m, Beck 4044 (LPB, NY); Prov. Sud Yungas, La Paz—Chulumani road, 12 km E of Chuspipata, 16°15’S, 67°10'W, 2260 m, Fay & Fay 2489 (MO, NY); Prov. J. Bautista Saavedra M., 15 km de Charazani hacia Apolo, 15°11'S, 68°46'W, Kessler 10412 (LPB, NY, UC). Santa Cruz. Prov. Valle- grande, 1 km de Loma Larga a Valle Grande, 18°45’S, 63°45’W, 2000 m, Kessler 6421 (LPB, NY, UC); 11 additional collections seen. Previously known only from Peru (Mickel in Tryon and Stolze, 1991) Zlelo'2\\eO ELAPHOGLOSSUM ANGUSTIUS MICKEL—La Paz. Prov. Nor Yungas, 37 km N de Caranavi, 1520 m, Fay & Fay 2153 (NY); Prov. Nor Yungas, 8 km de Chuspipata hacia Coroico, 16°23’S, 67°48’W, 2600, Kessler 12120 (LPB, NY, UC); Prov. Caranavi, Serranfa de Bellavista, 46 km de Caranavi hacia Sapecho, 15°39'S, 67°28’W, 1200 m, Kessler 11612 (LPB, NY, UC). Previously known only from Peru (Mickel in Tryon and Stolze, 1991). > "7 -olo\Z\49 — ELapHociossum ciuatum (C. PrEsL) T. MoorE—La Paz. Prov. Murillo, Zongo-Tal, Ortschaft Cahua, 1450 m, Feuerer 8750 a (NY). Previously known from Nicaragua to Panama, Colombia, Ecuador, and Peru (Mickel in Davidse et al., 1995). 2(o(9(2\Y3-—ELAPHOGLOssUM DICHROUM MICKEL—La Paz. Prov. J. Bautista Saavedra M., 15 km de Charazani hacia Chullina, 15°10’S, 68°53’W, 3400 m, Kessler 10641 (LPB, NY, UC); Prov. Nor Yungas, 5 km de Chuspipata hacia Coroico, 16°23’S, 67°48'W, 2750 m, Kessler 12001 (LPB, NY, UC). Previously known only from the type collection from n. Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM ENSIFORME MICKEL—Cochabamba. Prov, Chapare, Incachaca-Chusi, 2400 m, Stein- bach 9211 (NY); Prov. Chapare, Cochabamba 54 km hacia Villa Tunari, 2750 m, Beck 1426 (LPB, AMERICAN onda FERN 1999 JOURNAL QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Editor George Yatskievych Missouri Botanical Garden, P.O. Box 299, St. Louis, MO 63166-0299 Associate Editors Gerald J. Gastony, Department of Biology, Indiana University, Bloomington, IN 47405-6801 Christopher H. Haufler, Department of Botany, University of Kansas, Lawrence, KS 6604 Robbin C. Moran, New York Botanical Garden, Bronx, NY 10458-5126 James H. Peck, Department of Biology University of Arkansas-Little Rock, Little Rock, "AR 72204 The American Fern Society Council for 1998 DIANA B. STEIN, Dept. of Biological Sciences, Mount Holyoke College, South Hadley, MA 01075-6418 President BARBARA JOE HOSHIZAKI, 557 N. Westmoreland Ave., Los Angeles, CA 90004-2210. Vice-President W. CARL TAYLOR, 800 W. Wells St., Milwaukee Public Museum, Milwaukee, WI 53233-147 Sec redeay JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916- aon reasurer DAVID B. LELLINGER, 326 West St. NW., Vienna, VA 22180-4151. Membership esa JAMES D. MONTGOMERY, Ecology III, R.D. 1, Box 1795, Berwick, PA ears 9801. ack Issues Curator GEORGE YATSKIEVYCH, Missouri Botanical Garden, PO. Box 299, St. spk ‘MO 63166-0299. Journal Editor DAVID B. LELLINGER, U.S. National Herbarium MRC-166, Smithsonian Institution, Washington, DC 20560-0166. Memoir Editor CINDY JOHNSON-GROH, Dept. of Biology, Gustavus Adolphus College, 800 W. College Ave., St. Peter, MN 56082-1498 Bulletin Editor American Fern Journal EDITOR GEORGE YATSKIEV YCH ssouri Botanical Garden P.O. Box 299, fs Louis, MO 63166- 0299 ph. (314) 577-9522, e-mail: yeh o neer mobot.org ASSOCIATE EDITORS GERALD J. GASTONY.......... ne of ae. Indiana frre th Bloomington, IN 47405-6801 CHRISTOPHER H. HAUFLER ....Dept. of Botan me University of Kansas, Lawrence, KS 66045-2106 ROBBIN C. MORAN ew York Botanical Garden, Bronx, NY 10458-5126 JAMES H. PECK bone ee logy, Casters y of Arkansas—Little Rock, 1 S. University pine Little Rock, AR 72204 “American Fern Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the general st ay of ferns. It is owned by the American Fern Society, and published at a West St. NW., Vienna, VA 22180-4151. Periodicals postage paid at Vienna, VA, and additional Claims for missing issues, made 6 months (domestic) to 12 months (foreign) after the date of i a and orders for back issues should be sie to Dr. James D. Montgomery, Ecology III, R.D. Berwick, PA 18603-9801. Changes of address, dues, and applications for membership should be sent to the Membership bape inquiries concerning ferns should be addressed to the Secretar peroiertinis $20 hee gross, $19.50 net if paid through an agency (agency fee $0.50); sent free to members of the American Fern anere! (annual dues, $15.00 + $5.00 m Cae surcharge beyond U.S.A.; life membership, $300.00 + $140.00 mailing surcharge beyond U.S ack volumes are available for most years as printed issues or on ane. Please contact the Back Issues Ciieier for prices and availability. ena Send address changes to AMERICAN FERN JOURNAL, 326 West St. NW., Vienna, VA FIDDLEHEAD FORUM The editor .. the Bulletin of the American Fern Society welcomes contributions from members a members, including miscellaneous notes, offers to exchan ae or purchase materials, maonberes hiovticuloatal ae and reviews of non-technical books on fern SPORE EXCHANGE Wayne Baxter, 307 Riverdale rae a VA 22656-2120, is Director. Spores ex- euuaee and lists of available spores sent on req GIFTS AND BEQUESTS Gifts and bequests to the Society enable it to expand its services to members and to others interested in ferns. Back issues of the Journal and cash or other gifts are always welcomed and are tax-deductible Inquiries should be addressed to the Secretary. Table of Contents for Volume 89 (A list of articles arranged alphabetically by author) BRITTON, DONALD M., /soetes in Alaska and the Aleutians BRITTON, DONALD M.., (see Daniel E Brunton) BRUNTON, DANIEL FE (see Donald M. Britton) BRUNTON, DANIEL F, Rush Quillwort (/soetes junciformis, sp. nov.), a New Pterid- ophyte from Southern Georgia CAMLOH, MARJANA, Spore Age and Sterilization Affects Germination and Early Ga- metophyte Development of Platycerium bifurcatum CARLQUIST, SHERWIN (see Edward L. Schneider) GarciA-ALVAREZ, LORENA (see Emilia Pangua) GONZALES, JASIVIA (see Alan R. Smith) HOsHIZAKI, BARBARA Jog, The Cultivated Species of the Fern Genus Dryopteris in the United States IMPERATO, FILIPPO, 3-C-(6''’-O-Acetyl-B-cellobiosyl) Apigenin, a New Flavonoid from Pteris vittata IWATSUKI, KuUNIO (see Noriaki Murakami) KARRFALT, ERIC, Some Observations on the Reproductive Anatomy of /soetes an- KESSLER, MICHAEL (see Alan R. Smith) LEON, BLANCA, Blechnum penna-marina in Peru LorEA-HERNANDEZ, FRANCISCO G., Two New Fern Species from Southern Mexico Major, AGnes, Genet Composition of Diphasiastrum complanatum in Western Hun- gary: a Case Study Moran, RosBIN C., Salvinia adnata Desv. Versus S. molesta D.S. Mitch. ...........-. Murakami, Noriaki, Phylogeny of Aspleniaceae Inferred from rbcL Nucleotide Se- NocamI, SATORU, (see Noriaki Murakami) Ovor, PETER (see Agnes Major) PAJARON, SANTIAGO (see Emilia Pangua) PANGUA, EMILIA, Studies on Cryptogramma crispa Spore Germination ...........-.-++. Peck, JAMES H., Salvinia minima in Arkansas Peck, James H., Review: Ferns of the Tropics Peck, JAMES H., Review: The Ferns and Allied Plants of New England ............... PINTAUD, JEAN-CHRISTOPHE (see Dean P. Whittier) RANAL, MARL! A., Effects of Temperature on Spore Germination in Some Fern Spe- cies from Semideciduous Mesophytic Forest REDMAN, DONNELL E., Two Additional Stations for the Southern Woodfern Hybrid, Dryopteris Xaustralis in Maryland SCHNEIDER, EDWARD L., SEM Studies on Vessels in Ferns. 13. Nephrolepis .......... SMITH, ALAN R. (see Francisco G. Lorea-Hernandez) SMITH, ALAN R., New Records of Pteridophytes from Bolivia SMITH, ALAN R. (see Robbin C. Moran) STEVENS, RICHARD D., Interpopulational Comparison of Dose-Mediated Antheridi- ogen Response in Onoclea sensibilis TALBOT, STEPHEN S. (see Donald M. Britton) TELESCA, ANTONELLA (see Filippo Imperato) THOMAS, Barry A., Some Commercial Uses of Pteridophytes in Central America WATANABE, MIKIO, (see Noriaki Murakami) WERTH, CHARLES R. (see Richard D. Stevens) WHITTIER, DEAN P., Spore Germination and Early Gametophyte Development in Stro- matopteris WILSON, KENNETH A., Ontogeny of the Sporangia of Sphaeropteris cooperi .......... WILSON. KENNETH A. (see Barbara Joe Hoshizaki) YATSKIEVYCH, GEORGE, Review: Flora of Australia, Volume 48, Ferns, Gymno- sperms, and Allied Groups YATSKIEVYCH, GEORGE, Review: Illustrierter Leitfaden zum Bestimmen der Farne und farnverwandten Pflanzen der Schweiz und angrenzender Gebiete ................ Volume 89, Number 1, January—March, pages 1-100, issued 27 April 1999, Volume 89, Number 2, April—June, pages 101-180, issued 7 July 1999, Volume 89, Number 3, July-September, pages 181-220, issued 16 September 1999. Volume 89, Number 4, October-December, pages 221-281, issued 22 December 1999. 2b b(2i34 lobo WVO4 OOLOA7S lob \2IAp Lele bSo lo WUg Hell \ouyz blo yaa, SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 253 ~~ Prov. Ayopaya, 10 km Cocapata—Cotacajes, 16°38'S, 66°41’W, 2850 m, Kessler 9418 (LPB, NY, same general locality, 2700 oe — 9502 (LPB, NY, UC); same general locality, 3100 m, a 9548 (LPB, NY, UC). L . Prov. Nor Yungas, 1.9 km NE of Chuspipata on road to Coroico, 16°18'S, 67°48'W, wu m, rym 14929 (MOQ); Prov. Nor Yungas, trocha al valle = Coscapa, Parque Nacional Cotapata, 16°12'S, 67°53’W, 3250 m, Kessler 11808 (LPB, NY, UC); sam general locality, 3000 m, Kessler 11846 (LPB, NY, UC); Prov. Nor Yungas, 8 km de Ctasspipati hacia Coroico, 16°23’S, 67°4 8’W, 2650 m, Kessler 12057 (LPB, NY, UC); same general locality, 2500 m, Kessler 12172 (LPB, NY, UC). Previously known only from two collections from Cuzco, Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM GLOSSOPHYLLUM HIERON.