AMERICAN ic: FERN 1 JOURNAL PUBLISHED BY THE AMERICAN FERN SOCIETY EDITORS David W. Bierhorst Gerald J. Gastony David B. Lellinger John T. Mickel Terry R. Webster MERCURY PRESS, ROCKVILLE, MARYLAND 20852 CONTENTS Volume 73, Number 1, Pages 1-32, Issued March 31, 1983 Edgar T. Wherry and his Contributions to Pteridology W. H. WAGNER, JR 1 Stone Fort at Fort Totten: Last Habitat for Woodsia obtusa and Asplenium platyneuron in Queens County, Long Island, New York? ANDREW M. GRELLER and DAVID C. LOCKE 6 Starch Gel Electrophoresis of Ferns: A Compilation of Grinding Buffers, Gel and Electrode Buffers, and Staining Schedules DOUGLAS E. SOLTIS, CHRISTOPHER H. HAUFLER, DAVID C. DARROW, and GERALD J. GASTONY Shorter Notes: A New Combination in Asplenium; Adiantum furcatum and Microstaphyla furcata; Noteworthy Pteridotphyte Records for Nebraska; Spread of Marsilea quadrifolia in McDonough County, Illinois; Wood Ferns New to Maryland and ware; A New Combination for an Asplenosorus Hybrid 28 American Fern Journal 32 Index to Volume 73 122 Errata 124 Volume 73, Number 2, Pages 33-64, Issued June 16, 1983 Vittaria Gametophytes Discovered in a New Physiographic Province ALLISON W. CUSICK 33 Chapman’s Quillwort Reconsidered BRIAN M. BOOM 39 C-Glycosylxanthones in Diploid and Tissue Culture-induced Autotetraploid Davallia fejeensis P. MICK RICHARDSON and HARINDI.2 K. PALTA 43 The Distribution of Woodwardia areolata R. CRANFILL 46 Two Moonworts of the Rocky Mountains: Botrychium hesperium and a New Species Formerly Confused with It W. H. WAGNER, JR. and FLORENCE S. WAGNER 53 Shorter Notes: Lycopodium complanatum and L. annotinum Found in the Black Hills; Microfibrils in the Xylem of Blechnum viviparum; Bi i Sporophylis in Isoétes from Rajasthan 62 Reviews 12, 45, 52 Volume 73, Number 3, Pages 65-96, Issued September 29, 1983 Observations on the Structure and Function of Hydathodes in Blechnum Iehmannii JOHN S. SPERRY 65 A Reclassification of the Fern Genus Pyrrosia K. H. SHING 73 Notes on the Ecology and Development of Plagiogyria fialhoi PAULO G. WINDISCH and MARILIA PEREIRA-NORONHA 79 The Ferns of Elden Mountain, Arizona MICHAEL D. WINDHAM 85 Donovan S. Correll (1908-1983) 94 Reviews 93-96 Volume 73, Number 4, Pages 97-124, Issued December 29, 1983 Polyploidy and Aneuploidy in Hypolepis, and the Evolution of the Dennstaedtiales P. J. BROWNSEY 97 Pecluma, a New Tropical American Fern Genus MICHAEL G. PRICE 109 The Lady Fern, Athyrium filix-femina, in Saskatchewan VERNON L. HARMS 117 Shorter Note 121 American Fern Journal 122 Index to Volume 73 122 Errata 124 AMERICAN ee FERN 1 JOURNAL PUBLISHED BY THE AMERICAN FERN SOCIETY EDITORS David W. Bierhorst Gerald J. Gastony David B. Lellinger John T. Mickel Terry R. Webster MERCURY PRESS, ROCKVILLE, MARYLAND 20852 CONTENTS Volume 74, Number 1, Pages 1-32, Issued April 16, 1984 Promotion of Apogamy in Matteuccia struthiopteris, the Ostrich Fern P. VON ADERKAS A New Filmy Fern from Puerto Rico GEORGE R. PROCTOR Chromosome Numbers of Neotropical Isoétes R. JAMES HICKEY Two New Phenolic Glycosides in Asplenium septentrionale FILIPPO IMPERATO A Remarkable Cyathea Hybrid R. E. HOLTTUM The Ligule of Isoétes B. D. SHARMA and R. SINGH Shorter Notes: Two Species of — Newly Escaped in Florida; A New Statio Trichomanes petersii in ralecie ee geen Gametophytes at Bartholomew’s Cobble Reviews 6, 18, Volume 74, Number 2, Pages 33-64, Issued July 18, 1984 A Western Holly Fern, Polystichum x scopulinum in Newfoundland WARREN H. WAGNER, JR. and ERNEST ROULEAU Drymoglossum Under Water Stress C. S. HEW Problems in Asplenium, with Some New Species from Ecuador ROBERT G. STOLZE Frequency of Cyanogenesis in Bracken in Relation to Shading and volute Severity I. SCHREINER, D. NAFUS and D. PIMENTEL New Combinations and Some New Names in Ferns DAVID B. LELLINGER Gualterio Looser (1898-1982) Shorter Notes: Equisetum ramosissimum in Louisiana; Three New Combinations in Lox oxogramme; Notes on North American Ferns, II; Graves’ Spleenwort in Ohio 36, 50, Suggestions to Contributors Volume 74, Number 3, Pages 65-96, Issued December 6, 1984 The Habitat Characteristics and Abundance of Equisetum x ferrissii and its Parent Species, Equisetum hyemale and Equisetum laevigatum, in Iow LORENZ M. RUTZ and DONALD R. FARRAR 65 The Organic Nutrition of Botrychium Gametophytes Anti-microbial Activity ne Phenolic Acids in Pteridium aquilinu DEAN P. WHITTIER 77 MICHAEL SAN FRANCISCO and GILLIAN COOPER-DRIVER 87 Reviews 86, 96 Volume 74, Number 4, Pages 97-124, Issued December 28, 1984 The Identification of Hawaiian Tree Ferns of the Genus Cibotium Two New Tree Ferns from Panama Trunk Length and Frond Size in a Population of Nephelea tryoniana from El Salvador An Unusual New Elaphoglossum from Peru New Tropical American Ferns Shorter Note: aE Urban Locality for Asplenium euron 1985 A.1.B.S. Meeting—Call for Papers American Fern Journal Index to Volume 74 Errata RICH BECKER 97 ROBERT G. STOLZE RALPH L. SEILER ROLLA TRYON JOHN T. MICKEL 101 AMERICAN ae FERN _ JOURNAL | QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Edgar T. Wherry and his Contributions to Pteridology W. H. WAGNER, JR. 1 Stone Fort at Fort Totten: Last Habitat for Woodsia obtusa and Asplenium platyneuron in Queens County, Long Island, New York? ANDREW M. GRELLER and DAVID C. LOCKE 6 Starch Gel Electrophoresis of Ferns A Compilation of Grinding Buffers, Gel and Electrode Buffers, and Staining Schedules DOUGLAS E. SOLTIS, CHRISTOPHER H. HAUFLER, DAVID C. DARROW, and GERALD J. GASTONY 9 Shorter Notes: A New Combination in Asplenium; Adiantum furcatum and Microstaphyla furcata; *eoaaegeng Pteridophyte Records for Nebraska; Marsilea quadrifolia in McDonough Cea Illinois; Wood Ferns New to Maryland and Delaware; A New Combination for an Asplenosorus Hybrid 28 American Fern Journal 32 MissouR! BOTA APRIL4: The American Fern Society Council for 1983 DEAN P. WHITTIER, Dept. of Biology, Vanderbilt University, Nashville, TN 37235. President TERRY R. WEBSTER, Biological Sciences Group, University of Connecticut, Storrs, CT 06268. Vice-President MICHAEL I. COUSENS, Faculty of Biology, University of West Florida, Pensacola, FL 32504 See retary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Treasurer JUDITH E. SKOG, Dept. of Biology, George Mason University, Fairfax, VA 22030. Records Treasurer DAVID B. LELLINGER, Smithsonian Institution, Washington, DC 20560. Journal Editor ALAN R. SMITH, Dept. of Botany, University of California, Berkeley, CA 94720. Memoir Editor JOHN T. MICKEL, New York Botanical Garden, Bronx, NY 10458. Newsletter Editor setae Mire Journal DAVID B. LELLINGER U = cat Herbarium NHB-166, Smithsonian Institution, Washington, DC 20560. ASSOCIATE EDITORS DAVID W. BIERHORST Rt. 3, Box 188, Picayune, MS 39466. GERALD J. GASTONY Dept. of Biology, Indiana University, Bloomington, IN 47401. JOHN T. MICKEL New York Botanical Garden, Bronx, NY 10458. TERRY R. WEBSTER ....... Biological Sciences Group, University of Connecticut. Storis. CT 06268. The “American Fern Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the study of ferns. It is owned by the American Fern Society, and — at the Smithsonian instieutile Washington, DC 20560. Second-class postage paid at Washin Claims for missing issues, made 6 months (domestic) to 12 month (foreign) after the date of issue, and the matters for publication should be addressed to the Edito Changes of address, dues, and applications for membership aia be sent to Dr. Judith. E. Skog. Dept. of Biology, George Mason University, Fairfax, VA 22030. : Orders for back issues should be addressed to Dr. James D. Montgomery, Ichthyological Associates, — R.D. 1, Berwick, PA 18603. General inquiries concerning ferns should be addressed to the Secreta Subscriptions $9.00 gross, $8.50 net if paid through an = ( agency fee $0.50); sent free to — members of the American Fern Society (annual dues, $8.00 + $4.00 mailing surcharge beyond U.S.A., Canada, and Mexico; life membership, $160.00). A Back volumes 1910-1978 $5.00 to $6.25 each: single back numbers of 64 pages or less, $1.25; 65-80 pages, $2.00 each; over 80 pages, $2.50 each, plus shipping. Back volumes 1979 et seq. $8.00 each, — single back numbers $2.00 each, plus shipping. Ten percent discount on orders of six volumes or more. f Library : Dr. John T. Mickel, New York Botanical Garden, Bronx, NY 10458. is Librarian. Members . _ may borrow books at any time, the borrower paying all shipping costs. 4 Newsletter y Dr. John T. Mickel, New York Botanical Garden, Bronx, NY 10458. is editor of the newsletter. “Fiddlehead Forum.” The editor welcomes contributions from members and non-members. including — miscellaneous notes, offers to — or purchase materials, personalia. horticultural notes, and reviews, of non-technical books on Mr} Neill D. Hall, a 1230 None 8 Se le, W. a oe collection lists Sem, A 98115, is = Spores exchanges : in fers Botanical books, cs, back a OF the Journal, and cash or other ek i ways wel a are tax-deductible. - and her gifts are al < wekonel all a oe ni should be addressed to the Secretary. a ; AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) l Edgar T. Wherry and his Contributions to Pteridology' W. H. WAGNER, JR.* When he first became interested in ferns, Dr. Wherry was thirty years old. His background had been in chemistry and geology, and at the time he was an assistant curator in mineralogy at the U.S. National Museum. Some amateur botanists took him to see a patch of the walking fern, Camptosorus rhizophyllus, where it was growing on gneiss, and he became curious about its soil requirements. This led to his first publication on ferns “A Chemical Study of the Habitat of the Walking Fern” in 1916. After that Wherry became an expert on hydrogen-ion concentrations in the soils of eastern United States ferns, and he also broadened out into questions of their geography, ecology, and systematics. By the end of his career he had become an authority on North American pteridophytes north of Mexico. In 1917, Wherry transferred to the Department of Agriculture, where he came under the influence of F. V. Coville, the Chief Botanist, who encouraged him in the study of plants. By 1930, Wherry was prominent enough in his new profession to be invited to join the Botany Department of the University of Pennsylvania. He accepted this opportunity, which was ideal for him because it represented a return to his alma mater and to his home city of Philadelphia. Wherry taught at the University until his retirement in 1955. He stayed active for many years longer than most botanists: he continued to take field trips well into his 80’s and was still writing letters as late as his 90’s. Only in the last few years before his death at 97 on 19 May 1982 did he lose touch. Wherry was very active in the American Fern Society, and three-fourths of his pteridophyte publications were in the “American Fern Journal.” He was President of the Society from 1934 to 1939, and he also took charge of preparing a cumulative 25-year index to the “Journal.” When Maurice Broun wrote his “Index to the Ferns and Fern Allies of North America” (1938), Wherry supplied the habitat and range data for most of the species. After becoming discouraged by the poor quality of the manuals then available for identifying pteridophytes, he published one himself entitled “Guide to Eastern Ferns” (1937), which, together with the second edition, sold more than 6000 copies and did much to stimulate the popularity of ferns among naturalists and botanists. Later he wrote another valuable guide, this one for the southeastern United States. The royalties from his field manuals were donated to the American Fern Society. Wherry did not become a major botanist of his time: he was not involved in any of the “big ideas” such as biosystematics, cytotaxonomy, and phylogeny that burgeoned during his lifetime. His approach to the study of plants was deficient in such *Department of Botany and Herbarium, University of Michigan, Ann Arbor, MI 48109. 'A series of papers in tribute to Edgar T. Wherry (19 Sep 1885-19 May 1982) was published in Amer. Fern J. 66:33-80. 1976. Biographical notes have been published in Fiddlehead Forum 9:32-33. 1982 and by Prof. Wagner in Bull. Torrey Bot. Club 109:545-548. 1982. An obituary for Dr. Wherry has been published in Bartonia, 49:1—5. 1983, the journal of the Philadelphia Botanical Club. 2 AMERICAN FERN JOURNAL: VOLUME 73 (1983) subjects as morphology, anatomy, cytology, and physiology, and he used the laboratory and the microscope very little in his research. He functioned primarily as a “field naturalist,” as evidenced by his teaching and his publications. Over 40% of his fern bibliography dealt with geographical distributions, especially range extensions and reports of what he encountered on his numerous exploring trips. Among the many individuals who accompanied him on his pteridological field : expeditions were Harry W. Trudell, J. E. Benedict, and myself. For over a third of a © century, from high school student to university professor, I had the good fortune to go exploring from time to time with Wherry and to absorb his ideas of pteridophyte ecology. In the “Fern Guide” he wrote as follows: “Sometimes .. . a fern student may have the ‘thrill that comes once in a lifetime’ by discovering a species in an area where it had not previously been found.” Wherry passed his love for finding rare pteridophytes to others, and when the University of Virginia Mountain Lake Biological Station began its pteridology course in 1962, Wherry wrote to the class these inspiring words: “To the Mountain Lake Chapter, F. F. A. (Future Ferners of America) Greetings: At last a real study of the ferns of Mountain Lake is under way... . Many range extensions are to be expected there. Congratulations on yours!” He was so right—that first class and the subsequent ones (including the latest in 1982) made numerous range extensions of species in eight genera. The success of field pteridology courses everywhere in the United States, especially at Lake Itasca, MN, Highlands, NC, and at Flathead Lake, MT, owes a lot to Wherry’s teaching about the roles of soil requirements and indicator species. Wherry’s most significant contributions to the study of fern distribution and ecology are those concerned with pH requirements, the subject which originally stimulated his interest in ferns. He helped to design simple kits for making field — tests of soil acidity, and he showed, for example, that certain traditional reports of species growing on particular rock types were wrong. One author copied another that Asplenium bradleyi grows on limestone, and it took Wherry years to straighten this out and to convince pteridologists that this species is confined to acidic rocks. Thanks to his efforts to interest botanists in pH, it has become standard today for field workers to specify rock and soil types upon which plants grow. And if we wish to find a particular species, we first locate the proper substrate. We know now that it — would be practically impossible for Asplenium montanum to be found growing — beside A. ruta-muraria; their edaphic preferences are too different. Oddly, Wherry paid little attention to the positive effects of disturbance and — succession On pteridophytes, even though his viewpoint in general was broadly — ecological. He did not seem to appreciate how influential upsets such as floods, fires, and grazing may be in enhancing the abundance and spread of many species, — including botrychiums, certain dryopterids, and lycopods. Often, when a certain species disappeared from its habitat, Wherry would attribute it to vandalism or a over-collecting when it was more probably a matter of simple succession. When he tried, in his greenhouse, to duplicate the natural growth conditions of | saxicolous ferns by faithfully assembling the appropriate rocks, he almost invariably _ failed. Had he only known what we now know, namely that most rock ferns can f E ‘ W. H. WAGNER, JR.: EDGAR T. WHERRY 3 grow merely in soil-in pots, controlling the pH by simply mixing in limestone chips or sphagnum peat, he could have learned much more about the characters of rock ferns by culturing them side-by-side under uniform conditions. Practically everything he knew about the species he studied came from field observations. Wherry’s main contribution to pteridophyte taxonomy involved detecting hybrids in the wild. He was opposed to the wanton designation of trivial forms and varieties as illustrated by Fernald in “Gray’s Manual” (1950), and he was effective in steering serious fern students away from this. His argument in “The Fern Guide” (1961) was that “since these mostly occur in association with typical plants—indeed often arising from the same rootstocks—they are not named.” He distinguished between geographical varieties (what some botanists today call subspecies) and true species, as illustrated by his treatment of the decompound species of Dryopteris, such as D. spinulosa (= D. carthusiana) and D. intermedia, which by Fernald and his associates had been interpreted as varieties, D. spinulosa, typical variety, and D. spinulosa var. intermedia. He developed his concepts on the basis of studies of natural populations, long before modern experimental and biochemical techniques were available to confirm them. Together with W. D. Gray, Wherry presented illustrations of intermediates, which he interpreted as hybrids, between long recognized species of spleenworts. Appar- ently, however, neither Gray nor Wherry was aware, or even suspected, that two of the most important of the species then recognized, A. Xbradleyi and A. x pinnatifidum, were hybrids themselves (Wagner, W. H., Jr., Evolution 8:103—118. 1954). Both of these men, on the contrary, accepted the hybridity of the fertile A. x ebenoides from Alabama. The most interesting of the hybrids first recognized by Wherry is A. Xtrudellii (A. montanum X pinnatifidum, a beautiful example of hybrid vigor in a “sterile hybrid” which evidently can reproduce by unreduced spores. Wherry’s work on Asplenium is honored in the name A. Xwherryi D. Smith, the backcross of A. X bradleyi to A. montanum. Under Dryopteris he distinguished and/or named a number of hybrids which since have become important in the genome analysis of the basic species of eastern American wood ferns. Some of these are common ferns widespread in forests and swamps. They include D. x slossonae (D. cristata X marginalis), D. X triploidea (D. intermedia X carthusiana), and D. X benedictii (D. clintoniana X carthusiana). What he named D. /eedsii and interpreted to be D. goldiana X marginalis has since turned out to be D. celsa X marginalis (Wagner, W. H., Jr., Virginia Polytech. Inst. State Univ. Res. Div. Monogr. 2:147-192. 1970). His greatest difficulties with wood ferns involved the taxa resembling D. x clintoniana, the same ones that cause us trouble today. He nonetheless had a sharp eye for them and was the first person to recognize as distinct D. x australis (D. celsa x ludoviciana), although for good reasons at the time he initially interpreted it as a southern variety of D. clintoniana. His contributions to our knowledge of Dryopteris hybrids are commemorated in the name D Xneo-wherryi Wagner, which is the true hybrid of Goldie’s Wood Fern and the Marginal Wood Fern. Scientifically, Wherry’s influence on American pteridology was substantial. Many of today’s pteridologists are indebted to his teachings as well as his inspiration. His major contributions involved his investigations of the pH reactions of soils, distribu- tion patterns, and his studies of various taxa, especially spleenworts and wood ferns. The than three decades. For the general public, Wherry’s field manuals and his many lectures and guided trips for amateurs helped engender a widespread enthusiasm for the natural history of pteridophytes. 1916. 1917. 1920. 1920. published j . The soil reactions of certain rock ferns—II. Amer. Fern J. pasar : ag gravesii in Pennsylvania. Amer. Fern J. 10:119-121. : Wall ferns in Wilmington, North Carolina. Amer. Fern J. . Ferns of eastern West Virginia, I. Amer. Fern J. Pay ake . Some fern finds in Virginia. Amer. Fern J. 15:1-7. . The Appalachian aspleniums. Amer. Fern J. 15:46—54. . Soil reaction preferences of three adder’s-tongues. Amer. Fern J. 16:1—3. . The West Virginia locality of the southeastern relative of Woodsia scopulina. Amer. Fern J. ) ray . A fernless area. Amer. Fern J. 17:63-64. . Observations on the woolly lipfern. Amer. Fern J. 16:107-109. . Ferns of Dripping Spring, Oklahoma. Amer. Fern J. 18:61-63. . Notes on Asplenium Trudelli. Amer. Fern J. 17:135-138. . Further occurrence of the Alleghany cliff-fern. Amer. Fern J. 19:101—102. . The Asplenium ebenoides locality near Havana, Alabama. Amer. Fern J. 11:30-33. (with H. W. ll - Asplenium bradleyi erroneously reported on limestone again. Amer. Fern J. 21:111—113. 4 . Range-extensions and other observations, 1931-32. Amer. Fern J. 22:79-86. ; - Fern field notes, 1933. Amer. Fern J. 23:109-112. . Fern field notes, 1934. Amer. Fern J. 24 97-104 . Fern field notes, 1935. Amer. Fern J. 25: 123-126. . The northern limits of Polypodium polypodioides in the east. Claytonia 2:32. . Fern field notes, 1936. Amer. ren, 26:127- ae : Cand eae Aner Fem J. 28: 125-140, ot. Hz. . Recent fern finds in West Virginia. Castanea 4:14. : : 4 fs a 7A full aden Dr. Wherry, including his papers on flowering plants and —— has been a AMERICAN FERN JOURNAL: VOLUME 73 (1983) American Fern Society is indebted to him for his help in many ways over more ee ee eS eS Ne PTERIDOPHYTE BIBLIOGRAPHY OF EDGAR T. WHERRY’ A chemical study of the habitat of the walking fern, Camptosorus rhizophyllus (L.) Link. J. Washington Acad. Sci. 6:672-679. Observation on the habitat of certain ferns. Amer. Fern J. 7:110—-112. A fruitless search for Asplenium fontanum in ingore Amer. Fern J. 10:90-91. The soil reactions of certain rock ferns—I. Amer. Fern J. 10:15—22. a alia a clean soil reactions of the ferns of woods and swamps. a al J. 11:5-16. shel a al hale Ae le Ls el gbila ¥ Wood-ferns on Mt. Desert Island, Maine. Amer. Fern J. 16:3-7. 16:92-95. (with F. W. Gr. Be oP ot i acre tae de Trudell) eee ee pes ee ESR SRG eee Ce REE 3 in Bartonia, ‘9: 6-14. 1983, the journal of the Philadelphia Botanical Club | ‘ F 3 4 Ree ne AE LE Oe TT RR TT TO eA TMS PON Ne ETE CRM I Ge eR SON Ns PEE REE TAT eR EO CRE a ET IN NO CE ST EO A BE CE EN eT Te See ee W. H. WAGNER, JR.: EDGAR T. WHERRY 1940. 1941. . Two Virginia fern records. Virginia J. a — . A woodfern hybrid deserves a name. Bartonia 21:1—2, pl. 1. fig. 1 . The ferns and lycosphens of paras ‘Bertie 21:11-63, pl. 1, fig. 2. . Lycopodium sabinaefolium in Pennsylvania. — nip 9 32:111-113. . Observations on Florida ferns. Amer. Fern J. 32:139-145. . The discoveries of new Pennsylvania ferns. oo ae J. 32:148-149. . Guide to Eastern Ferns, 2nd ed. Science Press Printing Co.. Lancaster, PA. . Note on the southeastern relatives of oi iia inundatum. Amer. Fern J. 34:24. . Cystopteris bluff. Amer. Fern J. 34:92-9 . Osmunda cinnamomea f. ile Amer. Fern J. 34:94—95. . The indument of Cystopteris fragilis. Amer. Fern J. 35:54—55. . Notes on Illinois pteridophytes. Amer. Fern J. 35:92. . Our most-renamed native fern. Amer. Fern J. 35:128. . Notes on Muhlenberg’s ferns. Amer. Fern J. 36:54-58. . Selaginella rupestris in Pennsylvania. Amer. Fern J. 37:24-25. Five recent books on ferns. Bartonia 20:38. Asplenium adiantum-nigrum in Arizona. Amer. Fern J. 31:97—100. Our easternmost Cheilanthes species. Amer. Fern J. 37:77-79. . A crispate variant of the Christmas ne Amer. Fern J. 37:121. . Remarks on the American lady ferns. Amer. Fern J. 38:155—158. . Guide to Eastern Ferns. Univ. of Hasta Press, Philadelphia. . Geographic notes on the bog fern. Amer. Fern 9:18-19. 0. A new interpretation of the Dryopteris clintoniana ~ Amer. Fern J. 40:118—120. . Observations at Bartholomew’s Cobble. Amer. Fern J. 41:13-14. . Lowland lycopodiums in the New Jersey ay ore ai Torrey Bot. Club 81:364. 4. Nomenclature of the oak-ferns. Amer. Fern J. 44:85—86. How I became interested in ferns. Amer. Fern J. 50:225—228. . The Fern Guide. Doubleday, Garden Ci oe . Supplementary note on Dryopteris hybrids. Amer. Fern J. 51:33-35. . Native ferns in the garden. Plants and Garden 18: . The Southern Fern Guide. Doubleday, Garden City, NY, . Some new name-combinations for southeastern ferns. Amer. Fern J. 54: 143-146. . Southern records of Ophioglossum vulgatum. Amer. Fern J. 58:182-183. 6 AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) Stone Fort at Fort Totten: Last Habitat for Woodsia obtusa and Asplenium platyneuron in Queens County, Long Island, New York? ANDREW M. GRELLER* and DAVID C. LOCKE** In the course of documenting vegetation on the northeastern coast of Queens . County, Long Island, New York, the senior author discovered two ferns, Asplenium — platyneuron (L.) B.S.P. and Woodsia obtusa (Spreng.) Torrey, in what may well be — their last remaining habitat in Queens County (Borough of Queens, City of New York). They were found in the weathered cement between large blocks of granite © used to construct the walls of the “Stone Fort” at Fort Totten, a 19th Century coastal — fortification on the Willets Point peninsula (40°47'30" N, 73°46’40” W). The Stone Fort now forms Willets Point, rising from the shore level to a height of thirty feet. It was built in 1862-66 as part of the seacoast defenses of New York City; photographs and a brief history of the Stone Fort were published by Alperstein (1977). Reconnaissances of the site were undertaken in the spring of 1973, in the late spring — and early summer of 1978, and during the winter of 1981-82. Occasionally a few | plants were measured, but a census was not attempted. Soil samples were obtained adjacent to fern roots at two sites on the landward side — of the granite wall, for elemental analysis. Stones, sticks, and obvious plant matter — were separated, and a portion of well-mixed soil was ground in a mortar to a fine powder. A 1.00 g sample of each was dry-ashed at 450°C in a muffle furnace, anda — semi-quantitative elemental analysis conducted with a Baird Atomics Emission Spectrograph. A just-saturated slurry in distilled water was made of each raw soil — sample, and the pH measured with an Orion combination electrode and a Corning — pH meter. Further tests for sodium and chloride, indicators of saline spray, were made on samples taken near Woodsia roots, one from the seaward side of the granite wall, — and one from the landward side. Soil samples, 1.00 g, were extracted with 10.0 ml — of distilled, deionized water. The sodium content was determined by atomic — absorption spectroscopy using a Perkin-Elmer 303, and chloride content by ion- — selective electrode using an Orion electrode and a Beckman expanded scale pH ~ meter. Voucher specimens were deposited at the Herbarium of the Brooklyn Botanic | Garden (BKL). RESULTS Fewer than twenty-five individuals of A. platyneuron were observed during the — most recent visit (30 March 1982). These were concentrated around the central — stairway connecting the three levels of the fort, between the main and upper levels. — in a sheltered, moderately well lighted location. A few individuals were scattered — Over the rest of the main level. The plants grew from partially decomposed cement — “Department of Biology, Queens College, City University of New York, Flushing, NY 11367. Department of Chemistry, Queens College, City University of New York, Flushing, NY | 1367. —. GRELLER & LOCKE: STONE FORT AT FORT TOTTEN 7 in joints between granite blocks. They were well rooted in the joints, to perhaps 10 cm deep. Fronds seldom exceeded 10 cm in length. The population of W. obtusa numbered several hundred individuals. Plants occurred in the horizontal joints of the seaward granite wall of the fort, where sea spray and salt-laden air are features of the environment. More commonly, individu- als occurred in both vertical and horizontal joints in the complex of granite walls on the main, landward side. Plants of Woodsia appeared to be less deeply and securely rooted than those of Asplenium. Commonly, young sporophytes were seen attached to gametophytes. Woodsia plants in winter bear a rosette of sterile fronds approxi- mately 3-4 cm long. Fertile fronds are produced during the growing season; these regularly reach 30 cm or longer. The substrate of both ferns consisted of a mixture of black humus and light gray fragments of cement and sands. The soil evidently arises from the mortar decomposed through weathering and from organic materials deposited by the ferns and other organisms which have gained a foothold (Segal, 1969). This is supported by our chemical analyses. The results of the dry-ashing test show that organic materials account for nearly 40% by weight of the soil sample. In addition, the emission spectrographic elemental analyses of the inorganic portions of the wall soil samples reveal silicon, calcium, and aluminum to be the major constituents and an overall composition quite similar to that of Portland cement. Portland cement has been in widescale use since at least 1850 (Lea, 1971), and apparently was used as the mortar for the Stone Fort. Although raw, wet cement is highly alkaline (pH 11-12), acidic agents in rainwater and in humic substances from decayed plant materials act to neutralize the alkalinity. Our soil samples are neutral (pH 6.96—7.00); these pH values are typical of well-aged wall soils (Segal, 1969). Results of the sodium and chloride analyses, in wg/g of soil (ppm) are: landward, 250 ppm Na and 110 ppm Cl; seaward, 1200 ppm Na and 280 ppm Cl. The seaward wall soil has nearly five times as much sodium and more than twice as much chloride as the soil from the landward wall, confirming the influence of salt spray on the seaward side. DISCUSSION It is uncertain when A. platyneuron and W. obtusa first appeared in the mortar of the Stone Fort. Little of the mortar remains unweathered, and extensive sections are missing. It is possible that the latter sections once supported populations of the ferns. It cannot be decided on the basis of the available data whether it is the high calcium content or the neutral pH of the soil that accounts for the presence of the ferns; both conditions are uncommon on Long Island. We inspected herbaria at New York Botanical Garden, Brooklyn Botanic Garden, and Planting Fields Arboretum, and consulted with staff of the New York State Herbarium, Albany. Woodsia obtusa has been collected at Ridgewood, in Queens County (G. B. Brainerd in 1866, BKL 022985); on rocks near Greenport, in Suffolk County, Long Island (BKL 1862); and in “dry woods” there (R. Latham in 1918, NY 824). The Fort Totten collection (Greller in 1973, BKL) is the first Woodsia from stone walls on Long Island. Edward Frankel (pers. comm.) reports W. obtusa 8 AMERICAN FERN JOURNAL: VOLUME 73 (1983) as common on old walls in Westchester County, north of Long Island. Woodsia was — collected “on - of oe ice chute, south side” at Rockland Lake, Clarkstown Township, Rocklan , New York (J. H. Lehr in 1954, NY), also north of — Long Island. ee mee listed A. platyneuron as “frequent throughout Long — Island.” Nevertheless, Greller (1977, 1979) did not encounter it in the only remaining extensively forested sites in Queens County: Cunningham, Forest, and — Alley Parks. Until they are reported elsewhere, the Fort Totten populations of W. — obtusa and A. platyneuron are the only ones documented in Queens County. Their © absence from former terrestrial habitats may be caused by a decrease in soil pH, perhaps as a result of acid rain. Precipitation pH in Queens County since 1970 has — averaged about 4.0, monthly averages ranging from 3.5 (summer) to 4.5 (spring and fall) (D. C. Locke, unpubl.). i Occurrence of W. obtusa on the seaward wall is noteworthy because of the relatively high levels of sodium and chloride which presumably result from salt spray. Based on salinity measurements made in Hempstead Harbor, a similar bay a few miles east of Fort Totten, the salinity of Long Island Sound at Fort Totten is about 25%e (D. C. Locke, unpubl.), less than that of ocean water (35%c) but more than sufficient to produce saline spray. The relatively higher level of sodium than | chloride in both samples presumably reflects differential leaching of the two — elements; according to Lindsay (1979), the level of sodium in soils is typically — higher than that of chloride. The authors gratefully acknowledge the assistance of CWO-4 Jacob M. Fein U. S. Army (ret.), who acted as our guide and gave us historical information, reprints, and photographs of the Stone Fort; Thomas J. Delendick, Asst. Taxonomist and Curator — of the Herbarium, Brooklyn Botanic Garden, who helped us locate specimens; — Richard Posner, U. S. Testing Company, for emission spectrographic analyses; and Scott Marcus, for field assistance. LITERATURE CITED eS D. M. 1977. Fort Totten at Willets Point. Periodical (Council on Abandoned Military ee Arizona) 9(2):41—-52. : GRELLER, 5 M. 1977. A vascular Ha of the forested portion of Cunningham at Pie County, New York, with notes on the vegetation. Bull. Torrey Bot. Club 104:170- 7 - 1979. A vascular flora of the forested portion of Cunningham Park, oaecis County, be a York: Corrections and additions. Bull. Torrey Bot. Club 106:45. a JELLIFFE, W. E. 1899. The Flora of Long Island. Privately Printed, Lancaster, PA. LEA, F. M. 1971. The Chemistry of Cement and Concrete, 3rd ed. Chemical Publishing Company. — New York. LINDSAY, W. L. 1979. Chemical Equilibrium in Soil. Wiley, New York. SEGAL, S. 1969. Ecological Notes on Wall Vegetation. W. Junk, The Hague, Netherlands. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) 9 Starch Gel Electrophoresis of Ferns: A Compilation of Grinding Buffers, Gel and Electrode Buffers, and Staining Schedules DOUGLAS E. SOLTIS*, CHRISTOPHER H. HAUFLER**, DAVID C. DARROW***, and GERALD J. GASTONY*** The homosporous pteridophytes have been largely uninvestigated by electrophore- sis, despite the fact that they offer many exciting research possibilities (Soltis et al., 1980). The paucity of electrophoretic studies of ferns and fern allies may be due in large part to the high concentrations of condensed tannins that many species contain (Cooper-Driver, 1976 and pers. comm.). These compounds render enzymes inactive by binding with them following cellular disruption, thereby frustrating researchers who have attempted electrophoretic analysis utilizing standard methods of sample preparation. The method of sample preparation developed by Kelley and Adams (1977a, b) in their analysis of enzyme variation in Juniperus was an important procedural breakthrough in overcoming the difficulties that result from the liberation of large amounts of phenolic compounds during tissue preparation. Recently, a simplified version of that method was applied by Soltis et al. (1980) to fern leaf tissue, facilitating rapid preparation of active enzyme samples and thereby making electro- phoretic analyses of large numbers of individuals more feasible. In an attempt to improve methods of analysis of fern enzymes in starch gel electrophoresis, we have experimented with modifications of the method of sample preparation outlined by Soltis et al. (1980). We also have examined several different methods of sample preparation such as those of Gottlieb (1981a), Mitton et al. (1979), and Werth et al. (1982), and have evaluated the relative merits of each with fern tissue. Finally, during the course of our electrophoretic investigations of ferns we found that standard gel and electrode buffers and staining schedules, such as those of Brewer (1970) and Shaw and Prasad (1970), often provided unsatisfactory results when applied to ferns. We have determined gel and electrode buffers, as well as staining schedules, that provide clear starch gel enzyme banding for 22 enzyme systems in ferns. Requests for advice resulting from the recent surge of interest in fern enzyme electrophoresis have prompted us to compile our procedural data so that other researchers can take advantage of our experimentation. We hope that these data will stimulate more extensive electrophoretic investigation of pteridophytes and other electrophoretically difficult taxa. Gottlieb (1981b) recently reviewed aspects of enzyme electrophoresis primarily in gymnosperms and angiosperms. His discussion is equally relevant to understanding the potential applications and limitations of electrophoretic evidence in pterido- phytes. Since homosporous pteridophytes have high chromosome numbers, it is tempting to invoke polyploidy in interpreting their enzyme band patterns. It is well *Department of Biology, University of North Carolina, Greensboro, NC 27412. **Department of Botany, University of Kansas, Lawrence, KS ***D oy partment of Biology. Indiana University, Bloomington, IN 47405. TABLE |. ELECTRODE AND GEL BUFFER RECIPES USED SUCCESSFULLY IN ELECTROPHORETIC ANALYSIS OF FERNS (Gram ted). amounts are given for one liter final volume of buffer except where note Electrode Buffer 1. 0.400 M Citric Acid, trisodium salt; 117.64 g Citric Acid, trisodium salt dihydrate, 1.0 M HCI to pH 7.0 2, 0.135 M Tris, 0.032 M Citric Acid; 16.35 g Tris*, 6.10 g Citric Acid? 3. 0.135 M Tris, 0.017 M Citric Acid; 16.35 g Tris*, 3.35 g Citric Acid? 4. 0.223 M Tris, 0.086 M Citric Acid; 27.00 g Tris’, 4 52 g Citric Acid”, NaOH to pH 7 5. 0.223 M Tris, 0.069 M Citric Acid; 27.00 g Tris*, 13.33 g Citric Acid” 6. 0.100 M NaOH, 0.300 M Boric Acid; 4.00 g NaOH, 18.55 g Boric Acid? 7. 0.038 M LiOH, 0.188 M Boric Acid; 1.60 g LiOH-H,O, 11.60 g Boric Acid” Adjust to pH 8.3 with dry components pH @ 22°C 7.0 Gel Buffer 0.020 M Histidine-HCl; 4.19 g L-Histidine- HCI monohydrate, 1.0 M NaOH to pH 7.0 0.009 M Tris, 0.002 M Citric Acid; dilute 67 ml of electrode buffer to 1 liter 0.009 M Tris, 0.001 M Citric Acid; dilute 67 ml of electrode buffer to | liter 0.008 M Tris, 0.003 M Citric Acid; dilute 35 ml of electrode buffer to | liter 0.008 M Tris, 0.002 M Citric Acid; dilute 35 ml of electrode buffer to | liter 0.015 M Tris, 0.004 M Citric Acid; 1.84 g Tris*, 0.69 g Citric Acid” 0.045 M Tris, 0.007 M Citric Acid, 0.004 M LiOH, 0.019 M Boric Acid; Tris- yee: buffer (5.45 g Tris’, ric Acid”, bring volume to 900 ml), aad 100 ml electrode buffer to give 9:1 ratio, 1.0 M NaOH to pH 8.3 pH 22°C 7.0 (€861) £2 SWATOA “TWNYNO! NYA NVOININY 8. 0.039 M LiOH, 0.263 M Boric Acid; 8.0 1.64 g LiOH:H,O, 16.23 g Boric Acid” 9, 0.065 M L-Histidine free pis ca. Sc] 0.015 to 0.016 M Citric A 10.09 g L-Histidine free . Citric Acid? to pH 5.7 (= ca. 2.9 to 3.1 10. 0.180 M Tris, 0.004 M EDTA, 0.100 M 8.6 Boric Acid; 21.80 g Tris*, 1.52 g EDTA tetrasodium salt dihydrate, 6.18 g Boric Acid”, Boric Acid to pH 8.6 11. 0.400 M Citric Acid, trisodium salt; 7.0 117.64 g Citric Acid, trisodium salt 2 Syiree. 1.0 M HCI to pH 7.0 0.042 M Tris, 0.007 M Citric Acid; 0.004 M LiOH, 0.025 M Boric acid; 5.04 g Tris*, 1.25 g Citric Acid”, 0.16 g LiOH:H,O, 1.56 g Boric Acid”, 1.0 M HCI to pH 7.6 0.009 M L-Histidine, 0.002 M Citric Acid; dilute 140 ml of electrode buffer to | liter 0.045 M Tris, 0.001 M EDTA, 0.025 M Boric Acid; dilute 250 ml of electrode buffer to | liter 0.005 M Histidine-HCI; 1.05 g L-Histidine-HC! monohydrate, 1.0 M NaOH to pH 7.0 “Trizma-base (Sigma T1503; Sigma eh works rfl Ss oe is much less expens PA mounts of Citric Acid and Boric A re give ve). anhydrous free acid per liter. “Check pH of s sion after mixing; at of Tris buffers ty chatae with dilutio dit are from hal (198 1a). SNY34 40 SISINOHdOYLIIT3 139 HOYWIS “IW 13 SLNOS 3G 12 AMERICAN FERN JOURNAL: VOLUME 73 (1983) known, however, that multiple forms of a given enzyme may be coded by different alleles at a single locus (allozymes) or by genes at more than one locus (isozymes). Furthermore, many standardly assayed plant enzymes have isozymes located in two or more subcellular compartments (e.g., the cytosolic and chloroplastic isozymes of PGI and PGM discussed by Gottlieb, 1981b, 1982—for interpretation of enzyme symbols see Table 2). When found in pteridophytes, multiple, subcellularly com- partmentalized isozymes should not be misinterpreted as products of duplicated loci resulting from polyploidy. According to Chapman et al. (1979), attempts to determine the amount of heterozygosity per locus in homosporous pteridophytes are complicated by recombi- nation between homoeologous duplicated loci. An obvious prerequisite to recombina- tion of this kind is the actual presence of such loci. Gastony and Gottlieb (1982) developed a means of demonstrating whether duplicated loci are, in fact, present in pteridophytes. By electrophoretically analyzing sporophytes taken from nature and individual gametophytes grown from spores of these sporophytes, segregational analysis of sporophytic enzyme banding patterns can be conducted. This permits genetic interpretation of parental sporophytic enzyme phenotypes without the time- consuming crossing programs required by total reliance on sporophytic tissue. Use of this methodology enables the investigator to determine whether apparent heterozygosity of sporophytes is coded by alleles at a single locus or by genes at duplicated (homoeologous) loci. Our application of this methodology to several fern genera (e.g., Athyrium, Bommeria, and Pellaea) has demonstrated that the genetic variability observed in these taxa results from allelic diversity and segregation at single, not duplicated, loci. MATERIALS Living sporophytes of Athyrium filix-femina, Bommeria ehrenbergiana, B. hispida, B. pedata, B. subpaleacea, Botrychium virginianum, Ceratopteris thalictroides, Cystopteris bulbifera, C. dickieana, C. fragilis, C. laurentiana, C. protrusa, C. reevesiana, C. tennesseensis, C. tenuis, Isoétes butleri, I. engelmannii, Lygodium japonicum, Nephrolepis exaltata, Ophioglossum engelmannii, Pellaea andromedi- folia, P. atropurpurea, P. glabella, Polypodium polypodioides, P. virginianum, Polystichum acrostichoides, Pteridium aquilinum, Woodsia obtusa, and W. oregana were maintained in greenhouse culture and utilized in this investigation. Living gametophytes representing the species of Athyrium, Bommeria, Cystopteris, Pellaea and Preridium listed above were cultured on nutrient agar (Gastony & Haufler, 1976) and also provided material for this study. GRINDING BUFFER SOLUTIONS We routinely utilize modifications of either the phosphate grinding buffer—poly- vinylpyrrolidone (PVP) solution employed by Mitton et al. (1979), the Tris—maleate grinding buffer-PVP solution of Soltis et al. ( 1980), or the Tris-HCl grinding buffer-PVP solution of Gottlieb (1981a) with PVP substituted for polyvinylpoly- pyrrolidone (PVPP), as described below. Recipes for preparing these grinding buffer solutions are provided below; molarity or percent volume values are provided and D. E. SOLTIS ET AL.: STARCH GEL ELECTROPHORESIS OF FERNS 13 gram or milliliter amounts required to prepare 25 ml of buffer solution are given in parentheses. Phosphate grinding buffer-PVP solution.—0.029 M (0.28 g) sodium tetraborate, 0.017 M (0.08 g) sodium metabisulfite, 0.20 M (1.0 g) L-ascorbic acid sodium salt, 0.016 M (0.07 g) diethyldithiocar- bamic acid sodium salt, 4% w/v (1.0 g) PVP average molecular weight 40,000 (Sigma PVP 40T). or 36-40% w/v (9.0-10.0 g) PVP average molecular weight 10,000. Dissolve gram amounts in 25 ml of 0.10 M phosphate buffer pH 7.5 (to make 100 ml of phosphate buffer dissolve 1.36 g KH»PO, in H,O, add 9.0 ml IM NaOH, and bring volume to 100 ml with H,O) and then add 0.25 ml (1%) 2-mercaptoethanol. Tris—maleate grinding buffer-PVP solution.—0.20 M (1.91 g) sodium tetraborate, 0.02 M (0.095 sodium metabisulfite, 0.25 M (1.24 g) L-ascorbic acid sodium salt, 0.026 M (0.113 g) diethyldithiocarbamic acid sodium salt, 0.10 M (0.29 g) maleic acid, 0.10 M (0.30 g) Tris. 4% w/v (1.0 g) PVP average molecular weight 40,000 or 32-40% w/v (8.0-10.0 g) PVP average molecular weight 10,000. For 25 ml of buffer, dissolve amounts indicated in 19 ml distilled water; mix thoroughly and crush out lumps; adjust to pH 7.5 with 1.0 M HCI; add 0.025 ml (0.1%) 2-mercaptoethanol: add H,O to 25 ml. Originally (when we utilized PVP 10,000) it was necessary to allow the PVP to hydrate overnight before using the grinding buffer solution. When employing PVP 40,000, it is possible to prepare the grinding buffer solution immediately prior : sample preparation i stirring the PVP into solution. Tris-HCl grinding buffer-PVP solution.—0.1% v/v (0.025 ml) 2-mercaptoethanol, 0.001 M (0.010 g) EDTA (tetrasodium salt), jae M (0.019 g) potassium chloride, 0.010 M (0.050 g) magnesium chloride hexahydrate, 4 or 20% w/v (1 or 5 g) PVP 40,000, 25 ml 0.10 M Tris-HCl! buffer, pH 7.5. Stir the PVP into solution or allow it to hydrate in the buffer overnight. TABLE 2. GEL AND ELECTRODE BUFFER SYSTEMS THAT WE HAVE FOUND TO YIELD THE BEST BANDING IN FERNS WE HAVE ASSAYED. Enzyme Symbol Gel and Electrode Buffer System from Table | Acid phosphatase APH 6,7 onitase ACN 15,9 Aldolase ALD 1-5, 7, 10, 11 Aspartate aminotransferase AAT (or GOT) 6, 7,8 Catalase CAT 4, 7, $,.10 Esterase (Colorimetric) EST 6.7 Esterase (Fluorescent) FE 8 Fructose- 1 ,6-diphosphatas Fl ,6DP 1, 11 Glucose-6-phosphate deyropenae G6PDH 4, 5, 6,7 Glutamate dehydro GDH 3, 5,7 Glyceraldehyde-3- tl drogenase G3PDH 1, 11 Hexokinase HK 2, 35, 31 8 Isocitrate dehydrogenase IDH 1, 2,3, 455 Leucine aminopeptidase LAP 3, 7,8 Malate dehy n MDH 1, 4,5, 9 Malic enzyme ME 7,1 Peroxidase PER re Phosphoglucoisomerase PGI 5, 6,7 Phosphoglucomutase PGM 7.0. 1.5 6-Phosphogluconate dehydrogenase -6-PGD 1, 2, 4,5 Shikimate dehydrogenase SkDH 12, 4,3, 0 Triosephosphate isomerase TPI 2,5, 6, 7,8 14 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Comparison of these three grinding buffer solutions indicates that for most enzymes the staining results are highly comparable. For some enzymes, however, the Tris-HCI-PVP solution seems to improve enzyme band clarity (e.g., PGI, PGM, LAP), while for other enzymes the reverse is true. In Pellaea andromedifolia, for example, PGI banding was sharp with the Tris-HCI-PVP solution but was inhibited by the presence of ascorbic acid and sodium tetraborate in the Tris—maleate-PVP buffer. In Athyrium filix-femina, however, the Tris—maleate—PVP solution gives superior banding for PGI and one more observable EST locus when compared to the Tris-HCI-PVP solution. The phosphate-PVP solution often result- ed in reduced APH activity when compared to the other two grinding buffer solutions. In working with species of Asplenium, Werth et al. (1982) used a method of sample preparation that uses caffeine, but not PVP. Following this technique, equal weights of tissue and caffeine were ground in a 0.1 M HEPES pH 7.0 buffer with 0.2% 2-mercaptoethanol and 0.5% sodium metabisulfite to produce a slurry. We found that for virtually all enzyme systems investigated, the method of Werth et al. provided results roughly comparable to those obtained with grinding buffers contain- ing PVP. The presence of 0.5% sodium metabisulfite in the grinding buffer appears to be of great importance. When this ingredient is omitted from the buffer, activity is noticeably reduced or almost totally lost for many enzymes. Significantly, the full complement of PGI enzyme bands obtained with PVP was not expressed in most of the fern taxa investigated when sodium metabifsulfite was omitted. These results are in agreement with similar observations of Werth et al. (1982). It should be emphasized that to obtain the best results, the appropriate amount of PVP depends upon the taxon under investigation and in part upon the molecular weight of the PVP employed. Soltis et al. (1980) utilized a grade of PVP with an average molecular weight of 10,000 (although this is not stated in their report) and incorporated 40% w/v PVP (4 g PVP in 10 ml of Tris—maleate grinding buffer). At present, we routinely employ a grade of PVP with an average molecular weight of 40,000. Both molecular weights of PVP are suitable, but less PVP 40,000 is required than PVP 10,000. The amount of PVP used is an important consideration because, as reported by Soltis et al. (1980), use of excessive amounts of PVP in the preparation of grinding buffer-PVP solutions frequently results in a decrease or complete loss of enzyme activity. Although PVPP is effective with a wide range of — angiosperms, its substitution for PVP failed to produce banding for PGI in Pellaea andromedifolia whether the PVPP was hydrated in the grinding buffer or added — directly to the leaf tissue during grinding. This discussion stresses the importance of selecting optimal grinding buffer components and procedures for the taxon under investigation. As noted above, certain components of these buffers may inhibit band expression, whereas in other — | —. the full complement of components may be required to obtain clear banding. — : bt ave found that LAP activity is greater if sodium ascorbate, sodium tetraborate, sodium metabisulfite are eliminated from the grinding buffer. On the other — be very noticeably reduced when a buffer solution incorporating only PVP is hand, activity for some enzymes, (e.g., SDH, G6PDH. MDH. CAT. 6PGD) may D. £. SOLTIS ET AL: STARCH GEL ELECTROPHORESIS OF FERNS 15 utilized. A further complication in some taxa is that when a grinding buffer lacking 2-mercaptoethanol is employed, artifactual (“ghost”) bands proliferate for some enzymes and can hinder accurate interpretation of the band patterns. Given these examples, we strongly encourage comparative experimentation with grinding buffer solutions when initiating electrophoretic studies. SAMPLE PREPARATION AND ELECTROPHORESIS Leaf samples to be analyzed electrophoretically were taken from mature sporo- phytes. The methods discussed herein, however, are applicable to gametophytic as well as sporophytic tissue. In our original electrophoretic analyses of ferns, leaf samples were prepared according to the method of Soltis et al. (1980) in which small amounts of leaf tissue are ground under liquid nitrogen with a porcelain mortar and pestle until a fine dry powder is obtained. The powder is quickly mixed with grinding buffer to form a thick slurry. This slurry is absorbed directly into thick paper wicks (made from Whatmann 3 MM chromatography paper or some other suitable wick paper). It should be emphasized that the use of the grinding buffer-PVP solutions alone (outlined above), without the use of liquid nitrogen in the preparation of samples, provide clear enzyme banding in all of the fern taxa that we have examined so far. Therefore, at present we utilize a simplified method of sample preparation in which liquid nitrogen has been eliminated. In standardizing our grinding procedure by consistently homogenizing 100 mg of leaf material in 0.5 ml of grinding buffer, we obtained roughly equivalent staining intensity for all samples, which facilitates comparing individuals and scoring gels. If small amounts of tissue are being ground, it may be possible to substitute 43 mm7? plastic weigh boats for mortars and glass or plexiglass rods for pestles. As discussed by Gastony and Gottlieb (1982), it is possible to obtain enzyme banding from single gametophytes. Their technique involves smearing individual gametophytes onto wicks pre-moistened with extraction buffer-PVP solution. With all of the described grinding/extraction procedures, the amount of grinding buffer and the type and size of wicks should be tailored to the taxon and tissues under investigation. Once the plant material has been absorbed into wicks, the wicks are inserted into a vertical slit in the starch gel and subjected to horizontal electrophoresis at 4°C. Band definition may be improved by removing wicks from the gel after the first 10-20 minutes of electrophoresis. It is important to maintain electrical contact across the slit in the gel. We routinely place some sort of spacer (e.g., strips of Whatmann paper, a plastic straw, a thin piece of plexiglass) at the cathodal end of the gel to compress the gel slightly and to insure that the slit does not open. To permit comparison of banding patterns between gels, it is important to maintain a constant starch concentration throughout a study. We have experimented with a variety of starch concentrations throughout the practical range of 11.5 to 15% and have found concentrations of 12 to 13.2% most suitable, although in some instances alternative concentrations may improve band resolution. Connaught, Fish- er, and Sigma starch have been used and all provide clear enzyme banding, provided appropriate gel and electrode buffer systems and staining schedules are utilized. 16 AMERICAN FERN JOURNAL: VOLUME 73 (1983) In order to facilitate determining the location of the anodal front, it is useful to insert a wick bearing the marker dye Bromphenol Blue (Sulfone Form, Sigma BO126; 0.04% w/v in 95% ethanol). Depending on the gel and electrode buffer system employed, as well as the taxon under investigation, the enzyme bands will migrate varying distances behind the Bromphenol Blue marker. Optimal running times for each enzyme system will therefore have to be determined empirically. GEL AND ELECTRODE BUFFER RECIPES We have found that standard gel and electrode buffer systems, such as those of Shaw and Prasad (1970), and Brewer (1970), often yield unsatisfactory results with fern tissue. We have determined gel and electrode buffer systems that provide clear banding for 22 enzymes in ferns. Recipes for the gel and electrode buffers we most commonly employ are provided in Table /. Several of the gel and electrode buffer systems that we employ are modifications of those of Shaw and Prasad (1970). It should be noted, therefore, that errors are present in several of the recipes they provided. For example, in Shaw and Prasad buffer system I (see page 299, Table | of their report), utilization of 16.35 g of Tris yields an electrode buffer with a molarity of 0.135 (see present report, Table 1, electrode buffers 2 and 3), rather than 0.155 as reported by Shaw and Prasad. Similarly, use of 27.0 g of Tris in the preparation of the electrode buffer of Shaw and Prasad system XII yields a molarity of not 0.233 but 0.223 (see present report, Table 1, electrode buffers 4 and 5). Furthermore, use of the electrode buffer recipe of Shaw and Prasad system XII yields a buffer of approximately pH 5.5, which differs significantly from the estimate of pH 7.0 listed by Shaw and Prasad. Use of the amounts of Tris and citric acid given for system 4 in Table / of the present report will also yield an electrode buffer of pH 5.5; sodium hydroxide is then added to adjust the buffer to pH 7.5. Of the 11 gel and electrode buffer systems for which recipes are provided in Table /, some clearly are preferable to others for certain of the 22 enzymes for which we have obtained sharp banding. For example, best enzyme banding for MDH is obtained when system 1, 4, 5, or 9 is used. Which system works best is dependent, in part, on the taxon under investigation. We have found. for example, that the best systems for Athyrium and Cystopteris differ somewhat from those that supply clear enzyme banding for Bommeria. The systems provided here, however, should provide — an excellent framework for conducting electrophoretic analyses of most ferns. The — gel and electrode buffer systems that we have found to yield the clearest banding for each of the 22 enzymes with which we have experimented are listed in Table 2. STANDARD STAINING SCHEDULES During the course of our electrophoretic investigations of ferns, we found that — many published staining schedules, such as those of Shaw and Prasad (1970), — yielded unsatisfactory results. Enzyme banding often can be improved dramatically when these standard recipes are modified. One very important modification that we employ involves increasing the pH of the buffer in staining schedules for ALD, GDH, G6PDH, HK, IDH, MDH, ME, PGI, PGM, and 6PGD. Tris-HCI staining buffers of pH 7.0 or 7.1 typically are used in staining for these enzymes (Shaw & D. E. SOLTIS ET AL.: STARCH GEL ELECTROPHORESIS OF FERNS 17 Prasad, 1970). We have found, however, that in ferns the pH optima for these enzymes are in the 8.0-8.5 range, and we therefore use a Tris—HC] staining buffer of pH 8.0-8.5 for them. For some enzymes, this simple modification improves Staining dramatically. For example, standard MDH staining recipes use a buffer of pH 7.0. Application of such a protocol to ferns, however, often results in very little or inconsistent staining. Well stained bands are obtained consistently for MDH when a Tris-HCl staining buffer of pH 8.0-8.5 is used (provided an appropriate gel and electrode buffer system is employed; see Table 2). It also should be noted that we have consistently been unable to obtain observable activity with fern leaf tissue for some enzymes, despite considerable experimenta- tion. These include Alcohol dehydrogenase, Alkaline phosphatase, a-Glycerophos- phate dehydrogenase and Lactate dehydrogenase. There are several enzymes for which we have obtained observable activity with ferns, but have not yet obtained clear banding, such as Diaphorase and Peptidase, and these are therefore not included in this report. In addition, there are a number of enzyme staining schedules that we have not yet attempted. We hope that the staining schedules provided below will stimulate additional experimentation in this regard. The staining schedules that we utilize for the 22 enzymes for which we obtain clear banding are provided below in alphabetical order. The final volume for all stain recipes is 100 ml. Depending on gel size and staining container capacity, it may be possible to assay two gel slices simultaneously with each of the following 22 solutions. Alternatively, when assaying single gel slices, the recipes may be halved to 50 ml final volume. Readers are encouraged to consult Enzyme Nomenclature /978 (International Union of Biochemistry, Nomenclature Committee, 1979 [here- after referred to as I.U.B., 1979]) for details of enzyme specificity and Gottlieb (1981b) for a recent review of the electrophoretic technique and its application to plant populations. It should be noted that enzymes such as peroxidases, esterases, and acid phosphatases operate on a large number of substrates in vitro: therefore, standard staining protocols for these enzymes result in the staining of various numbers of isozymes whose homologies are not apparent. Although the utility of these isozymes in between-species comparisons is therefore limited, their banding patterns can be useful in assessing variability within populations. Several different organisms serve as the source for commercial preparations of Glucose-6-phosphate dehydrogenase. This enzyme typically is specific for NADP, but some forms are capable of utilizing either NADP or NAD. It should be emphasized that NADP is much more expensive than NAD, hence selecting a Glucose-6-phosphate dehydrogenase type that can use NAD will save considerable money. Therefore, in those recipes that call for added Glucose-6-phosphate dehydro- genase, the notation NAD(P)* reflects this option. Acid phosphatase (EC 3.1.3.2) 0.05 M sodium acetate buffer, pH 5.0 100 ml 1.0 M MgCl, 0.5 mi @-naphthyl acid phosphate, sodium salt 100 mg Fast garnet GBC salt aan We have obtained results with buffer molarities ranging from 0.05 M to 0.2 M and within a pH range of 5.0 to 6.0. Stain at room temperature; a modification of 18 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Scandalios (1969). Gottlieb (1981b) noted that plants may have a dozen or more acid phosphatases. We consistently have observed one very intense zone of acid phosphatase activity, with several fainter bands occasionally evident. Aconitase (EC 4.2.1.3.; = Aconitate hydratase in I.U.B., 1979) 1.0 M Tris-HCI buffer, pH 8.5 10 ml H,0 90 ml cis-aconitic acid 70 mg Isocitrate (= Isocitric) dehydrogenase 7 units 1.0 M MgCl, 1 ml NADP 10 mg MTT 5 mg PMS Stain in the dark at 30°C; a modification of Shaw and Prasad (1970). Aldolase (EC 4.1.2.13; = Fructose-biphosphate aldolase in I.U.B., 1979) 1.0 M Tris-HCI buffer, pH 8.5 10m HO 90 ml Fructose-1 ,6-diphosphate, tetra(cyclohexylammonium) salt 500 mg or trisodium salt 200 mg 1.0 M arsenic acid, sodium salt 1 ml Glyceraldehyde-3-phosphate dehydrogenase 200 units NAD MTT 20 mg PMS Stain at room temperature; a modification of Shaw and Prasad (1970). Aspartate aminotransferase (EC 2.6.1.1.; = Glutamate oxaloacetate trans- aminase 1.0 M Tris-HCI buffer, pH 8.0 10 ml HO 90 ml L-aspartic acid 100 mg a-ketoglutaric acid 100 mg Adjust pH to 8.0 with 1.0 M NaOH as necessary, then add: Pyridoxal-5’-phosphate 5 mg Fast blue BB salt 100 mg Stain in the dark at room temperature; a modification of Gottlieb (1973a) and Selander et al. (1971, appendix). Our results show that this enzyme is very sensitive to the pH of the staining buffer; very little staining is observed at pH 7.0 or at pH 9.0. The number of loci coding for proteins that are capable of this aminotransferase reaction is known to vary in plants. Although we typically encounter a single locus, Gottlieb (1981b) has reported the likelihood of specific subcellular localizations for the several dimeric isozymes. Catalase (EC 1.11.1.6) 3% © KIQV2 we 0.1 M phosphate buffer, pH 7.0 10 ml 0.06 M Na,S,03°5H O aes : 7 ml D. E. SOLIS ET AL.: STARCH GEL ELECTROPHORESIS OF FERNS 19 Incubate in this solution at room temperature for 1-2(30) minutes: pour off the solution, rinse several times with distilled water, then add: 0.09 M KI 50 ml H,O ml Catalase activity will appear as white bands on a dark blue background: a modification of Shaw and Prasad (1970). For some of the buffer systems in Table /, it may be necessary to add several drops (approximately 2 ml) of glacial acetic acid in order to induce staining. An alternative method is to substitute 50 ml 0.05 M sodium acetate buffer (pH 5.0) for the 50 ml H;0. The gel may turn completely blue in a very short time. Be prepared either to photograph the gel or score it while staining. One locus has been observed in ferns. The protein is reported to be tetrameric (Scandalios, 1969). Esterase (Colorimetric; EC 3.1.1.-) a-naphthy! acetate 40 mg 8-naphthy! acetate 40 mg dissolved in acetone 2 mi 1.0M phosphate buffer, pH 6.0 10 ml H,0 90 ml Fast blue RR salt 100 mg Stain at room temperature; a modification of Gottlieb (1974). Esterase (Fluorescent; EC 3.1.1.-) 4-methylumbelliferyl acetate 42 mg dissolved in acetone 25m 1.0 M sodium acetate buffer, pH 5.0 18 ml H,0 Stain in the dark at room temperature and observe under long-wave ultraviolet light; Staining schedule of Mitton et al. (1979). Observe immediately after staining (bands fade quickly). Caution should be exercised in scoring gels because flavonoid bands” may also be visualized under UV light. To determine whether flavonoid “bands” are present, the gel slice should be observed under UV light prior to Staining, Fructose-1,6-diphosphatase (EC 3.1.3.11; =Fructose-biphosphatase in LUB., 1979) 1.0 M Tris-HCI buffer, pH 8.0 Aer H,0 90 ml Fructose- | ,6-diphosphate, tetra(cyclohexylammonium) salt 250 mg or trisodium salt in ae 10M MgCl, ss Grosphoglucoisomerase paced UCose-6-phosphate dehydrogenase seat NAD(P)* “i i ie MTT 5 mg PMs ime Stain in the dark at 30° C; a modification of Shaw and Prasad (1970). 