—Cochabamba. Prov. Ayopaya, 10 km Cocapata—Cotacajes, 6°38'S, 66°41’W, 3000 m, Kessler 9348 (LPB, NY, UC); Prov. José Carrasco Torrico, 107 km antigua carretera Cochabamba-Villa Tunari, 17°10’S, 65°38'W, 3050 m, Kessler 6662 (LPB, NY, UC); Prov. José Carrasco Torrico, km 104 al Chapare, 3100 m, Steinbach 583 (NY). La Paz. Prov. J. Bautista Saavedra M., 15 km de Charazani hacia Chullina, 15°10’S, 68°53’W, 3400 m, Kessler 10604 (LPB, NY, UC); Prov. Nor Yungas, Unduavi, on old road, 16°19'S, 67°50’W, 3300 m, Solomon 4904 (MO); Prov. Nor Yungas, Cotapata, north on trail on side of mountain, 16°15'S, 67°50’W, 3200 m, Fay & Fay 2607 (MO); 25 ddtiicisat collections seen. Previously known from Colombia, Ecuador and Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM HERPESTES MickEL—Cochabamba. Prov. José Carrasco Torrico, 116 km antigua car- etera Cochabamba—Villa Tunari, 17°08’S, 65°38’W, 2350 m, Kessler 7065 (LPB, UC); Prov. José Carrasco Torrico, 130 km antigua carretera Cochabamba-—Villa Tunari, 17°07’S, 65°36’W, 2000 m etme 7215 (LPB, UC); Prov. José Carrasco Torrico, 3 km de Siberia hacia Karahuasi, 17°48’S, 41'W, 2400 m, Kessler 9146 (LPB, UC). La Paz. Prov. Nor Yungas, 19.8 km from Yolosa toward Chuspipata, 16°15'S, 67°45'W, 2280 m, Fay & Fay 2192 (MO), 2197 (NY, MO); Prov. Nor Yungas, of Yolosa on road to Chuspipata, 16°16’S, 67°47'W, 2400 m, Solomon 11750 (MO). cena known only from Ecuador (Mickel, 1985). EL (KLoTzscH) T. Moore, VEL AFF.—La Paz. Prov. J. Bautista Saavedra M., 12 km de Charazani hacia Apolo, 15°11'S, 68°46’W, 2500 m, Kessler 10512 (LPB, NY, UC); Prov. Nor Yungas, 2 km de Chuspipata hacia Coroico, 16°22’S, 67°49’W, 2900 m, Kessler 11934 (LPB, NY, UC). Previously known from Colombia, w. Venezuela, and Ecuador. ELAPHOGLOSSUM HICKENI (SopIRO) C. CHR., VEL AFF.—La Paz. Prov. J. Bautista Saavedra M., 15 km de Charazani hacia Apolo, 15°11'S, 68°46’W, 2400 m, Kessler 10442 (LPB, NY, UC); Prov. Murillo, Valle de Zongo, 23.8 km N de la cumbre, 16°08’S, 68°07'W, 2900 m, Solomon 16374 (LPB, NY, UC). Previously known from Ecuador and Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM KILLIPI! MICKEL—La Paz. Prov. Nor Yungas, del desvio a Suapi, entrando 21 km, pasando Suapi, 1290 m, Beck 13661 (LPB, NY). Previously known from Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM (Bory) T. MoorE—La Paz. Prov. J. Bautista Saavedra M., Pauji-Yuyo, entre Apolo y Charazani, 15°03 S, 68°29’W, Kessler 10147 (LPB, NY, UC). Previously known from Venezuela, Ecuador, and Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM LECHLERIANUM (METT.) T. MoorE—Cochabamba. Prov. José Carrasco Torrico, 116 km antigua carretera Cochabamba-Villa Tunari, 17°08’S, 65°38'W, 2350 m, Kessler 7061 (LPB, UC); Cochabamba, Prov. Chapare, 1700 m, Steinbach 9403 (NY). La Paz. Huancane, 2450 m, Fay & Fay 8 | Coroico, 16°24'S, 67°47'W, 2500 m Kessler 12174 (LPB, NY, UC). Dpto. unknown. Bang s.n. (NY) Previously known from Ecuador and Peru (Mickel in Tryon and bois: 1991). ELAPHOGLOSSUM LITANUM (SopIRo) C, CHR.—Beni. Hess Ballividn, 25 km from Yucumo on Yucumo-— uiquibey road, in the Pilén Lajas, 15°17’S, 4'W, 950 m, Fay & Fay 2767 (MO, NY, UC). Cochabamba. Prov. José Carrasco Torrico, i aa antigua carretera Cochabamba-Villa Tunari, Z(olol (LAY Nolo \\lo 3 FA lo \| (32 a ZlobkO 1537 2 \olo0] | Zlob \1 62S 2 lo 11621 254 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) 17°07'S, 65°34'W, 1300 m, Kessler 7611 (LPB, UC); Prov. José Carrasco Torrico, 134 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34'W, 1650 m, Kessler 7796 (LPB, UC); Prov. José Carrasco Torrico, 145 km antigua carretera Cochabamba—Villa Tunari, 17°07’S, 65°34’W, 1200 m, —— 7967 (LPB, UC). Santa Cruz. Prov. Ichilo, 4 km al Sw del Campamento Macufiuci, 17°44’S, 450 m, Kessler 8674 (LPB, NY, UC). Erroneously cited as endemic to Peru by Mickel (in ae si Stolze, 1991), but seoviously already known from northwestern Ecuador and western Colombia, and also now known from eastern Ecuador (e.g., Moran & Rohrbach 5292, MO, UC). ELAPHOGLOSSUM MACILENTUM MIcKEL—Cochabamba. Prov. José Carrasco Torrico, Cavernas del Re- pechon, Parque Nacional Carrasco, 17°02'S, 65°26’W, 550 m, Kessler 8300 (LPB, NY, UC). Previ- ously known only from the type collection from s. Peru (Mickel in Tryon and Stolze, 1991). ee MEGALURUM MICKEL—La Paz. Prov. J. Bautista Saavedra M., Pauji-Yuyo, entre Apolo Char: 5°02'S, 68°29'W, 1050 m, Kessler 9942 (LPB, NY. , UC). Previously known only from the type sorrel from n. Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM MELADENIUM MICKEL—Cochabamba. Prov. Ayopaya, 2 km ’ SE de Saila Pata, 16°54'S, 66°55’W, 3800 m, Kessler 12464 (LPB, NY, UC); Prov. José Carrasco Torrico, 100 km 3500 m, Kessler 5900, 5901 (AAU, NY). Previously known only from Peru (Mickel in Tryon and Stolze, 1991) ELAPHOGLOSSUM MELANOPUS (KUNZE) T. MoorE—La Paz. Unduavi, 3300 m, Buchtien 886 (US); Quia- ELAPHOGLOSSUM MUSCOSUM (Sw.) T. MoorE—La Paz. Prov. J. Bautista Saavedra M., 18 km de Char- Ichilo, summit of cerro Amboré, 17°45’S, 63°39'W, 1470 m, Nee 39131 (NY). Previously known from Mexico and the West Indies south to Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM NIGRESCENS (HOOK.) DigLs—Cochabamba. Prov. José Carrasco Torrico, 143 km an- tigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 1300 m, Kessler 7525, 7536b, 7842 (LPB, NY, UC); Prov. José Carrasco Torrico, 137 km antigua carretera Cochabamba—Villa Tunari, 17°06'S, 65°35'W, 1600 m, Kessler 7395 (LPB, NY, UC). La Paz. Prov. Nor Yungas, Puerto Linares 39 km hacia Caranavi, después de la cumbre, 1410 m, Beck 465 (LPB, NY). Previously known from Venezuela south to Peru (Mickel in Tryon and Stolze 1991). ELAPHOGLOSSUM OBTUSUM MICKEL—Beni. 5 km NW of Guayaramerin, Anderson 11818 (NY). Previ- ously known only from Peru (Mickel in Tryon and Stolze, 1991), but now also from Ecuador (Morona-Santiago, Ollgaard 1919, AAU) ELAPHOGLOSSUM OCULATUM MICKEL—Cochabamba. Prov. José Carrasco Torrico, 108 km antigua Car- retera Cochabamba-Villa Tunari, 17°09’S, 65°38’ W, 2950 m, Kessler 6619 (LPB, UC). La Paz. Prov. Inquisivi, along road between Loma Linda and Turculi, 16°38’S, 67°10’W, 1850 m, Lewis 36881 (LPB, MO, UC); Prov. Sud Yungas, Huancané 7,5 km hacia el sudeste sobre el camino nuevo, 2410 m, Beck 3134A (LPB, NY); Prov. Nor Yungas, 10 km de Chuspipata hacia Coroico, 16°24'S, e Caranavi hacia Sapecho, 15°40’S, 67°29’W, 1500 m, Kessler 11362 (LPB, NY, UC). Santa Cruz. Prov. Valle Grande, 4 km de Loma Larga a Masicuri, 18°47’S, 63°53 W, 1600 m, Kessler 6289 (LPB, UC); Prov. Manuel Maria Caballero, 50 km al norte de Mataral, 2000 m, Smith et al. 13319 (LPB, MO, UC). Previously known only from Peru (Mickel in Tryon and Stolze, 1991). < 0103803 ZI 104 ZOLORSY2Z hole ISA Lleol S545 Zeb Sey 26S 18) ZlklA\S Fe ZOGIO94Y SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 255 ELAPHOGLOSSUM PERUVIANUM (L.D, GOMEZ) MickEL—Cochabamba. Prov. José Carrasco Torrico, 113 km antigua carretera Cochabamba-Villa Tunari, 17°07'S, 65°38’W, 2650 m, Kessler 6934 (LPB, UC); Prov. José Carrasco Torrico, 117 km antigua carretera Cochabamba-Villa Tunari, 17°08'S, 65°38’ W, 2350 m, Kessler 7078 (LPB, UC). Previously known only from Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM PILOSIUS MICKEL—Cochabamba. Prov. José Carrasco Torrico, 85 km antigua carretera Seep: —Villa Tunari, 17° =a 'S, 65°43’W, prin m, Kessler 6583 (LPB, UC); Prov. José Carrasco Torrico, 63 km antigua carretera Cochabamba-Villa Tunari, 17°15'S, 65°43’ W, 3700 m, Kessler 6879 (LPB, UC). Previously known from Costa Rica, Panama, Colombia, Venezuela, Ecuador, and Peru (Mickel in Tryon and Stolze, 1991; Mickel in Davidse a al., 1995). ELAPHOGLOSSUM PLUMOSUM (FEE) T. MOORE, VEL. AFF.—Cochabamba. Prov. José Carrasco Torrico, 112 km antigua carretera Cochabamba-Villa Tunari, 17°07'S, 65°38'W, 2700 m, Kessler 6923 (LPB, NY, UC); Prov. José Carrasco Torrico, 114 km antigua carretera Cochabamba-—Villa Tunari, 17°07’S, 65°36'W, 2200 m, Kessler 6991 (LPB, NY, UC); Prov. José Carrasco Torrico, 130 km antigua carretera Cochabamba-Villa Tunari, 17°07'S, 65°36’W, 2200 m, Kessler 7249 (LPB, NY, UC), same locality, Kessler 7287 (LPB, NY, UC). Previously known from the Guianas, Venezuela, Colombia, Ecuador, Peru, and Brazil (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM PUMILIO MiCKEL—La Paz. Prov. J. Bautista Saavedra M., Charazani-Tal, bei den Fel- sen auf der linken Talseite unmittelbar bei der Furt bei Cilij, 2750 m, Feuerer 6727a (NY); Prov. Larecaja, 56 km de Sorata a Consata, 2600 m, Kessler 4351 (AAU, LPB). Previously known only from Peru (Mickel in Tryon and Stolze, 1991). ELAPHOGLOSSUM RAYWAENSE (JENMAN) ALSTON—Cochabamba. Prov. Chapare, El Palmar, 155 km an- tigua carretera Cochabamba-Villa Tunari, 17°05'S, 65°32'W, 750 m, Kessler 8107 (LPB, UC); Prov. José Carrasco Torrico, Valle del Sajta, 17°08’S, 64°50'W, 220 m, Kessler 8836 (LPB, UC). Previously own from Venezuela, Guianas, Ecuador, Peru, and Amazonian Brazil (Mickel in Tryon an Stolze, 1991). ELAPHOGLOSSUM SETIGERUM (SOpDIRO) DieLs—Cochabamba. Prov. José Carrasco Torrico, 111 km a tigua carretera Cochabamba-Villa Tunari, 17°08’S, 65°38’W, 2750 m, Kessler 6845 (LPB, NY, UC): bamba-Villa Tunari, 17°07'S, 65°36’W, 2000 m, Kessler 7262 (LPB, NY, UC). La Paz. Chuspipata, 2450 m, Gentry 44713 (MO); Huarinillas, 2800 m, Beck 21896 (LPB, NY); Prov. Nor Yungas, Trocha al Valle de Coscapa, Parque Nacional Cotapata, 16°12'S, 67°53'W, 3000 m, Kessler 11877 (LPB, NY, UC); Prov. Nor Yungas, 10 km de pn ne oat 16°24’S, 67°47'W, 2500 m, Kessler 12145 (LPB, NY, UC). Previously known from Ecuador and Peru (Mickel in Tryon and Stolze, 1991). Bolivian specimens of this species differ from Ecuadorian ones by the unequal-sided, at- tenuate bases on the fertile blades. ELAPHOGLOSSUM SMITHI (BAKER) H. CHRIST, VEL AFF.—La Paz. Prov. Caranavi, 37 km Caranavi-—Sa- pecho, 15°40’S, 67°29’W, 1500 m, Kessler 11270 (LPB, NY, UC). Previously known from Hispan- iola, Lesser Antilles, Costa Rica, and Panama (Mickel in Davidse et al., 1995) but recently also collected in Ecuador (Pichincha, @ligaard 1014, AAU; Morona-Santiago, Oligaard 2703, AAU). ELAPHOGLOSSUM ZEBRINUM MICKEL, VEL AFF.—Cochabamba. Prov. Chapare, Parque Machia, 1 km al E de Villa Tunari, 16°58’S, 65°24'W, 350 m, Kessler 8489 (LPB, NY, UC). Previously known only from Colombia and Peru (Mickel in Tryon and Stolze, 1991), and now also Ecuador (Fay & Fay 2745, UC). This is related to a group of white-streaked species with blades long-attenuate at the base, such as E. oblanceolatum C. Chr. and E. lellingeri Mickel from Colombia and Ecuador. Ela- phoglossum zebrinum appears to grow at much lower elevations than its relatives (mostly 1000— 2900 m) and has more slender rhizomes. ERIOSORUS NOVOGRANATENSIS A.F. TRYON, VEL AFF.—Cochabamba. Prov. José Carrasco Torrico, 108 Zbl\Y2\2 ZOO 2157 500SU%32 2 (oleO06 229 lo0727 2 bool YFu> 18) | & 4 if ] ' 256 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) km antigua carretera Cochabamba-Villa Tunari, 17°09’S, 65°38’ W, 2950 m, Kessler 6598 (LPB, UC). Previously known from Colombia (Tryon, 1970). Attributed a wider range by Moran (in Davidse et al., 1995), in Nicaragua, Costa Rica, Panama, Venezuela, Ecuador, Peru, and Bolivia, but the identification of the specimens upon which these records are based is questionable. HUPERZIA CAPILLARIS (SODIRO) HOLUB—La Paz. Prov. Caranavi, Serranfa Bellavista, 37 km Caranavi— Sapecho, 15°40’S, 67°29’W, 1500 m, Kessler 11294 (AAU, GOET, LPB). Previously known from s. Mexico to Panama, Colombia, Venezuela, Ecuador, and Amazonian Brazil (Ollgaard, 1992; Ollgaard in Davidse et al., 1995). — SUBULATA (PoIR.) HOLUB—Cochabamba. Prov. Chapare, Serranfa de Callejas NE Colomi, ar Aguirre-E] Palmar road, 3000 m, Miiller 6624 (GOET, LPB). Previously known from Honduras, st op Colombia, Ecuador, and Peru (Qllgaard in Tryon and Stolze, 1994; Ollgaard in Davidse et al., 1995). HUPERZIA TENUIS (WILLD.) TREVIS.—Cochabamba. Sailapata [ = Saila Pata], Ayopaya, 3500 m, Car- denas 3221 (GH). Previously known from Costa Rica, Colombia, Venezuela, Ecuador, and Peru (Oligaard in Tryon and Stolze, 1994; Ollgaard in Davidse et al., 1995; B. Ollgaard, pers. comm.). HUPERZIA WILSONII (UNDERW. & F.E. LLoyD) B. @LLG.—Cochabamba. Prov. José Carrasco Torrico, 137 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°35’W, 1500 m, Kessler 7772 (AAU, LPB). Previously known from Antilles, saey Costa Rica, Panama, Colombia, Venezuela, Ec- uador, and Peru (Ollgaard in Davidse et al., 1995) HYMENOPHYLLUM (SUBG. SPHAEROCIONIUM) AMABILE C.V. MORTON—La Paz. Prov. Nor Yungas, Trocha al Valle de Coscapa, Parque Nacional Cotapata, 16°12’S, 67°53’W, 3450 m, Kessler 11738, (LPB, UC). Previously known from Ecuador and Peru (Morton, 1947: Tryon and Stolze, 1989a). HYMENOPHYLLUM (SUBG. SPHAEROCIONIUM) HEMIDIMORPHUM R.C. MORAN & B. @LLG.—Cochabamba. Prov. José Carrasco Torrico, 130 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°36'W, osta Rica and Ecuador (Moran and @llgaard, 1995; Rojas-Alvarado, 1996), also now Colombia (Silverstone-Sopkin 4297, UC). HYMENOPHYLLUM (SUBG. MECcopIUM) MATHEWSII BoscH—Cochab ov. Ayopaya, Pujyani, 10 km Cocapata—Cotacajes, 16°38’S, 66°41'W, 2900 m, Kessler 9315, 9330 eine UC). Previously known from Ecuador and Peru (Tryon and Stolze, 1989a). HYMENOPHYLLUM (SUBG. SPHAEROCIONIUM) TRAPEZOIDALE LIEBM.—Cochabamba. Prov. José Carrasco Torrico, 7 km de Siberia hacia Karahuasi, 17°47’S, 64°41’W, 2200 m, Kessler 9101 (LPB, UC). La Paz. Prov. Sud Yungas, Choquetanga, ca. 2980 m, Lewis 86-334 (F, UC). Previously known from s. Mexico, Guatemala to Panama, Colombia, hasan and Surinam (Morton, 1947; Pacheco in Davidse et al., 1995). HYPOLEPIS POEPPIGH (KUNZE) R.A. RODR., VEL AFF.—Cochabamba. Prov. Ayopaya, 10 km Cocapata— Cotacajes, 16°38’S, 66°41'W, 3100 m, Kessler 9535 (LPB, UC). Previously known from Chile (in- cluding Juan Fernandez), and s. Argentina (Rodriguez in Marticorena and Rodriguez, 1995). sbepraitig viscosA METT.—La Paz. Prov. Nor Yungas, 5 km de Chuspipata hacia Coroico, 16°23’S, 7°48'W, 2700 m, Kessler poe (LPB, UC). Previously known from Guatemala, Costa Rica, Pan- ama, Colombia, and Venezuela (Moran in Davidse et al., 1995). Moran also reported the species from s. Mexico, apparently erroneously. ISOETES GARDNERIANA KUNZE EX METT., VEL AFF.—Santa Cruz. Nuflo de Chavez, Lomerio, ca. 63 km S of Concepcién to La Trancas, then ca. 10 km N, ca. 16°30’S, 61°53’W, ca. 500 m, Abbott 16374 (UC, USZ); Nuflo de Chavez, Lomerfo, ca. 63 km S of Concepci6n to La Trancas, then ca. 0.5 km SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 257 a. 16°35’S, 61°52’W, ca. 500 m, Abbott 16383 (UC, USZ). Previously reported from Brazil and ait (Pfeiffer, 1922). 2660529 a GouDOTI (HIERON.) C. CHR.?—La Paz. Prov. P. D. Murillo, entre Pongo y Unduavi, Mina 0, subiendo hacia la Mina San Luis, 3960 m, Beck 21524, 21525 (LPB, UC). Previously known Colombia, Ecuador, and Peru (Tryon, 1962; Tryon and Stolze, 1989b). LASTREOPSIS KILLIPI (MAXON) TINDALE—La Paz. Prov. Nor Yungas, 10 km de Chuspipata hacia Co- 2ob\22 32, roico, 16°24’S, 67°49'W, 2500 m, Kessler 12133 (LPB, UC). Previously known from Costa Rica, Panama, Colombia, s. Venezuela, Ecuador, and Peru (Tindale, 1965; Tryon and Stolze, 1991; Moran in Davidse et al., 1995; Smith in Steyermark et al., 1995) 26l0141747 7 — Major (COPEL.) A.R. SM. & R.C. MORAN, VEL AFF.—Cochabamba. Prov. José Carrasco , 107 km antigua carretera Cochabamba-Villa Tunari, 17°12'S, 65°42’W, 3300 m, Kessler em nLee UC). Previously known from Venezuela, Colombia, Ecuador, and Peru (Smith et al., 1991; Tryon and Stolze, 1993). POLYSTICHUM TURRIALBAE H. Curist—Cochabamba. Prov. Ayopaya, 10 km Cocapata—Cotacajes, ~ 16°38’S, 66°41'W, 2850 m, Kessler 9443 (LPB, UC); same general locality, 3100 m, Kessler et al. soe (LPB, UC); Prov. Ayopaya, 2 km al N de Saila Pata, 16°54’S, 66°56’W, 3100 m, Kessler 12403 (LPB, UC). Previously known from Mexico, Costa Rica, and Panama (Mickel and Smith, unpub- lished data). This is one of the few indusiate species of Polystichum in Andean regions. sileon CONSANGUINEA METT. EX KUHN—Cochabamba. Prov. José Carrasco Torrico, 147 km antigua rretera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 2100 m, Kessler 7931 (LPB, UC). La Paz. Pr rov. Caranavi, Serranfa Bellavista, 38 km de Caranavi hacia Sapecho, 15°40'S, 67°29'W, 1500 m, Kessler 11395 (LPB, UC). Previously known from Venezuela (Smith, 1985; Tryon and Stolze, Zlolo02 ULY 26613645 2600337 ) J A LN S$O70y S8 SON\G37A4 2(olo24UOL0 260 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) 1989b), but also in Colombia (e.g., van der Werff 9732, UC) and Ecuador (e.g., Moran & Rohrbach 5251, UC). PTERIS GRANDIFOLIA L.—La Paz. Prov. Sud Yungas, Alto Beni, 500 m, Seidel & Vaquiata 7773 (LPB, UC). Previously known from the Antilles, s. Mexico, Guatemala to Panama, Colombia, Venezuela, Trinidad, Ecuador, Peru, and Brazil (Moran in Davidse et al., 1995). PTERIS PEARCE! BAKER—Beni. Prov. Ballivian, 25 km from Yucumo on Yucumo—Quiquibey road, in the Pil6n Lajas, 15°17’S, 67°04’W, 950 m, Fay & Fay 2711 (UC). Previously known from Colombia, Venezuela, Peru, and n. Brazil (Tryon and Stolze [1989b] treating the species under the synonym P. petiolulata R.M. Tryon) (Smith in Steyermark et al., 1995) PTERIS TRIPARTITA Sw.—Cochabamba. Prov. José Carrasco Torrico, Projec to Valle del Sacta, 241 km W of Santa Cruz, 219 km E of Cochabamba, 17°12’S, 64°43’W, 290 m, 8, 9, and 12 Jul 1989, Fay & Fay 2285, 2303, 2348 (LPB, MO, UC); Prov. José Carrasco Torrico, Valle de Sajta, 17°08’S, 64°50'W, 220 m, 5 Oct 1996, Kessler 8829 (LPB, UC). In the Neotropics, previously known from Antilles, Costa Rica, Panama, Colombia, Venezuela, Guianas, Ecuador, and Peru; introduced and naturalized from the Old World (Moran in Davidse et al., 1995). SELAGINELLA CF. CALOSTICHA SPRINGC—Cochabamba. Prov. José Carrasco Torrico, 136 km antigua carretera Cochabamba-Villa Tunari, 17°06’S, 65°34'W, 1700 m, Kessler 7370 (LPB, UC). Previously own from Venezuela and Peru (Alston et al., 1981). SELAGINELLA CAVIFOLIA A. BRAUN—La Paz. Prov. Nor Yungas, Trocha al Valle de Coscapa, Parque Nacional Cotapata, 16°12’S, 67°53’W, 3000 m, Kessler et al. 11893 (UC). Previously known from Colombia, Venezuela, and Ecuador (Alston et al., 1981; Valdespino, 1995). SELAGINELLA MORITZIANA SPRING VAR. MORITZIANA—Cochabamba. Prov. José Carrasco Torrico, 108 km antigua carretera Cochabamba-Villa Tunari, 17°09’S, 65°38'W, 2950 m, Kessler 6550 (LPB, UC); Prov. José Carrasco Torrico, 107 km antigua carretera Cochabamba-Villa Tunari, 17°10’S, 65°38’W, 3050 m, Kessler 6677 (LPB, UC); Prov. José Carrasco Torrico, 115 km antigua carretera Cochabam- ba-Villa Tunari, 17°08’S, 65°38’W, 2500 m, Kessler 6962 (LPB, UC); Prov. José Carrasco Torrico, . UC); Prov. José Carrasco Torrico, 108 km antigua carretera Cochabamba—Villa Tunari, 17°06’S, m, Kessler 12202 (LPB, UC). Previously known from s. Mexico, Guatemala, Honduras, Costa Rica, anama, Colombia, Venezuela, Ecuador, and Peru (Valdespino, 1995); Alston et al. (1981) circum- scribed the species much more narrowly, with a range only in Venezuela and Ecuador. Valdespino recognized three varieties, and the most widespread one, var. moritziana, appears to be the one occurring in Bolivia. Selaginella moritziana often produces sobols from axils on proximal branches. SELAGINELLA REVOLUTA BAKER—Cochabamba. Prov. José Carrasco Torrico, Valle de Sajta, 17°08’S, 64°50’W, 220 m, Kessler 8820 (LPB, UC). Previously known from Panama, Colombia, Venezuela, Guianas, Peru, and Amazonian Brazil (Alston et al., 1981; Fraile in Davidse et al. [1995] cited Bolivia in the range without documentation). STICHERUS PERUVIANUS (MAXON) A.R. SM., M. KESSLER & J. GONZALES, COMB. NOV.—Dicranopteris peruviana Maxon, Amer. Fern J. 33:133. 1943.—Cochabamba. Prov. Chapare, 163 km W of El Sacta, 56 km E of Cochabamba, 17°20’S, 65°50’W, 2460 m, Fay & Fay 2386 (LPB, MO, UC). La Paz. Prov. Nor Yungas, Cotapata, roadside behind gas station, 16°15'S, 67°50’W, 3225 m, Fay & Fay 2465 (LPB, MO, UC); Prov. Sud Yungas, La Paz—~Chulumani road, 15.1 km W of Chulumani, oe © tico Zbl 2240 Zob224 2 SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 261 9.3 km from Huancané, 16°15’S, 67°30’W, 2450 m, Fay & Fay 2584 (LPB, MO, UC). Previously known only from Peru (Tryon and Stolze, 1989a). frie asiabislate LECHLERI (METT.) C. CHR.—La Paz. Prov. Prov. Caranavi, Serrania Bellavista, 47 km vi hacia Sapecho, 15°29'S, 67°28’W, 1150 m, Kessler 11645 (LPB, UC). Previously known Snap ae Rica, Colombia, w. Venezuela, Ecuador, and Peru (Moran, 1991). TECTARIA ANTIOQUOIANA (BAKER) C. CHR., VEL AFF.