20 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Glucose-6-phosphate eee serene (EC 1.1.1.49) 1.0 M Tris-HCI! buffer, pH 8.0 10 ml H,0 90 ml Glucose-6-phosphate, disodium salt 100 mg NADP 20 mg MTT (or NBT) 10 mg PMS mg Stain in the dark at 37° C; a modification of Shaw and Prasad (1970). Glutamate pe lscceroe! (EC 1.4.1.2) 1.0 M Tris-HC1 buffer, pH 8 10 ml HO 70 ml 1.0 M L-glutamic acid, pH 8.0 (use free acid or monosodium salt and add NaOH to pH 8.0) 20 ml NAD 20 mg MTT (or NBT) 10 mg PMS 2 mg Stain in the dark at room temperature; a modification of Gottlieb (1973b) and of Shaw and Prasad (1970). Glyceraldehyde-3-phosphate ee (EC 1.2.1.12;: = Glyceraldehyde- phosphate dehydrogenase in I.U. B., 1979) 1.0 M Tris-HCI buffer, pH 8.0 10 ml H,0 90 ml Fructose-1,6-diphosphate, trisodium salt 100 mg Aldolase 10 units Incubate above mixture approx. 30 min. at 30-37° C, then add: 1.0 M arsenic acid, sodium salt 1 ml NAD 10 mg MTT 5 mg PMS | mg Stain in the dark at 30° C; a modification of Shaw and Prasad (1970). Hexokinase (EC 2.7.1 1) 1.0 M Tris-HCI buffer, pH 8.5 10 ml HO 90 ml Glucose 90 mg 1.0 M MgCl, 5 ml EDTA, tetrasodium salt, dihydrate 40 mg NAD(P)* 10 mg MTT (or NBT) 15 mg Pee Phosphate dehydrogenase 40 units ATP sa Stain in the dark at room temperature; a modification of Shaw and Prasad (1970). Isocitrate dehydrogenase [NADP* ne M Tris-HC] buffer, pH 8 i ie a < 30 10 ml ee | Isocitric acid, trisodium salt a wd 10M MgCl, pa NADP D. E. SOLTIS ET AL.: STARCH GEL ELECTROPHORESIS OF FERNS 21 MTT (or NBT) 15 mg PMS 2 mg Depending on the taxon being analyzed, results have been obtained by using buffers ranging from pH 7.2 to pH 8.5. Stain in the dark at room temperature: a modification of Shaw and Prasad (1970). Leucine aminopeptidase (EC 3.4.11.-) L-leucine-B-naphthylamide (free base or acid salt) 20 mg dissolved in dimethyl formamide 5 ml 1.0 M phosphate buffer, pH 6.0 10 ml H,0 90 ml Black K salt or fast black K salt 50 mg Stain in the dark at room temperature; a modification of Gottlieb (1973c). This enzyme has broad activity; it can cleave a number of N-terminal amino acids. Although commonly referred to as Leucine aminopeptidase (“LAP”), it is more precisely referred to as Aminopeptidase. There are a number of aminopeptidases, some of which are specific for certain N-terminal amino acids. Aminopeptidase (cytosol; EC 3.4.11.1) is activated by heavy metals, whereas Aminopeptidase (microsomal; EC 3.4.11.2) is not activated by heavy metals (I.U.B., 1979). It may be necessary to modify the pH of the stain in order to increase staining intensity. One isozyme is usually observed and a second is occasionally apparent; both are monomeric. Malate dehydrogenase (EC 1:4:1.37) 8.5 1.0 M Tris-HC1 buffer, pH 8.0 or 10 ml 2.0 M DL-malic acid, pH 8.0 (add NaOH to pH 8.0) 10 ml HO 80 ml NAD 10 mg MTT (or NBT) 10 mg PMS 2 Stain in the dark at room temperature; a modification of Shaw and Prasad (1970). Some investigators add 38 mg EDTA (tetrasodium salt, dihydrate) to this recipe. We use 2.0 M DL-malic acid rather than 1.0 M L-malic acid as most standard staining schedules require because DL-malic acid is much less expensive than purified L-malic acid, which is the actual substrate. There are at least four malate NAD* dehydrogenases reported (I.U.B., 1979). The one most frequently reported in routine electrophoresis is L-Malate: NAD+ oxidoreductase (EC 1.1.1.37), of which at least two putative isozymes have been observed. The protein is dimeric; subcellu- lar compartmentalization of the isozymes is discussed by Gottlieb (1981b). Malic enzyme (EC 1.1.1.40; = Malate dehydrogenase [Oxaloacetate-decar- boxylating, NADP*] in I.U.B., 1979) 8.5 1.0 M Tris-HCI buffer, pH 8.0 or 8. 10 mi 20 80 ml 2.0 M DL-malic acid, pH 8.0 (prepare as in Malate dehydrogenase) 10 ml 10M MgCl, 2 ml NADP 20 mg MTT (or NBT) 20 mg PMS 2 mg AMERICAN FERN JOURNAL: VOLUME 73 (1983) Stain in the dark at room temperature; a modification of Richmond (1972 and pers. comm.). Peroxidase (EC 1.11.1.7) 3-amino-9-ethy! carbazole 65 mg dissolved in dimethyl formamide 5 ml 0.05 M sodium acetate buffer, pH 5.0 95 ml 0.1 M CaCl, 2 ml 2 ml Incubate the gel in a refrigerator until the bands appear (30-60 min); a modification of Shaw and Prasad (1970) and of Gottlieb (1973b). Phosphoglucoisomerase (EC 5.3.1.9; = Glucosephosphate isomerase in I.U.B., 1.0 M Tris-HC1 buffer, pH 8.0 10 ml > 90 ml 1.0 M MgCl, 1 ml Fructose-6-phosphate, disodium salt 30 mg Glucose-6-phosphate dehydrogenase 40 units NAD(P)* MTT (or NBT) 20 mg PMS 2 mg Stain in the dark at 37°C or at room temperature; a modification of Shaw and Prasad (1970). This dimeric enzyme has numerous synonyms (I.U.B., 1979) including: Phosphohexose isomerase, Phosphohexomutase, Oxoisomerase, Hexose phosphate isomerase, Glucose-6-phosphate isomerase, Phosphosaccharomutase, and Phosphohexoisomerase. Cytosolic and chloroplastic isozymes should be expected (Gottlieb, 1981b, 1982). Phosphoglucomutase (EC eit) 1.0 M Tris-HCI buffer, pH 8.0 or 8.5 10 ml H,0 90 ml 1.0 M MgCl, 2 ml Glucose-1-phosphate, disodium (Sigma G7000) or dipotassium (Sigma G6875) salt 100 mg 1.7 x 104M «-D-glucose-1,6-diphosphate, tetra(cyclohexylammonium) salt 5 mi Glucose-6-phosphate dehydrogenase 40 units NAD(P)* 10 mg MTT (or NBT) 20 mg PMS ; : 2 mg A modification of Shaw and Prasad (1970). An equally suitable alternate recipe which eliminates the need to purchase a-D-glucose- 1 ,6-diphosphate is: - M Tris-HC1 buffer, pH 8.0 oe ; l Glucose- |-phosphate, disodium salt Sj Se€-6-phosphate dehydrogenase peste NAD(P)* aD 10 mg in 10 mg 2 mg Stain in the dark at 37°C or at room temperature. TG EL PADS SE D. E. SOLTIS ET AL.: STARCH GEL ELECTROPHORESIS OF FERNS 23 6-Phosphogluconate dehydrogenase (EC 1.1.1.44) 1.0 M Tris-HC1 buffer, pH 8.0 10 ml 2 90 ml 6-phosphogluconic acid, barium salt 40 mg 1.0 M MgCl, 2 ml NADP 10 mg MTT 10 mg PMS 2 mg Stain in the dark at room temperature or at 37°C: a modification of Shaw and Prasad (1970). Shikimate dehydrogenase (EC 1.1.1.25) 1.0 M Tris—HC1 buffer, pH 8.5 10 ml H,0 90 ml Shikimic acid 100 mg NADP 10 mg MTT (or NBT) 20 mg PMS 2 mg Stain in the dark at room temperature or at 37°C. Triosephosphate isomerase (EC 5.3.1.1) 1.0 M Tris-HCI buffer, pH 8.0 10 ml H,0 90 ml Dihydroxyacetone phosphate, lithium salt 10 mg EDTA, tetrasodium salt, dihydrate 38 mg NAD 30 mg MTT 10 mg PMS 2 mg Arsenic acid, sodium salt 460 mg Glyceraldehyde-3-phosphate dehydrogenase 300 units Stain in the dark at 37°C; Gottlieb (pers. comm.). An alternative and much less expensive method of staining for TPI follows (also see agarose recipe): 1.0 Tris-HCI buffer, pH 8.0 —— H,0 90 ml DL-a-glycerophosphate 200mg Pyruvic acid, sodium salt 100 mg a-Glycerophosphate dehydrogenase 100 units Lactate dehydrogenase 100 ‘units Incubate the above solution for 2 hours at 30-37°C. At the end of the incubation period, inactivate the enzymes by adjusting the solution to pH 2.0 with 1.0 M HCI (taking care not to go below pH 2.0) and then re-adjust to pH 8.0 with 1.0 M NaOH. Add: 1.0 M arsenic acid, sodium salt | ml Glyceraldehyde-3-phosphate dehydrogenase 100 units NAD 20 m MTT 5 mg PMS mg Stain in the dark at 30°C. The bands will be dark blue on a light blue background. Readers who follow this protocol must first show the lack of a-Glycerophosphate dehydrogenase activity in their taxa. This can be done simply by staining a gel slice with: 24 AMERICAN FERN JOURNAL: VOLUME 73 (1983) 1.0 M Tris-HC1 buffer, pH 8.0 10 ml 90 ml DL-a-glycerophosphate 200 mg NAD 20 mg MTT 5 mg PMS | mg AGAROSE STAINING SCHEDULES Several of the reagents commonly required in enzyme electrophoresis, such as NADP, are expensive. Costs may be reduced by using staining schedules employing agarose (modified from Mitton et al., 1979 and Gaines, pers. comm.), which require much smaller quantities of most ingredients than do standard staining schedules but yield comparable results. Agarose staining schedules are provided below for ALD, G6PDH, GDH, IDH, MDH, PGI, PGM, 6-PGD, SkDH, and TPI. The following general procedure is employed to stain one starch gel slice using the agarose technique: dissolve all ingredients required for enzyme staining (see below) in 6 ml of Tris-HC1 buffer; in a second flask mix 0.06 g agarose (Agarose type II, Sigma A6877) with 6 ml distilled water (a 1% agarose solution) and bring this solution to a boil while stirring constantly; combine the two solutions and very quickly apply to surface of one gel slice (it may be necessary to increase ml amounts of buffer and 1% agarose depending on the size of the gel slice); stain all slices in the dark at room temperature. Aldolase (EC 4.1.2.13) PMS 1 mg MTT 5 mg NAD 10 mg Fructose-1 ,6-diphosphate, trisodium salt or tetra(cyclohexylammonium) salt 75 mg Arsenic aci 25 mg Glyceraldehyde-3-phosphate dehydrogenase 40 units 1 M Tris-HCI buffer, pH 8.5 6 ml 1% agarose 6 ml Glucose-6-phosphate dehydrogenase (EC 1.1.1.49) PMS I mg MTT 4 mg NADP 4 mg EDTA, tetrasodium salt, dihydrate 6 mg Glucose-6-phosphate, disodium salt 5 mg 0.1 M Tris-HCI buffer, pH 8.0 6 ml 1% agarose 6 ml Glutamate dehydrogenase (EC 1.4.1.2) PMS 1 mg MTT 4 mg NAD 3 mg L-glutamic acid, monosodium salt 100 mg 0.1 M Tris-HCI buffer, pH 8.0 6 ml 1% agarose D. E. SOLTIS ET AL.; STARCH GEL ELECTROPHORESIS OF FERNS 25 Isocitrate dehydrogenase [NADP* ] (EC 1.1.1.42) I mg MTT 4 mg NADP 4 mg 2 40 mg Isocitric acid, trisodium salt 10 mg 0.1 M Tris-HCl! buffer, pH 8.0 1% agarose 6 ml Malate dehydrogenase (EC 1.1.1.37) PMS 2 mg MTT 7 mg NAD 7 mg L-malic acid 16 mg 0.25 M Tris-HC1 buffer, pH 8.6 6 ml 1% agarose 6 ml Keep PMS and L-malic acid powders separate until adding the buffer. Phosphoglucoisomerase (EC 5.3.1.9) PMS | mg MTT 4 mg NAD(P)* | mg 0.1 M Tris-HCI buffer, pH 8.0 4 ml 10% MgCl, 1 ml 0.018 M fructose-6-phosphate, eine salt Glucose- _ = toe dehydrogena 10 units 1% agaro 6 ml Phosphoglucomutase (EC 2.7.5.1) PMS | mg MTT 5 mg NAD(P)* 4 mg 0.1 M Tris-HCI buffer, pH 8.0 iy 0.05 M glucose-1-phosphate, sala salt (Sigma G7000) 2.5 ml Glucose-6-phosphate dehydrogen 10 units 0.001 M fructose-1 ,6- diphaghae: "eounies salt 1 ml 10% MgCl, 1 ml 1% agarose 6 ml 6-Phosphogluconate dehydrogenase (EC 1.1.1.4.4) PMS I mg MTT 4 mg NADP 4 mg 6-phosphogluconic acid, barium salt 10 mg 10% MgCl, 0.5 ml 0.1M Te HCl buffer, pH 8.5 5 ml 1% agar 6 ml Dissolve the 6- -phosphogluconic acid barium salt in 1.0 ml of 0.1 M Tris—HC1 buffer (pH 8.5) approximately 15 minutes before mixing in other ingredients. 6 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Shikimate dehydrogenase (EC 1.1.1.25) PMS I mg MTT 4 mg 4 mg Shikimic acid 10 mg 0.1 M Tris-HC1 buffer, pH 8.0 6 ml 1% agarose 6 ml Triosephosphate isomerase (EC 5.3.1.1) PMS I mg MTT 4 mg NAD 10 mg EDTA, tetrasodium salt, dihydrate 10 mg Arsenic acid 150 mg Dihydroxyacetone phosphate 1.5 mg Glyceraldehyde-3-phosphate dehydrogenase 75 units 0.1 M Tris-HC1 buffer, pH 8.0 6 ml 1% agarose 6 ml GEL FIXATION AND DOCUMENTATION AAT and PER gel slices are fixed in 50% glycerol; all other gel slices are fixed in 50% ethanol. In order to keep a permanent record of results, we routinely photograph gel slices using Kodak Technical Pan Film 2415 following the exposure index and developing procedure recommended for high contrast. ACKNOWLEDGEMENTS D.E.S. and C.H.H. thank Jeff Atwood, Harriet Blanton, Leslie Gottlieb, James Hamrick, Terry Lastovicka, and Andrew Torres for laboratory assistance and helpful comments and Charles Werth for providing details of his sample preparation University of Kansas, and to D.C.D. from the Department of Biology of Indiana University, NSF Doctoral Dissertation Improvement Grant DEB-8117239, and the Society of the Sigma Xi are gratefully acknowledged. LITERATURE CITED Be One T. and R. J. JOLY. 1980. Genetics of allozyme variants in loblolly pine. J. Hered. aa G. J. 1970. An Introduction to Isozyme Techniques. Academic Press, New York. AN, R. H., E. J. KLEKOWSKI, Jr. and R. K. SELANDER. 1979. Homoeologous Cone eee and recombination in the fern Pteridium aquilinum. Science 204:1207—1209. mIVER, G. 1976. Chemotaxonomy and phytochemical ecology of bracken. J. Linn. Soc., Bot. 73:3 GASTONY, G. J. and L. D. GOTTLIEB. 1982. Evidence for genetic heterozygosity in a homosporous fern. Amer. J. Bot. 69:634-637. » and C. H. HAUFLER. 1976. Chromosomes and apomixis in the fern genus Bommeria (Gymnogrammaceae). Biotropica 8:1—1] 4, D. E. SOLTIS ET AL.:; STARCH GEL ELECTROPHORESIS OF FERNS 27 GOTTLIEB, L. D. 1973a. Genetic control of Glutamate oxaloacetate transaminase isozymes in the diploid plant Stephanomeria exigua and its allotetraploid derivative. Biochem. Genet. 9:97—107. 73b. Genetic differentiation, sympatric speciation, and the origin of a diploid species of Fass Amer. J. Bot. 60:545-553. . 1973c. Enzyme differentiation and phylogeny in Clarkia franciscana, C. rubicunda and C. amoena. Evolution 27:205—214. . 1974. Genetic confirmation of the origin of Clarkia lingulata. Evolution 28:244-250. . 1981a. Gene numbers in species of Astereae that have different chromosome numbers. Proc. Natl. Acad. Sci. USA 78:3726—3729. . 1981b. Electrophoretic evidence and plant populations. Progress in Si beaticphae sh 7:1-45. ——. 1982. Conservation and duplication of isozymes in plants. Science 216 380. INTERNATIONAL UNION OF BIOCHEMISTRY, NOMENCLATURE he: 1979. Enzyme Nomenclature 1978. Academic Press, New York. KELLEY, W. A. and R. P. ADAMS. 1977a. Preparation of extracts from Juniper leaves for electrophoresis. Anbeeaser reel 16:513-516 and R. P. AD 77b. Seasonal variation of isozymes in Juniperus scopulorum: systematic signa ied J. Bot. 64:1092—10 MITTON, J. B., Y. B. LINHART, J. L. HAMRICK, and J. + BECKMAN. 1977. Observations on the genetic structure kee mating system of ponderosa pine in the Colorado front range. Theor Appl. Genet. 51:5-13. INHART, K. B. STURGEON, and J. L. HAMRICK. 1979. Allozyme polymorphism detected 1 in mature needle tissue of ponderosa pine. J. Hered. 70:86-89. RICHMOND, R. C. 1972. Enzyme variability in the Drosophila willistoni group. I]. Amounts of variability in the superspecies, D. paulistorum. Genetics 70:87-112. SCANDALIOS, J. G. 1969. Genetic control of multiple molecular forms of enzymes in plants: a review. eae Genet. 3:37-79. SELANDER, R. K., M. H. SMITH, S. Y. YANG. W. E. JOHNSON, and J. B. GENTRY. 1971. IV. Biochemical polymorphism and systematics in the genus Peromyscus. I. Variation in the old-field mouse (Peromyscus polionotus). Studies in as VI. Univ. Texas Publ. 7103:49-90. SHAW, “ R. and R. PRASAD. 1970. Starch gel electrophoresis of enzymes—a compilation of recipes. ochem. ay 4:297-320. SOLTIS, 3 a ap HAUFLER, and G. J. GASTONY. 1980. gi ie enzyme variation in the fern genus Bonnet an analysis of nie Syst. Bot. 5:30- WERTH, C. R., . KARLIN, and S. I. GUTTMAN. 1982. ane extraction from phenolic- esata i tissues using ae Isozyme Bull. 15:139. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) SHORTER NOTES A NEW COMBINATION IN ASPLENIUM.—A new fern was recently added to the flora of Alabama with the naming of the backcross hybrid x Asplenosorus boydstonae Walter (Amer. Fern J. 72:62. 1982.). The type material was collected in Hale County, Alabama, in a locality famous for its fertile alloploid population of Scott’s Spleenwort, which arose through hybridization of the Walking Fern and the Ebony Spleenwort. The new hybrid has Scott’s Spleenwort and the Ebony Spleen- wort as its parents. If the view is held that the Walking Fern belongs in the genus Asplenium rather than Camptosorus, the “hybrid genus” X Asplenosorus is taxonom- ically superfluous. The new hybrid then needs to be placed in Asplenium and the epithet corrected in conformity with Art. 73.10 of the “International Code of Botanical Nomenclature”: Asplenium x boydstoniae (Walter) Short, comb. nov. * Asplenosorus boydstonae K. §. Walter, Amer. Fern J. 72:62. 1982. TYPE: Havana Glen, Hale Co., Alabama, Wagner & Walter 70011 (MICH not seen). John W. Short, 217 Cook St., Auburn, AL 36830. ADIANTUM FURCATUM AND MICROSTAPHYLA F URCATA.—The is- land of St. Helena, outside the Gulf of Guinea off the west coast of Africa, is not only famous because it was the residence in exile of Napoleon Bonaparte. It has also produced a fern, Microstaphyla bifurcata, which stands out of proportion when its diminutive size (ca. 10-20 cm) is compared with the long list of names it has been called. Since the establishment of the genus Microstaphyla in 1851, it has been placed in no fewer than nine genera! The last revisionary treatment of Microstaphyla was by Maxon (J. Washington Acad. Sci. 13:28-31. 1923), who included among the synonyms the name Adiantum furcatum L. f. In the “Index Fil icum,” Christensen (1905) accepted that epithet as Elaphoglossum furcatum (L. f.) Christ. That basionym also accounts for the citation Microstaphyla furcata (L. f.) Presl, which has been used by some authors, including Maxon, who have overlooked Presl’s explicit citation of Osmunda bifurcata Jacq. as the basionym of his combination. In any event, accepting A. furcatum as a synonym for the sole species of Microstaphyla is an error which originated when the younger Linnaeus included a Sously interpreted by the younger Linnaeus to be the same as his A. furcatum (Sp. nmeaenals 1881), a plant he reported as “in Cap. Bonae Spei et insula so : hunberg. Thouin.” Although Thunberg collected in the Cape and Thouin in Reunion, neither of them set foot on St. Helena, where Microstaphyla is Pe. Furthermore, A. furcatum is not an Adiantum at all, but is Asplenium ee (Lam.) C. Chr. (syn. A. rutaefolium (Berg.) Kunze). For this reason, . innaean epithet should be deleted as a synonym of M. bifurcata (Jacq.) Presl, correct name.—Luis Diego Gémez P., Museo Nacional de Costa Rica, Apartado AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) 29 NOTEWORTHY PTERIDOPHYTE RECORDS FOR NEBRASKA.— Field studies in the Niobrara River Valley of north-central Nebraska during the summer of 1982 yielded several significant finds. Field work was carried out in connection with a floristic survey of the 54,000-acre Niobrara Valley Preserve, owned by The Nature Conservancy, and in areas peripheral to the Preserve. Botrychium matricariifolium A. Br. was collected for the first time in Nebraska. In the Great Plains it is otherwise known only from Custer County, South Dakota (Brooks, R. E. Amer. Fern J. 70:91. 1980) and from Lac Qui Parle County, Minnesota (Atlas of the Flora of the Great Plains, lowa State Univ. Press. 1977). Three plants were observed in a juniper-oak woodland on the south side of the Niobrara River about 15 miles NNE of Johnstown, Brown County. The plants were growing in a sandy, partially shaded soil along the floodplain of the river close to an access road to a center-pivot irrigation pump along the river. Much of the area had been cleared for the road and for an irrigation pipe up a steep, north-facing slope. No additional plants were found during subsequent searches. The plants were photographed and, after the fronds senesced, collected (Freeman 15/7, KANU). Ophioglossum vulgatum var. pseudopodum (Blake) Farw. was first collected in Nebraska in 1913 by J. M. Bates in the vicinity of Kennedy, Cherry County. Clausen (Mem. Torrey Bot. Club 19:126. 1938) cited the specimen without infraspecific reference; Brooks (Amer. Fern J. 70:92. 1980) referred it to var. pseudopodum. A thriving population was found on the south side of the Niobrara River ca. 55 yds east of the Berry Bridge, 5 miles west and 3 miles south of Sparks, Cherry County. The plants covered an area of about 7 x 2 yds at the west end of a beaver pond. They were growing in wet, loamy soil at the edge of a paper birch forest along with Cicuta bulbifera, Onoclea sensibilis, and Thelypteris palustris. The area showed obvious signs of human manipulation, as a dike situated between the river and the west end of the pond was fitted with a pipe to supplement water in the pond during dry periods, possibly as part of a fish hatchery built many years ago. Photographs were taken and voucher specimens (Freeman & Churchill 1672) were distributed to various herbaria, including KANU, KSC, NEB, and NY. Dryopteris carthusiana (Villars) H. P. Fuchs was collected around the turn of the century in Brown, Cass, Cherry, Lancaster, and Thomas counties in Nebraska (A. J. Petrik-Ott, Beih. Nova Hedw. 61:141. 1979). Field studies conducted by a number of researchers in 1980 resulted in the discovery of one population along Little Cedar Creek Canyon in Cherry County (Sutherland, Boner, Lamp & Joern 5053, NEB; Sutherland & Harrison 5314, NEB). During the course of our survey, we found four populations, all in rich paper birch forests along spring-fed canyons on the south side of the river. Two populations were in Cherry County. One was photographed and collected along Little Cedar Creek Canyon, 5.2 miles south of Sparks (Freeman & Churchill 1200, KSC, NEB). A second was just east of Smith Falls, 3 miles east and 4 miles south of Sparks (Churchill & Brogie 12424, NEB). The latter was the largest of the populations. Two populations were found in Brown County. One was along Dutch Creek Canyon, 16 miles north of Ainsworth (Churchill 12300, NEB). The second was in Kantak Coulee, 3 miles west of Meadville (Churchill 12408, NEB). AMERICAN FERN JOURNAL: VOLUME 73 (1983) We wish to thank Dr. W. H. Wagner, Jr. and Ralph Brooks for verifying the specimen of Botrychium and Dr. T. M. Barkley and Ralph Brooks for reviewing the manuscript.—Craig C. Freeman, Division of Biology, Kansas State University, Manhattan, KS 66506 and Steven P. Churchill, Division of Biological Sciences, University of Kansas, Lawrence, KS 66045. SPREAD OF MARSILEA QUADRIFOLIA IN MCDONOUGH COUNTY, ILLINOIS.—Thirty-five years ago Myers found Marsilea quadrifolia L. in Spring Lake, McDonough County, Illinois. It is of interest to record how much this alien, potentially weedy plant has spread in that time. Myers (Amer. Fern J. 40:256. 1950) stated that during the years following 1947 Marsilea did not become abundant, but he did recognize that it “has persisted following partial draining of the lake, dredging, and a considerable increase in the water level when a higher dam was built” (Annotated Catalogue and Index for the Illinois Flora. Western Illinois Univ. Ser. Biol. Sci. 10:58. 1972). At present, the plant has not spread much in the lake and is still not abundant. Water from the lake flows downstream via Spring Creek for about 4.3 miles where it enters the LaMoine River, which is a tributary of the Illinois River. In 35 years, M. quadrifolia has migrated down Spring Creek only one mile. The quantity and distribution of the plants vary, progressing from many plants in the upper part of the creek below the lake and dam to a very few, widely scattered plants beyond one-half mile to finally one plant one mile downstream. The plants are usually at the edge of the creek, particularly near the lake, but further downstream are found characteristically in small “cut-offs” at the edge of the creek. Downstream migration of Marsilea has not been rapid, which seems surprising considering the numerous times it could have been dispersed by propagules (includ- ing rafts of vegetation), especially during the rush of high waters in spring and fall. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 1 (1983) 31 WOOD FERNS NEW TO MARYLAND AND DELAWARE.— While collect- ing specimens for Towson State University, I discovered four plants of the Crested Log Fern Hybrid, Drvopteris celsa x cristata, growing as a fairy ring near Creswell, Harford County, Maryland (Redman 3547, BALT. MICH). This discovery represents the fifth state known for this rare hybrid. the closest known locality being about 150 miles northeast in northeastern New Jersey. The plants grew on alluvial soils derived from the James Run gneiss formation in a Beech-Oak woods. The parents were not growing nearby: D. celsa is known trom 5 miles away and D. cristata from 7 miles away. Unfortunately, the hybrids’ locality is being destroyed for development, but the plants have been moved into cultivation. | am indebted to Drs. Warren and Florence Wagner for verifying the identity of the specimens. I also discovered three plants of Miss Slosson’s Hybrid Woodfern, D. x slossonae Wherry (D cristata X marginalis), near Kenton, Kent County, Delaware (Redman 2814, BALT). The habitat is alluvial woods under a canopy of oak trees. Plants of both parents grow nearby. Apparently this is the first record for this hybrid in Delaware.—Donn E. Redman, 2615 Harwood Road, Baltimore, MD 21234. A NEW COMBINATION FOR AN ASPLENOSORUS HYBRID.—Tradi- tionally, the Walking Fern has been segregated from the spleenworts (Asplenium) as Camptosorus rhizophyllus (L.) Link. A number of pteridologists, however, question the distinction, pointing out the morphological similarity of Walking Fern to Asplenium, its comparable diploid chromosome number (n= 41), its preference for rock substrates, and its northeastern distribution. A close alliance with the spleen- worts is further demonstrated by its tendency to cross with representatives of the genus Asplenium to produce sterile hybrids, some of which have become fertile through chromosome doubling and now behave as distinct species. In the Appala- chian Mountains such hybridization and ploidy level changes have led to an extensive allopolyploid complex which has undergone considerable study. Those Who maintain the Walking Fern in Camptosorus place hybrids with Asplenium and their derivatives in the hybrid genus Asplenosorus. Most of the hybrids were Originally described as spleenworts, and therefore have combinations available under Asplenium. One new Walking Fern—Spleenwort hybrid, however, was recently de- scribed as an Asplenosorus. Its incorporation into the genus Asplenium may be accommodated by the following combination: Asplenium x shawneense (R. C. Moran) H. E. Ballard, comb. nov. Asplenosorus x shawneensis R. C. Moran, Amer. Fern J. 71:85, 1981. This is the hybrid of Asplenium rhizophyllum and A. trichomanes, and is known So far only from southern Illinois.—Harvey E. Ballard, Jr., Department of Biology, Western Michigan University, Kalamazoo, MI 49008. 32 AMERICAN FERN JOURNAL: VOLUME 73 (1983) AMERICAN FERN JOURNAL Terry R. Webster, of the Biological Sciences Group, University of Connecticut, Storrs, CT 06268, has been appointed to the editorial staff of the JOURNAL. Professor Webster is a morphologist. His special interest is the morphology and reproductive biology of Selaginella. He will be taking on some manuscript reviewing and proofreading duties that are so necessary to the functioning of the JOURNAL —D.B.L (Required by 38 U.S.C. 3685) 1 STATEMENT OF OWNERSHIP, M, MANAGEMENT AND CIRCULATION (OF PUBLICA’ A. PUBLICATION AMERICAN FERN JOURNAL quarterly $8 mem; $9 subscr U. S. Nat'l, Herbarium, 10th & Constitution Ave. N.W., Washington, DC 20560 U. S, Nat'l. 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Full members receive THE FERN GAZETTE and the BULLETIN OF THE BRITISH PTERIDOLOGICAL SOCIETY. Membership subscriptions are £ 7 for full members, £ 5 for ordinary members (not receiving the GAZETTE), and £5 for student members (under 25 years of age). For particulars, U. S. residents should apply to Dr. J. Skog, Biology Department, George Mason University, Fairfax, VA 22030. Non-U. S. residents should apply to Lt. Col. P. G. Coke, Robin Hill, Stinchcombe, Dursley, Gloucestershire, England. _.. from TRIARCH We are grateful for the purchase orders and collec- tions of material received during Triarch’s 56 years of service. We try to say thank you by maintaining quality of product, variety of listings, and by supporting biolog- ical societies with memberships, donations, and advertising. Having completed 25 years in the management of Triarch, | add this written word of thanks for your support, without which Triarch would not exist! Paul L. Conant, President TRIARCH PREPARED MICROSCOPE SLIDES P.O. Box 98 Ripon, Wisconsin 54971 Sahih Sr aoe St acacia a tango eae cenmee nae Ferns and Vhed Plants With Special Reference to Tropical America ROLLA M. TRYON and ALICE F. TRYON, Harvard University MORE THAN 850 PAGES This exceptional volume is the first modern, in-depth treatment of ferns and related plants, particularly in tropical America. Written by Rolla M. and Alice F. Tryon, well known for their work in this area, FERNS AND ALLIED own wide-ranging research. Extensive bibliographies of modern literature on fern biology are provided. In addition to tropical American genera, this volume also has extensive information on ferns of temperate an paleotropical regions, making it of worldwide perspective. Each of the 127 tropical and temperate American genera, represented by 29 families, is organized by accounts of its: O ecology O nomenclature and typification O geograph es O generic characteristics O cytology MORE THAN 2,000 ILLUSTRATIONS AND PHOTOGRAPHS Carefully compiled, magnificently produced, and superbly illustrated, FERNS AND ALLIED PLANTS has more than 2,000 individual illustrations and photographs prepared especially for this volume, including: © photographs of living plants in their natural habitats O photographs and illustrations of biological and systematic features O distribution ma © more than 700 electron micrographs of fern spores Geologists who use palynological records in hydrocarbon exploration will find the spore micrographs of immense practical value. Unrivaled in its Coverage, FERNS AND ALLIED PLANTS will serve as the authoritative reference for years to come. 1982/approx. 896 pp. /2022 individual illustrations and photographs in 639 figures/ISBN 0-387-90672-X/cloth $148.00 For more information, or to order, please write to: SPRINGER-VERLAG NEW YORK INC. , Dept. $5490, 175 Fifth Avenue, New York, New York 10010 a SPRINGER-VERLAG New York Heidelberg Berlin 14 NEE AGRE AES, Sa LIEN rhea ete 5 IE A Bee ERENT Se gg ee ee aaa ee Se Le AMERICAN 33 FERN — JOURNAL | QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Vittaria Gametophytes Discovered in a New Physiographic Province ALLISON W. CUSICK Chapman’s Quillwort Reconsidered BRIAN M. BOOM C-Glycosylxanthones in Diploid and Tissue Culture-induced Autotetraploid Davallia fejeensis P. MICK RICHARDSON and HARINDER K. PALTA The Distribution of Woodwardia areolata R. CRANFILL Two Moonworts of the Rocky Mountains: Botrychium hesperium and a New Species Formerly Confused with It W. H. WAGNER, JR. and FLORENCE S. WAGNER Shorter Notes: Lycopodium complanatum and - annotinum Found in the Black Hills; Microfibrils in the Xylem of Blechnum viviparum; Bisporangiate Anomalous Sporophylls in Isoétes from Rajasthan Reviews 42, 45, Ww we 52 MISSOURI BOTANICAL os ~ The American Fern Society Council for 1983 DEAN P. WHITTIER, Dept. of Biology, Vanderbilt University, Nashville, TN 37235. President TERRY R. WEBSTER, Biological Sciences Group, University of Connecticut, Storrs, CT 06268. Vice-President MICHAEL I. COUSENS, Faculty of Biology, University of West Florida, Pensacola, FL 32504. Secretary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Treasurer JUDITH E. SKOG, Dept. of Biology, George Mason University, Fairfax, VA 22030. Records Treasurer DAVID B. LELLINGER, Smithsonian Institution, Washington, DC 20560. Journal Editor ALAN R. SMITH, Dept. of Botany, University of California, Berkeley, CA 94720. Memoir Editor JOHN T. MICKEL, New York Botanical Garden, Bronx, NY 10458. Newsletter Editor American Fern Journal EDITO DAVID B. LELLINGER U.S. Nat’l Herbarium NHB-166, Smithsonian Institution, Washington, DC 20560. ASSOCIATE EDITORS DAVID W. BIERHORST Rt. 3, Box 188, Picayune, MS 39466. GERALD J. GASTONY Dept. of Biology, Indiana University, Bloomington, IN 47401. JOHN T. MICKEL New York Botanical Garden, Bronx, NY 10458. TERRY R. WEBSTER ....... Biological Sciences Group, University of Connecticut, Storrs, CT 06268. The “American Fern Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the general study of ferns. It is owned by the American Fern Society, and published at the Smithsonian Institution, Washington, DC 20560. Second-class postage paid at Washington. Claims for missing issues, made 6 months (domestic) to 12 months (foreign) after the date of issue, and the matters for publication should be addressed to the Editor. Changes of address, dues, and applications for membership should be sent to Dr. Judith. E. Skog, Dept. of Biology, George Mason University, Fairfax, VA 22030. Orders for back issues should be addressed to Dr. James D. Montgomery, Ichthyological Associates, R.D. 1, Berwick, PA 18603. General inquiries concerning ferns should be addressed to the Secretary. Subscriptions $9.00 gross, $8.50 net if paid through an agency (agency fee $0.50); sent free to members of the American Fern Society (annual dues, $8.00 + $4.00 mailing surcharge beyond U.S.A., Canada, and Mexico; life membership, $160.00). Back volumes 1910-1978 $5.00 to $6.25 each; single back numbers of 64 pages or less, $1.25; 65-80 pages, $2.00 each; over 80 pages, $2.50 each, plus shipping. Back volumes 1979 et seq. $8.00 each; single back numbers $2.00 each, plus shipping. Ten percent discount on orders of six volumes or more. Library Dr. John T. Mickel, New York Botanical Garden, Bronx, NY 10458, is Librarian. Members may borrow books at any time, the borrower paying all shipping costs. Newsletter John T. Mickel, New York Botanical Garden, Bronx, NY 10458, is editor of the newsletter ‘iddiehead Forum.” The editor welcomes contributions from members and non-members, including miscellaneous notes, offers to exchange or purchase materials, personalia, horticultural notes, and reviews of non-technical books on ferns. Spore Exchange Mr. Neill D. Hall, 1230 Northeast 88th Street, Seattle, WA 98] 15, is Director. Spores exchanged and collection lists sent on request. Gifts and Bequests s = = bequests to the Society enable it to expand its services to members and to others interested rag ana ee books, back issues of the Journal, and cash or other gifts are always welcomed, and are tax-deductible. Inquiries should be addressed to the Secretary . AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) 33 Vittaria Gametophytes Discovered in a New Physiographic Province ALLISON W. CUSICK* Since their recognition over 20 years ago, knowledge of the distribution and taxonomic affinities of the perennial Vittaria gametophytes found in the eastern United States has expanded greatly. Once thought to be extremely rare and local, they now are known to be common in selected habitats over a broad region of the Appalachians. Farrar (1978) summarized the history of this taxon and presented a map of its distribution as then understood. Vittaria gametophytes have been discov- ered in the unglaciated Appalachian Plateau, Blue Ridge, Ridge and Valley, and Upland Piedmont provinces, all uplifted bedrock areas which presumably have been in continuous existence since Tertiary times. Farrar et al. (1983) discussed these recent finds and reported the gametophytes from sites in Ohio and Pennsylvania near the limits of Pleistocene glaciation. This article describes the presence of Vittaria gametophytes beyond these limits. This unusual fern now has been discovered on the glaciated Appalachian Plateau nearly 90 km north of the southern limit of Wisconsinan glaciation. This also is the first report of Vittaria gametophytes from the watershed of the Great Lakes. The Vittaria gametophytes were found 10 June 1982 on Little Mountain, Geauga and Lake counties, Ohio, on property of the Holden Arboretum, Mentor, Ohio (Fig. 1). Accompanying the author were Brian Parsons and Thomas Yates, field natural- ists of the Arboretum, and Robert McCance of the Division of Natural Areas and Preserves, Ohio Department of Natural Resources. To say that we were surprised by this discovery would be an understatement. Only a small collection was made at this time (Cusick 21673, ISC). The author revisited the site on 24 August, searching more thoroughly for gametophytes, and again preparing a voucher (C usick 22000, OS). Colonies of Vittaria were found to occur discontinuously in a linear band over approximately 0.5 km of the slope of Little Mountain. Identity of the gametophytes was verified by Dr. Donald R. Farrar of Iowa State University. The site of this discovery is an extraordinary area not at all typical of glaciated Ohio and a habitat highly suitable for the growth of Vittaria. Little Mountain is at the extreme edge of the glaciated Appalachian Plateau, being one of the localized bedrock exposures that mark the escarpment between the elevated Plateau and the Lake Plains of the Central Interior Lowlands province. The mountain is shaped roughly like a dumbbell, oriented north to south, with knobs at either end. The northern knob rises to 380 m, the southern knob to 386 m. Between these summits is a flattened saddle only slightly lower in elevation. The northern knob lies partly in Concord Township, Lake County, while the bulk of the mountain is in Chardon Township, Geauga County. The shoreline of Lake Erie is only 6.5 km north of the mountain. The approximate mean elevation of the lake at that point is 174 m. Thus, *Division of Natural Areas and Preserves, Ohio Department of Natural Resources, Fountain Square, Columbus, OH 43224. Volume 73, number 1, of the JOURNAL, was issued March 31, 1983. 34 AMERICAN FERN JOURNAL: VOLUME 73 (1983) TauMe@uLL PORTAGE ft MAHOMING STARK COLUMBAN A a ae 2h YH % + TUSCARAWAS A HOPPE RSON 7 - FIG. 1. The northeastern quarter of Ohio. Solid line indicates the northern limit of the glaciated Appalachian Plateau; dashed line, the southern limit of Wisconsinan glaciation. Solid dot is location of Little Mountain. there is a difference in elevation of 206 m between the mountain and Lake Erie (U.S. Geological Survey, 1970 This dramatic change in elevation between lake and summit has a significant effect on local weather patterns. Winds moving south across Lake Erie pick up moisture which is quickly lost once this escarpment is reached. In the lee of these summits, then, is the locally-famous “snow belt” of northeastern Ohio. This local phenomenon is clearly shown on the snowfall map in Gordon (1969) on which the site of Little Mountain may be seen in the pattern of isopleths. Snowfall in the Little Mountain region averages nearly 3 m yearly, far in excess of Ohio’s annual average snowfall of about 1 m. The summits of Little Mountain and the other elevations at the edge of the glaciated Appalachian Plateau in Ohio are capped by the Sharon Conglomerate formation of the Pottsville group of the Pennsylvanian System of sedimentary rocks. Throughout northeastern Ohio, the Sharon Conglomerate is noted for the formation of massive cliff faces, rock shelters, and slump blocks (Rau, 1970). The designation Differential weathering of this cement results often in honeycomb patterns in the cliff faces (Heimlich et al., 1970). A. W. CUSICK: VITTARIA GAMETOPHYTES IN A NEW PROVINCE 35 At the base of the Sharon formation lies a major disconformity with the Meadville Shale of the Cuyahoga group of Mississippian age sediments. This disconformity is responsible for two phenomena which contribute to the habitat supporting the populations of Vittaria, namely, the origination of springs and the formation of slump blocks. The Sharon Conglomerate is highly permeable, whereas the underly- ing shale is relatively impervious. This disconformity, then, is marked by a zone of seeps and springs (Heimlich et al., 1970; Rau, 1970). The jointed conglomerate tends to slip along this lubricated shale surface, splitting the rock, which slowly moves downslope. On Little Mountain this joint-controlled slippage forms a pictur- esque and complex system of blocks and chasms ranging from | to 3 m wide, occasionally narrower, 10 to 15 m deep, and often as long as 50 m (Aronson, 1974). The springs which flow from the disconformity on Little Mountain are excep- tional in that they average 8-9°C year-round (Parsons, pers. comm.). This cold temperature allied with the depth and narrowness of the fissures on the mountain permits snow and ice to persist long into the warmer months. The writer noted the remains of ice on 10 June 1982. Even in mid-summer one’s breath condenses before one’s face while walking through the chasms on the mountainside. The environment within the crevasses of Little Mountain conforms in every particular to the ecological requirements of Vittaria as outlined by Farrar (1978, p. 4): “a low light intensity of 100 ft-c or less, relatively high humidity, and protection from temperature extremes . overhanging rock outcrops, dense forest canopy, and nearness to running water.” Floristically, Little Mountain is a part of the White Pine-Hemlock—Northern ardwoods community (Braun, 1950), a forest association of very limited occur- rence in Ohio. The first botanist to visit the mountain may have been John L. Riddell, who collected several species from this “pine-clad, rubblestone knob” (Riddell, 1836, p. 567). Read (1873a, b) listed the major forest trees of the mountain, particularly noting the Hemlock (7suga canadensis (L.) Carr.) and White Pine (Pinus strobus L.) on the northern end of the mountain, the extensive growth of American Chestnut (Castanea dentata (Marsh.) Borkh.) throughout the summit, and “Rock Oak” (Quercus prinus L.) on the southern knob. All these species are yet extant on the mountain, although the chestnut is represented only by fallen, dead trees and root sprouts. Ferris (1887) presented an extensive list of vascular species for the mountain. While his list contains some obvious inaccuracies, it also provides further evidence that most of the herbaceous and woody species found on Little Mountain in the past still grow there today and that the original vegetation conformed to Braun’s concept of the White Pine—-Hemlock—Northern Hardwoods community. In most of its known range, the Vittaria gametophyte seems associated with the Mixed Mesophytic forest as defined by Braun (1950). This is the case, for instance, with other known Ohio populations. However, the Warren County, Pennsylvania Station (Farrar et al., 1983), like Little Mountain, is within the White Pine- Hemlock-Northern Hardwoods floristic province. The occurrence of Vittaria ga- metophytes probably is more closely related to appropriate physical and geological setting than to floristic province. 36 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Little Mountain was a prominent landmark to the first settlers of northeastern Ohio. As early as 1831, a hotel was built on the mountaintop, and within the following 50 years Little Mountain became a celebrated resort. At least three hotels were built on the summit, together with a complex of summer cottages, gazebos, churches, schoolhouses, and bowling alleys. The largest and best-known of the hotels, the Pinecrest, built in the late 1880’s, featured a Western Union telegraph line and a post office. The cool springs also induced developers to advertise water cures for “invalids.” Guests clambered through the chasms on sweltering summer days, enjoyed the cool breezes through the white pines, and gazed at the dramatic view of Lake Erie from the northern knob. It was fashionable to carve one’s name on the conglomerate in the “Devil’s Kitchen,” as one large grotto was dubbed. Many of these graffiti are still legible today, a sort of permanent guest register. The resort era ended by 1920 and the Pinecrest hotel was torn down in 1941. Little Mountain ceased to be the playground of the public and instead became the private preserve of the Little Mountain Club, an elite group of wealthy Clevelanders (Ahlstrom, 1961; Ferris, 1887). The mountain could then recover from the years of wanton vandalism. The present composition of the vegetation and the beautiful stand of mature White Pine are tributes to the ability of plant communities to survive and recover from severe disturbance. Other than the graffiti on the rocks, now largely cloaked by bryophytes, the most notable evidence of the former resort is the abundance of Vinca minor L. on the mountaintop. Little of the disturbance described above would have had a direct effect on the Vittaria population. However, it is difficult to be certain how common or widespread the gametophytes might have been prior to the time of the hotels. Today, the Vittaria, although occurring over a considerable linear area of the mountainside, grows in isolated pockets, often only a few square cm in extent, on the walls of the Ohio. An occasional colony of the gametophytes occurs in the cast left by the fall of a particularly large quartz pebble. Most often the plants grow in recesses or narrow clefts formed in zones of finer pebble size. The plants definitely are less frequent in (1980, 1982) maps thin layers of Hiram Till of Wisconsinan age over even the highest summits of the limits of the Appalachian Plateau in Ohio. Clearly, Little Mountain was not a refugium on which individuals of Vittaria could have survived glaciation. The means of dispersal of this species, even in the heart of its range, is little Dies § Although Vittaria gametophytes prolifically produce vegetative gemmae, there seems to be no obvious mechanical means for transporting these gemmae across the 90 km gap between Little Mountain and the nearest limit of the rs A. W. CUSICK: VITTARIA GAMETOPHYTES IN A NEW PROVINCE 37 Wisconsinan ice. It seems unlikely that the few-celled gemmae could survive wind-borne dispersal over such a distance without fatal desiccation. Also, it is difficult to understand how wind currents can lift these gemmae from the sheltered grottos and canyons in which they occur and convey them into the upper air. A possibility, which seems highly unlikely at least to this writer, is that the plants were introduced by an animal or human visitor who had accidentally picked up the gemmae in southeastern Ohio only a short time before. Both Farrar and this writer have found Vittaria gametophytes at the limit of Wisconsinan glaciation in Fairfield County, Ohio, yet have failed to locate the species in similar habitats and on the same rock strata in areas only a few kilometers beyond the glacial boundary in other Ohio counties. Farrar’s two western Pennsylva- nia stations for Vittaria gametophytes are located in Lawrence and Warren counties. The Lawrence County site is about 80 km southeast of Little Mountain and is located just north of the Illinoian glacial limit and south of the Wisconsinan boundary. The Warren County population is about 160 km east-northeast of Little Mountain and is directly south of the Illinoian limit. At present, the Warren County Station is the northernmost known occurrence of these gametophytes (Farrar et al., 1983). It is probably useless to speculate overmuch on the origin of the Little Mountain population of Vittaria until other, similar sites on the glaciated Appalachian Plateau of Ohio and nearby ‘states are examined for this species. Appropriate habitats elsewhere in glaciated regions may well harbor populations of this gametophyte. Indeed, the perennial gametophyte of an as yet unknown species of Trichomanes has been found on the glaciated Appalachian Plateau from central Ohio to New Hampshire (Farrar et al., 1983; McAlpin et al., 1978). Perhaps this find of Vittaria should not come as such a surprise after all. In glaciated Ohio, suitable habitats are very limited in extent and highly disturbed. If the case of Little Mountain is considered, however, disturbance which may have extirpated other types of vegetation may not have had so severe an effect on Vittaria. As of this writing, Vittaria gametophytes have not been found elsewhere in glaciated Ohio. But much field work remains to be done before any firm conclusions can be reached as to the true distribution of the gametophytes, either nationally or statewide. This find opens new realms for fern enthusiasts to explore. The author is indebted to R. Henry Norweb, Jr., Director of the Holden Arboretum, for access to the Arboretum property and for permission to collect vouchers. Brian Parsons and Thomas Yates of the Arboretum staff have provided field assistance as well as access to unpublished data on Little Mountain and its history. The Arboretum and its staff should be congratulated for their commitment to the protection of natural areas. Mr. and Mrs. Hugh Pallister of Willoughby, Ohio, also have provided the author with information on the history of Little Mountain. My special thanks are owed to Dr. Donald R. Farrar of Iowa State University, Ames, lowa, for verifying the identity of the Vittaria gametophytes, for his sharing of data, and for his encouragement. This research was supported by the Division of Natural Areas and Preserves, Ohio Department of Natural Resources. 38 AMERICAN FERN JOURNAL: VOLUME 73 (1983) LITERATURE CITED AHLSTROM, J. M. 1961. The Little Mountain story. The Historical Society Quarterly, Lake Co., Ohio 3:43-46. ARONSON, J. L. 1974. Mass wasting on Little es Lake and Geauga counties, Ohio (abstr.). Geol. Soc. America Abs. with Programs BRAUN, E. L. 1950. Deciduous Forests of Eastern North America. Blakiston, Philadelphia. COOGAN, A. H., R. M. FELDMAN, E. J. SZMUC, and J. V. MRAKOVICH. 1974. Sedimentary environments of the Lower Pennsylvanian Sharon Conglomerate near Akron, Ohio. Pp. 19-41 in: R. A. Heimlich and R. M. Feldman, eds. Guidebook No. 2: Selected Field Trips in staribediets Ohio. Ohio Geological Survey, Columbus FARRAR, D. R. 1978. Problems in the identity and origin 0b the Appalachian gametophyte, a fpornye fern of the eastern United States. Amer. J. Bot. 65:1—12. PARKS, and B. W. McALPIN. 1983. The fern genera Vittaria and Trichomanes in the northeastern United States. Rhodora 85:83-92. FERRIS, 'E. J. 1887. History of the Little Mountain from 1810 to 1887. Painesville Advertiser, Painesville, OH GORDON, R. B. 1969. The natural vegetation of Ohio in pioneer days. Ohio Biol. Surv. Bull., 3(2):xi + 113 HEIMLICH, R. A., J. V. MRAKOVICH, and G. W. FRANK. 1970. The Sharon Conglomerate. Pp. 125-133 in: P. O. Banks and R. M. Feldman, eds. Guide to the Geology of Northeastern Ohio. North. Ohio Geol. Soc., Cleveland. MCcALPIN, B., and D. R. FARRAR. 1978. Trichomanes gametophytes in Massachusetts. Amer. Fern J. 68:97-98. RAU, J. L. pl Pennsylvanian System of northeast Ohio. Pp. 69-124 im: P. O. Banks and R. M. Fe n, eds. Guide to the geology of northeastern Ohio. North. Ohio Geol. Soc., Cleveland. READ, M. ee Geology of Geauga County. Rept. Ohio Geol. Surv. 1(1):520—533. —————. 1873b. Geology of Lake County. Rept. Ohio Geol. Surv. 1(1):510-519. RIDDELL, J. 1836. Supplementary catalogue of Ohio plants. Western J. Med. Phys. Sci. 35 US. Ss Rerind SURVEY. see Mentor Quadrangle, 7.5 minute series (topographic). U. S. Dept. Interior, Washingto WHITE, G. W. 1980. Glacial pelea of Lake County, Ohio. Ohio Geol. Invest. Rept. No. 117. ————.. 1982. Glacial geology of northeastern Ohio. Ohio Geol. Surv. Bull. 68. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) 39 Chapman’s Quillwort Reconsidered BRIAN M. BOOM* Certainly one of the least known pteridophytes in the southeastern United States is Chapman’s Quillwort. Described by Engelmann (1882) as /soétes flaccida Shuttlew. var. chapmanii Engelm., the taxon has been collected only a few times since its initial collection by A. W. Chapman in 1848. The collections are always from the same general locality in Jackson County, Florida. Larger megaspores with smoother ornamentation were said by Engelmann to distinguish it from the typical variety. Underwood (1900) and Clute (1928) both recognized the distinctness of the taxon, and Small (1932) even elevated it to specific status. Lakela and Long (1976) maintained J. chapmanii (Engelm.) Small, and in their key distinguished it from /. flaccida on the basis of megaspore ornamentation and sporangium shape. On the other hand, Pfeiffer (1922) and Boom (1979, 1982) were not convinced of the distinctness of J. chapmanii, and they synonymized it with I. flaccida. Recently I was able to study the spores of these plants more carefully by means of scanning electron microscopy (SEM), and I now regard the evidence as sufficient for maintaining J. flaccida var. chapmanii as a rare, but distinct element of Florida’s pteridophyte flora. Spores were coated by a two-step procedure consisting of the deposition from a low voltage arc of an initial carbon layer onto which was subsequently sputtered a 60:40 gold-palladium alloy using a 15 watt DC diode coater. Specimens were examined with a Cambridge S4-10 SEM. Photographs were taken with Polaroid Type 665 Positive/Negative Land Film. Spores of some thirty specimens of /soétes flaccida were examined (vouchers cited in Boom, 1979), but the photomicrographs of only three are presented herein: var. flaccida: Lake Flirt, near Lake Okeechobee, Florida, Aug 1878, A. P. Garber s. n. (US 240797); cypress pond near Cobb, Sumter Co., Georgia, R. M. Harper 1046 (US 400120); and var. chapmanii: Tributary of the Chipola River at Marianna, Jackson Co., Florida, R. K. Godfrey 61963 (US 2424571). : Both megaspores and microspores of the two varieties show differences In perine ornamentation which are taxonomically significant (Figs. / -6). Additionally, the differences in megaspore size provide a useful means of distinguishing the varieties. The ornamentation of J. flaccida var. chapmanii megaspores is low tuberculate, densely so on the distal hemisphere and sparsely so on the proximal (Fig. /). Greater magnification shows the perine to be minutely echinate (Fig. 3). In var. flaccida, the megaspores are also low tuberculate, but densely so on both hemt- spheres (Fig. 2). Greater magnification reveals a finely rugulate perine, the muri anastomosing so as to give a spongy appearance (Fig. 4). The diagnostic perine characters (extent and distribution of tubercules) can be seen at 40X, and the size difference detected with a dissecting microsc Although the microspores of the varietie perine ornamentation differs: that of var. chapmanii b ope. s are essentially the same size, their eing echinate with an arach- *New York Botanical Garden, Bronx, NY 10458. 40 AMERICAN FERN JOURNAL: VOLUME 73 (1983) 6 FIGS. 1-6. Isoétes flaccida spores. FIG. . Megaspore of var. chapmanii (US itll ds scale = 100 um. FIG. 2. Megaspore of var. flaccida ‘ie sieht scale = um. F 3. Megaspore perine of var. chapmanii (US 2424571); scale = 7 wm. FIG. 4. Meg aspore perine of var. _Placelue 240797); scale = Hm. FIG. 5. Microspore pe var. chapman (US 2424571); scale = § . FIG. 6. Microspore of var. flaccida (US 400120): scale = Lm. 8 S ———— B. M. BOOM: CHAPMAN'S QUILLWORT RECONSIDERED 41 noid perine between echinae (Fig. 5), and that of var. flaccida being papillate with a laevigate perine between papillae (Fig. 6). Given their average size of ca. 30 ym, the microspores do not provide useful taxonomic characters for the field or herbarium botanist, but the ornamentation differences help confirm their distinctness. Aside from the spore characters, there are no other apparent morphological, chemical, or ecological differences that separate the varieties. The orbicular or obovoid sporangium ascribed to var. chapmanii by some authors (Engelmann, 1882; Small, 1932; Lakela & Long, 1976) as a feature distinguishing it from var. flaccida (with an oval, ellipsoid, or subglobose sporangium) is not a good character, as this structure varies too much depending on plant size and ecological conditions. The flavonoid chemistry of the two varieties was examined by Boom (1979), but the chromatographic profiles were identical. Ecologically, there appears to be nothing striking about the habitat of var. chapmanii. A topotype collection (Boom 333, TENN) reports “sandy/peaty substrate, clear water in deep bald cypress swamp;” the stream is underlain by limestone. This general description could apply to many areas in Florida where var. flaccida occurs, but perhaps there are significant microhabitat factors involved which have not yet been detected. A distribution map of I. flaccida is given in Boom (1982). Just as Taylor et al. (1975) used SEM to confirm the distinctness of Isoétes butleri Engelm. from J. melanopoda Gay & Dur. based on spore ornamentation, it has been possible to show that two entities are encompassed by /. flaccida. | prefer to recognize these two taxa at the varietal level because, unlike the case of J. butleri and J. melanopoda, the two taxa involved here are not ecologically or geographically differentiated. I do not regard I. flaccida var. chapmanii as a hybrid because it meets none of the criteria used in the detection of several instances of hybridization in other southeastern U.S. quillworts (Boom, 1982). I consider I. flaccida var. flaccida to encompass J. alata Small and /. flaccida vat. rigida Engelm. The following key separates the two recognized varieties of the species: KEY TO VARIETIES OF ISOETES FLACCIDA Megaspores 300-400(440) zm diam., densely low tuberculate on both hemispheres, papillate; common throughout Florida and southern Georgia ........:.ssssseereeeeressee es Megaspores 440-540 xm diam., densely low tuberculate on distal hemisphere, sparingly so on proximal; microspores echinate; rare in northwestern Florida (Jackson Co.) ......-- var. chapmani Appreciation is extended to David Lellinger, Andrea Sessions, and Walter Brown for assistance with /soétes spore studies at US. The present work was materially aided by the Botany Department of The University of Tennessee, Knoxville, by a Grant-In-Aid of Research from Sigma Xi, and by a travel grant from The Smithsonian Institution. microspores var. flaccida LITERATURE CITED BOOM, B. M. 1979. Systematic studies of the genus Isoétes in the southeastern United States. M.S. Thesis, Univ. of Tennessee, Knoxville, TN. . 1982. Synopsis of Isoétes in the southeast RS aegy ern United States. Castanea 47:38-5 CLUTE, W. N. 1928. The Fern Allies of North America 9. North of Mexico. Willard N. Clute & Co., - Joliet, IL. : : ENGELMANN, G. 1882. The genus Isoétes in North America. Trans. St. Louis Acad. Sci. 4:358-390. LAKELA, O. and R. W. LONG. 1976. Ferns of Florida. Banyan Books, Miami, FL. V 42 AMERICAN FERN JOURNAL: VOLUME 73 (1983) PFEIFFER, N. E. 1922. Monograph of the Isoétaceae. Ann. Missouri Bot. Gard. 9:79-233. SMALL, J. K. 1932. Ferns of Florida. Science Press, New York, NY. TAYLOR, W. C., R. H. MOHLENBROCK, and J. A. MURPHY. 1975. The spores and taxonomy of Isoétes butleri and I. melanopoda. Amer. Fern J. 65:33-38. UNDERWOOD, L. M. 1900. Our Native Ferns and Their Allies. Henry Holt, New York, NY. REVIEW Flora Malesiana, Series II—-Pteridophyta, volume 2, part 5, Thelypteridaceae, by R. E. Holttum. Pp. 331-599, including index and addenda for entire volume, plus pp. 1-20, contents and dedication. 1982. Martinus Nijhoff, BV, The Hague, Netherlands. $63.00.—Historically, no other group of ferns has been subjected to such varied circumscriptions, both of the family and genera; moreover, species have been notoriously under-collected, ill-defined and confused, and their relationships misunderstood. Part of the reason is their mundane (cringe) and superficially similar look that causes collectors to sample one or a few and pass by the rest. Another reason is that few taxonomists have bothered to look at them closely. The first who did look was Carl Christensen, to whom Holttum appropriately dedicates this volume; but Christensen concerned himself mostly with Neotropical species. In 1963, Ching presented a revised classification of Old World genera that provided a springboard for Holttum’s many precursory revisions culminating in the present work. The sheer size and complexity of Thelypteridaceae have made Holttum’s task formidable. This family is probably the largest in ferns, with almost 1000 species. In the Flora Malesiana region alone Holttum treats 440 species in 22 genera. The number of new taxa (75 species, 18 varieties) is staggering and reminds us how poorly known are the tropical floras and how urgent it is to preserve what little is left. In Sphaerostephanos, with 152 Malesian species, 43 are known only from the type and many additional species are not much better represented in herbaria. But Holttum’s most important contribution is not the number of species named and redefined; it is the understanding of the large natural groups of species. Here, he has brought order from chaos. Doubtless there will be taxonomists, including myself, who will prefer to assign lower rank to some of Holttum’s genera. But his circumscriptions of natural groups are likely to survive intact or with only minor redefinition. At least 13 new chromosome counts are embedded in the work: they may well be overlooked by compilers of chromosome indices. The most interesting, n= 66 for a species of Coryphopteris, is a new base number (x= 33) for Thelypteridaceae and may lend support to Holttum’s contention of a relationship between this family and Cyatheaceae. The magnitude, originality, and scholarship shown in this work are truly impres- sive and culminate a brilliant career. But Holttum is already turning to other ahi — and one wishes him many more years of taxonomic insight. —Atan RK. Smith, Herbarium, Departme iversi ifornia, Berkeley, CX oF Dp nt of Botany, University of Califo AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) 43 C-Glycosylxanthones in Diploid and Tissue Culture-induced Autotetraploid Davallia fejeensis P. MICK RICHARDSON and HARINDER K. PALTA* 1,3,6,7-tetrahydroxy-C-glycosylxanthones are phenolic compounds which have been found relatively infrequently in ferns. A recent survey (Wallace et al., 1982) reported their occurrence in seven genera: Asplenium, Athyrium, Elaphoglossum, Cardiomanes, Hymenophyllum, Trichomanes, and Marsilea. They have also been reported in Ctenitis decomposita (Bohm, 1975). The present communication reports C-glycosylxanthones in Davallia fejeensis Hooker. Diploid and tetraploid samples of both the sporophytic and gametophytic generations were examined. MATERIALS AND METHODS Rhizome tips (about 2 cm long) of diploid Davallia fejeensis were collected in the Enid A. Haupt Conservatory of the New York Botanical Garden, cleaned, rinsed in tap water and further trimmed. Voucher specimens are at NY. Explants (6-8 mm long) were cultured on 1% agar-gelled sterile nutrient medium of Knudson as modified by Steeves et al. (1955) and supplemented with 2% sucrose in order to provide control diploid sporophytes growing in axenic culture. Roots, leaves, and thizomes of the resulting plants were excised and induced to differentiate into aposporous diploid gametophytes. Stock cultures of diploid gametophytes were multiplied and further grown on liquid media supplemented with 0.6% agar to effect fertilization and raise tetraploid sporophytes. Tetraploid gametophytes were isolated from this tissue culture-induced tetraploid material by repeating the procedure followed at the diploid level. In this way, both diploid and tetraploid sporophytes and gametophytes were made available for this study. Phenolic compounds were isolated by two-dimensional paper chromatography of an 80% methanol extract of green material in t-BuOH-HOAc-H,0, 3:1:1 (TBA) and 15% aqueous acetic acid (HOAc), followed by one-dimensional paper chromatogra- phy in water. C-glycosylxanthones appeared as orange compounds in ultraviolet light and turned yellow when fumed with ammonia. Both purified compounds were co-chromatographed with mangiferin and isomangiferin isolated from Asplenium montanum Willd. (Bozeman & Radford 11552, NY). Rr values in TBA, BAW, HOAc, and H,O were: mangiferin, 0.31, 0.43, 0.43, 0.12; isomangiferin, 0.19, 0.29, 0.23, 0.04; Asplenium mangiferin, 0.34, 0.42, 0.42, 0.11; Asplenium isomangiferin, 0.21, 0.32, 0.23, 0.03; and rutin standard, 0.43, 0.45, 0,56; 0:25. The BAW and HOAc values are very similar to those of Smith and Harborne (1971), but the TBA and HOAc values differ from those of Markham and Wallace (1980). Absorption spectra (MeOH, nm) were: mangiferin, 232, 260, 270sh, 310, 364; isomangiferin, 240, 258, 270sh, 312, 365. The compounds were unaffected by acid hydrolysis (1 hr, 2N HCI, 100°C). “Harding Laboratory, New York Botanical Garden, Bronx, NY 10458. 