—Cochabamba. Prov. Chapare, Parque Ecoturis- km al E de Villa Tunari, 16°58'S, 65°24’W, 400 m, Kessler 8540 (LPB, UC). Previ- ously known from Nicaragua, Costa Rica, Panama, Colombia, and Venezuela (Moran in Davidse et al., 1995). RIA LIZARZABURUI (SOpIRO) C. CHR., VEL AFF.—La Paz. Prov. J. ee Saavedra M., 15 km de en hacia Apolo, 15°11’S, 68°46’W, 2400 m, Kessler 11443 (LPB, UC). Previously known from Colombia, Venezuela, Ecuador, and Peru (Tryon and Stolze, 1991; Smith in Steyermark et al., 1995). 2OolS ) \2 Tecraria Pitosa (FEE) R. C. MoRAN?—Beni. Prov. ag haere 12 km por el camino maderero, al SW del km 12 Yucumo-Rurrenabaque, 15°04’'S, 67°07’W, 450 m, Kessler 10692 (LPB, UC). Cochabamba. Prov. Chapare, Cavernas del Repechén, sl Nacional Carrasco, 17°02’S, 65°26’W, 550 m, Kessler 8352 (LPB, UC); Prov. Chapare, 78 km W of El Sacta, Projecto Valle del Sacta, 241 km W of Santa Cruz, 17°00’S, 65°25'W, 355 m, Fay & Fay 2378 (LPB, MO, UC). La Paz. Prov. Prov. aranavi, Serrania Bellavista, 37 km de Caranavi hacia Sapecho, 15°40'S, 67°29' W, 1500 m, Kessler 11298 (LPB, UC). Previously known from Jamaica, Costa Rica, Panama, Colombia, Ecuador, Peru, and s. Brazil (Moran in Davidse et al., 1995) Phe 17226 ecmbaratae MOLLISSIMA (FEE) A.R. SM., VEL AFF.—La Paz. Prov. Caranavi, Serrania Bellavista, 47 km de Car; ZLLORS FY ZaelS1bS 2607493 Zl 1y 4, blo . Q avi hacia Sapecho, 15°39’S, 67°28’W, 1150 m, Kessler 11676 (LPB). Previously known from Aaoiitlae s. Mexico to Panama, Colombia, Venezuela, Guianas, and Ecuador (Smith and Mor- an in Davidse et al., 1995). This species is peculiar in lacking rhizome scales, and is most similar and closely related to T. senilis (Fée) A.R. Sm. (also known from Bolivia), which has rhizome THELYPTERIS (SUBG. AMAUROPELTA) AMPHIOXYPTERIS (Soprro) A.R. SM.—La Paz. Prov. J. Bautista Saa- vedra M., Pauji-Yuyo, entre Apolo y Charazani, 15°02'S, 68°29'W, 1200 m, Kessler 9974 (LPB, UC). eeiousls known from Panama, Colombia, aa Ecuador (Smith 1983; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. AMAUROPELTA) ANDICOLA A. R. SM.—Cochabamba. Prov. José Carrasco Torrico, . km antigua carretera Cochabamba-Villa Tunari, 17°14'S, 65°13'W, 3750 m, Kessler 6894 (LPB, UC); hg Ayopaya, 10 km cnr sap 16°38'S, 66°41’W, 3050 m, Kessler 9389 (LPB, UC). La Paz. Prov. J. Bautista Saavedra M., 15 km de Charazani ne Chullina, 15°10’S, 68°53, 3400 m, cada 10647, 10652 (LPB, UC); mid Nor Yungas, Unduavi a Chulumani, 3125 m, Schmit 169B (LPB, UC); Prov. Nor Yungas, Unduavi, 3300 m, Buchtien ass 2710 (UC). Previously known only from Peru (Smith in Tryon and Stolze, 1992). cepa (suBG. GONIOPTERIS) BIOLLEYI (H. CHRIST) PROCTOR—La Paz. Prov. J. Bautista Saavedra M. ji-Yuyo, entre Apolo y Charazani, 15°02’S, 68°29'W, 1450 m, Kessler 9897 (LPB, UC); Prov. J. Sansa Saavedra M., 10 km de Camata hacia Apolo, 15°13’S, 68°41'W, 1300 m, Kessler 10344 (LPB, UC); Prov. Caranavi, Serrania Bellavista, 36 km de Caranavi hacia Sapecho, 15°41’S, 67°30'W, 1300 m, Kessler 1 1268 (LPB, UC). Previously known from Jamaica, s. Mexico to Panama, and Colombia to Peru, Venezuela, and n. Brazil (Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. STEIROPTERIS) DECUSSATA (L.) PROCTOR VAR. pECUsSsATA—Cochabamba. Prov. José Carrasco Torrico, 141 km antigua carretera Cochabamba-Villa Tunari, 17°07'S, 65°33'W, 1400 m, Kessler 7739 (LPB, UC); Prov. Chapare, 151 km antigua carretera Cochabamba-Villa Tunari, Z2C\lo0 79 26007439 i G 262 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) 17°05'S, 65°34'’W, 900 m, Kessler 8073 (LPB, UC). La Paz. Prov. J. Bautista Saavedra M., Pauji- Yuyo, entre Apolo y Charazani, 15°03’S, 68°29’W, 1350 m, Kessler 10113 (LPB, UC); Prov. Caranavi, Serrania de Bellavista, 47 km de Caranavi hacia Sapecho, 15°29’S, 67°28’W, 1150 m, Kessler 11644 (LPB). Previously known from Antilles, Guatemala to Panama, Colombia, Venezuela, Guyana, French Guiana, Ecuador, and Peru (Smith in Davidse et al., 1995). THELYPTERIS (SUBG. STEIROPTERIS) DECUSSATA (L.) PROCTOR VAR. VELUTINA (SopIRO) A.R. SM.—Cocha- bamba. Prov. José Carrasco Torrico, 136 km antigua carretera Cochabamba—Villa Tunari, 17°06’S, 65°35'W, 1900 m, Kessler 7286 (LPB, UC). Previously known only from Colombia and Ecuador (Smith, 1983). THELYPTERIS (SUBG. AMAUROPELTA) DEFLEXA (C. PRESL) R.M. TRYON—Cochabamba. Prov José Carrasco Torrico, 111 km antigua carretera Cochabamba—Villa Tunari, 17°08’S, 65°38’W, 2750 m, Kessler 6846 (LPB, UC); Prov. José Carrasco Torrico, 113 km ant 17°07'S, 65°38’W, 2650 m, Kessler 6935 (LPB, UC); Prov. José Carrasco Torrico, 115 km antigua carretera Cochabamba-Villa Tunari, 17°08’S, 65°38’W, 2500 m, Kessler 7016 (LPB, UC); Prov. José Carrasco Torrico, 116 km antigua carretera Cochabamba-Villa Tunari, 17°08’S, 65°38’W, 2400 m, Kessler 7035 (LPB, UC). La Paz. Prov. Nor Yungas, 2 km de Chuspipata a Coroico, 16°22’S, 67°49’W, 2900 m, Kessler 11903 (LPB, UC). Previously known from s. Mexico to Panama, Colom- bia, Venezuela, Ecuador, and Peru (Smith, 1983; Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. AMAUROPELTA) DEMISSA A.R. SM.—Cochabamba. Prov. Ayopaya, 10 km Cocapata— otacajes, 16°38’S, 66°41'W, 2900 m, Kessler 9366 (LPB, UC); same general locality, 3100 m, Kes- sler 9534 (LPB, UC). La Paz. Prov. J. Bautista Saavedra M., 12 km de Charazani hacia Apolo, 15°11’'S, 68°46’W, 2500 m, Kessler 10486 (LPB, UC); Prov. Nor Yungas, Trocha al Valle de Coscapa, Parque Nacional Cotapata, 16°12’S, 67°53’W, 3250 m, Kessler 11817 (LPB, UC); Prov. Nor Yungas, 5 km de Chuspipata hacia Coroico, 16°23'S, 67°48’W, 2750 m, Kessler 11975 (LPB, UC). Previously known only from Peru (Smith in Tryon and Stolze, 1992), from the type and one other collection. THELYPTERIS (SUBG. GONIOPTERIS) EGGERSII (HIERON.) C.F. REED—Cochabamba. Prov. José Carrasco Torrico, 149 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34’W, 1000 m, Kessler 8038 (LPB, UC). Previously known from Costa Rica, Panama, w. Colombia, w. Ecuador, and Peru (Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. MENISCIUM) ENSIFORMIS (C. Cur.) R.M. TRYoN—Cochabamba. Prov. José Carrasco Torrico, 143 km antigua carretera Cochabamba-Villa Tunari, 17°07’S, 65°34'W, 1300 m, Kessler 7642 (LPB, UC). La Paz. Prov. Caranavi, Serrania Bellavista, 42 km de Caranavi hacia Sapecho, 15°40'S, 67°29'W, 1400 m, Kessler 11520 (LPB, UC). Previously known from Costa Rica, Colombia, w. Venezuela, Ecuador, and Peru (Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. AMAUROPELTA) EXUTA A.R. SM., VEL AFF.—La Paz. Prov. Caranavi, Serrania Bel- lavista, 36 km de Caranavi hacia Sapecho, 15°41’S, 67°30'W, 1300 m, Kessler 11266 (LPB, UC). Previously known from two collections in Ecuador (Smith, 1983). THELYPTERIS (SUBG. AMAUROPELTA) FUNCKII (METT.) ALSTON—La Paz. Prov. Nor Yungas, Trocha al Valle de Coscapa, Parque Nacional Cotapata, 16°12'S, 67°53'W, 3350 m, Kessler 11781 (LPB, UC); same general locality, 3000 m, Kessler 11847 (LPB, UC); Prov. Nor Yungas, 5 km de Chuspipata hacia Coroico, 16°23'S, 67°48’W, 2700 m, Kessler 12011 (LPB, UC). Previously known from Costa Rica, Colombia, Venezuela, Guyana, and Ecuador (Smith 1983; Smith in Davidse et al., 1995). THELYPTERIS (SUBG. AMAUROPELTA) LEPIDULA (HIERON.) ALSTON, VEL AFF.—Cochabamba. Prov. José arrasco Torrico, 109 antigua carretera Cochabamba-Villa Tunari, 17°09’S, 65°38’W, 2950 m, Kessler 6643B (LPB, UC). Previously known from Costa Rica, Colombia, Venezuela (Smith in Dav- idse et al., 1995), and also, now, Ecuador (Prov. Carchi, Hoover et al. 3531, 3595, MO). The sole Bolivian specimen differs from others elsewhere in the range by the very shortly setulose recep- SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 263 tacles, and from Venezuelan and Costa Rican specimens by the short, adpressed hairs on the blades adaxially. THELYPTERIS (SUBG, AMAUROPELTA) uals (Sopiro) C.F. REED—La Paz. Prov. Nor Yungas, 5 km e Chuspipata hacia Coroico, 16°23'S, 67°48’W, 2700 m, Kessler 12012 (LPB, UC); Prov. Nor Yun- Zlole\SO49 gas, 8 km de Chuspipata hacia Coroico, rts S, 67°48'W, 2650 m, Kessler 12056 (LPB, UC). Pre- viously known from Costa Rica, Panama, w. Colombia, w. Ecuador, and Peru (Smith in Davidse et al., 1995). THELYPTERIS (SUBG. MENISCIUM) MEMBRANACEA (METT.) R.M. TRYON—Cochabamba. Prov. Chapare, 2bbLb967 Cavernas del Repechén, Parque Nacional Carrasco, 17°02'S, 65°26’W, 550 m, Kessler 8293 (LPB, 2666957) UC). Previously known from Colombia, Ecuador, and Peru (Smith, 1983; Smith in Tryon and Stolze, 1992). THELYPTERIS (SUBG. AMAUROPELTA) NITENS (DESV.) R.M. TRYON—La Paz. Prov. Bautista Saavedra, un- Z(o009 0.2, terhalb Chajaya am Beginn des Weges nach Inca, 3540 m, Feuerer 6369a (F, UC). Previously known from Ecuador and Peru (Smith, 1983; Smith in Tryon and Stolze, 1992). oti (SUBG. AMAUROPELTA) NUBICOLA DE LA SOTA—Chuquisaca. Prov. Sud Cinti, camino entre amento Rinconada del Bufete y la cumbre del Cerro Bufete, 20°49'49"S, 64°22'28”W, 1900- ae Om, a0 rano et al. 1285, ai USZ). Santa Cruz. Prov. Valle Grande, 1 km de Loma Larga a DOWD 7 ssasteul 18°46’S, 63°54 W, 1900 m, shite 6175, 6208 (LPB, UC); Prov. Valle Grande, 5 km de Loma Larga a Valle Grande, ree S, 63°54 W, 2100 m, Kessler 6353, 6382 (LPB, UC). Previously known only from Prov. Jujuy, es va ote 1987). THELYPTERIS (SUBG. AMPHINEURON) OPULENTA (KAULF.) FOSBERG—Beni. Prov. Gral. Ballividn, 12 km por el camino maderero, al SW del km 12 Yucumo-Rurrenabaque, 15°04’S, 67°07’W, 450 m, Kes- Dolo bd b\9 sler 10853 (LPB, UC); Prov. ee Ballividn, 20 km por el camino maderero, 12 km de Yucumo— 2 Gale Rurrenabaque, 15°08’S, 67°07’W, 950 m, Kessler 11060 (LPB, UC). La Paz. Prov. Abel Iturralde, Rio San Antonio, 46 km de Ixiamas a Alto Madidi, 13°38’S, 68°26’W, 300 m, Kessler 11139 (LPB, UC). Previously known from the Antilles, Costa Rica, Panama, Colombia to Surinam, Ecuador, pian ee to se. Asia and Pacific islands (Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). 5 (lepine prone (suBG. GONIOPTERIS) POITEANA (BORY) PROCTOR—Beni. Prov. Gral. Ballivian, 12 km por ‘’\*e"" el camino maderero, al SW del km 12 Yucumo-Rurrenabaque, 15°04'S, 67°07'W, 450 m, Kessler 10852 (LPB, UC). Previously known from the Antilles, s. Mexico to Panama, Colombia, Venezuela, Guianas, Ecuador, Peru, and n. Brazil (Smith, 1983; Smith in Tryon and lan 1992; Smith in Davidse et al., 1995). SOV2L 42 pl eecgried oe AMAUROPELTA) TAMANDAREI (ROSENST.) PONCE, —Santa Cruz. Prov. Valle Grande, ma Larga a Valle Grande, 18°39'S, 63°55'W, 2300 m, Kessler 6479 (LPB, UC). Previ- orale seen only from s. Brazil (Christensen, 1920). 2.0607 4\y\ THELYPTERIS (SUBG. cra pan THOMSONI (JENMAN) PRocTOR—Cochabamba. Prov. José Carrasco rrico, 116 Cochabamba-Villa Tunari, 17°08’S, 65°38’W, 2400 m, Kessler 7038, 7039 (LPB, UC). La Paz. “it Nor Yungas, 4.7 km NE de Chuspipata, 16°17'S, 67°47'W, 2 m, Solomon 17337 (MO, UC). Previously known from Jamaica, Hispaniola, s. Mexico to Panama, Colombia, Ecuador, and Peru (Smith, 1983; Smith in Tryon and Stolze, 1992; Smith in Davidse et al., 1995). g = — is} =} f=? gg =] p 2(624S2S THELYPTERIS (SUBG. AMAUROPELTA) VATTUONE! ( ) ABBIATTI—Chuquisaca. Prov. Jaime Mendoza, 68 km de Monteagudo a Padilla, 19°34’S, 64°09’ Ww, ie m, Kessler 5002 2 (AAU, LPB). Previously known from nw. Argentina (Ponce, 1987). 2'olo (600 TRICHOMANES (SECT. NEUROPHYLLUM) HOSTMANNIANUM (KLOTZSCH) KUNZE—Beni. Prov. Vaca Diez, 13 ~ km E of Riberalta on road to Guayaramerin, then 3 km N of side road, 10°58’S, 65°58'W, 230 m, 264 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Solomon 7821 (MO, UC). Previously known from e. Colombia, s. Venezuela, Guianas, Ecuador, e. eru, and n. Brazil (Tryon and Stolze, 1989a; Lellinger, 1994). TRICHOMANES (SECT. DIDYMOGLOSSUM) OVALE (E. FOURN.) WESS. BOER—Santa Cruz. Prov. Chiquitos, errania Santiago along Rio Roboré, 18°18'S, 59°44’W, ca. 300 m, Lewis 85-1217 (F, UC); Prov. Chiquitos, below summit of Cerro Tataraqui, 13 km NE of Roboré, 18°16’S, 59°39’W, ca. 790 m, Lewis 85-1271 (F, UC). Previously known from the Greater Antilles, s. Mexico to Panama, Colom- bia, Venezuela, Surinam, and Brazil (Wessels Boer, 1962; Pacheco in Davidse et al., 1995). ACKNOWLEDGMENTS Kessler and Gonzales thank all their field companions during three years of botanical investi- gation in Bolivia, especially Amparo Acebey, Kerstin Bach, Sebastian K. Herzog, Stephan Hohn- wald, Ivan Jimenez, Thorsten Krémer, Ana Portugal, Elke Rapp, and Michaela Sonnentag. This study would have been impossible without the support of the staff of the Herbario Nacional de sum), Dr. Robbin C. Moran (Asplenium subg. Hymenasplenium, Polybotrya), Dr. Benjamin Ollgaard (Huperzia), and Dr. Ivan Valdespino (Selaginella). Field work by Kessler and coworkers was financed by the Schimper Foundation (1995) and by the Deutsche Forschungsgemeinschaft (German Science Foundation) (1996-7). Gonzales was further supported by the Centre for Research on the Cultural and Biological Diversity of Andean Rainforests (DIVA) under the Danish Environ- mental Programme. LITERATURE CITED ADOLFO Maria, H. 1966. Nomina de la plantas recolectadas en el Valle de Cochabamba. Fasc. 2: 1-86. Editorial Canelas, Cochabamba, Bolivia. ALSTON, A. H. G., A. C. JeRMy, and - RANKIN. 1981. The genus Selaginella in tropical South America. Bull. Brit. Mus. Nat Hist., Bot. 9:233-330. ASPLUND, E. 1926. Contributions to the flora of the Bolivian Andes. I. Pteridophyta, Gymnosper- mae, Helobiae. Ark. Bot. 20A(7):1-38. BARRINGTON, D. S. 1978. A revision of the genus Trichipteris. Contr. Gray Herb. 208:3-93. CHRISTENSEN, C. 1913. A monograph of the genus Dryopteris. Part I. The tropical American pin- natifid-bipinnatifid species. Kongel. Danske Vidensk. Selsk. Skr., Naturvidensk. Math. Afd. ser. 7, 10:55—282 192 - A monograph of the genus Dryopteris. Part II. The tropical American bipinnate- decompound species. Kongel. Danske Vidensk. Selsk. Skr., Naturvidensk. Math. Afd. ser. 8, 6:3—132. Conant, D. S. 1983. A revision of the genus Alsophila (Cyatheaceae) in the Americas. J. Arnold Arbor. 64:333-382. DA SiLva, J. M. C. 1995. Biogeographic analysis of the South American Cerrado avifauna. Steen- strupia 21:49-67. DavibsE, G., M. Sousa S., and S. Knapp, general eds. 1995. Flora Mesoamericana. Vol. 1. Psilo- taceae a Salviniaceae. R. C. Moran and R. Riba, pteridophyte eds. Universidad Nacional Auténoma de México, Mexico City. DE LA Sora, E. R. 1973. A new species of Microgramma from Argentina. Amer. Fern J. 63:61-64. ———. 1977. Pteridophyta. Pp. 1-275 in A. L. Cabrera, ed. Flora de la Provincia de Jujuy, Parte 2. Coleccion Cientifica del INTA, Vol. 13(2), Instituto Nacional de Tecnologia Agropecuaria, Buenos Aires, Brazil. Foster, R. C. 1958. A catalogue of ferns and flowering plants of Bolivia. Contr. Gray Herb. 184:1— GasTONy, G. J. 1973. A revision of the fern genus Nephelea. Contr. Gray Herb. 203:81-148. SMITH ET AL.: BOLIVIAN PTERIDOPHYTE RECORDS 265 HANAGARTH, W. 1993. Acerca de la geoecologia de las sabanas del Beni en el noreste de Bolivia. Instituto de Pastis La Paz, Bolivia. HENNIPMAN, E. 1977. A monograph of the fern genus Bolbitis (Lomariopsidaceae). Leiden Bot. Ser. 2:i—xili, 1- ne IBIscH, P. L. 1996. iets ue Epiphytendiversitét—das Beispiel Bolivien. Martina Galunder- ype Wiehl, G Bolivia is a F nuadibeisin country and a developing county. Pp. ae in W. shen a M. Winiger, eds. Biodiversity. A challenge for d } olicy. Springer-Verlag, Berlin. IpiscH, P. L., C. NEINHUIS, and P. Rojas N. 1996. On the biology, biogeography, and taxonomy of Arachnitis Phil. nom. cons. (Corsiaceae) in respect to a new record from Bolivia. Willde- nowia 26:321-332. KRAMER, K. U. 1957. A revision “8 . genus Lindsaea in the New World with notes on allied genera. Acta Bot. Neerl. 6:97— . 1978. The pteridophytes of tibet Uitgaven Natuurw. Studiekring Suriname Ned. An- KROMER, T., B. K. Hotst, M. KESSLER, H. E. LUTHER, P. L. IBISCH, R. VASQUEZ, W. TILL, a uDA. In press. Ghockiees of oa Bolivian bromeliads with notes on species abiaicn by altitudinal ranges, states and levels of endemism. Selbyana. LELLINGER, 55 B. 1972. A revision of the fern genus Niphidium. Amer. Fern J. 64:101—120. ——. 19 props and confusing bipinnate-dimidiate adiantums of tropical America. Amer. die y 81:99-10 994. eet Fasc. 3:1-66 in A. R. A. Gorts-van Rijn, ed. Flora of the Guianas, series B: ferns and fern allies. Koeltz Scientific Books, Kénigstein, Germany. MarTICORENA, C., and R. RODRIGUEZ. 1995. Flora de Chile. Vol. 1. Pteridophyta—Gymnospermae. Universidad de Concepcion, Concepcion, Chile. MICKEL, J. T. 1962. A peneeePnt study of the fern genus Anemia, subgenus Coptophyllum. Iowa ane J. Sei. 36:4 985. The ales species of Elaphoglossum (Elaphoglossaceae) and their relatives. Brittonia 37:261—278. Moran, R. C. 1987. Monograph of the neotropical fern genus Polybotrya (Dryopteridaceae). Bull. Illinois Nat. Hist. Surv. 34:1-138. ———. 1991. Monograph of the errsurcs fern genus Stigmatopteris (Dryopteridaceae). Ann. Missouri Bot. Gard. 78:857-9 Moran, R. C., and B. OLLGAARD. tt Six new species of ferns (Polypodiopsida) from Ecuador. Nordic J. Bot. 15:177-185. Morton, C. V. 1947. The American species of Hymenophyllum, section Sphaerocionium. Contr. S. Natl. Herb. 29:139-201. Sean N., and R. C. Moran. 1993. Monograph of the on . species of Asplenium sect. Hyme verges (Aspleniaceae). Ann. Missouri Bot. Gard. 80:1—-38. MURILLO, M. T. 1 Blechnum subgénero Blechnum en Sur edie con especial referencia a las especies ae Colombia. Nova Hedwigia 16:329-366. OLLGAARD, B. 1992. Neotropical Lycopodiaceae—an overview. Ann. Missouri Bot. Gard. 79:687— Tite PFEIFFER, N. E. 1922. Monograph of the Hanetarean. uaa Missouri ee Gere 9:79-232 Ponce, M. M. 1987 [1988]. Revisién de la Thelypt ) Argentinas alone 7-390. ROJAS-ALVARADO, A. eh 1996 [1998]. Aportes a la flora Pteridophyta Costarricense. I. Informes. B re SEHNEM, A. 1968 a Fasc. ASPL:3—96 in P. R. Reitz, ed. Flora Ilustrada Catarinense. Ita aia share Catarina, Brazi 5 ee Fasc. ASPI: 3-356 in P. R. Reitz, ed. Flora Ilustrada Catarinense. Itajai, San a Catan, Bra SMITH, a R. 19 a ialay Spite pediasissThotypwccidens. Fasc. 18:1-148 in G. Harling and B. Sparre, ids ide of Ecuador. Swedish Research Council, Stockholm 266 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) - 1985. Pteridophytes of Venezuela, an annotated list. Published by the author, Berkeley, ee 2. A review of the fern genus Micropolypodium {Grammitidacene), Novon a rosie sseiie 4 eg and R. C. Moran. 1987. New combinations y mer rm J. 77:124-130. : get Melpomene, a new genus of Grammitidaceae (Pteridophyta). Novon 2:426—432 SMITH, A. R., R. C. Moran, and L. E. BisHop, 1991. Lellingeria, a new genus of Grammitidaceae. Amer. scar 7 76-88. STEYERMARK, J. A., P. E. BERRY, and B. K. Hoist, general eds. 1995. Flora of the Serie Guayana. Vol. 2. Pteridophytes, Spermatophytes: Acanthaceae-Araceae. P. E Berry, B. K. ag and K. Yatskievych, volume eds. Timber Press, Portland, OR [Pteridophyta, pp. 1-334. A. R. Smith and collaborators]. STOLZE, R. - 1974, A taxonomic revision of the genus Cnemidaria (Cyatheaceae). Fieldiana, Bot. 37:1 STOLZE, R. - L. PACHECO, and B. OLLGAARD. 1994. 14 (5B). Polypodiaceae-Dryopteridoideae- -Phy- sematieae. Fasc. 49:1—-108 in G. Harling and L. Andersson, eds. Flora of Ecuador. Swedish Research — Stockholm. TINDALE, M. D, 1965. A wet ual of the genus Lastreopsis Ching. Contr. New South Wales Natl. Herb. 3: pis pl. 1- TRYON, He - 1962. A SS of the fern genus Jamesonia. Contr. Gray Herb. oe bite 0. A monograph of the fern genus casas — Gray Herb. 200:54— TRYON, k 1942. A revision of the genus Doryopte tr. Gray Herb. 143:1 964. The ferns of Peru, iil Asahigon a to Oleandreae). Cis: Gray Herb. - 1-253 TRYON, R. M., and R. G. STOLZE. 1989a. Pteridophyta of Peru. Part I. 1. Ophioglossaceae—12. Cy- pe ei Fieldiana, Bot., n.s. 20:1-145. ———.. 1989b. Pteridophyta of Peru. Part II. 13. Pteridaceae—15. Dennstaedtiaceae. Fieldiana, Bot., ns. 22:1—-128 - 1991. Pteridophyta of Peru. Part IV. 17. Dryopteridaceae. Fieldiana, Bot., n.s. 27:1-176. . 1992. Pte oe of Peru. Part III. 16: Thelypteridaceae. (contributed by A. R. Smith). Fieldinns: Bot., . 