44 AMERICAN FERN JOURNAL: VOLUME 73 (1983) RESULTS AND DISCUSSION The C-glycosylxanthones mangiferin and isomangiferin were the only phenolic compounds appearing on the two-dimensional chromatograms of an 80% methanol extract of each sample. The diploid and tetraploid sporophytes contained relatively large amounts of both compounds, but in both cases the mangiferin spot was much larger than the isomangiferin spot. Gametophyte extracts contained very small amounts of each compound. Further identification was performed only on com- pounds from the sporophytes. Spots from the chromatograms were eluted and then chromatographed in water to purify them sufficiently for ultraviolet spectrophotome- try and co-chromatographic testing. The purified compounds showed reduced Ry values in comparison to those measured on the two-dimensional chromatograms, strongly suggesting that Ry values based on such chromatograms may be misleading. These results suggest that the same C-glycosylxanthones are produced in both sporophytes and gametophytes. This is comparable to the only similar published study of fern flavonoids (Petersen & Fairbrothers, 1980), where the same flavonol glycosides appeared in both generations. Secondly, it appears that an induced doubling of the chromosome number in Davallia has no effect on the production of C-glycosylxanthones in either generation. If we accept that the C-glycosylxanthones in Davallia are equivalent to the flavonoids in flowering plants, then the effects of induced chromosome doubling on phenolic production in ferns may be compared with the effect of a similar chromosome doubling in angiosperms. Mears (1980) has reviewed the chemistry of polyploids. In some cases of induced autotetraploids in Phlox (Levin, 1968), no qualitative differences in phenolics were detected; in another study in the same genus (Levy, 1976), such differences did occur. In Briza (Murray & Williams, 1976) qualitative differences in phenolic production some- times occurred in induced tetraploids. A more complex situation occurred in autotetraploids of Gibasis (del Pero de Martinez & Swain, 1977), where polyploidy is correlated with Robertsonian Fusion, a phenomenon which may be responsible for changes in phenolic expression. This is the first study of the chemistry of induced autoploidy in fern gametophytes and sporophytes. The tissue culture method followed in this study is an excellent way to effect such changes in chromosome numbers. A survey for the presence of C-glycosylxanthones in other fern genera is being undertaken in our laboratory; it is pees that these compounds may not be as rare as they were previously thought to LITERATURE CITED BOHM, B. A. 1975. Xanthones in the fern Ctenitis decomposita. Phytochemistry 14:287—288. LEVIN, D. A. 1968. The genome constitutions of eastern North American Phlox amphidiploids. Evolution 22:612-632. LEVY, M. 1976. Altered piycofiavane expression in induced autotetraploids of Phlox drummondii. Biochem. Syst. Ecol. 4:249-254. MARKHAM, K. R. and J. W. WA ALLACE. 1980. C-glycosylxanthone ~ flavonoid variation within the filmy ferns (Hymenophyllaceae). Phytochemistry 19:415—4 MEARS, J. A. 1980. Chemistry of polyploids: a summary with comments on Parthenium (Asteraceae- Ambrosiinae). In W. H. Lewis, ed. Polyploidy. Plenum Press, New York and London. RICHARDSON & PALTA: C-GLYCOSYLXANTHONES IN DAVALLIA FEJEENSIS 45 MURRAY, B. G. and C. A. WILLIAMS. 1976. Chromosome number and flavonoid biosynthesis in Briza L. (Gramineae). Biochem. Genet. 14:897—904. PERO de MARTINEZ, M. A. del and T. SWAIN. 1977. Variation in flavonoid patterns in relation to chromosome changes in Gibasis schiedeana. Biochem. Syst. Ecol. 5:37-43. PETERSEN, R. L. and D. E. FAIRBROTHERS. 1980. Flavonoid synthesis and antheridium initiation in Dryopteris gametophytes. Amer. Fern. J. 70:93-95. SMITH, D. M. and J. B. HARBORNE. 1971. Xanthones in the Appalachian Asplenium complex. Phytochemistry 10:2117-2119. STEEVES, T. A., I. M. SUSSEX and C. R. PARTANEN. 1955. In vitro studies on abnormal growth of prothalli of the bracken fern. Amer. J. Bot. 42:232-245. WALLACE, J. W., K. R. MARKHAM, D. E. GIANNASI, J. T. MICKEL, D. L. YOPP, L. D. GOMEZ, J. D. PITTILLO and R. SOEDER. 1982. A survey for 1,3,6,7-tetrahydroxy-C- glycosylxanthones emphasizing the “primitive” leptosporangiate ferns and their allies. Amer. J. Bot. 69:356—-362. REVIEW / THE GENUS SELAGINELLA IN TROPICAL SOUTH AMERICA, by A. H. G. Alston, A. C. Jermy, and J. M. Rankin. Bull. Brit. Mus. (Nat. Hist.), bot. ser. 9(4):233-330. 1981. £14.00 postpaid.—A. H. G. Alston was the premier Selaginella authority in the middle of this century. He published extensively on the genus in the Old World and in southern South America, Brazil, the West Indies, and Central America in the New World. At the time of his death, he had begun to study Selaginella from the Andean countries and the Guyanas. The present paper com- pletes and extends Alston’s work and includes Brazil, thus accounting for virtually all of tropical South America. The region contains 133 species and six infraspecific taxa of Selaginella. A table of species distribution by countries shows that Colombia has by far the greatest number of species. The bracketed key to the species can be read forward or backward, a decided advantage because there are no species descriptions. Each taxon treated has a synonymy, a statement of range, a list of specimens, and, sometimes, additional notes. The authors have uncovered some overlooked synonyms in rare literature, always a pleasure to see. However, most synonyms state only the country where the type was collected, rather than the type locality, the collector and number, and the herbarium of deposit. A list of references and an index to accepted names and synonyms concludes the volume. New taxa are illustrated by a photograph of the herbarium sheet and by SEM photographs of the leaves, which range in magnification upwards from a mere eight times natural size, a useful technique for illustrating clearly the cilia and other small details of the leaves. This paper is indispensible for identifying Selaginellas from tropical America and Will be the standard reference for many years to come.—D.B.L. 46 AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) The Distribution of Woodwardia areolata R. CRANFILL* Phytogeographical studies in North America, as compared with those in Europe, are still in their infancy. Lack of detailed atlases and the relative paucity of extensive paleo- and neoecological studies in North America hamper the botanist who wishes to investigate the factors affecting the distribution of a particular species. A few botanical studies of this sort are available (Hocker, 1956; Koevenig, 1976; Salisbury, 1926), although most are limited to local areas or to single factors. The distribution of a particular plant is determined by several interrelated factors, including: (1) climate and soils, both present and past, (2) interactions with other organisms, (3) production and dissemination of propagules, and (4) evolutionary history, including time and place of origin (Billings, 1952; Cox et al., 1976; Krebs, 1972; Pielou, 1979). In general, physical parameters of the environment, such as moisture availability or extremes of temperature, place absolute limits on_ the distribution of an organism. Interactions with other species (including man!), soils, dispersability, and other historical factors (e.g., hurricanes, stream piracy, and glaciation) constrain the organism within these bounds (Billings, 1952; Gorham, 1954). As many have pointed out (Cox et al., 1976; Koevenig, 1976), the study of the interaction of limiting factors is problematic because of our inability to deal quantitatively with several seemingly inextricably linked variables at once. Analysis of widespread or narrowly restricted species is often easiest because in these Situations one or a small number of variables contributes differentially to the distribution. Even so, the implications of such correlations are not always as clear as they may seem. In the absence of corroborative experimental evidence, caution should always be exercised in the interpretation of distributional correlations. Woodwardia areolata (L.) Moore is ideal for this type of study because it is widespread and common throughout the southeastern and Atlantic United States and is well collected. One of the three species in subgenus Lorinseria, it is characteristic of acidic mucky, sandy, and peaty bogs throughout the southeastern United States (Wherry, 1921, 1964). The aim of this paper is to make some ecological inferences from the distribution of W. areolata and to discuss some of the problems inherent in this sort of induction. MATERIALS AND METHODS The distribution of W. areolata was compiled from herbarium specimens depos- ited at the following institutions: DHL, F, FLAS, FSU, GH, KY, MEM, MICH, MO, NCU, NY, SMU, TAES, TENN, TEX, TNS, UC, USF, VDB (abbreviations of Holmgren et al., 1981). Over 700 sheets were examined and plotted accurately on a base map of the eastern United States and Canada. Climatic and edaphic parameters were drawn from a variety of sources. Isotherms were composed and redrawn from individual state maps (National Atmospheric and Oceanographic Institute, 1980). *Department of Botany, University of California, Berkeley, CA 94720. —e — R. CRANFILL: DISTRIBUTION OF WOODWARDIA AREOLATA 47 RESULTS AND DISCUSSION The distribution of W. areolata is shown in Fig. 1. A specimen found at USF (Cheever s. n., 25 June 1917) verifies the occurrence of this species in Maine (previously questioned by Ogden et al., 1948). It also seems likely that the species may occur in southern Indiana and extreme eastern Kansas. Associations of the distribution with several edaphic and climatic factors were found and are discussed below. Climatic Factors.—The climatic feature that appears to have the greatest influ- ence on W. areolata is minimum winter temperature. Of the three isotherms examined (0°C, —2.0°C, and —4.5°C), the —4.5°C mean made the closest fit. All stations, save five, fell to the south of the line, with especially close correspondence in New England. Of the sites north of the isotherm, three are no longer extant (Kitfield, 1974; Price, pers. comm.), while another lies close to the Atlantic Ocean which may have a moderating influence not reflected in the climatic data. It is interesting to note the absence of this species from the higher Appalachians of Virginia and West Virginia (Wagner, 1963), even though suitable acidic bogs and seeps exist in the area. It is apparent that mean temperature in itself is not a very useful index. As an indicator of potential climatic severity it can be important, although analysis is nearly impossible in the absence of values for variance. Thus, survival of W. areolata may not depend as much on its tolerance of —4.5°C as on its ability to withstand the occasional much colder winter. Of course the problem may be even more complex. Winter minima may reflect other associated parameters, such as the length of the growing season, that more directly affect the ability of the plant to grow, reproduce, and compete with its neighbors. Competition can be critical in the distribution of organisms (Hynes, 1954; Jaeger, 1970). Since competition varies as a function of community composition, which changes over the distribution of any particular species, a perfect or even very close fit between abiotic influences and distributional limits often does not obtain. Distributions of weedy species, therefore, probably more closely parallel absolute climatic limits than do distributions of plants of stable habitats, where interactions with other organisms are more structured. The recent northward spread of Asplenium platyneuron (L.) B.S.P., primarily a southern species, is a result of an increase in the disturbed habitats that provide this species with suitable sites in which to become established (Wagner and Johnson, 1981) and demonstrates that the Previous exclusion of this spleenwort was more a result of its competition with established communities than a result of its tenderness. A similar explanation may account for the presence of W. areolata on siliceous cliffs and ledges at the northern and western limits of its range, even though seemingly suitable bogs and seeps exist in these areas. At some point, the species composition of the typical habitat of this Species changes sufficiently so that it no longer is competitive in such situations. Sandstone cliffs and ledges in the same vicinity, which present physiological demands similar to those of acidic bogs (in the form of nutrient inavailability and drought stress), possess less plant cover and are characterized by a much lower diversity of species. It seems likely, therefore, that lithophily in W. areolata may be a AMERICAN FERN JOURNAL: VOLUME 73 (1983) 48 FIG. 1. Distribution of Woodwardia areolata. Dots represent terrestrial and unspecified neha while stars indicate epipetric stations. The ragged line represents the Janu —4.5°C isotherm. ert shows Canadian province of Nova Scotia. X indicates extirpated population. Arrows indicate outlying populations. R. CRANFILL: DISTRIBUTION OF WOODWARDIA AREOLATA 49 direct response to competition. Such cases would be parallel to but more subtle than Billings’ (1950, 1952) “Compensation Effect,” in which edaphic factors of certain soils and rock types compensated for the negative effects of other environmental factors. Transplants of individuals of W. areolata from Kentucky to Michigan survived over two winters (until 1981 when the experiment was discontinued) and indicate that winter minima alone are insufficient to account for the distribution of this fern. Further demographic and transplant experiments are needed to adequately test the hypothesis that competition is the controlling factor. Edaphic Factors.—The distribution of W. areolata fitted that of certain soil types even better than it did temperature. My measurements and those of Wherry (1921) indicate that the sporophyte of this fern is extremely acidophilous. The pH of several sites checked in Kentucky and Tennessee all fell below 5.5, with one reading of 4.0. It is not surprising then that the species is entirely absent from calcareous regions within its range. No stations are known in the limestone regions of the Interior Low Plateaus or in the calcareous Black Belt region of Alabama and Mississippi. Absence of the species from glaciated areas in southern Illinois and southeastern Ohio can be ascribed to the predominance of calcareous glacial drift in these areas (Fig. 2). The alluvial plain of the lower Mississippi River is also avoided. As a region of heavy basic to circumneutral clays (Kellogg, 1936; Braun, 1950), the Mississippi Alluvial Plain is devoid of most species characteristic of the sandy uplands and acidic seeps to the east and west. Correspondence of the distributions of W. areolata and Pinus echinata Miller (Little, 1971), another calcifuge, is striking. The best fit to soils occurs in eastern Texas (see inset, Fig. 2). The absence of the Chain Fern from the regions of the Blackland Prairie and Coast Prairie soils, both predominantly calcareous clays (Arbingast et al., 1976), is striking. Because of their greater matrix potential, clay soils must be hydrated to a higher percentage than coarser grained soils to exceed the permanent wilting percentage. As a result, clay soils tend to be droughtier in more arid regions and may become critical for W. areolata as it reaches its western limit. The patchwork distribution of these soils has made it impossible, therefore, to assess the relative importance of moisture availabil- ity (for an index, see Thornthwaite, 1948) and relative humidity as limiting factors; no good correlation was found between these factors and the western limit of W. areolata. As with the consideration of climate, distributional evidence is insufficient to demonstrate that heavy clay soils or pH are in themselves limiting. The indefinite Survival of adult sporophytes on the heavy, calcareous, nutrient-rich soils of the Inner Bluegrass region of Kentucky indicates that competition may actually consti- tute the limiting factor. The absence of this fern from much of the Allegheny Plateau cannot be explained by the climatic or edaphic criteria just discussed. Contrary to its name, the plateau region is highly dissected and affords little of the seep and bog habitat necessary for the species. It also lacks extensive deposits of resistant sandstone and conglomerate that form the secondary habitat of the fern. Although the previous discussion concerned the sporophyte, other stages 1n the life history may be important in controlling the distribution. Pteridophytes are like 0 AMERICAN FERN JOURNAL: VOLUME 73 (1983) bs) FIG. 2. Distribution of Woodwardia areolata in relation to soils; a = Mississippi Alluvial Plain, b = region of Interior Low Plateaus with calcareous soils, c = Ridge and Valley Province, d = Black blackland prairie soils while crosshatched area represents the extent of coastal prairie soils. Hatched line depicts extent of Wisconsin Glaciation. X indicates extirpated population. R. CRANFILL: DISTRIBUTION OF WOODWARDIA AREOLATA 5] lower plants in which the alternation of generations can have drastic consequences on the distribution of the organism (see the example of Laminaria, Dixon, 1965, pp. 109-115). In summary, the present distribution of W. areolata is clearly associated with the mean —4.5°C minimum January temperature to the north. The distribution of calcareous soils, especially clays, effectively limits the distribution to the south and may be important, in concert with increasing aridity, to the west. No correlation was found between the distribution of this fern and the indices of moisture availability and relative humidity. In all cases it appears likely that competition narrows the limits imposed by abiotic influences since preliminary experiments demonstrate the ability of this species to survive in more rigorous environments when competition is removed. The change from bogs and seeps to cliffs and ledges at the range limit is also compatible with this hypothesis. This “competitional compensation point” is probably more important in limiting the distributions of non-weedy species, which occupy more stable habitats than do weedy species. Experimental studies of problems of distribution are greatly needed. Through nar- rowing and refining our investigations, they may aid materially in the study of the effects of competition on community structure. Ferns make excellent subjects because they are small and fairly easy to grow and because the production and dissemination of propagules, critical in many flowering plants, is not so limiting in pteridophytes. I am grateful to A. R. Smith and Herbert Baker who provided comments on a draft of this paper. I am indebted to D. L. Melroy who provided assistance and criticism in numerous ways. LITERATURE CITED ARBINGAST, S. A., L. G. KENNAMER, and R. H. RYAN. 1976. Atlas of Texas. Bureau of Business Research, Univ. of Texas, Austin, TX. BILLINGS, W. D. 1950. Vegetation and plant growth as affected by chemically altered rocks in the western Great Basin. Ecology 31:62—74 . 1952. The environmental complex in Biol. 27:251-265. BRAUN, E. L. 1950. Deciduous Forests of Eastern North America. Free Press, New York, NY. COX, C. B., I. N. HEALEY, and P. D. MOORE. 1976. Biogeography, an Ecological and Evolutionary Approach. Blackwell Scientific Publications, Oxford, U.K. DIXON, P. S. 1965. Changing Patterns of Distribution in Marine Algae. Pp. 109-1 15 in C. G. Johnson and L. P. Smith, eds. The Biological Significance of Climatic Changes in Britain. Academic Press, London, U.K. : GORHAM, E. 1954. An early view of the relationship between plant distribution and environmental factors. Ecology 35:97—98. ae HOCKER, H. W. 1956. Certain aspects of climate as related to the distribution of Loblolly Pine. Ecology 37:824-834. : HOLMGREN, P., W. J. KEUKEN, and E. SCHOFIELD. 1981. Index Herbariorum, Part I. Guide to the Herbaria of the World, 7th ed. W. Junk, The Hague, Netherlands. HYNES, H. B. N. 1954. The Ecology of Gammerus duebeni Lilljeborg and its occurrence in fresh water in western Britain. J. Animal Ecol. 23:38-84. , R. C. 1970. Potential extinction through compe salamanders. Evolution 24:632-642. relation to plant growth and distribution. Quart. Rev. JAEGER tition between two species of terrestrial 52 AMERICAN FERN JOURNAL: VOLUME 73 (1983) KELLOGG, C. E. 1936. Development and Significance of the Great Soil Groups in the United States. . S. Dept. Agr. Misc. Pub. : KITFIELD, A. 1974. A range extension for Woodwardia areolata. Rhodora 76:312. KOEVENIG, J. L. 1976. Effect of climate, soil, physiography and seed germination on the distribution of the River Birch (Betula nigra). Rhodora 78:420-437. KREBS, C. J. 1972. Ecology, the Experimental Analysis of Distribution and Abundance. Harper and Row, New York, NY. LITTLE, E. L. 1971. Atlas of United States Trees, vol. I. Conifers and Important Hardwoods. U. S. Dept. Agr. Misc. Pub. 1146. NATIONAL ATMOSPHERIC AND OCEANOGRAPHIC INSTITUTE. 1980. Climate of the States, 2 vols. Dept. of Commerce, Washington, : OGDEN, E. C., F. H. STEINMETZ, and F. HYLAND. 1948. Checklist of the Vascular Plants of Maine. Bull. Josselyn Bot. Soc. Maine 8. PIELOU, E. C. 1979. Biogeography. Wiley and Sons, New York, NY. SALISBURY, E. J. 1926. The geographical distribution of plants in relation to climatic factors. Geogr. . 67:312-335. THORNTHWAITE, C. W. 1948. An approach to a rational classification of climate. Geogr. Rev. 38:55—94., WAGNER, W. H., Jr. 1963. Pteridophytes of the Mountain Lake area, Giles County, Virginia, including notes from Whitetop Mountain. Castanea 28:113-150. ————, and D. JOHNSON. 1981. Natural history of the Ebony Spleenwort, Asplenium platyneuron (Aspleniaceae) in the Great Lakes area. Canad. Field Nat. 95:156-166. WHERRY, E. T. 1921. The soil reactions of ferns of woods and swamps. Amer. Fern J. 11:5-16. —————. 1964. The Southern Fern Guide. Doubleday Nature Guide Series. Doubleday, Garden City, NY. REVIEW FERNS AND FERN ALLIES OF THE DRIFTLESS AREA OF ILLINOIS, IOWA, MINNESOTA AND WISCONSIN, by James H. Peck. Milwaukee Public Museum Contributions in Biology and Geology 53:1-140. 1982. $13.50 postpaid. —The driftless area, a pocket in the upper midwestern landscape which escaped at least the more recent glaciations, holds, by virtue of its topography and age of its habitats, a diverse and interesting array of plants. It has been the subject of botanical Study for many years. The present account is by far the most complete and useful of any we have had for the pteridophytes, which number 73 species, 13 hybrids, and 6 infraspecific taxa. Peck’s treatment includes a useful introduction, a discussion of the affinities of the flora with special reference to the interesting disjunct Thelypteris simulata, a systematic list with Synonyms, statement of habitat, and specimen citations, a key to the genera and species of pteridophytes found in the driftless area, and a large and useful section of literature cited. Ninety. pages are devoted to full-page county distribution maps for the driftless area ferns that include all of Minnesota, Wisconsin, Iowa, Illinois, and Missouri. The maps not only place the ferns of the driftless area in the context of the surrounding region, but they are sure to stimulate searches for pteridophytes in counties where the first records are yet to be obtained.—D.B.L. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) 53 Two Moonworts of the Rocky Mountains; Botrychium hesperium and a New Species Formerly Confused with It W. H. WAGNER, JR. and FLORENCE S. WAGNER* The taxonomy of western North American botrychiums still needs much research. Interpretations of the past were based largely upon scanty and poorly prepared collections. For over 30 years only two B. matricariifolium-like moonworts have usually been accepted for this region—B. boreale subsp. obtusilobum (Rupr.) Clausen and B. matricariifolium subsp. hesperium Maxon and Clausen (Clausen, 1938). As to the former, we conclude that the western North American plant is not closely related to B. boreale, as will be discussed in a monograph of this genus currently in preparation. The correct name for taxon obtusilobum is B. pinnatum St. John (Fig. 1, a—g). Taxon hesperium also proves to be a distinct species, readily distinguished from B. matricariifolium A. Br. subsp. matricariifolium, which occurs in North America only east of the Great Plains (Fig. 2, h-n). With our recent opportunity to investigate large populations of these plants in the field in numerous localities, we are now confident of the distinctness of not only B. pinnatum and B. hesperium, but of a third element as well, which is described here for the first time. It is no surprise to discover a new species related to B. hesperium in western North America, where the rate of endemism among moonworts is the highest in the world. With some ten out of 14 of the described and undescribed species known only there, western North America is clearly the metropolis for this subgenus (Botrychium subg. Botrychium). Using primarily the sterile lamina as a basis, we provide the following key to its major groups. KEY TO THE GROUPS OF BOTRYCHIUM SUBG. BOTRYCHIUM IN NORTH AMERICA 1. Sterile lamina absent; frond composed of two sporophores........--+.+++++s00sser007 B. paradoxum group 1. Sterile lamina present; frond with a single sporophore. 2. Larger pinnae mostly fan-shaped or wedge-shaped, not pinnatifid; costa absent or poorly developed. 3. Pinnae and lobes, especially the distal ones, commonly irregularly confluent; basal pinnae often SPONBAY GRammer ated. 22. Sin cccateage ss ste = swe ennennnenderseostengyeossienesstrees®* B. simplex group 3. Pinnae and lobes regularly separated; basal pinnae conform or only slightly exaggerated. B. lunaria group 2. Larger pinnae mostly oblong to lanceolate, pinnatifid, costa usually present, at least in basal i B. lanceolatum group Early in our studies we did not notice that the original collections of B. hesperium were mixtures of two species. Although certain specimens were different in a we simply believed that some specimens had narrow, ere an *Herbarium and Department of Botany, University of Michigan, Ann Arbor, MI 48109. 54 AMERICAN FERN JOURNAL: VOLUME 73 (1983) shag 1. Silhouettes of moonworts similar to B. echo and B. hesperium. FIGS. a-g. B. pinnatum : ae ae MICH). FIGS. h-n. B. matricariifolium (h, j, n, Wagner 81003, MICH; others Wagner WAGNER & WAGNER: TWO ROCKY MOUNTAIN MOONWORTS 55 and that its apparent variability resulted from this. Our field studies, however, soon dispelled this interpretation. The type specimen of B. hesperium includes only individuals with rounded segments. It was taken at Glacier Lake in Rocky Mountain National Park by a high school teacher, E. Bethel, in 1914. Bethel and others took numerous specimens there during the period 1911 to 1921. Bethel was aware that there were actually two elements involved, and he and Ira Clokey separated their 1921 collection at Glacier Lake into two groups—Bethel & Clokey 3987, typical, and 3987a, with “segments narrow, more acute.” W. R. Maxon was also aware of these distinctive plants as shown by his annotations of one of the herbarium sheets (US984959). Our new interpretation is based upon over 200 specimens of B. hesperium studied in the field in seven localities, and over 300 of the new species, B. echo, in ten localities. The two species grow in such similar habitats that one description of the habitat will suffice for both. They tend to occur together, often side-by-side. Also associated in genus communities with them in the southern Rockies are B. lanceolatum, B. lunaria, and B. minganense. In Montana and Alberta, where B. echo is not known, B. hesperium occurs also with B. paradoxum and B. pinnatum. Most of the localities that we studied had only a few plants. To illustrate, for B. echo, ten localities yielded respectively 1, 2, 9233, 9, al, 25, 9, and 150+ plants. The two most productive localities were in Arizona at Mount Baldy and at San Francisco Peaks. In the former locality, B. hesperium was absent, but at the latter we recorded 88 individuals of this species. At only one locality in the southern Rockies did we find only B. hesperium (Wagner 81158, 3 individuals in company with B. lanceolatum), but in the northern Rockies it apparently regularly occurs alone. The two species grow on grassy slopes, roadsides, and at edges of lakes. The soil is usually rocky, the substrate including decomposed granite as well as other rock types. In the southern Rockies the plants grow at elevations between 8,500 and 11,500 ft. The easiest way to find them is to drive along roads at proper altitudes and to seek flat roadside ditches with gravelly soil and scattered shrubby vegetation, Picea saplings and Salix shrubs dominating. Plants are sometimes found growing even in the gravel of the road shoulder! In addition to spruces and willows, other woody associates encountered are Lonicera involucrata, Potentilla fruticosa, and species of Abies, Juniperus, and Ribes. The herbaceous associates include weeds and involve such genera as Achillaea, Antennaria, Arenaria, Carex, Cerastium, Epilobium, Festuca, Fragaria, Frasera, Mertensia, Penstemon, Potentilla, Saxifraga, Sedum, Selaginella, Setaria, Solidago, Trifolium, Valeriana, and Zygadenus. Seek- ers of these plants should be warned that only one out of ten or twenty seemingly appropriate habitats yield these botrychiums. It is therefore necessary to sample many likely sites. : Rather than distinguish only B. hespertum and B. echo, all of the North American taxa of the B. lanceolatum group that might be confused with them are keyed. For more details of the differences between B. hesperium and B. echo themselves, the reader is referred to the descriptions and to Figure 4. This key is based upon medium and large individuals. AMERICAN FERN JOURNAL: VOLUME 73 (1983) uw mn uch FIG. 2. Silhouettes of moonworts. FIGS. a—i. B. echo (Wagner 82107b, MICH). FIG. ee wae divided form of B. echo (Bethel & Clokey 3987a, US; see also Fig. 3, a—e). FIGS. k-o. B. hesp (k-n, Wagner 82107a, MICH; 0, Bethel in 1914, US—type). WAGNER & WAGNER: TWO ROCKY MOUNTAIN MOONWORTS 57 KEY TO THE BOTRYCHIUM LANCEOLATUM GROUP IN NORTH AMERICA 1. Sterile lamina broadly deltate, sessile to subsessile; sporophores of full-sized plants usually composed of several major upright axes; plants of northern North America :.....B. lanceolatum 1. Sterile lamina mostly oblong to oblong-deltate, subsessile to stalked; sporophores of full-sized plants usually with one major upright axis, sometimes with one or two upright laterals. 2. Sterile segment mostly conspicuously stalked, the stalk 20-30% of the blade length; segment tips usually serrulate or crenulate; living lamina pale blue-green, dull; sporophore commonly twice as long as sterile lamina; plants of eastern North America B. matricariifolium 2. Sterile segment short-stalked or subsessile, the stalk 5-20% of the blade length; segment tips usually entire, repand, or pointed; living lamina color various; sporophore usually only 1.5 times as long as sterile segment (except in B. hesperium); plants of western North America. 3. Pinnae and lobes well separated, not approximate or overlapping, mostly more or less parallel- sided, linear to oblanceolate; pinna tips pointed; basal pinnae, except in the smallest and largest fronds, usually deeply cleft into a single lower projection and larger upper projection; lamina shiny green in life............--.s:sceecsseeeerersernnenenrerseeeesees ech 3. Pinnae and lobes usually approximate or overlapping, the large ones abruptly contracted at base, oblong-lanceolate to ovate to deltate; pinna tips blunted or rounded; basal pinnae not cleft into two projections; color and luster various. 4. Pinnae with few lobes, these mainly on the basal side; lowest pinnae exaggerated, ascending and subclasping, strongly asymmetrical, the lower side with coarse basiscopic lobe; segments broadly adnate at base; lamina gray-green, dull in POS Baga gr el een ORES B. hesperium _ Pinnae with numerous lobes, these roughly equal in number on the upper and basal sides; lowest pinnae mostly equal to or slightly larger or smaller than next distal pair, not ascending or clasping, nearly symmetrical, the lower side with small lobes subopposite to those on upper side; segments narrowly adnate at base; lamina bright green, shiny in life. B. pinnatum The following descriptions have been condensed to emphasize the most important differences between the species. Botrychium echo W. H. Wagner, sp. nov. Figs. 2-5. Sporophorum segmento sterili plerumque sesquilongius; segmentum sterile vivum supra vivide viride nitidumque, fere sessile vel brevissime stipitatum, lamina late oblonga 2.2 (1—4.5) cm longa; pinnae bene separatae, non imbricatae, oblanceolatae, , icl asibus subzygomorphis; pinnarum par infimum longitudine par proximum aequans vel paulo superans, patens, non amplectens; sporae tenelle verrucatae. Plants exclusive of their roots 9.5 (3-15) cm tall, the common stalk 6 (2-10) cm - g , short-stalked, broadly oblong, 2.2 (I-4.5) cm long; pinnae narrowly attached to a telatively narrow rachis, remote to approximate, not overlapping, lanceolate e aminar margins nearly entire; basal pinna pair not exaggerated in length, equal to or somewhat longer than the adjacent pair, spreading or only moderately ascending, not clasping; spores 37 (27-53) pm in maximum diameter, irregularly and finely verrucate, the warts small, low, and separated by narrow, shallow channels. TYPE: Glacier Lake, Boulder Co., Colorado, 2800 m. alt, E. Bethel & I. W. Clokey 3937a (US; isotypes CAS, WTU). PARATYPES: ARIZONA: Apache Co.: White Mts., Mt. Baldy, on open bald, Wagner 82101 (MICH). Coconino Co.: San Francisco Mt., Inner Basin, E. L. Little, Jr. 4741 (US—mixed with B. lunaria), NE slope of 58 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Fey Me g FIG. 3. Silhouettes of moonworts. FIGS. a-—e. Unusually dissected forms of B. echo resembling B. matricariifolium (Wagner 82101, MICH—see also Fig. 1, j). FIG. f. Probable B. lanceolatum X minganense hybrid; note the few, long segments (Wagner 82/2], MICH). FIGS. g—i. Probable B. echo < minganense hybrids; note the relatively more numerous, shorter segments (Wagner 82104, MICH). Mt. Doyle, Wagner 82107b (MICH). COLORADO: Boulder Co: Glacier Lake, July 1914, E. Bethel (US), 13 July 1912, E. R. Cross (US), Bethel & Clokey 3987 (UC—2 sheets, both mixed with B. hesperium); Arapahoe Moraine, E slope, 1 mi S of University Camp, W. A. Weber 3431 (WTU). Clear Creek Co.: Roadside near Echo Lake, Wagner 80136b (MICH), 8/153b (MICH); Warren Mt. Picnic Ground, 2.8 mi E of CO-515, Wagner 81158 (MICH); US-6, 0.8 mi S of I-70, Wagner 81160 (MICH). El Paso Co.: Pikes Peak below moraine, H. L. Shantz 52 (US). Gunnison Co.: Monarch Pass, Wagner 82118 (MICH); 0.5 mi E of Monarch Pass on CO-50, Wagner 82123 (MICH). Lake Co.: Road to Independence Pass, 3.1 mi E of Hairpin Turn, Wagner 82/27 (MICH). Summit Co.: Near Breckenridge, K. K. Mackenzie 99 (NY—mixed with B. lanceolatum); CO-91, 2-3 mi S of 1-70, Wagner 81164 (MICH). UTAH: Summit Co.: 1.5 mi S of Spirit Lake, A. H. Holmgren et al. 7130 (UC—mixed with B. lunaria). The Greek specific epithet echo is used here in apposition. It was chosen to reflect the fact that this moonwort seems to repeat the characteristics of other, similar species. One of the best areas to study it is where we first recognized its distinctions from B. hesperium, namely Echo Lake on the slopes of Mount Evans, olorado WAGNER & WAGNER: TWO ROCKY MOUNTAIN MOONWORTS 59 A 10 MM BR FIG. 4. Comparison of B. hesperium (A) and B. echo (B) from a mixed population at San Francisco P eaks, AZ. Tracings of cleared sterile blades. The characters are: 1=ovate, broadly attached vs. linear-lanceolate, narrowly attached pinnae. 2= Overlapping vs. separated pinnae. 3=Repand vs. vs. short In certain respects, especially the shape of the pinnae and the glossy laminar surfaces, B. echo resembles B. lanceolatum. We have not yet detected any hybrids between the two species, but they might readily be confused with one or the other of the parents. A distinctive but uncommon form of B. echo with more dissected pinnae than normal is known (Fig. /, j; Fig. 3, a-e). The basal pinnae and sometimes the medial pinnae possess 3 or 4 lobes on the lower side and 2 or 3 lobes on the upper side. Comparison of the outline of these with B. matricariifolium (Fig. 1, h—-n) shows considerable resemblance. Clausen (1938), who combined B. echo and B. hesperium as a single taxon, surely had such plants as these in mind when he wrote that some specimens “can be matched almost exactly by material of [typical matricariifolium).” The dissected form is especially well developed at Mount Baldy, White Mountains, Arizona. It is connected by intermediates to the normal form. Botrychium hesperium (Maxon & Clausen) Wagner & Lellinger, 71:92. 1981, pro hybr. BS bes ny matricariifolium subsp. hesperium Maxon & Clausen, Me 38. Amer. Fern J. m. Torrey Bot. Club 19:88. 60 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FIG. 5. Spores of B. echo (A) and B. hesperium (B). SEM photographs x 1,100. Note the differences in size and form of the verrucae. Plants exclusive of their roots 12 (5-20) cm tall, the common stalk 7 (3-13) cm tall, sporophore relatively tall, 5 (3-10) cm tall, nearly twice as long as sterile segment, 80 percent with | or more basal branches 4 or more as long as main axis of sporangial cluster; sterile segment gray-green and dull in life, mostly short-stalked, subdeltate, 2.5 (1-5) cm long; pinnae broadly attached to a relatively wide rachis, crowded to commonly overlapping, ovate to lanceolate with rounded apices, the pinna bases asymmetrical, the lamina margins finely repand; basal pinnae com- monly exaggerated, up to twice as long as the adjacent ones, often upright and commonly clasping; spores 37 (29-50) um in maximum diameter, irregularly and coarsely verrucate, the warts large, prominent, and separated by wide, deep channels. TYPE: Glacier Lake, Boulder Co., Colorado, 8500 ft, July 1914, E. Bethel (US—S isotypes in addition). OTHER COLLECTIONS EXAMINED: CANADA: Alberta: Waterton Lakes National Park, W of Red Rock Canyon Parking Area, Wagner 81103a-d (MICH). ARIZONA: Coconino Co.: San Francisco Peaks. Inner Basin, NE slope of Doyle Mt., Wagner 82107a (MICH). COLORADO: Boulder Co.: Glacier Lake, Bethel & Clokey 3987 (CAN, CAS, US); Rocky Mt. National Park, Loch Vail Trail, Glacier Gorge, A. E. Porsild & B. E. Willard 23102 (CAN). Clear Creek Co.: Roadside near Echo Lake, Wagner 80136a (MICH). 8/153a (MICH). Lake Co.: Road to Independence Pass, 3.1 mi E of Hairpin Turn, Wagner 82128 (MICH). MONTANA: Deer ge Co.: Flint Ridge Mts., Storm Lake, S of Georgetown Lake, Wagner 80/29a (MICH), 81116b (MICH). It is interesting to note that Maxon’s original, unpublished interpretation of this taxon was as given here. On the type specimen he had written the label to read “Botrychium hesperium Maxon sp. nov.” The occurrences that we have found so far WAGNER & WAGNER: TWO ROCKY MOUNTAIN MOONWORTS 6l in Montana and Alberta contain plants much smaller on the average than those in Colorado and Arizona. The northern plants resemble young stages of the southern ones, and they may be dwarfed by the climate. The coarsely sculptured spores that we observed in B. hesperium (Fig. 5, B) are like those of B. lanceolatum. For B. echo, our collection records range from 13 July to 20 September, with most specimens taken in August. Botrychium hesperium may be similar, but there is some evidence that it may appear earlier and die down earlier. Judging from the condition of the large sample of plants taken at San Francisco Peaks, B. hesperium shows more effects of ageing. Seventy percent of the specimens of B. hesperium collected on 21 August 1982 showed evidence of damage—browned margins and broken, eaten, or otherwise tattered pinnae—compared to only 30% of B. echo collected at the same place and time. The leaves of B. echo appeared to be fresher. s would be expected, sterile interspecific hybrids involving both B. hesperium and B. echo have been encountered. Their hybrid origin is deduced by their association with parents, occurrence as usually one or a few plants, morphological intermediacy, and abortion of spores. Because only a single leaf is produced per year, it is difficult to obtain cytological observations as a rule; however, Sahashi (1979) has documented irregular meiosis in a Japanese hybrid. The most obvious interspecific hybrids are those in which the parents differ strongly (e.g., the Scandinavian hybrid of B. boreale and B. lunaria). Botrychium hesperium hybridizes readily with B. paradoxum to produce very obvious intermediates, striking because one of the two sporophores is “half sterile” (Wagner & Wagner 1981; Wagner et al., 1982). We have encountered a few definite hybrids involving B. echo. These involve B. minganense Victorin. Some are from Mount Baldy, Arizona, where the parents occur together locally in abundance. At San Francisco Peaks, we found a number of sterile plants involving what appear to be combinations of B. echo, hesperium, and lunaria, but these will require more study to separate. Some hybrids involve yet other species, for instance, a specimen from Monarch Pass, Colorado, we first thought was B. echo X minganense (Fig. 3 f). However, the cutting is somewhat different; also B. echo is rare and sporadic at this locality, while both B. lanceolatum and minganense are abundant and the latter species probably are the parents. This is part of an investigation of the evolution and systematics of Botrychium made possible by NSF grants DEB 800536 and DEB 8202768. We are indebted to a number of individuals who have helped us, especially W. R. Anderson, R. Eccleston, R. H. Hevly, J. Kuijt, J. D. Montgomery, A. Neas, W. A. Weber, M. D. Windham, and G. Yatskievych. The following herbaria have kindly supplied speci- mens of the species considered here: ASC, BRY, CAN, CAS, DAO, MO, NY, UC, US, and WTU. ‘Confirmation of spore abortion is accomplished in Botrychium using only a dissecting microscope at 30-60X magnification. The spores of specimens that were dried while the sporangia still had not completely discharged tend to be released and become attached (electrostatically?) to the sporophore axes and opened sporangial walls. If the spores are abortive, the sizes are extremely irregular—very small, normal, and very large. The largest spores tend to be more or less spherical and not tetrahedral. 62 AMERICAN FERN JOURNAL: VOLUME 73 (1983) LITERATURE CITED CLAUSEN, R. T. 1938. A monograph of the Ophioglossaceae. Mem. Torrey Bot. Club 19:1—-177. SAHASHI, N. 1979. Morphological and taxonomical studies on Ophioglossales in Japan and the adjacent regions. Identity of Sceptridium Lyon in the Izu Islands. J. Jap. Bot. 54:273—281. WAGNER, W. H., Jr. and F. S. WAGNER. 1981. New species of moonworts, Botrychium subg. Botrychium (Ophioglossaceae), from North America. Amer. Fern J. 71:20-30. , F. S. WAGNER, and C. HAUFLER. 1982. A hybrid population involving the “all-fertile” Botrychium paradoxum and the hemidimorphic B. heSperium (Ophioglossaceae). (Abstract). Bot. Soc. Amer. Misc. Publ. 162:77-—78. SHORTER NOTES LYCOPODIUM COMPLANATUM AND L. ANNOTINUM FOUND IN THE BLACK HILLS.—Lycopodium complanatum L. and L. annotinum L. were found growing together in the Black Hills of Wyoming on July 27, 1982. The location is in Crook County, Upper Sand Creek at confluence with Spottedtail Gulch, T51N R60W Section line of 20-21, elevation 5600 feet. The plants were growing under White Spruce, Picea glauca (Moench) Voss, and Hazelnut, Corylus cornuta Marsh. along with Low Red Huckleberry, Vaccinium scoparium Leiberg. Lycopo- dium complanatum was found only at this location, but L. annotinum extended on down the canyon at several more localities. Specimens are deposited at the Univer- sity of Wyoming (RM) and the New York Botanical Garden (NY) (Dorn 3793, 3792). The closest known locality for L. complanatum is about 450 miles to the northwest in Lewis and Clark County, Montana. The closest known locality for L. annotinum is about 270 miles to the west in Park County, Wyoming. Several other pteridophytes were collected within two miles of the Lycopodium location along the same creek: Equisetum sylvaticum L. (Dorn 3777, collected for the first time in Wyoming one day earlier by E. F. Evert in the Big Horn Mountains), Athyrium filix-femina (L.) Roth (Dorn 3 778), and Equisetum scirpoides Michx. (Dorn 3809, another first record for Wyoming).—Robert D. Dorn, Box 1471, Cheyenne, WY 82003. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 2 (1983) 63 MICROFIBRILS IN THE XYLEM OF BLECHNUM VIVIPARUM. — Cellulose forms the framework of plant cell walls by a system of microfibrils (as well as other molecular shapes) in which various substances are incrusted. The presence of microfibrils can be demonstrated by chemical removal of the incrusting substances or, rarely, by their actual appearance in the lumen of pits, particularly in relation to tori and plasmodesmata of some angiosperms and conifers. After the formation of secondary walls in some Gnetaceae and Coniferae (Warchop et al., Holzforsch. 13:115-120. 1959), a sort of “tertiary growth” appears in the shape of warts or of fibrils, either isolated, at random, or in a margo (a marginal reticule that fuses at or near the center of the torus), and sometimes obliterates the lumen (Butterfield & Meyland, Three-dimensional Structure of Wood, p. 72. 1980). FIG. 1. Isolated, sporadic xylem elements in Blechnum viviparum interpreted as secondary vessels. Two pits showing a loose net of microfibrils, = 10,000. In the material of Blechnum viviparum (Broadh.) C. Chr. studied and reported by Montiel and Guevara (Rev. Biol. Trop. 27:171-176. 1979) and reinterpreted by Gomez (Brenesia 18:253—258. 1980), some of the pits show what in my opinion are microfibrils (Fig. 1) which, to my knowledge, have either been overlooked or have not yet been reported from vascular cryptogams where xylem elements or secondary growth have been detected. It must be pointed out that the microfibrils reported here seem somewhat thicker than those usually found in higher plants and, unlike those, ours are sporadic and quite scattered throughout the material. The presence of microfibrils in ferns is interesting from a phylogenetic point of view inasmuch as they represent another intermediate character between the tracheid and the coniferophyte-type element. Furthermore, the presence of loose microfibrils Partially obliterating the lumina between crassulae, projecting from the middle lamellae to the secondary walls in a pteridophyte, seems to support the idea of a coherent, organized constitution of cellulose in the primary membranes as proposed by Frey-Wyssling et al. (Planta 47:1 15-126. 1956) and Miihlethaler (Biochem. Biophys. Acta 5:1-9. 1950), at least in these primitive plants.—Luis D. Gomez P., Museo Nacional de Costa Rica, Apartado 749, San José, Costa Rica. 64 AMERICAN FERN JOURNAL: VOLUME 73 (1983) BISPORANGIATE ANOMALOUS SPOROPHYLLS IN ISOETES FROM RAJASTHAN—The presence of a solitary, adaxial sporangium on or in association with a sporophyll is the fundamental feature of all lycopods. Only two exceptions have been reported. The fossil Lycostachys protostelicus was described by Pant and Walton (Palaeontographica 108:1—10. 1961) as having two subarchesporial pads in a sporangium; and in /soétes indica, Pant and Srivastava (Proc. Natl. Inst. Sci., India, B, Biol. 23:242-280. 1962) described sporophylls having two, almost equally developed sporangia. The discovery of bisporangiate sporophylls in two additional species of Isoétes, I. coromandelina L. f. collected at Poasa near Jaipur, Rajasthan, and in an apparently new species of /soétes from Mt. Abu, Rajasthan, India, are additions to this list. FIG. 1. Bisporangiate, abnormal sporophylls of /soétes coromandelina, X 1.2. FIG. 2. Longisection of bisporangiate sporophyll of /soétes sp. from Mt. Abu showing two unequal sporangia, X 20. A number of abnormal sporophylls were discovered in J. coromandelina which possess double sporangia (Fig. 1). These seem to be produced by fusion between adjacent sporophylls during their development. The proximal portions of the sporophylls are united and bear two sporangia; the distal, acicular portions of the sporophylls are free. The ligules and upper labia are also double. However, in other details of morphology and anatomy the sporophylls are normal. In the Isoétes from Mt. Abu, an abnormal sporophyll bearing two sporangia was discovered during sectioning of the material. The two sporangia are unequal and are Separated by a partitioning wall (Fig. 2). The smaller sporangium is empty and seems to be a bud-like, sterile structure; the bigger one is normal and contains numerous megaspores. From the present discovery, it seems that double sporangia are not so rare in Isoétes. Further investigations are needed to establish any possible phylogenetic conclusions.—B. D. Sharma, R. Singh, and D. R. Bohra, Department of Botany, University of Jodhpur, Jodhpur 342001, India. ——— ee LI NT NE Ny Re BRITISH PTERIDOLOGICAL SOCIETY Open to all who are interested in growing and studying ferns and fern-allies. Full members receive THE FERN GAZETTE and the BULLETIN OF THE BRITISH PTERIDOLOGICAL SOCIETY. Membership subscriptions are £7 for full members, £ 5 for ordinary members (not receiving the GAZETTE), and £5 for student members (under 25 years of age). For particulars, U. S. residents should apply to Dr. J. Skog, Biology Department, George Mason University, Fairfax, VA 22030. Non-U. S. residents should apply to Lt. Col. P. G. Coke, Robin Hill, Stinchcombe, Dursley, Gloucestershire, England. 4 ho? yr _.. from TRIARCH We are grateful for the purchase orders and collec- tions of material received during Triarch’s 56 years of service. We try to say thank you by maintaining quality of product, variety of listings, and by supporting biolog- ical societies with memberships, donations, and advertising. Having completed 25 years in the management of Triarch, | add this written word of thanks for your support, without which Triarch would not exist! Paul L. Conant, President TRIARCH PREPARED MICROSCOPE SLIDES P. O. Box 98 =e Ripon, Wisconsin 54971 AMERICAN ae FERN ee mber, 19 JOURNAL QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Observations on the Structure and Function of Hydathodes in Blechnum lehmannii JOHN S. SPERRY 65 A Reclassification of the Fern Genus Pyrrosia K. H. SHING 73 Notes on the Ecology and Development of Plagiogyria fialhoi PAULO G. WINDISCH and MARILIA PEREIRA-NORONHA 79 The Ferns of Elden Mountain, Arizona MICHAEL D. WINDHAM 85 Donovan S. Correll (1908-1983) 94 Reviews 93-96 RSSOURT BOTANICAL OcT 6 1983 GARDEN LIBRARY The American Fern Society Council for 1983 DEAN P. WHITTIER, Dept. of Biology, Vanderbilt University, Nashville, TN 37235. President TERRY R. WEBSTER, Biological Sciences Group, University of Connecticut, Storrs, CT 06268. Vice-President MICHAEL I. COUSENS, Faculty of Biology, University of West Florida, Pensacola, FL 32504 Sec retary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Treasurer JUDITH E. SKOG, Dept. of Biology, George Mason University, Fairfax, VA 22030. Records Treasurer DAVID B. LELLINGER, Smithsonian Institution, Washington, DC 20560. Journal Editor ALAN R. SMITH, Dept. of Botany, University of California, Berkeley, CA 94720. Memoir Editor JOHN T. MICKEL, New York Botanical Garden, Bronx, NY 10458. Newsletter Editor American Fern Journal EDITOR DAVID B. LELLINGER U.S. Nat’l Herbarium NHB-166, Smithsonian Institution, Washington, DC 20560. ASSOCIATE EDITORS DAVID W. BIERHORST Rt. 3, Box 188, Picayune, MS 39466. GERALD J. GASTONY Dept. of Biology, Indiana University, Bloomington, IN 47401. JOHN T. MICKEL New York Botanical Garden, Bronx, NY 10458. TERRY R. WEBSTER ....... Biological Sciences Group, University of Connecticut, Storrs, CT 06268. The “American Fern Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the general study of ferns. It is owned by the American Fern Society, and published at the Smithsonian Institution, Washington, DC 20560. Second-class postage paid at Washington. Claims for missing issues, made 6 months (domestic) to 12 months (foreign) after the date of issue, and the matters for publication should be addressed to the Editor. Changes of address, dues, and applications for membership should be sent to Dr. Leslie G. Hickok, Degaiunels of Botany, University of Tennessee, Knoxville, TN 37916. Orders for back issues should be addressed to Dr. James D. Montgomery, Ichthyological Associates, R.D. 1, Berwick, PA 18603. General inquiries cree ferns should be addressed to the Secretary. Subscriptions $9.00 gross, $8.50 net if paid Re an agency (agency fee $0.50); sent free to members of the Sea eats Society Siem — $8.00 + $4.00 mailing surcharge beyond U.S. A Canada, and Mexico; life membership, $160 Back volumes 1910-1978 $5.00 to $6.25 ste ‘single back numbers of 64 pages or less, $1.25; 65-80 pages, $2.00 each; over 80 pages, $2.50 each, plus shipping. Back volumes 1979 et seq. $8.00 each; single back numbers $2.00 each, plus shipping. Ten percent discount on orders of six volumes or more. Library Dr. John T. Mickel, New York Botanical Garden, Bronx, NY 10458, is Librarian. Members may borrow books at any time, the borrower paying all shipping costs. Newsletter Dr. John T. Mickel, New York Botanical Garden. Bronx, NY 10458, is editor of the newsletter “Fiddlehead Forum.” The editor welcomes contributions from members and non-members, including miscellaneous notes, offers to exchange or purchase materials, personalia, horticultural notes, an nd reviews of non-technical books on ferns Spore Exchange Mr. Neill D. Hall, 1230 oe 88th Street, Seattle, WA 98115, is Director. Spores exchanged and collection lists sent on reque Gifts and Bequests Gifts and bequests to the Society enable it to expand its services to members and to others ee in ferns. Botanical books, back issues of the Journal, and cash or other gifts are always welcomed, a are tax-deductible. Inquiries should be addressed to the Secretary. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 3 (1983) 65 Observations on the Structure and Function of Hydathodes in Blechnum lehmannii JOHN S. SPERRY* The fronds of many ferns in the Polypodiaceae (Ogura, 1972) and Cyatheaceae (Weiler, cited in Lippmann, 1925) possess swollen vein endings associated with specialized adaxial epidermal cells. Their structure is similar in all ferns (Gardiner, 1883: Potonié, 1892; Poirault, 1893; Goebel, 1930; Guttenburg, 1934), including Blechnum lehmannii Hieron. (Figs. 1-3). The depressed epidermis (Fig. 3) sug- gested the term “Wassergriibchen” for the vein endings to European anatomists of the nineteenth century (e.g., Potonié, 1892). Vein endings on developing fronds secrete water (Fig. /) when conditions reduce or stop transpiration (Mettenius, 1856; Haberlandt, 1894a; Goebel, 1926, 1930). In some species, the secreted water contains salts in solution which form “chalk scales” after the water has evaporated (Mettenius, 1856; Potonié, 1892; Lippmann, 1925). Most sources report that water secretion is dependent on the metabolic activity of the specialized epidermal cells adaxial to the vein ending (Fig. 3, e); it is thus regarded as an active, or glandular, process (Gardiner, 1883: Haberlandt, 1894a, 1914; Lepeshkin, 1906; Stocking, 1956; Fahn, 1979). This conclusion is reflected in various references to the vein endings as water glands, salt glands, or chalk glands. There is, however, very little evidence to support any conclusions on the mechanism of secretion. All work on the subject was performed in the last century, and the results obtained are questionable due to the bias in experimental design which prevented testing of alternative hypotheses and to the possibility of artifacts in technique (see comments in Spanjer, 1898; Haberlandt, 1898; Lepeshkin, 1923). Moreover, the results are contradictory, since they have been interpreted as support- ing either an active (Gardiner, 1883; Haberlandt, 1894a, 1914) or a passive (Spanjer, 1898) mechanism of secretion. Finally, all experiments were performed on a single species, Polypodium aureum L., and need not represent the situation in other ferns. Because the mechanism of secretion from vein terminations is unknown, they will be referred to in this paper by the term “hydathode” which, as originally defined by Haberlandt (1894b), did not presuppose a mechanism of secretion. Haberlandt used his term to refer to structures on above-ground portions of the plant, particularly foliage leaves, which function in water transfer to and from the plant surface. Previous work on the mechanism of hydathode secretion in ferns neglected important aspects of the problem. Despite the fact that root pressure could play a Significant role in the secretion process, the relationship of root pressure to secretion in intact plants was not studied. In fact, there is no well documented account of root Pressure in ferns. In addition, investigators could not analyze secretion from a structural standpoint because anatomical studies were confined to the mature, non-secreting hydathodes; the structure of the secreting hydathodes on developing leaves was unknown. In this study, the relationship of root pressure to secretion as well as developmental changes in hydathode structure were investigated in B. oe Forest, Petersham, MA 01366. olume 73, number 2, of the JOURNAL was issued 16 June 1983. 66 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FIG. 1. Hydathodes secreting water on a developing frond; arrow indicates a droplet of water above a hydathode. FIG. 2. Epidermal peel of hydathode photographed with dark field illumination. There 1s no intercellular space in the epidermal layer; the separation of cells marked by the arrow is an artifact of preparation due to the weak adherence between epidermal cells. FIG. 3. Transverse section of a hydathode on a mature frond made parallel to the leaf margin and treated with I-KI/H,SO,. e = epidermal cells of hydathode, en =endodermis; arrows indicate Casparian strip. lehmannii in its native locality, the Central American cloud forest. Observations Suggest that one mechanism of secretion from fern hydathodes is the passive transmission of xylem sap from vein endings to the plant surface through the apoplast of the hydathode under a pressure gradient induced by root pressure. MATERIALS AND METHODS Plants growing in the cloud forests near Xalapa in the state of Veracruz, Mexico, provided material for both experimental and anatomical work. Individual plants consist of a single, erect rhizome with short internodes. ; Root pressure was measured with a bubble manometer attached to the cut stipe (Fig. 4A) and a thermometer. The manometer was made with surgical tubing (ca. J. S, SPERRY: OBSERVATIONS ON BLECHNUM HYDATHODES 67 © 1.204 S = : = oO O = S ® 2 | z= = 0b 2 3 © 5 | ep) Yt | © 1.00 C5, | hos ee oikece rater, CEL ae 05 10 2.0 B time (hrs.) FIG. 4A. Manometer attachment for measuring root pressure. B = clear plastic bag enclosing the entire crown of an individual plant, S=cut stipe to which the manometer is attached, C=clamp, Y = forked connector, BB = bubble at sealed end of the manometer tube. FIG. 4B. Root pressure vs. time for plants 1, 2, 3. S=onset of hydathode secretion. 0.8 mm inside diameter) partially filled with acid fuchsin and sealed at one end so as to leave a bubble at the sealed end (Fig. 44, BB). Bubble length was assumed to be proportional to bubble volume. At the beginning of the experiment, bubble length at ambient pressure and air temperature was determined by releasing the clamp (Fig. 4A, C). After the clamp was closed, subsequent readings of bubble length and air temperature were made, and exudation pressure relative to ambient was calculated according to Gay-Lussac’s law (Weast & Astle, 1981). Clamping itself did not affect bubble length. Care was taken when attaching the manometer so that a continuous column of liquid was present from stipe to bubble, and the attachment was gently secured with twist-ties to help guard against leakage. The relationship between root pressure and hydathode secretion was studied in three plants growing along a shaded stream in the cloud forest on a clear day. The entire above-ground portion of each individual plant was enclosed in a clear plastic bag and root pressure was monitored with a manometer attached to a single cut frond for each plant (Fig. 4A). Observations of xylem pressure and hydathode secretion were then made. Anatomical differences between the secreting and non-secreting hydathodes on alcoholic sodium hydroxide followed by lactic acid. Observations were made on Preparations left unstained or stained in toluidine blue. The distribution of lignin was determined from hand sections stained in phloroglucinol and concentrated HCI. Suberized and cutinized cell walls were identified using the I-KI/H2SO,j test. 68 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FIGS. 5-13. Development of Casparian strip. All preparations were treated in I-KI/H,SO,. FIGS. 5-8, 10, and 12 are from cleared preparations; FIGS. 9, 11, and 13 are transverse sections made parallel t0 the leaf margin. Arrows = Casparian strip. FIGS. 5~9. Non-secreting hydathodes on mature frond. FIG. 5. Vein subtending a hydathode and surrounded by endodermis with fully developed Casparian strip. FIG. 6. Hydathode viewed from the adaxial epidermis and focussed on the Casparian stip bordering the edge of the vein ending. FIG. 7. Higher magnification of the Casparian strip as viewed 9 J. S. SPERRY: OBSERVATIONS ON BLECHNUM HYDATHODES 69 OBSERVATIONS Secretion.—Secretion was observed on developing, unenclosed fronds in foggy and rainy weather in the cloud forest and when individual crowns were enclosed in plastic bags. In bagged plants, secretion was initiated on successive pinnae in an acropetal direction so that the last hydathodes to begin secreting on developing fronds were those on the most distal pinnae. Based on qualitative observations, distal hydathodes appeared to have a greater rate of secretion than more proximal hydathodes, which on developing fronds sometimes showed no secretion. Observa- tions of fronds at different stages of development as judged by relative size, color, and texture suggested that the permanent loss of secreting capability by hydathodes occurs acropetally. No salt deposits of any kind were observed to accumulate above the hydathodes oot pressure.