1993. Na a ue Por. Part V. 18: Aspleniaceae—21. Polypodiaceae. Fieldiana, Bot., n.s. 32:1-190. - 1994. Pteridophyta of Peru. Part VI. 22: Marsileaceae—28. Isoetaceae. Fieldiana, en ns, 4:1-123. VALDESPINO Q., I. A. 1995. A monographic revision of Selaginella P. Beauv. subgenus candid aker in Central and South America. Ph.D. dissertation, City University of New WESSELS Boer. J. G. 1962. The New World species of Trichomanes sect. i ne aon and Mi- crogonium. Acta Bot. Neerl. 11:277-330. American Fern Journal 89(4):267—269 (1999) SHORTER NOTES Blechnum penna-marina in Peru.—Nineteen species of Blechnum were rec- ognized for Peru by Tryon and Stolze (Fieldiana Bot., n.s. 32:56-68. 1993). One of the species, B. penna-marina (Poir.) Kuhn was included based on a personal communication reporting a specimen from Cusco (Leon et al. 2757, CUZ, USM.) Reexamination of this specimen, however, revealed that it had been misidentified. In the central Andes, only B. andinum (Baker) C. Chr. and B. penna-marina have dimorphic leaves and stoloniferous rhizomes. These two species can be separated by the following key: 1. Lamina with 1-4 reduced proximal pinnae, these distant from distal pinnae; veins simple, rarely furcate on the proximal acroscopic side; indusia erose; laminae herbaceous, less SIAN, 10; CiteReader SW se eee ae ee all) Wali aie B. andinum 1. Lamina gradually reduced, all pinnae or segments contiguous; veins furcate; indusia en- tire, slightly crenate; laminae usually coriaceous, more than 10 cm long .. . B. penna-marina The Peruvian specimen cited by Tryon and Stolze was collected at 3390 m in a very humid montane forest on the eastern slopes of the Peruvian Andes (approx. 13°14’S, 71°32'W), growing on a rock among mosses and forming colonies. The specimen has a short, erect rhizome, with stoloniferous axes, fasciculate, dimorphic leaves less than 10 cm long, indusia dentate, and well- spaced, reduced proximal pinnae. Based on these characters this specimen is B. andinum, a species otherwise known in Peru from two collections made by Biies in Cusco during the 1930's. Because this was the only specimen cited for B. penna-marina by Tryon and Stolze (1993), the question remains whether this species occurs in Peru. Under B. penna-marina, Tryon and Stolze commented on the name B. alpinum var. elongatum Mett. They did not see the type and suggested that it might be another species. Recently, however, Chambers and Farrant (Fern Gaz. 15:92. 1996) considered this name a synonym of B. penna-marina ssp. penna-marina, although they did not examine the type and did not list Peru within the range of its distribution. Mettenius (Fil. Lech]. 2:15. 1859) named B. alpinum var. elongatum based on a Lechler collection from Agapata (Ayapata), located in the Province of Carabaya, Department of Puno, Peru, approx. at 13°52’S, 70°19’W, 3600 m. From his diagnosis and discussion it is clear that Lechler’s specimen is di- morphic, with numerous and contiguous pinnae, leaves 30-40 cm long, the fertile one longer than the sterile, and the rhizome stoloniferous with ovate scales. These features describe B. penna-marina. Two Blechnum specimens collected by Lechler in ““Agapata” in June 1854 are accessioned at B; both are B. penna-marina ssp. boliviana (Rosenst.) T.C. Chambers & P.A. Farrant. In Peru, other dimorphic species of Blechnum with contiguous reduced proximal pinnae are B. binervatum (Poir.) C.V. Morton & Lellinger ssp. fragile 268 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) (Liebm.) R.M. Tryon & Stolze and B. lehmannii Hieron. Neither of these has a stoloniferous rhizome. Both species also grow only in forested areas below 3500 m. In conclusion, both B. andinum and B. penna-marina ssp. boliviana have been collected in Peru. Records for the latter are based on a single collection made during the nineteenth century, and for the former from three collections made during the last 60 years. Thanks to Harald Komposch for the translation of German comments of Met- tenius. I also thank David B. Lellinger, Robbin C. Moran, Alan R. Smith, and Kenneth R. Young for reviews and comments of the manuscript. Thanks also to Alan R. Smith, Tyana Wachter, and Brigitte Zimmer for providing me with photocopies of the studied taxa.—BLANCA LEON, Museo de Historia Natural, Av. Arenales 1256, Apartado 14-0434, Lima-14, Peru, and Botany Department, Field Museum, Chicago, IL 60605-2496. Salvinia adnata Desv. Versus S. molesta D.S. Mitch.—The name Salvinia mo- lesta D.S. Mitch. (Brit. Fern Gaz. 10:251—252. 1972) has been widely used for an aquatic fern native to the New World tropics but introduced and weedy in the Old World tropics. Recently, de la Sota (Darwiniana 33:309-313. 1995) proposed replacing this name with an earlier one, S. adnata Desv. (Prodromus, 177. 1827). The subject of this note is our differing interpretation from that of de la Sota concerning the provenance of the type of S. adnata and whether it can be proven conspecific with S. molesta. The name S. adnata is based on a specimen that, according to its label, was collected on Réunion (‘‘Habitat in insula Borboniae’’). This locality was ac- cepted by de la Sota (1995); however, it conflicts with what is known about the plant’s weediness, geographic distribution, and insect enemies. Evidence from several sources indicates that S. molesta is native to southern Brazil (Forno, Aquat. Bot. 17:71-83. 1983; Mitchell, Brit. Fern Gaz. 10:251— 252. 1972). It is never weedy in South America and has several insect herbi- vores that feed exclusively upon it. In contrast, in the Old World it is an ag- gressive weed, in some cases carpeting thousands of hectares of water, and has no native insect enemies that attack it (Thomas and Room, Nature 320: 581-584. 1986). The species was first recorded from the Old World (India) in 1939. Presumably, had it been native, it would have been collected there before that date. Moreover, if it had been native to the Old World, why would it have become a weed only in the 1950s and not before? All of these observations argue that the plant is native to the New World, not the Old World. Why, then, is the type of S. adnata, which de la Sota claims is conspecific with S. molesta, reportedly from Réunion? The most likely explanation is a label error. Christensen first pointed this out in his work on the pteridophytes of Madagascar (Dansk. Bot. Ark. 7:203. 1932) and annotated the type of S. adnata accordingly with “patria certe er- ronea.” In the original description, Desvaux wrote “Hab. in aquosis insularum SHORTER NOTES 269 Africae orientali’” which is less specific than Réunion, perhaps further indi- cating his uncertainty about the provenance of the type. Christensen also pointed out that Desvaux queried with a “?” the localities of several other of his type specimens reportedly from Africa, Madagascar, and the Mascarenes, and that in several cases these localities were untrustworthy. It is highly un- likely that S. adnata would be known today natively only from southern Brazil and have a type from Réunion, where it does not occur (Baker, 1877, Flora of Mauritius and the Seychelles). A more likely explanation is that the type of S. adnata came from southern Brazil and a label mix-up occurred or that Desvaux himself guessed wrongly about the provenance of the specimen. Thus, the type should be cited as coming from Brazil, not Réunion. Provenance aside (and more importantly), it cannot be proven that the type of S. adnata is the same as S. molesta. De la Sota did not discuss the char- acteristics he used to equate the type of S. adnata with S. molesta. The type of S. adnata (pictured by de la Sota, Darwiniana 33:309-313. 1995. Fig. 3) is vegetative, and therefore cannot be distinguished from another closely related species that also grows in southern Brazil: S. biloba Raddi (Forno, Aquat. Bot. 17:71—83. 1983). As far as we know, it is impossible to distinguish specimens of S. biloba from S. adnata if the plants lack fertile axes bearing sporocarps. Therefore, we cannot be certain whether S. adnata is the same as S. molesta or S. biloba. For these reasons, S. adnata should be treated as a name of uncertain ap- plication. The name S. molesta, which is well-established in the literature, should continue to be used for this economically important, highly weedy, and widely known fern.—RosBIN C. Moran, New York Botanical Garden, Bronx, NY 10458-5126; ALAN R. SMITH, University Herbarium, University of California at Berkeley, 1001 Valley Life Science Bldg., #2465, Berkeley, CA 94720-2465. American Fern Journal 89(4):270—271 (1999) REVIEWS Flora of Australia, Volume 48, Ferns, Gymnosperms, and Allied Groups, by A. E. Orchard, Executive Editor, P. M. McCarthy, Volume Editor, and 21 contributors. 1998. CSIRO Publishing, P.O. Box 1139 (150 Oxford Street), Col- lingwood, Victoria 3066, Australia. xxi, 766 pp. Hardcover (ISBN 0 643 05971 7) $US 94.95; softcover (ISBN 0 643 05972 5) $59.95. May be ordered directly from http://www.publish.csiro/au or by e-mail: sales@publish.csiro.au. Although Australia has had many fine local and regional pteridophyte floras, this is the first comprehensive treatment for the entire continent. It treats 456 species of pteridophytes, these classified into 112 genera and 35 families. For each species there is given nomenclatural and type information for the ac- cepted name and its common synonyms, a short description, geographic dis- tribution (with maps provided at the back of the book), specimens examined, and comments. There are 157 figures of pteridophytes, many of them color photographs, and the rest line drawings. Particularly helpful to users will be the illustrations by P. J. Edwards showing the indument characteristics of tree ferns. All of the illustrations are of high quality. The introductory matter includes a helpful review by Mary D. Tindale of fern morphology, terminology, cytology, biogeography, ecology, and history of Australian fern floristics. Andrew Drinnan provides a well-researched over- view of the history of fern phylogeny and classification, and Robert S. Hill and Gregory J. Jordan summarize the fossil record for Australian pteridophytes. A key to families is provided by P. M. McCarthy, and in the text the families are arranged by a phylogenetic, not alphabetical, sequence. The keys are of the indented type, not bracketed. A glossay of specialized pteridophyte terms, compiled mostly by Mary D. Tindale, is given toward the back of the book. It is immensely satisfying to see so much information brought together for the entire Australian pteridophyte flora. This book is a major contribution to pteridology, and anyone seriously interested in pteridophytes will want a copy. Congratulations to our pteridological mates down under for a job well done!—RossIN C. Moran, New York Botanical Garden, Bronx, NY 10458-5126. Illustrierter Leitfaden zum Bestimmen der Farne und farnverwandten Pflanzen der Schweiz und angrenzender Gebiete, by Eugen Kopp and Ruth Schneebeli-Graf. 1998. Schweizerische Vereinigung der Farnfreunde. 226 pp.. Softcover (ISBN 3-9521349-0-2) CHF 45.00. May be ordered from Publications, Natural History Museum of Luzern, Kasernenplatz 5, CH-6003 Luzern, Swit- zerland (http://www.luzern@naturmuseum.ch). [In German]. They say that imitation is the sincerest form of flattery. In this case, the Swiss Association of “Fern-friends” really liked ‘The Illustrated Guide to Ferns and Allied Plants of the British Isles” by Clive Jermy and Josephine Camus (1991, REVIEWS 74 Natural History Museum Publications, London). They liked it so well, in fact, that the group undertook to translate this book from English into German and to modify the contents to cover the pteridophytes growing in and around Swit- zerland. The result is a book that is quite similar in appearance to the Jermy and Camus guide, but contains a number of changes and additions to tailor it to a new audience. Kopp and Schneebeli-Graf have not only translated most of the original text, but have written new treatments for 20 of the 88 native taxa treated. The group also contracted with Peter Edwards, who provided the elegant illustrations in the original work, to provide drawings of the additional species, so it is difficult to tell what is old and what is new without noting the little indicators of the translated parts. This compact book contains all the elements of a flora: dichotomous keys, descriptions, statements of habitat and distribution, and supplementary dis- cussions of taxonomy and conservation status. The illustrations are a mixture of silhouettes of plants and fronds and line drawings of details. The introduc- tory material provides a glossary and a nice summary of morphology, life cycle, hybridization, and how to use the book. A separate section at the end discusses five non-native species encountered in the region. In spite of this completeness of coverage, the book is layed out in a “user-friendly” format. Complaints about this excellent guide are few. Buyers should note the ex- istence of an erratum sheet that provides several missing couplets for the key to Aspleniaceae. A few of the drawings from the original work suffer slightly in reprinting. For those who have difficulty handling German text, a copy of the Jermy and Camus book might prove useful as a guide for translating the less obvious technical terms (although it apparently has recently gone out of print). All in all, this book is a handy work for any “fern friend” who might be visiting northern Europe and wish a pocket guide to his or her favorite plants.—GEORGE YATSKIEVYCH, Missouri Botanical Garden, P.O. Box 299 St. Louis, MO 63166-0299. American Fern Journal 89(4):272 (1999) Referees for 1999 Maintaining the quality of papers published in the American Fern Journal i is perhaps | one of the Fern Society is indebted to these individuals for their service, as is the editor—GEORGE YATSKIE- EMMET JUDZIEWICZ MASAHIRO KATO RICHARD C. KEATING ROBIN C. KENNEDY JEFFERSON PRADO . MICK GERALD J. GASTONY PETER M. eects iget WARREN HA ROBBIN C. M LIsA A. eerie DONALD YOUNG JOAN N. HUDSON JAMES H. PECK Announcement: New Editor The present issue is the last one to be published under the present editor, who has completed his five-year appointment to the post. Working on the American Fern Journal has been a rewarding and satisfying experience, and the readership hopefully will agree that the journal has not faltered too badly during the past five volumes. We are pleased to announce that we have located an excellent candidate to lead our publication into he new millenium. Effective immediately, all new neg oad and correspondence concerning the American Fern Journal should be directed to the new editor Dr. R. James Hickey e-mail: hickeyrj@muohio.edu INDEX TO VOLUME 89 Index to Volume 89 (New taxa and combinations are in boldface) 3-C-(6"’’-O-Acetyl-B-cellobiosyl) Apigenin, a Asplenium (continued) New Flavonoid from Pteris vittata, 2 delitescens, 248 Acanthaceae, 245 Acer m, 104 Adiantopsis, 149-158 cajennense, 247 ruizianum, 247 posse © 247 ten peach a 248 as Alsophila, 211, 248 2 ; ape Anarlirioe 205, 211 Announcement: new editor, 272 Bein Onoclea, 221-231 Type A, 2 Type PT, re 1-231 presets Pteris vittata, 217-220 niodes ochro were 248 pba uniflora, 2 Arkansas, Salvinia hie 215 pers 245 um spinulosu ee yee a 232-243 Asplenium, 104, 168, 232, _ 241; classification, 232-2 antiquum, 232-235, = sh 242 3. cardiophyllum, 234 cataractarum, cheilosorum, 234 ensiforme, 233, 234 extensum, 248 eifianan, 234, 235, 238, 240 heterochroum, 24 inaequilaterale, 248 incisum, 234, 241 Xkenzoi, 232, 233, 235. 236, 241, 242 2 pu 234, 238, 240 var. boreale, 234, 241 var. normale, 234, 241 var. shimurae, 234 obliquissimum, 234 oligophlebiom, 234, 238 prolongatum, 232-235, 238, 240-242 pseudo-wilfordii, 234, 238, 241 riparium, 234 ventas 234, 241 i, 234 sarelii, oak 238, 241, 241 posi? canary : 104 Xhioboma, bh e, 234, Po aay seg nh ne 241 tripteropus, 234, 238, 241 viviparum, 220 wilfordii, 234, 235, 238 wrightii, 232, 234, 235, 241 yoshinagae, 234, 238, 241 Astrolepis, 171, 177 PP RAPE Peres fy Acolla mavicone, 215 Barba de jolote, 104 Belize, medicinal plants, 104 Bird’s nest fern, 23 — 245, 249, 267, 268 binervatum, 267 274 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Blech bi: tum (continued) ssp. fragile, 267 Con oe 106-123 blechnoides, 245, 249 emidaria uleana, 250 laevigatum, 249 var. abitaguensis, 250 lehmannii, 268 var. uleana, 250 obtusifolium, 249 Cola de caballo, 104 penna-marina, 267, 268; in Peru, 267, 268 Corsiaceae, 245 ssp. boliviana, 267, 268 Costa Rica, commercial fern cultivation, 101-105 ssp. penna-marina, 267 Bias oth ma ane spore germination, Blechnum penna-marina in Peru, 267 9-170 Bolbitis nicotianifolia, 249 ae 250 Bolivia, 244-266 nigrovenia, 250 New pteridophyte records, 244-266 pedicellata, 250 Ss diversity, 244, 245 refulgens, 250 mmeria sloanei, 250 Boniniella, a 240 Culantrillo, 104 enoi, Cyathea Botrychium, 146 cooperi, 205 i pore en 149-158 delgadii, 152 BRITT ALD M. 7 ‘a, 25 Iso soins ka and the Aleutians, 133 Cyatheaceae, 152, 205 ‘Dadial F, Brunton), 187 Cycads, in tree fern pots, 104 Bromeliads, 245 Gpstopberis: BIE SE UNTON, DANIEL F Rush Quillwort (Isoetes junciformis, sp. nov.), Davalliaceae, 172 ‘w Pteridophyte from Southern Danaea nodosa, 251 Georgia, 187 Dennstaedtia sprucei, 251 (see Donald M. Britton), 133 Deparia ve 236 Bryophytes, 220 DIFUR, 1 Boke 106-123 Cacti, 245 complanatum, 107, 108 Calaguala, 103 Breeding system, 119, 120 CAMLOH, MARJANA, Spore Age and Sterilization Genet composition, 106-123 Affects Germination and Earl Spacial autocorrelation of genotypes, Gametophyte Development of Platycerium 115-118 bifurcatum, 124 igitatum, 107 Campyloneurum solutum, 249 Diplazium, 181-184, 251 Camptosorus, 232, a altissimum, 183 , 234, 240, 2 tatum, 1 peers SHERWIN rile Edward L. Schneider), drepanolobium, 183 entecnum, ae 250 errans, 181-184 ta, 250 lentum, 236 triphylla, 250 expansum, 251 Cellobiose, 220 lonchophyllum, 183, 251 Ceratopteris, 221 moccenianum, 251 pteridoides, 152 — ~ richardii, 129, 221, 229 obscuru thalictroides, 152 Hetgnion, 183, 184 Cheilanthes, 250 riedelian micropteris, 250 uidion: ths tweediana, 250 — se Cheiropleuria, 205 vera-pax, 183, — DNA, 232 werckleanum, ed 184 Ciervo, 104 Dipteris, 205 INDEX TO VOLUME 89 Doradilla, 104 Dryopteris (continued) Doryopteris, 250, 251 darjeelingensis, 9 collina, 251 decipiens, 76 Dryopteris, 1-98 dickinsii, 10, 12, 95, 236 abbreviata, 14, 41, 44 dilatata, 66, 69, 72, 96 aemula, 58, 61 ‘Crispa Whiteside,’ 66 affinis, 14, 17, 18, 23, 38, 39, 44 ‘Cristata,’ 66 ‘Congesta Cristie , 18 ‘Grandiceps,’ 66 ‘Congesta Griepa?. 18 ‘Jimmy Dyce,’ 69 ‘Crispa onpentay 18 ‘Lepidota Crista 69 ‘Crispa,’ 18 ‘Recurv ‘Crisp Gracilis,’ 18 erythrosora, 73, 76, "77, 79, 81, 84, 86, 88, *Cristata,’ 18 94,95 ‘Cristata aed 18 f. prolifica, 77, 79 ‘Grandiceps,’ 18 - viridosora, 7 ‘Pinderi,’ 18 ar. cystolepidota, 73 ‘Polydactyla,’ 18 var. dilatata, pariah var. Age - ‘Stabler expansa, 46, 63, 6 ‘Stableri oer 18 filix-mas, 16-18, z ‘a 39, 42, 94 ssp. affinis, 16, 17, 18, 46 ‘Barnesii,’ 38 ssp. borreri, 16, 17, 18, 25 ‘Crispa Cristata,’ 39 ssp. cambrensis, 1 ‘Cristata,’ 39 amurensis, 61, 62 ‘Cristata Martindale,’ 39 arguta, ecomposita,’ 3 yates . ‘Grandice 9 atrata, 95 : foo Lamia 216, 217 ‘Linearis Congesta,’ 39 austriaca, 66, ‘Linearis Cristata,’ 39 azorica, 66, 72 ‘Linearis Polydactyla,’ 39 bissetiana, ea 86, 87, 88, 91, 95 ‘Polydactyla,’ 39 blandfordii, 9 ‘Ramo-cristata,’ 39 Map Ost oe ‘Robust,” 17 b ‘Undulata Robusta,’ 39 campo 63, 66, ye 71 formosana, 88 carthusiana, 63, =i 6,71 fragrans, 39, 41 ica, 35, var. remotiuscula, 41 celsa, 27, 2 fructuosa, 94 ns int 216 fuscipes, 79 celsa X cristata, 216 gamblei, 9 championii, . goeringiana, 555 clayton niana goldiana, Zi, 41 clintowlana; cai 30, 41, 94 guanchica, 9 a fs gymnosora, 81 commixta, 10 hangchowensis, 12 comps, 17: 38,40 hikoensis, 88 coreano, 9 hirtipes, 7, 9, 10 ssp. SS coreano-montana, 44, 45, 94 hondoensis, 77, 81, 84 crassirhizoma, 18, f. c var. setosa, 44 —- 2,3 cristata, 27, 30, 41, 94, 216 esos — 3; gts 84 7, 9, 10, 95 intermedia, 63, 64, 4, 66, 69, 70, 94 cy 8 ti 73, 76 juxtaposita, 48, 51 276 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Dryopteris — Dryopteris (continued) koidzumiana, 9 submontana, 54, 55 page “ga a 95 tokyoensis, 33 lacera, undulata, 17 eck 2 uniformis, 58 ludoviciana, >, 30, 216 “‘Crispata,’ 58 marginalis, 51 ‘Cristata,’ 58 megalodus, 95 monstrosity crispata, 58 mindshelkensis, 54, 55, 96 var. crispata, 58 munchii, 95 varia, 92 namegatae, 10, 95 var, E nigra, 20 var, hikoensis, 95 Ae var. setosa, 86 odontoloma, 95 villarii, 55, 96 oreades, 38, 41, 44 ssp. , 54 ispata,’ 44 wallichiana, 20, 25, 96 “Cristata,’ 44 yigongensis, 9 ‘Incisa Crispa,’ 44 pacifica, 88, 95 Effects of Temperature on Spore Germina paleacea, 25, 96 Some Fern Species from Semideciduous pallida, 55 Mesophytic Forest, 149 ssp. pallida, 55 Elaphoglossum, 104, 252-255 parallelogramma, 25 amaz , polylepis, 20, 23 7 , 25 ps erythrosora, 73 amphioxys, 252 ORB 16, 20, 23 amplum, 252 angustius, 252 purpurella, eo 84, 86 ciliatum, 252 pycnopteroides, 10, 12 dichroum, 252 remota, 46 ensiforme, sacrosancta, 88, 95 glossophyllum, 253 saxifraga, 91, 92 herpestes, 253 scottii, 12 heteromorphum, 253 . Aemulae, 58 hickenii, 253 Cinnamomeae, 95 killipii, 253 Sect. Dryopteris, 33 , 253 Sect. Erythrovariae, 72, 95 latifolium, 104 Sect. Fibrillosae, 12, 96 lechlerianum, 253 Sect. Hirtipedes, 7, “p lellingeri, 255 Sect. Lophodium, 58, 61, 94 litanum, 253 Sect. Pallidae, 46, . * 95, 96 macilentum, 254 Sect. P. megalurum, 254 Sect. Remo : Sect. Variae, 76, 86, 95, 96 melanopus, 254 sichotensis, , 254 sieboldii, a 7 nigrescens, 254 sordidipes, 96 oblanceolatum, 255 spinulosa, 63 obtusum, 254 var. americana, 63 oculatum, 254 intermedia, 69 peruvianum, 255 steno 3 pilosius, 255 tewartii, 55, 66, 94 plumosum, 255 Subg. Dryopteris, 7 pumilio, 255 Subg. Erythrovariae, 4,72 Be AGy Subg. Pycnopteris, 5 setigerum, 255 sublacera, 56, 58 smithii, 255 INDEX TO VOLUME 89 Elaphoglossum (continued) Equisetum, bogotense, 104 Eriosorus sowogrenatunile, 255 Ferns of the Tropics (Review), 179 Flora of Australia, Volume 48, Ferns, ymnosperms, and Allied Groups 8 RENA (see Emilia Pangua), Genet Composition of Diphasiastrum ‘um in Western Hungary: a Case Georgia, Isoetes, 187-197 Gleicheniaceae, 146, 147, 213 colhtdedcied fern cultivation, 101-105 Hemionitis palmata, 221 Hemitelia capensis, 211 Hoja de ciervo, 104 HOSHIZAKI, BARBARA JOE, The Cultivated Species of the Fern Genus Dryopteris in the United States, 1 Honduras, commercial fern cultivation, 101-105 ungary Clonal plants, 106-123 0s enium, 232, 233, 240, 241 cardiophyllum, 234, 240 catar 234 iquissimum, 233, 234 riparium, 234, 240 Iberian Peninsula, 159 Illustrierter Leitfaden zum Bestimmen der Farne und farnverwandten Pflanzen der Schweiz ), 270 IMPERATO, FILIPPO, 3-C-(6" ’’ -O-Acetyl-B-cello- biosyl) in, a New Pteris vi 217 T ] Ip 4 £ Diweee. Med ateA An en Response in Onoclea 1 is, Isoetes, i in ego 133-141, in Georgia, 187-197 la, 198-203 ream it yte development, 199-203 sie pth oom 198-203 pore germination, 1 akakeun 187, 188, ste 195 asiatica, 133, 139 Spore ology, 189-191 echinospora, 133-135, 139, 140 number, 134, 135 Spores, 135, 137, 138 echinospora X occidentalis, 138 a 187, 195 engel mannii 195 evivedhiis 195 flaccida, 187, 188, 194-196 flaccida x melanopoda, 195 gardneriana, 256 georgiana, 187, 195 ea srt melanopoda, 195 hickeyi, 19 hyemalis, ns 188, 194 iformis, 193-196 Spore Morphology, 189-191 louisianensis, 194, 195 278 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Isoetes onicaseis macounii, 133 gusribieen: 133-135, 139, 140 cried 187, 194-196 muricata, ssp. aries, 133 var. braunii, 133 occidentalis, 133, 139, 140 Chromosome oan 134, 135 Spores, 135, sss Rg pie 140 Chromosome number, 134, 135 Spores, 137, 138 Isozymes, in Diphasiastrum, 106-123 IWATSUKI, KUNIO (see Noriaki Murakami), 232 Jack pine, Jamesonia ik 257 KARRFALT, ERIC, Some Observations on the Reproductive Anatomy of Isoetes andicola, 198 KESSLER, MICHAEL (see Alan R. Smith), 244 Lagua de ciervo, 104 Lastrea, 35, 41, 55 filix-mas, 35 i ES 41 Lasteops lip 257 Lelling uspensa, 257 tungurahuae, 257 Lemma spp., 2 LEON, BLANCA, Blechnum penna-marina in Peru, 267 Leptosporangia, 204, 212, 213 SP eee ferns, 232 Lindsaea isms par Hl 216 Lophosoriaceae, 152 LOREA-HERNANDEZ, FRANCISCO G., Two New Fern Species from Southern Mexico, 181 Luteolin L rn at 107, 146, 147 Lygodium, 104 venustum, 104 Major, AGNES, Genet Composition of we trum Cee i in Western ngary: a Cas 106 Ma: Pers Dryopteris ae 215, 216 Masiow 257 aden ji cog bccn 258 Me OD 245 Melpomene, assurgens, 258 vernicosa, 258 Mexico, Guerrero, two new ferns, 181-186 Microgramma, 149-158, 258 lindbergii, oe cama 149-158 megalophylla, 2 vaciniifolia, 25 ncicola, MORAN, RosBIN C., rey adnata Desv. Versus molesta D.S. Mi MUBARIEIL NorIAKI, cay of Aspleniaceae Inferred from bck, Nucleotide Sequences, Lae Neottopteris, 232, 238-241 , 235, 238 Nephrodium, 81 gymnosorum, 81 Nephrolepidaceae, 172 cordifolia, vessels, 171- ig exaltata, vessels, 171 New Records of nt He from Bolivia, Niphidium rufosquamatum NOGAMI, SATORU, (see conde Murakami), 232 Opor, PETER (see Agnes Major), 106 INDEX TO VOLUME 89 Onoclea sensibilis, 129, 222, 223 Orchids, in ee ye pots, 104 Osmundaceae, 171, 213 PAJARON, SANTIAGO (see Emilia Pangua), 159 Palms, 245 PANGUA, EMILIA, Studies on Cryptogramma a Spore Germination, 159 Pata de sante, 104 PECK, JAM aed Boris of the Tropics, 179 Review: The Ferns and Allied Plants of New England, 178 alvinia — in Arkansas, 215 Peru, Blechnum Phishodiaen, 102, 104 171, 175, B77, 205: aureum, 102, 2 decumanum, Ha ps reum, 102 Phibalura flavirostris, 245 Phyllitis, 232, 240, 241 ium, 234 Phylloglossum, 146 priest of Aspleniaceae Inferred from rbcL ucleotide Sequences, 232 Mytohemisy, Pteris, 217-220 Pine, jack, 1 PINTAUD, ects (see Dean P. hittier), 142 Pinus banksiana, 120 Spore germination and gametophyte development, eo a age and viability, 12 ore sterilization sg 124-132 Sobiboity a botryoi 3 Air bar aaa 204, 205, 221 Polypodium, 102, 129 eta spore germination, 149-158 latipes, spore germination, 149-158 leucatomas, 10 pleopeltidifolium, spore germination, 149-158 polypodioides, spore germination, 149-158 rare, 129, Polystichoid ferns, 221 Polystichum, 91, 92, 171 distans, 186 hartwegii, 186 schizophyllum, 184-186 speciosissimum, 18 2/2 soso - 259 varium, Portugal, 15 Palseadead 146, 147 Psilotum, 146, 147, 202 Pteridaceae, 217 Pteridophytes, oS horticulture, 101-105 Pteridium, 171, 1 regscrmausy me 221-223 Pteris, 217, packs 259 denticulata, 2 eh a 149-158 grandifolia, 21 pearcei, 260 tripartita, 260 vittata, 129, 130 Phytochemistry, 217-220 Pyrrosia, 20 Quercetin, 217 RANAL, MARLI A., Effects of Temperature on Spore Germination in Some Fern Species oes Semideciduous Mesophytic Forest, — casio, 232-243 ELL E., Two Additional Stations ioe — Boolian Woodfern Hybrid, Dryopteris Xaustralis in Maryland, 216 Referees for 1999, 272 ospora sobitaria, 195 Rosa de dire oO. 104 adiantiformis, 101 Rush Phe ae (Isoetes junciformis, sp. nov.), a New Pteridophyte from Southern Georgia, 187 Salvia uliginosa, 220 Salvinia, 215 Salvinia adnata Desv. Versus S. molesta D.S. itch., 268 Salvinia minima in Arkansas, 215 somrmtnag 474, 213 SCHNEIDER, EDWARD L., SEM Studies on Vessels ms ral 13. Nephrolepis, 171 cotland, spore gta 159-170 Selaginell, 104, 2 calo cavifolia, 260 lepidophylla, 104 moritziana, 260 var. salaibchion; 260 280 Selaginella (continued) pallescens, 104 revoluta 260 SEM Studies on Vessels in Ferns. 13. Nephro- 244 (see Francisco G. Lorea- Hernandez), 181 (see Robbin Cc. Moran), 2 268 Central Ass America, 101 Some Observations on the uctive Anatomy of Isoetes andicola, 198 —o 204-214 Spirodela spp pa Viorelopaea, Sphaeropteris, 204-214 and Early Gametophyte agg of Platycerium bifercamam, 1 Spore germination in Cryptogramma crispa, 159-170 Ferns of semideciduous forests, 149-158 Tsoetes, 198-203 Platycerium, 124-132 Stromatopteris, 142-148 Spore Germination and Antheridiogen Response in Onoclea sensibilis, 221 ticherus peruvianus, 260 pester lechleri, 261 Stromatopteris, = 202 moniliform 142, Spore germina tion and gametophyte ceca, 142-148 gemmifera, 198 TALBOT, STEPHEN S. (see Donald M. Britton), 3 pilosa TELESCA, ANTONELLA (see Filippo Imperato), 217 AMERICAN FERN JOURNAL: VOLUME 89 NUMBER 4 (1999) Terpsichore mollissima, 261 The Cultivated Species of the Fern Genus Dryopteris in the United States, 1 The Ferns and Allied Plants of New England iew), 178 Thelypteris, 63, 169, 261-263 amphioxipteris, 261 261 biolleyi, 261 decussata spinulosa _ cabeains 261-263 263 ineuron, pag Goniopteris, oe Subg. Meniscium, 262, 2 Subg. sevetiet 261, rn tamandarei, Us 263 THOMAS, BARRY A., Some Commercial Uses of idophytes in Central America, 101 Tmesipteris, 146, 147 Tree ferns, collection of for pots, 103, 104 adensis, 1 Didymoglossum, 264 Sect. Neurophyllum, 263 sum, 220 Two Additional Stations for the Southern Woodfern Hybrid, Dryopteris xaustralis in Two New F. Speci South Mexico, Vessels, in Nephrolepis, 171-177 Vittariaceae, 204, 205 Walking fern, 241 WATANABE, MIKIO, (see Noriaki Murakami), 232 | a INDEX TO VOLUME 89 15 WERTH, CHARLES R. (see Richard D. Stevens), 221 WHITTIER, DEAN P., Spore Germination and Development in cooperi, 204 oe see Barbara Joe Hoshizaki), 1 ilvensis, 171, 175 scopulina, 171, 175 281 Woodwardia virginica, 169 Xiphopteris, 205 YATSKIEVYCH, GEORGE Review: Flora of Australia, Volume 48, Ferns, mnosperms, and Allied Groups, 270 Review: IIlustrierter Leitfaden zum Besti: Farne und farnverwandten der Schweiz und angrenzender Gebicte, a Authors are encouraged to submit manuscripts pertinent to pteridology for pub- lication in the American Fern Journal. Manuscripts should be sent to the Editor. Acceptance of papers for publication depends on merit as judged by two or more referees. Authors are encouraged to contribute toward publishing costs; however, the payment or non-payment of page charges will affect neither the acceptability of manuscripts nor the date of publication. Authors should adhere to the following guidelines; manuscripts not so prepared may be returned for revision prior to review. 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