—Root pressure was determined by examining the bleeding behavior of cut stipes (Fig. 4A, S) on plants growing in conditions which were assumed to severely reduce or stop transpiration. Such conditions were either natural, as on rainy or foggy days, or induced by enclosing the entire above-ground portion of an individual plant in a clear plastic bag (Fig. 4A, B). Root pressure was assumed to be present when the attached stipes showed continual bleeding after being cut. Initially, some of this bleeding was due to mucilage flow as judged by the viscous nature of the exudate. However, after continual blotting over a period o several minutes, the viscosity of the bleeding fluid decreased considerably, and with the aid of a hand lens, xylem sap could be seen issuing from the stele. Plants in which transpiration was not inhibited did not show this bleeding behavior from cut stipes; here the only exudation noted was a small amount of mucilaginous material. Although the secretion of mucilage from the cut end of the stipe was a potential problem for root pressure measurements, the results indicate that the pressure produced by the secretion is insignificant. Certain plants (e.g., plant 1, Fig. 4B) did not show root pressure even when transpiration was reduced by enclosure of the crown in a plastic bag. The absence of root pressure in these plants was apparently due to insufficient soil moisture. Despite the fact that the stipes of these plants showed mucilage secretion when cut, no significant increase in pressure above ambient was noted from attached manometers. The relationship between root pressure and hydathode secretion studied in the three bagged plants (labelled 1, 2, 3) is summarized in the graph of Figure 4B. In plants 2 and 3, the buildup of root pressure in the xylem correlated positively with the observation of water secretion from hydathodes of developing fronds. In bot plants, the measured pressures were sufficient to support a column of water taller than the height of the plant (1.8 m for plant 2, 1.2 m for plant 3). Plant | developed directly on the tracheids of the vein ending. FIG. 8. Casparian strip adaxial to the vein ending in surface Probably phenolic vacuoles. 70 AMERICAN FERN JOURNAL: VOLUME 73 (1983) no root pressure, and no secretion was observed from its hydathodes. Variation in root pressure in the three plants is correlated with position relative to the stream flowing under the bank where they were growing. Plant 2, which showed the most dramatic rise in pressure, was growing roughly 0.3 m above the stream level. Plant 1, which showed no root pressure, was 2.5 m above the stream. Plant 3 was intermediate in pressure and bank position. Possibly the explanation for this correlation is decreasing soil moisture with increasing height above the stream; greater soil moisture would favor the generation of higher root pressures. Hydathode development.—The ability of hydathodes to secrete is correlated with specific features of their development. Hydathode cells reach their mature size and are capable of secreting water long before the cells of the surrounding leaf tissue have finished expanding. Hydathodes capable of secreting protrude above the surrounding epidermal cells (Fig. 13). Only after the leaf cells reach their full size and the leaf achieves its mature thickness do hydathodes become the depressed “Wassergriibchen” consistently described by early anatomists (Figs. 3 and 9). In the recessed stage, hydathodes were observed to be incapable of water secretion. The structural change that appears to be most closely related to secretory function is the progressive suberization of the hydathode endodermis. The Casparian strip surrounding the hydathode (Figs. 3 and 9) is the distal extremity of a network which surrounds the entire mature vascular system of the plant. The development of this continuous Casparian strip is acropetal, and lags considerably behind the maturation of protoxylem and hydathode cells (Figs. /0, 12, and 13). Thus, hydathodes in developing tissues do not possess a suberized endodermis. The acropetal extension of the Casparian strip to the hydathode is first evident in the endodermis abaxial to the vein ending (Fig. //, arrows). The vein subtending the hydathode is completely surrounded by a Casparian strip (Fig. 5). At this time, the Casparian strip around the vein ending is like a mitten with the abaxially turned palm cut out. Development of the Casparian strip proceeds with the first steps represented by thin bands of suberin extending from the well developed strip below the vein. Eventually, the abaxial endodermis of the vein ending possesses 4 Casparian strip as well developed as anywhere else in the frond (Fig. 9). The degree of Casparian strip development at the hydathode correlates closely with the capability of the hydathode to secrete water. Examination of developing fronds from plants 2 and 3, which possessed distal secreting hydathodes and proximal non-secreting hydathodes, indicated that secretion only occurred from hydathodes where the Casparian strip was either absent or incompletely developed. In one frond, for example, distal secreting hydathodes which from qualitative observation showed the highest secretion rate possessed no Casparian strip (Figs. 10 and /3). Hydathodes on pinnae from the middle part of the frond, which appeared to have a lower rate of secretion, possessed an incompletely developed Casparian strip adaxial to the vein ending (Fig. //). Hydathodes at the base of the frond showed no secretion and were found to be completely surrounded by a thick Casparian strip (Figs. 6-9). In addition, examination of fronds at different stages of Casparian strip development indicated that the acropetal progression of complete hydathode suber- ization correlates with the acropetal loss of secretion capability in the hydathodes. : { J. S. SPERRY: OBSERVATIONS ON BLECHNUM HYDATHODES 71 DISCUSSION The correlation of root pressure with secretion from hydathodes on the one hand, and the correlation between Casparian strip development and the cessation of hydathode secretion on the other, suggest a simple explanation for the occurrence of water secretion from hydathodes in B. lehmannii. Root pressure is directly responsi- ble for secretion by supplying a pressure gradient sufficient to drive the passive movement of xylem sap from the vein ending of the hydathode, through the apoplast of the sub-epidermal and epidermal cells, and through the thin cuticle to the outside of the plant. When the Casparian strip is completely formed around the vein ending of the hydathode, the apoplast is sealed and secretion from the hydathode is blocked. Preliminary results from Polypodium and Nephrolepis species indicate that this hypothesis can also apply to hydathodes in these genera. Root pressure is known to be directly responsible for secretion from the epidermal- pore hydathodes of angiosperms (Fahn, 1979). Although pores are not present in the epidermis of fern hydathodes, root pressure can move water to the plant surface through the anticlinal walls of the epidermal cells. According to Meidner (1977), root pressure causes the secretion of water from the epidermal cell walls of Gladiolus leaves which lack epidermal-pore hydathodes, and the same explanation may account for Goebel’s observation (1930) of secretion from the blade margins of ferns without hydathodes. The results presented in this paper do not preclude the possibility that water secretion is partially or wholly active, and more research is required to decide the question. Hydathode epidermal cells could play a role in the modification of the solute content of the secreted xylem sap. Thus, they would be analogous to glandular cells found associated with epidermal-pore hydathodes of the angiosperms (Dieffenbach, et al., 1980). If the epidermal cells are capable of secreting salts, there may also be a metabolically dependent osmotic mechanism which contributes to water secretion, at least for those fern species that characteristically secrete large amounts of salts and form “scales” (see Lepeshkin, 1906). Regardless of the specific mechanism of secretion, however, the correlation of root pressure with hydathode secretion suggests the functional significance of the fern hydathode is to be sought in its relationship with root pressure. This project was initiated while I was enrolled in the Harvard course Biology 247, which included a field trip to Mexico. I would like to thank the instructors, Rolla and Alice Tryon, for organizing the trip, which was financially supported by the Atkins Fund of Harvard University. I also thank the Tryons for advice during early Phases of the project and P. B. Tomlinson and M. H. Zimmerman for their critical review of the manuscript. Frank Lang provided the photograph in Figure 1; Figure 2 was photographed with the assistance of George Wilder. LITERATURE CITED DIEFFENBACH, H., U. LUTTGE, and M. G. PITMAN. 1980. Release of guttation fluid from passive hydathodes of intact barley plants. Il. The effects of abscisic acid and cytokinins. Ann. Bot. —712 FAHN, A. 1979. Secretory Tissues in Plants. Academic Press, London. 12 AMERICAN FERN JOURNAL: VOLUME 73 (1983) GARDINER, W. 1883. On the physiological significance of water glands and nectaries. Proc. Camb. Phil. Soc. 5:35-50. GOEBEL, K. 1926. Morphologische und biologische Stunden. IX. Beitrage zur Kenntnis der Verwandschaftsverhaltnisse einiger Javanischer Farne. Ann. Jard. Bot. Buitenz. 26:107-160. ________. 1930. Archegoniatenstudien. XX. Farne mit punktierten Blattern. Flora 124:410-422. GUTTENBERG, H. 1934. Studien an Pflanzen der Sunda-Inseln. Ann. Jard. Bot. Buitenz. 34:1-62. HABERLANDT, G. 1894a. Ueber Bau und Function der Hydathoden. Ber. Deutsch. Bot. Ges. 12:367-378. _ 1894b. Anatomisch-physiologische Untersuchungen iiber das tropische Laubblatt. II. Uber wassersecernirende und -absorbirende Organe. Sitzungsber. Akad. Wiss. Wien., Math-Nat. KI. 103:489-583. _ 1898. Bemerkungen zur Abhandlung von Otto Spanjer “Untersuchungen tiber die Wasserapparate der Gefisspflanzen.” Bot. Zeit. 61:177-181. —_——.. 1914. Physiological Plant Anatomy. (Translated from the 4th German edition by Montague Drummond.) Macmillan, London. LEPESHKIN, W. W. 1906. Zur Kenntniss des Mechanismus der aktiven Wasserausscheidung der Pflanzen. Beih. Bot. Zentralbl. 19:409—452. _ 1923. Uber active und passive Wasserdrusen und Wasserspalten. Ber. Deutsch. Bot. Ges. 41:298-300. LIPPMANN, E. 1925. Ueber das Vorkommen der verschiedenen Arten der Guttation und einige physiologische und 6kologische Beziehungen. Bot. Arch. 11:361—464. MEIDNER, H. 1977. Sap exudation via the epidermis of leaves. J. Exp. Bot. 28:1408-1416. METTENIUS, G. H. 1856. Filices Horti Botanici Lipsiensis. Leopold Voss, Leipzig. OGURA, Y. 1972. Comparative Anatomy of Vegetative Organs of the Pteridophytes. Gebriider Born- traeger, Berlin. POIRAULT, G. M. 1893. Recherches anatomiques sur les cryptogames vasculaires. Ann. Sci. Nat. Bot., VII, 8:113-256. POTONIE, H. 1892. Uber die den Wasserspalten physiologisch entsprechenden Organe bei fossilen und recenten Farnarten. Sitzungsber. Ges. Naturf. Freunde Berlin 1892:1 17-124. SPANJER, O. 1898. Untersuchungen iiber die Wasserapparate der Gefasspflanzen. Bot. Zeit. 61:6-81. STOCKING, R. C. 1956. Guttation and bleeding. Pp. 489-502 in: W. Ruhland, ed. Encyclopedia of Plant Physiology, vol. 3. Springer-Verlag, Berlin. WEAST, R. C. and M. J. ASTLE, eds. 1981. CRC Handbook of Chemistry and Physics. CRC Press, Inc., Boca Raton, Florida. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 3 (1983) 73 A Reclassification of the Fern Genus Pyrrosia' K. H. SHING* The name Pyrrosia, although published by Mirbel in 1803, was overlooked by pteridologists until relatively recently. Instead, the synonyms Cyclophorus Desv., 1811, or Niphobolus Kaulf., 1824, were used. Giesenhagen (1901) published the only monograph, treating 50 species in Niphobolus and describing their venation, epidermis, and indument. Farwell (1931, pp. 241-246) recognized that Pyrrosia is the correct name of the genus. R. C. Ching (1935) studied the species of the Asian mainland, Japan, and Taiwan Province of China. He transferred 49 known species to Pyrrosia and described five new species. This has been the only extensive system- atic paper on the genus. Nayar and others (1961, 1965, 1967) did detailed work on the morphology and anatomy of the Indian species of Pyrrosia in which they divided the 13 Indian species into six groups, some of which are apparently unnatural and inconvenient to use. For example, they placed P. lingua (Thunb.) Farw., which has stellate hairs with only lanceolate arms in the P. heteractis group, although each stellate hair of P. heteractis (Mett. ex Kuhn) Ching has 1-3 longer, acicular arms in addition to the lanceolate arms. Van Alderwerelt van Rosenburg (1908, pp. 678-696) divided 34 Malesian species into two sections under the generic name Cyclophorus. Section Niphobolus had venation similar to that of Campyloneurum (more or less regular, subquadrangular areolae with two or more simple or forked, included, or less irregularly netted with single, free, included, variously oriented veinlets). This arrangement has something to recommend it, but since the blade of Pyrrosia is very thick and in most species the veins cannot be seen without clearing, venation is difficult to use for identifications. In China, Pyrrosia is an herb of traditional use. Since the laminae are shaped like a dagger with a leathery texture and usually trail on rocks, it was called “Rocky Leather” or “Flying Dagger” in old Chinese herbals. It is a diuretic which can be used to clean the lungs and to alleviate fever. Now in the pharmacy there are two kinds of medicine on the market, “Shiwei Tablet” and “Instant Shiwei Powder” (Shiwei in Chinese means Pyrrosia), made from Pyrrosia plants and said to have curative effects for nephritis and chronic tracheitis. Since special care is unnecessary for the survival of these plants, except for good drainage, they may be cultivated in hed i iji hina, but during the writer's visit to the United States, he received a grant from the “cpap tee e Foundation (020138) and through the kindness of the curators had ae city to examine more specimens in the ipllowing herbaria: GH, MICH, NY, and US. es woul like to extend his thanks to Prof. R. C. Ching and Prof. W- H. Wagner, Jr. for their kind guidance, to Mr. M. G. Price for his helpful discussions, and to Dr. A. F. Tryon for her helpful discussion of spore characters and for providing SEM photograph for Figure 2. 74 AMERICAN FERN JOURNAL: VOLUME 73 (1983) PE). FIG. 7. Spore of P. polydactilis, x 1200 (Taiwan, Tanaka 140, PE). FIG. 8. Spore of P. borneensis, 1100 (Borneo, H. F. Sun 198, PE). FIG. 9. Lower epidermis of P. clavata. FIG. 10. Upper epidermis of P. clavata. FIG. 11. Lower epidermis of P. angustata. K. H. SHING: RECLASSIFICATION OF PYRROSIA 75 FIG. 12. Types of stellate hairs in Pyrrosia. a= broad arm type, b=acicular arm type, c, d= dimorphic arm types, e=crispate-lanose arm type. a number of Pyrrosia species have been imported into the United States from the East Indies, the Philippines, Japan, and New Guinea (Hoshizaki, 1981). Pyrrosia includes about 100 species, mostly in Asia, with a few in Australia, New Zealand, Oceania, and Africa. Fifty species occur in China. Because of their more or less uniform appearance, identification may be quite difficult. During the past several years, the writer has observed their spore morphology, epidermises, stomata, and indument, and has tried to establish their taxonomic relationships. The 2010 8 2010 (%) 16 AMERICAN FERN JOURNAL: VOLUME 73 (1983) present paper will discuss these characters and will propose a new classification scheme for the genus. MORPHOLOGICAL CHARACTERISTICS Spores.—Almost all species of the genus have a uniform, tubercular perine, which varies only slightly in size and thickness (Figs. 2—8). However, spore ornamentation of P. angustata (Swartz) Ching is sharply different from most other members of the genus, and appears ribbed under the SEM (Fig. /). Since the fronds are dimorphic and firmly coriaceous, the sori very large, round, deeply sunk, and uniseriate on each side of the costa, J. Smith (1857, p. 6) considered this species to be a separate genus, Niphopsis. But in my opinion, considering all important features, it is really a member of Pyrrosia. Another similar species, P. samarensis (Mett. ex Presl) Ching, has the same spore ornamentation. I propose to use the rank of subgenus to indicate their advanced position in Pyrrosia. Rhizome scales.—The rhizome scales of Pyrrosia are either entire or are ciliolate with the hairs spread along the sides or sometimes with a tuft of hairs at the apex. Since the scales are easily broken and their tips lost on most specimens, it 1s difficult to use this feature for identification. However, since the scales of different species may also be very different in form, size, and texture, they still are useful for distinguishing some species. Epidermis and stomata.—The arrangement of the upper epidermal cells in the genus is uniform (Fig. /0). There are, however, mainly two types of stomata in the lower epidermis. Most are pericytic, with the stomata completely surrounded by a single subsidiary cell with guard cells and subsidiary cell not linked together by any anticlinal walls (Figs. 9 and //). Some are polycytic, having the stomata in large part surrounded by a single, U-shaped subsidiary cell with the anticlinal cell walls of the guard cells and the subsidiary cell linked together toward the distal end. A few are desmocytic or copericytic (Sen & Hennipman, 1981). Indument.—There are four kinds of stellate hairs in different groups of Pyrrosia: the broad arm type has 6-9 lanceolate arms (Fig. /2, a), the acicular arm type has 8-12 acicular arms (Fig. 12, b), the dimorphic arm type has 6—9 lanceolate arms and I-3 acicular arms (Figs. 12, c and d), and the crispate-lanose arm type has the arms crisped and intertwined like cotton velvet, mostly on the bottom layer of the dense, thick indument (Fig. /2, e). Hair types are a constant and reliable character for subdividing the genus. CONSPECTUS OF THE GENUS PYRROSIA Pyrrosia subg. Pyrrosia TYPE: Acrostichum lingua Thunb. [= Pyrrosia lingua (Thunb.) Farw.]. 4! Pyrrosia sect. Pyrrosia YPE: Acrostichum lingua Thunb. | =Pyrrosia lingua (Thunb.) Farw.]. Pyrrosia ser. Pyrrosia bes Acrostichum lingua Thunb. [= Pyrrosia lingua (Thunb.) Farw.]. : is is the largest series in the genus, with nearly 40 species, and is widespread 10 the Asian tropics and subtropics. Two species occur in Australia and one in Africa. K. H. SHING: RECLASSIFICATION OF PYRROSIA 77 204 \3 Pyrrosia ser. Heteractides Ching & Shing, ser. nov. Pilis stellatis e brachiis lanceolatis necnon setis paucioribus (1-3) aciculatibus longioribusque compositis. TYPE: Polypodium heteractis Mett. ex Kuhn [=Pyrrosia heteractis (Mett. ex Kuhn) Ching]. This series includes only P. heteractis, which is found in southwestern China, Vietnam, and the Himalayan area, and P. eberhardtii (Christ) Ching, distributed in r southern China and Vietnam. 0° Pyrrosia ser. Drakeanae Ching & Shing, ser. nov. Pilis stellatis e brachiis 8-12 aciculatis compositis. TYPE: Polypodium drakeanum Franch. [ = Pyrrosia drakeana (Franch. ) Ching]. This series includes about 20 species from eastern Asia to southeastern Asia, with one species in Africa. ,oo\4 Pyrrosia sect. Dichlamys Ching & Shing, sect. nov. Frons subtus indumento bistrato e pilis stellatis stipitatis dimorphis laxe composito praedita, superne e pilis aciformibus, et inferne e pilis lanosis crispatis compositis. TYPE: Niphobolus mollis Kunze [= Pyrrosia mollis (Kunze) Ching]. 90° Pyrrosia ser. Costatae Ching & Shing, ser. nov. Pilis strati superi e brachiis lanceolatis, inferi e filamentis lanosis crispatisque compositis. TYPE: Apalophlebia costata Pres| [= Pyrrosia costata (Presl) Tag. & Iwats.]. This series includes, besides the type species, P. nummularifolia (Swartz) Ching and P. strigosa (Swartz) Ching; the species are limited to the Himalayas and southeastern Asia. 1o\-© Pyrrosia ser. Molles Ching & Shing, ser. nov. Pilis strati superi e brachiis longe aciculatis compositis, inferi crispatis lanosisque. TYPE: Niphobolus mollis Kunze | = Pyrrosia mollis (Kunze) Ching]. This series contains more than 20 species, with one in Africa. +o Pyrrosia subg. Niphopsis (J. Smith) Shing, comb. & stat. nov. Niphopsis J. Smith, Cat. Cult. Ferns 6. 1857. TYPE: Polypodium angustatum Swartz [= Pyrrosia angustata (Swartz) Ching]. Rhizome thick, long-creeping, the scales caudate-lanceolate, grayish brown to- ward the apex and broken off in age, dark brown toward the persistent base, leaves fully dimorphic, the sterile blades shorter, lanceolate or elliptic-lanceolate, the fertile blades longer, linear-lanceolate or often narrowed to a linear apex; sori large, round or oblong, in a single series along the costae; spores bilateral, with many linear ridges, contracted at each end to an abrupt beak. This subgenus includes two species distributed in southeastern Asia, Singapore, Malaya, Indonesia, and the Philippines to New Guinea and New Zealand. Morpho- logically this subgenus is advanced in the genus. Its perispore is similar to that of some species of Dryopteris. Pyrrosia samarensis (Mett. ex Presl) Ching has the same spore ornamentation as P. angustata, but its sterile leaf is narrower and its sor! are confluent at maturity. Its laminae bear many, dark brown, stellate hairs with aciculate arms covering the hairs of the basal layer of indument, which are of the crispate-lanose type. This species is endemic to the Philippines. 78 AMERICAN FERN JOURNAL: VOLUME 73 (1983) NEW COMBINATIONS IN PYRROSIA According to my studies, the following species require names in Pyrrosia: Pyrrosia dispar (Christ) Shing, comb. nov. Cyclophorus dispar Christ, Nova Guinea 8:155. 1909.—New Guinea. Pyrrosia intermedia (Goy) Shing, comb. nov. Cyclophorus intermedia Goy, Queensl. Nat. 10:48, t. 6. 1937.—Queensland. Pyrrosia macrocarpa (Copel.) Shing, comb. nov. pee gion macrocarpa Copel. Univ. Calif. Publ. Bot. 12:381. 1931.—Pacific Islands. Thi cies is somewhat similar to P. angustata in outline, and Christensen (Ind. Fil. mht 3:65. 1934) reduced it to a variety. It differs from P. angustata in that the plants are smaller, the spore ornamentation is closely tubercular (Fig. 2), and the sori are never impressed. It is found only in several islands of the Pacific, such as the Cook Islands, Pitcairn Island, and the Austral Islands. Pyrrosia distichocarpa (Mett.) Shing, comb. nov. Polypodium distichocarpum Mett. Ann. Lugd. Bat. 2:231. 1866.—Sumatra. Cyclophorus winckleri Rosenst. Repert. Sp. Nov. Fedde 7:149. 1909.—Sumatra. Pyrrosia winckleri (Rosenst.) Tagawa, Acta Phytotax Geobot. 25:180. 1973. Pyrrosia winckleri and P. distichocarpa are here united for the first time. I have seen several specimens of the latter species, but no type material. Pyrrosia borneensis (Copel.) Shing, comb. nov. Cyclophorus borneensis Copel. Phil. J. Sci. 12C:64. 1917.—Borneo. Pyrrosia rasamalae (Racib.) Shing, comb. nov. Polypodium rasamalae Racib. Pterid. Buit. 99. 1899.—Java. LITERATURE CITED ALDERWERELT van ROSENBURGH, C. R. W. K. van 1908. Malayan Ferns. Dept. Agr. Netherl. Indies, Batavia. CHING, R. C. 1935. On the genus Pyrrosia Mirbel from the mainland of Asia including Japan and Formosa. Bull. Chin. Bot. Soc. 1:1-72. FARWELL, O. A. 1931. Fern notes II. Amer. Midl. Nat. 12:233- oe GIESENHAGEN, K. 1901. Die Farngattung Niphobolus. Fischer, Jen HOSHIZAKI, B. J. 1981. The genus Pyrrosia in cultivation Pobjnediseaet Baileya 21:53-76. NAYAR, B. K. 1961. Studies in Polypodiaceae. VII. Pyrrosia Mirbel. J. Indian Bot. Soc. 40: 164-183. ————, and S. CHANDRA. 1965. Ferns of India~XV, Pyrrosia Mirbel. Bull. Natl. Bot. Gard. Lucknow 117:1-98. ———, and S. CHANDRA. 1967. Morphological series within the genus Pyrrosia, and their phyloge- netic interpretation. Canad. J. Bot. 45:615—634. SEN, U. and E. HENNIPMAN. 1981. Structure and ontogeny of stomata in Polypodiaceae. Blumea SMITH, J. 1857. Cultivated Ferns. Wm. Pamplin, London. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 3 (1983) 79 Notes on the Ecology and Development of Plagiogyria fialhoi PAULO G. WINDISCH* and MARILIA PEREIRA-NORONHA** Plagiogyria fialhoi (Fée & Glaziou) Copel. is a shade-loving fern that can be found in cloud forests of elevated regions in the Brazilian states of Rio de Janeiro, Espirito Santo, Minas Gerais, S40 Paulo, Santa Catarina, Rio Grande do Sul, and probably also Parana (Brade, 1956; Sehnem, 1967). Some authors (e.g., Tryon & Tryon, 1982) consider Plagiogyria in America to be a single species, P. semicordata (Presl) Christ, but in this study the separation into six species by Lellinger (1971) was adopted. Some aspects of the ecology and development of the gametophytes and sporo- phytes of a population of P. fialhoi occurring in a cloud forest in the Serra da Mantiqueira in southeastern Brazil (45°25'S, 22°40’W, ca. 2,000 m alt.) were studied. The locality is on a small ridge next to the Campos do Jordao State Park, in the state of Sado Paulo. Cloud forests occur mainly on the eastern slopes of the mountains, above 1,600 and up to 2,000 m alt., and are composed of low (ca. 6 m) trees covered by a great quantity of epiphytes. Frequent drizzle, fog, and low clouds keep the humidity extremely high. The rainfall at the locality is thought to be about 2,000 mm per year. Climatological data collected at considerably lower altitudes in the nearby state park (Seibert et al., 1975) indicate July as the driest month (ca. 30 mm rainfall) and January as the wettest (more than 300 mm). The warmest month is February (17.7°C average, absolute maximum 27.2°C); the coldest is July (9.5°C average, absolute minimum —4.4°C). A short description of the floristic composition of these cloud forests is presented by Seibert et al. (1975), together with the detailed climatic data for the region. MATERIALS AND METHODS Gametophytes and ca. 60 young sporophytes with fronds up to 3 cm long were collected and preserved in weak chromo-acetic fixative (Sass, 1951); 36 larger specimens were pressed (voucher specimens at HRCB). Frond longevity was studied in 15 plants by marking their youngest fronds and following their development during visits 240 and 374 days later (February and June, 1982). The complete underground parts (rhizome, roots, and stipe bases) of ten well developed specimens were carefully removed and the shape and position of the rhizome correlated with the topography. The average number of fronds per adult plant was calculated from data from the ten removed specimens (April, 1981) and from the 15 marked plants (June, 1981). Average size of the fronds was obtained based on those of the 10 removed specimens and confirmed later (March, 1983) by measuring 128 adult fronds. The distance between the elements of 55 pairs (nearest neighbors) of plants was measured. *Instituto de Biociéncias, Letras e Ciéncias Exatas, Universidade Estadual Paulista UNESP, Caixa Postal 136, 15100 S. José do Rio Preto - SP, Brazil. **Curso de Pés-graduacao em Ciéncias Bioldgicas, Universi Postal 178, 13500 Rio Claro - SP, Brazil. dade Estadual Paulista—UNESP, Caixa 80 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FIGS. 1-14. Gametophytes and heteroblastic development of the sporophyte of Plagiogyria fialhoi. Figs. 1-4. Gametophytes attached to young sporophytes. FIGS. 5-12. Sporophytes in diverse stages of development. FIG. 13. Lamina of a frond 15.5 cm long. FIG. 14. Pinna from the middle of a frond 40 cm long. y a > Se — i oe ar i ee ee ee ee WINDISCH & PEREIRA-NORONHA: NOTES ON PLAGIOGYRIA FIALHOI 81 Humus and soil samples were taken from ten points (March, 1983), keeping the top material (O—5 cm depth) separated from the humus-soil mixture below (S—10 cm depth). Spores were collected (June, 1981 and February, 1982) and germinated in the laboratory after storage at 3-5°C for three and 120 days, using a solid agar medium (technique described by Dyer, 1979). Gametophyte cultures were kept for 100 days growing under 850 lux (fluorescent plus incandescent bulbs, 12 hours/day) at ca. 24°C. RESULTS AND DISCUSSION Plants of P. fialhoi were found in great numbers on the steep, humid slopes (30-50% declivity) of the southeastern side of the ridge, which rises ca. 70 m above a small stream. The density of plants increases closer to the water. The distance (nearest neighbor) between the elements of 21 pairs of plants growing more than ca. 10 m from the stream was 45-520 (ave. 189.4) cm, whereas the distance between the elements of 34 pairs growing within 10 m of the stream was 18-300 (ave. 95.9) cm. The plants grew in the rich humus layer that covers the ground. Humus samples from the top layer (5 cm) had a pH of 3.2 (in CaCly); 48g/cm? P (resin method) and K, Ca, Mg, H+Al values (m. eq. 100 cm’) of 0.5, 0.8, 0.6, and 24.4. Samples from the humus and soil mixture (19.6% organic matter) from the layer below (5-10 cm) had a pH of 3.1; 43 pg/cm? P; and K, Ca, Mg, H+Al values of 0.29, 0.2, 0.3, and 46.0, respectively. In a few places on the slope, associations of gametophytes and juvenile sporo- phytes of Plagiogyria in diverse stages of heteroblastic development, together with fern gametophytes and young sporophytes of other species, mosses, liverworts, and angiosperm seedlings were found. In contrast with the surrounding area, where the adult fern sporophytes occurred, the amount of organic matter on the ground at those spots was very small, indicating that these associations were of a pioneer nature, probably covering soil exposed by running water or fallen trees. In nature, the first frond of the young sporophytes (still attached to the gameto- phytes) measured 5.5—8.0 (ave. 7.0) mm long, with stipes 3.0-5.2 (ave. 3.9) mm long and laminae 1.3—4.3 (ave. 3.1) mm long. These laminae were symmetric and bilobed or asymmetric with one of the segments once more lobed (Figs. 1-5). The margin of the laminae in plants of all developmental stages presented a characteristic row of more or less rectangular cells (Fig. 15), which was of basic importance In the identification of juvenile specimens. Fronds ca. 14-35 mm long had a central vascular strand and tended to form pinnatifid laminae (Figs. 6-7). Pinnatisect to pinnate laminae were observed on plants with fronds longer than 35 mm (Figs. 8-13). These fronds (up to 80 mm long) had segments with serrate margins and simple veins. The lower pinnae of fronds ca. 85-150 mm long had biserrate margins (Fig. 13) and bifurcate proximal veins. Free veins that reach the lamina margins Was a consistent character in all stages of development. The sterile laminae of adult sporophytes had biserrate margins (Fig. /4), and most of the veins were bifurcate, so that a veinlet ended in each tooth. In well developed specimens (with rhizomes longer than 6 cm) the fronds were 43-117 (ave. 75-68) cm long, the stipe representing about 1/3 of the total length. 82 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FIGS. 15-23. Detail of the lamina and rhizome of the sporophyte and earl ages of gametophyte Placi arly st development of Plagiogyria fialhoi. FIG. 15. Laminar margin of a young sporophyte. FIG. 16. pee curved rhizome (old stipe bases removed). FIGS. 17-23. Early developmental stages of gametophy in the laboratory. WINDISCH & PEREIRA-NORONHA: NOTES ON PLAGIOGYRIA FIALHOI 83 Fertile fronds were observed from February to June. It seems that they are formed around January and probably persist until July. Viable spores were collected in June, 1981. A visit in February, 1982 revealed many young fronds, but the spores collected from the most developed of these did not germinate. These data indicate a definite periodicity of fertility. Fertile fronds measured 70-117.5 (ave. 103.0) cm long (the stipe representing about 1/2 of the total length) and were usually longer than the sterile fronds of a given plant. The minimum ratio observed for numbers of sterile to fertile fronds observed was 10:4, the maximum 21:1. The rhizome, covered with the bases of fallen fronds and adventitious roots, was usually erect if no longer than 4 cm, but curved in larger specimens so that the vegetative parts usually were level with or barely above the litter on the ground. The older parts remained buried. The apex of each rhizome bore the fronds in a rosette protecting frond primordia and young fronds. Litter carried down the slope accumu- lated against the frond-covered rhizome apices. The resulting pressure on the uphill side and perhaps a more intense washing of the ground on the other side, together with apical rhizome growth, promoted a slow and gradual movement of the whole plant downhill. The rhizome and its semi-decayed remains are curved, with the main axis oriented down the slope. In a large specimen, the angle of deviation between the alignment of the rhizome apex and that of the oldest remains was ca. 155° (Fig. 16), indicating the change of relative position of the old parts in the ground due to the downhill movement. The one year-long observation of marked fronds revealed that fronds persisted for about one year. The number of fronds on adult plants (of different ages) was 9-21 (ave. 12). Therefore, the minimum age of a plant can be estimated by dividing the total number of stipe bases by 12. On very old specimens, the semi-decayed parts still present are of large size, indicating that at the time those were formed the plant was already well developed. One specimen with a rhizome 21 cm long had a minimum age of 24 years. The youngest fertile specimen collected had a rhizome 12 cm long; its age was estimated to be more than 15 years, indicating a minimum Spore-to-spore life cycle of considerable duration. Vegetative reproduction was not observed, although Bower (1926) cited the occurrence of stolons in P. pycnophylla (Kunze) Mett. and bifurcation of the rhizome in P. semicordata. Spores sown on culture media three days after collection did not germinate up to 30 days after sowing, but after 50 days, germinating spores and gametophytes with 3, 4, 9 and 12 cells were seen. Slow germination occurring at irregular intervals was also observed in cultures of P. glauca (Blume) Mett. and P. semicordata by Stokey and Atkinson (1956). This kind of gradative germination can explain the finding in nature of gametophytes together with young sporophytes in diverse stages of development in a single small area. Spores stored for 120 days showed an extremely low germination rate; a single spore in about 2,000 germinated after 30 days, reconfirming the short viability of the spores of this genus as discussed by Stokey and Atkinson (1956) and Nayar and Kazmi (1962). The first stages of gametophyte development (Figs. 17-20) are very similar to those described by Nayar and Kazmi (1962) for P. triquetra Mett. No apical meristematic cell was observed. The development of a heart-shaped apex (Figs. 84 AMERICAN FERN JOURNAL: VOLUME 73 (1983) 21-23) started earlier (65 days after sowing) than in the Old World species studied by Nayar and Kazmi (1962). Elongated young plate stages and filamentous stages as cited for P. glauca and P. semicordata by Stokey and Atkinson (1956) were not observed; perhaps they were a consequence of the physical conditions under which Stokey and Atkinson’s material was grown. They considered the probability of competition with algae as being the cause of these elongated plate stages, but our cultures were also heavily contaminated. Unfortunately, the algae were so numerous that further development of the gametophytes could not be followed in the laboratory. In nature, we found that mature gametophytes were cordiform (up to 4.4 mm wide, 5.3 mm long at the larger wing), generally with one of the wings overlapping the other (Figs. 1-4), and had a thick central region. is study was made possible by the cooperation of the officers of the Forestry Institute of the State of Sao Paulo, Campos do Jordao State Park and by support rom the Conselho Nacional de Desenvolvimento Cientifico e Tecnolégico—CNPq (Proc. 30.1339/77 and Proc. 10.6254/79), the Universidade Estadual Paulista Julio de Mesquita Filho—UNESP, and the Fundagao de Amparo a Pesquisa do Estado de Sao Paulo—FAPESP (Proc. 81/1369-5). LITERATURE CITED BOWER, F. O. 1926. The Ferns (Filicales), vol. Il. Cambridge Univ. Press, Cambridge. BRADE, A. C. 1956. A flora do Parque Nacional do Itatiaia. Bol. Parque Nac. Itatiaia 5. Ministério da Agricultura, Rio de Janeiro. DYER, A. F. 1979. The culture of fern gametophytes for experimental investigation. /n Dyer, A. F, ed. The Experimental Biology of Ferns. Academic Press, London. are D. B. 1971. The American species of Plagiogyria sect. Carinatae. Amer. Fern. J. 7110-118. NAYAR, B. K. and F. KAZMI. 1962. Morphology of the spores and prothalli of five species of Plagiogyria. Bull. Soc. Bot. Bengal 16:3-8. SASS, J. E. 1951. Botanical Microtechnique. Iowa State Coll. Press, Ames. SEHNEM, A. 1967. Plagiogiridceas. Jn R. Reitz, ed. Flora Ilustrada Catarinense, part I. Itajat. SEIBERT, P. et al. 1975. Plano de manejo do Parque Estadual de Campos do Jordao. Bol. Tec. Inst. Flor. Sao Paulo. 19:1-153. STOKEY, A. G. and L. R. ATKINSON. 1956. The gametophytes of Plagiogyria glauca (BI.) Mett. and P. semicordata (Pr.) Christ. Phytomorphology 6:239-249. TRYON, R. M., Jr. and A. F. TRYON. 1982. Ferns and Allied Plants. Springer Verlag, New York. AMERICAN FERN JOURNAL: VOLUME 73 NUMBER 3 (1983) 85 The Ferns of Elden Mountain, Arizona MICHAEL D. WINDHAM* Elden Mountain, located immediately northeast of Flagstaff in north-central Arizona, is an unusual volcanic peak with a diverse fern flora. The earliest pteridophyte collections from the area are those taken by L. N. Goodding in 1913. Whiting and Bradey collected extensively on the mountain during the 1930’s and were apparently the first to encounter Asplenium adiantum-nigrum in 1935. In a short article discussing the occurrence of this species in Arizona, Wherry (1941) men- tioned five other ferns casually observed during a visit to the locality. Phillips (1946, 1947) listed five additional taxa for the mountain, bringing the number of reported species to 11. The flora of Elden Mountain received little attention during subse- quent years, and much of the area remained unexplored when a forest fire inflicted heavy damage in June, 1976. Concerned about the fate of several rare species, I began a comprehensive survey of surviving fern populations in 1978. Elden Mountain is a massive dacite dome located near the center of the San Francisco volcanic field. Although precise dates are not yet available, geologists agree that the dome was formed sometime during the last million years (Kluth & Kluth, 1974). The dacitic magma was quite viscous, and well-defined lava flows are evident on all slopes. Juxtaposition of lava flows created a highly complex topogra- phy, and erosion has produced innumerable cracks and crevices that provide favorable habitats for a variety of unusual plants. The elevation at the base of the mountain averages 7000 feet; the summit falls just short of 9300 feet. A lengthy meteorological record is available for nearby Flagstaff at 6900 feet elevation (Sellers & Hill, 1974); much of the general climatic data is applicable to Elden Mountain. At Flagstaff, the lowest average monthly temperature (21,3: ¥) occurs in January, the highest (65.5° F) in July. The mean frost-free period extends from June 8 to September 26, a total of 110 days. Annual precipitation averages 18.31 inches, and seasonal rainfall patterns are distinctly bimodal. The greatest amount of rainfall occurs in July and August, when storm systems enter the region from the Gulf of Mexico. Pacific-based storms produce a second precipitation peak in January and February, but the months of May, June, and November are character- ized by drought, which was a major factor in the disastrous fire of 1976. The distinct vegetation zones observed on the San Francisco Peaks (Merriam, 1890) are all but absent on Elden Mountain. Abrupt topography brings diverse habitats into close proximity, and plant associations complex. The areas surrounding the base of the mountain are i ’ ponderosa, which is frequently associated with Quercus gambelii. South-facing s, Berberis fremontit, Juniperus deppeana, J. osteosperma, Pinus edulis, Opuntia phaeacantha, and Yucca baccata. The northern slope and crest of the mountain support a mixed conifer forest in which Pseudotsuga menziesii, Abies concolor, and Pinus flexilis dominate. Small meadows and stands of Populus tremuloides occupy 4 limited area near the summit. Some areas show a haphazard mixture of all the species mentioned above. *Department of Biological Sciences, Northern Arizona University. Flagstaff, AZ 86011. 86 AMERICAN FERN JOURNAL: VOLUME 73 (1983) METHODS AND MATERIALS The data presented herein result from intensive fieldwork undertaken between February, 1978 and March, 1983. Considering the size and complexity of Elden Mountain, it would be unwise to suggest that the pteridological survey is complete. However, most suitable habitats have been examined, and recent visits to the area have failed to yield additional, unreported species. Detailed field notes were kept concerning the distribution and habitat preference of each taxon, and population estimates were derived through a census of individual adult plants. Ten mature sporangia were gathered from each species, and their contents were examined in glycerol to determine spore number. Most cytological materials were field fixed in May and June using Farmer’s solution (3 parts ethanol: | part glacial acetic acid). Plants not demonstrating the proper meiotic condition were transferred to a green- house to stimulate the production of fertile fronds. Fixed sporangia were hydrolyzed for approximately 15 minutes in IN HCI prior to staining with aceto-orcein. Hoyer’s solution was used as the squashing and mounting medium in all chromosome preparations. Counts were derived from cells at late diplotene or diakinesis and were documented using a camera lucida. Voucher specimens for all phases of this study have been deposited at the Deaver Herbarium, Northern Arizona University (ASC). RESULTS AND DISCUSSION All fern species previously reported for Elden Mountain were relocated during this survey, proving that none was lost as a direct result of the 1976 fire. In fact, the number of recognized species was nearly doubled by the discovery of two unreported herbarium collections and seven taxa new to the mountain. None of the additions were found in fire-damaged areas, and it is apparent that they have existed on the mountain for quite some time. Nearly all are characterized by low population densities, which may explain why they were overlooked by previous collectors. The survey has revealed that the Elden Mountain fern flora comprises 20 species representing 11 genera. Chromosome counts have been obtained for 15, and spore count data are available for all but one. Adiantum pedatum L.—Discovered in 1979, the colony consists of ca. 25 plants growing on the western slope at 8100 ft elevation. The plants are confined to a small area of permanent seepage on the back wall of a narrow cliff recess. The length/width ratio of the laminae is unusually high compared to other Arizona collections, but elongate fronds are occasionally observed throughout the range of the species. Mature sporangia consistently produced 64 normally developed spores. Asplenium adiantum-nigrum L.—The Elden Mountain population comprises at least 100 individuals scattered over the southern and eastern slopes at 7100-8100 ft elevation. The plants favor shaded cracks, crevices, and ledges on relatively dry. south-facing cliffs. All sampled sporangia yielded 64 normal spores, and a chromo- some count of n=72 II (Fig. 1A) agrees with previous determinations from Europe and Colorado (Shivas, 1969) Colorado specimens were originally described as Asplenium andrewsii A. Nels., and Chihuahuan collections provided the type materials of A. chihuahuense J. G. Baker and A. dubiosum Davenp. (Knobloch & Correll, 1962). ‘The proliferation of names in the New World resulted from the M. D. WINDHAM: FERNS OF ELDEN MOUNTAIN 87 sae hes eid Bie LY & Pak ] ‘ a, Whee ra fans 2 ae ee a »” a 2 Sen 42 ~ + + af ex% * ws ty avs be 4 s ‘+ « ty wa = ite Woy hg 7x8 AN B C oF oe Ls aad x a + ) eo « .) x * ee eas bad + ye ate Ms 4< x ite PG! s 2 “ + by rat * roe + ro od 1+ Ax, % - *+ Sige + gs xe c+ a Mex a ¥% ‘xr 0 az e * +3° * D E F Cad *) <5 - + = + oo +, x0 ee oR RS Vass, ¥ ~ \ yr ag = en cons Ue ea g CS of ag % Aste an ty. et? ty ot eH LS hehe an , Cy +, '° 2 > 4 ee bar) 4749 ¥ .) E Pa si ha oF sf ~ hee, G H | prod ay * ++ “ee an os « . * hy SS 3 ay * > gy" 4 * me eal a = ig i ie aay % 44 4 ~~ * ont vm o*~. » > aoe” . 0 fw Oot 7 . uP Fine ee vote, SR He A - oe is = | J K + & 3 a | » = % (98 } ye >) H.punctata® c. 5spp. 150. 1Sp. FIG. 4, Phyletic scheme deriving the Hypolepidaceae from the Dennstaedtiaceae. Solid lines and numbers in unbroken circles indicate probable lines of evolution and established chromosome numbers. roxen lines and numbers in broken circles indicate possible lines of evolution and doubtful or ke Postulated cytotypes. 104 AMERICAN FERN JOURNAL: VOLUME 73 (1983) PHYLOGENETIC CONSIDERATIONS Mickel (pers. comm.) has pointed out that the spores of H. distans are more similar in their perispore pattern to those of Paesia than to any species of Hypolepis, including H. nigrescens. He recognized some similarity in frond form between H. distans and the species of Paesia and suggested that it could be part of an aneuploid series in Paesia which has resulted in the loss of the characteristic inner indusium. Despite the similarity of its spores to Paesia, I do not believe that this alone provides sufficient evidence for regarding H. distans as a species of Paesia. In all other characters it is consistent with Hypolepis rather than Paesia: the inner indusium is absent: the sorus terminates a single vein; the veins are conspicuous (in Paesia they are difficult to see); the veins end in emarginations (rare in Hypolepis, but unknown in Paesia); the hairs are scarce and non-glandular (in Paesia they are usually abundant and glandular); and the rachis is straight, bearing pinnae which arise at right angles in more or less opposite pairs (in Paesia it is characteristically zig-zag bearing pinnae which arise alternately at an acute angle). Holttum (1958) related the shape of the rachis to periodic dormancy of the apex, pointing out that in P. elmeri the rachis rests while a single pinna develops in turn, but that in H. brooksiae and other genera showing periodic dormancy, a pair of pinnae develop together while the rachis rests. Although, in my opinion, H. distans is closer to the polyploid species of Hypolepis (with n=52 or 104) than to species of Paesia, there is no doubt that all three groups are very closely related. Nevertheless, H. distans, with or without H. nigrescens and H. brooksiae, constitutes a distinctive element. Smith (1981, P. 136) has already noted the cytological heterogeneity of the genus and suggests that “Hypolepis as presently circumscribed may be unnatural.” There is certainly a strong case for distinguishing the aneuploid species from the polyploid, but whether such taxonomic recognition should be at the generic or subgeneric level, and whether the aneuploid species themselves constitute a natural grouping, are very much more difficult questions to answer. For the moment, no change is proposed, and Hypolepis is interpreted here in a broad sense. The question arises as to whether the aneuploid species of Hypolepis provide a link with other allied genera, or whether they are merely the end products of divergent evolution from an ancestral stock with n= 26. Paesia and Dennstaedtia are central to any consideration of this possibility. Copeland (1947, p. 57), Holttum (1949), and Mickel (1973) have pointed to the similarity between Hypolepis and Dennstaedtia, Copeland in particular remarking “The more primitive element In [Hypolepis] .. . is hardly distinguishable from a similar element in Dennstaedtia.” Species of the two genera share many vegetative characters, although in my experience the indusial and spore characters will always distinguish them. Neverthe- less, the presence of an aneuploid line in Hypolepis including the numbers 28 and 29 is of considerable interest since it ties in remarkably with the numbers 30-34, 46, and 47 so far known in Dennstaedtia (Lovis 1977, p. 275; Léve et al. 1977, P- 184). These numbers have already been cited by Lovis (1977, p. 303) in postulating that the evolution of the Dennstaedtiaceae (i.e., Pichi Sermolli’s Dennstaedtiales) !8 the result of a long series of aneuploid reductions from a base in the Cyatheaceae. P. J. BROWNSEY: POLYPLOIDY AND ANEUPLOIDY IN HYPOLEPIS 105 It may be more than mere coincidence, therefore, that Hypolepis brooksiae bears a quite extraordinary morphological similarity to Dennstaedtia scandens (Blume) Moore. In the field both are scrambling, thicket-formers with periodic dormancy of the rachis (Holttum, 1958); in the herbarium they can be distinguished only by a very careful scrutiny of the indusium or by the spores. It is highly significant that Tsai (1973) has reported the chromosome number of D. scandens in Taiwan to be n= 30. Unfortunately, the chromosome number of H. brooksiae is not yet known, but if, as its morphological similarity to H. distans and H. nigrescens suggests, the number proves to be in the high 20’s, this would constitute good evidence of an evolutionary link between Dennstaedtia and Hypolepis. Some features of H. distans, notably the pronounced emarginations, the thin, highly polished stipes, and the virtually glabrous fronds, are not typical of Dennstaedtia. One possible answer to this paradox may be that H. nigrescens, H. distans, and perhaps H. brooksiae, represent remnants of unstable karyotype combinations thrown up in the course of a series of aneuploid reductions from a source in Dennstaedtia extending back to at least x=34 and possibly as far as x=47. An unbroken series of base numbers from 34 in Dennstaedtia punctilobula, through 33 or 32 in D. scabra, 31 in D. wilfordii, 30 in D. hirsuta and D. scandens, 29 in Hypolepis nigrescens, and 28 in H. distans, ultimately giving rise to a felicitous combination of 26 chromosomes in Paesia, Pteridium, and the polyploid species of Hypolepis, is an entirely plausible evolutionary pathway (Fig. 4). The very fact that many of the same characters appear in different combinations in the four genera reinforces the belief that these genera do share a common origin. For example, Dennstaedtia, Paesia and Pteridium have an inner indusium, but Hypolepis does not; Dennstaedtia and Hypolepis have sori terminating a single vein, whereas Paesia and Pteridium have sori linking several veins; Dennstaedtia and Pteridium have trilete spores, whereas Hypolepis and Paesia have monolete spores. LIMITS OF THE HYPOLEPIDACEAE The possibility that members of the Hypolepidaceae were derived from a source in the Dennstaedtiaceae necessitates a more careful consideration of the genera which constitute the former family. There is no doubt in my mind that Hypolepis, Paesia, and Pteridium form a natural group of genera with a common ancestry based on x= 26 and that they are related to—and possibly derived from—Dennstaedtia, but I find it much more difficult to ally them with some of the other genera which have been referred to the Hypolepidaceae. ae : The family was first circumscribed and its relationships outlined by Pichi Sermolli (1970, 1977, p. 431). He included in it Hypolepis, Paesia, Preridium, Histiopteris, Lonchitis, and Blotiella. To these (as subfamily Hypolepidoideae of the Denn- Staedtiaceae) Lovis (1977, p. 276) added Monachosorum, Taenitis, and /diopteris, although most workers would now probably agree that the latter two genera are better placed with the Gymnogrammeoid and Pteridoid ferns, respectively. The true affinities of Monachosorum have never been satisfactorily established, but it fits no better in the Hypolepidaceae than in any of the other families to which it has been assigned (Christensen, 1938; Holttum, 1947; Copeland, 1947, p. 51; 106 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Mickel, 1973). Lovis’s main reason for putting it with the Hypolepidaceae was the belief that 7 =56 in M. maximowiczii (Bak.) Hayata might have been derived from x=28, which relates to known numbers in Hypolepis. Lonchitis and Blotiella have also been regarded as somewhat discordant elements in the Hypolepidaceae, both by Mickel (1973) and even by Pichi Sermolli (1977, p. 432). Their chromosome numbers (7 =50 and 38) do not fit at all comfortably. There remains the genus Histiopteris, which Mickel (1973) and Pichi Sermolli (1977, p. 432) found no difficulty in allying to Hypolepis, Paesia, and Preridium. However, Copeland (1947, p. 60) considered it related to Preris, and Holttum (1973) included it in his list of genera whose relationships need “fresh examination.” My own experience of Histiopteris in New Zealand suggests that it is not closely allied to other members of the Hypolepidaceae. The presence of scales on the rhizome, glaucous and virtually glabrous fronds, anastomosing veins, and sessile pinnae are all characters alien to Hypolepis, Paesia, and Preridium. Most importantly, the known chromosome numbers of n=48 and 96 (Love et al. 1977, p. 191) cannot easily be related to a base of 26 in the other genera. Assuming that Monachosorum, Lonchitis, Blotiella, and Histiopteris, if not members of the Hypolepidaceae, are at least Dennstaedtialean, room must obviously be found for them elsewhere. Crabbe et al. (1975, p. 155) have already proposed for Monachosorum a monogeneric subfamily, Monachosoroideae, within their Dennstaedtiaceae (equivalent to Pichi Sermolli’s Dennstaedtiales), and a solution of this nature may be necessary for the other genera. EVOLUTION OF THE DENNSTAEDTIALES SENSU PICHI SERMOLLI Looking beyond the Hypolepidaceae, Wagner (1980) has recently proposed that the neotropical genus Loxsomopsis might have an “affinity to either the dennstaedtioid or lindsaeoid ferns” on account of its chromosome number of n=46. This contrasts with n= 50 reported for Loxsoma (Brownsey, 1975), a genus previously considered very close to Loxsomopsis but whose relation to other genera has always been very uncertain. Investigation of its gametophytes by Stokey and Atkinson (1956) and of its stomata by van Cotthem (1970, 1973) suggested an affinity between Loxsomaceae and Cyatheaceae, but the cytological evidence indicates that any such relationship must be fairly remote. However, Lovis’s (1977, p. 303) suggestion that the Dennstaedtiaceae (i.e., Pichi Sermolli’s Dennstaedtiales) originated by a long series of aneuploid reductions from a source in the Cyatheaceae provides a means 0 reconciling these differences. If Loxsoma and Loxsomopsis are admitted to be Dennstaedtialean ferns, as at least some of their morphological characters and certainly their chromosome numbers suggest, then the distant affinity with the Cyatheaceae, hinted at by the gametophyte and stomatal characters, lends some support to Lovis’s hypothesis. It may be that the troublesome Monachosorum also belongs on the evolutionary line from the Cyatheaceae to the Dennstaedtiales. Its position here could help to reconcile the views of Copeland (1947), Crabbe et al. (1975), and Pichi Sermolli (1977, p. 430), who allied it with the Dennstaedtiaceae, with those of Christensen (1938), who allied it with the Thelypteridaceae; in both Holttum’s (1973) and P. J. BROWNSEY: POLYPLOIDY AND ANEUPLOIDY IN HYPOLEPIS 107 Lovis’s (1977, fig. 3) scheme, the latter family is itself an offshoot of the Cyatheaceae. Certainly the chromosome number of Monachosorum (n=56) fits better here than in the Hypolepidaceae. Moreover, the numbers 46, 50, and 56 now _ in Loxsomopsis, Loxsoma, and Monachosorum go some way to bridging e gap between the highest confirmed number of n=48 in the Dennstaedtiales . 1977, p. 304) and the range of 56—69 (excepting 95 or 96 in Metaxya) in the Cyatheaceae sensu Lovis (1977, p. 273) This leaves an awkward group of genera, Blotiella (n= 38), Histiopteris (n= 48), and Lonchitis (n=50), whose affinities are far from clear. Blotiella and Histiopteris have much in common morphologically, but cytologically the latter genus is closer to Lonchitis. Tryon (1962) suggests that Blotiella is related to Hypolepis, but that Lonchitis is closer to Pteris. Histiopteris has also been allied with both the Dennstaedtialean and Pteridoid ferns, but its cytology fits comfortably into neither group; the Pteridoid ferns are clearly based on x =29 or 30, albeit with an aberration of n=27 in Idiopteris, while the known range in Dennstaedtialean ferns (26—c. 50) would require Histiopteris to be placed at the higher end of the cytological spectrum where its morphology is even more out of place than at the lower end with the Hypolepidaceae. The affinities of these three genera need to be more thoroughly investigated; meanwhile, I regard their placement as uncertai I am extremely grateful to Dr. J. E. Braggins, University of Auckland, New Zealand, for supplying me with the photograph illustrated in Figure 2, to Dr. J. T. Mickel, New York Botanical Garden, for his unpublished observations on New World species of Hypolepis and Paesia, and to Dr. Mickel and Professor J. D. Lovis, University of Canterbury, New Zealand, for their critical and extensive comments on the manuscript. LITERATURE CITED ALLAN, H. H. 1961. Flora of New Zealand, vol. 1. Government Printer, Wellington BOWER, F. O. 1928. The ferns, vol. III. The Leptosporangiate Ferns. Cambridge ‘University Press, Cambridg BROWNLIE, G. 1954. Introductory note to taxonomic studies of New Zealand ferns. Trans. & Proc. Roy. Soc. New Zealand 82:665—666. . 1957. Cytotaxonomic studies on New Zealand Pteridaceae. New Phytol. 56:207-209. ————. 1958. Chromosome numbers in New Zealand ferns. Trans. & Proc. Roy. Soc. New Zealand 85:213-216. . 1961. aes chromosome numbers—New Zealand ferns. Trans. Zealand. Bot. 1: BROWNSEY. Fes, o . chromosome count in Loxsoma. New Zealand J. Bot. 13:355-360. CARSE, H. 1929. Botanical notes and new varieties. Trans. & Proc. New Zealand Inst. 60: 305-307. CHRISTENSEN, C. 1938. Filicinae. Pp. 520-550 in F. Verdoorn (ed.), Manual of Pteridology. Nijhoff, The Hague. COCK KAYNE, L. and H. H. ALLAN. 1934. An annotated list of groups of wild hybrids in the New Zealand flora. Ann. Bot. (London) 48:1— COPELAND, = B. oe Genera Filicum. Chronica ‘Botanica, Waltham, MA COTTHEM, W. R. J. n 1970. Comparative morphological study of the stomata in the Filicopsida. Bull. Jard. Soe. Etat. 40:81-239. n,n 1973. Stomatal types and systematics. Bot. J. Linn. Soc. 67, Suppl. 1:59-71. CRABBE, J. A., A. C. JERMY, and J. T. MICKEL. 1975. A new generic sequence for the Pteridophyte herbarium. Brit. Fern Gaz. 11:141-162. Roy. Soc. New 108 AMERICAN FERN JOURNAL: VOLUME 73 (1983) FABBRI, F. 1965. Secondo supplemento alle Tavole cromosomiche delle Pteridophyta di Alberto Chiarugi. Caryologia 18:675—731. HOLTTUM, R. E. 1947. A revised classification of leptosporangiate ferns. J. Linn. Soc. (Bot.) 53:123-158. . 1949. The classification of ferns. Biol. Rev. 24:267—296 _ 1954. A revised flora of Malaya, vol. II. Ferns of Malaya. Government Printing Office, Singapore. ees The genus Paesia in eae — Bull. 13:454—- ae —_—_—. 1973. Posing the problems. Bot. J. . Soc. 67, Suppl. 1 KURITA, j 1962. Chromosome bee ae of some +, ee ferns III. i ‘est Arts Sci., i eis Ser. 3:463—46 Chromosome a of Japanese species of Pteridophyta. Ann. Rep. Foreign as College, Chiba Univ. 2:41-56. . 1972. Chromosome numbers of some Japanese ferns (8). Ann. Rep. Foreign Students College, Chiba Univ. 7:47-53. LOVE, A., D. LOVE, and R. E. G. PICHI SERMOLLI. 1977. Cytotaxonomical Atlas of the Pteridophyta. Cramer, Vaduz and O. T. SOLBRIG. 1964. IOPB chromosome number reports I. Taxon 13: i 110. LOVIS, ‘ D. 1977. Evolutionary patterns and processes in ferns. Adv. Bot. Res. 4:229-4 MANTON, I. and W. A. SLEDGE. 1954. Observations on the cytology and i of the pteridophyte flora of Ceylon. Phil. Trans., B, 238:127—185. and DA. 1968. Cytology of the fern flora of Tristan da Cunha. Proc. Roy. Soc. Laie B, 170:361-379. MEHRA, P. N. and K. R. KHANNA. 1959. Cytology of some Himalayan ferns. J. Genet. 56:1-14, 311-313 Chiba Univ., , and S. C. VERMA. 1960. Cytotaxonomic observations on some west Himalayan Pteridaceae. Caryologia 13:619—650. MICKEL, J. T. 1973. The classification and phylogenetic position of the Dennstaedtiaceae. Bot. J. Linn. Soc. 67, Suppl. 1:135-144. ; WAGNER, Jr., and K. L. CHEN. 1966. Chromosome observations on the ferns of Mexico. Caryologia 19:95-102. MITUI, K. 1968. Chromosomes and speciation in ferns. Sci. Rep. Tokyo Kyoiku Daigaku, B, 13:285-333. tigi Chromosome numbers of Japanese pteridophytes. Bull. Nippon Dental College, Gen. 4:221-271 - 1976. Chromosome studies on Japanese ferns (5). Bull. Nippon Dental College, Gen. Ed. 5: 133-140. PICHI SERMOLLI, R. E. G. 1970. Fragmenta pteridologiae II. Webbia 24:699-722. er Tentamen Pteridophytorum genera in taxonomicum ordinem redigendi. Webbia —512. ROY, R. P., B. M. B. SINHA, and A. R. SAKYA. 1971. Cytology of some ferns of the Kathmandu Valley. Brit. Fern Gaz. 10:193-199. SMITH, A. R. 1981. Flora of Chiapas, Part 2. Pteridophytes. California Academy of Sciences, San sow ———, and J. T. MICKEL. 1977. Chromosome counts for Mexican ferns. Brittonia 29:391- 398. STOKEY, ‘% G. and L. R. ATKINSON. 1956. The gametophytes of Loxsoma cunninghamii R. Br. an xsomopsis costaricensis Christ. Phytomorphology 6:249-261. STOLZE, + G. 1981. Ferns and Fern Allies of Guatemala, Part II. Polypodiaceae. Fieldiana Bot., 1.5. 5 TRYON, R. M., jr 1962. Taxonomic fern notes. III. Contr. Gray Herb. 191:91-107. TSAI, J. 1973. Chromosome numbers of some Formosan ferns (2). J. Sci. Eng. 10:261- WAGNER, F. S. 1980. New basic chromosome numbers for genera of neotropical ferns. Amer. J. Bot. 67:733-738. WALKER, T. G. 1966. A cytotaxonomic survey of the pteridophytes of Jamaica. Trans. Roy. Soc. Edinburgh 66: 169-237. AMERICAN FERN JOURNAL: VOLUME 73 EER F (1983) 109 Pecluma, a New Tropical American Fern Genus MICHAEL G. PRICE* The species of Pecluma were submerged in Polypodium until modern taxonomists separated Grammitids from Polypods. Then, Copeland (1956) included some members of Pecluma in a generic treatment of the Grammitid Ctenopteris. That provided the impetus for three papers which supplied evidence that Pecluma species indeed belonged with Polypodium (Stokey, 1959; Wilson, 1959; Sota, 1963). A later revision of the individual species by Evans (1969) has facilitated a new examination of the relationships of the group, and I have come to the conclusion that those authors who reaffirmed an alliance with Polypodium are correct, but that Copeland’s error may be partly excused on the grounds that Pecluma also has many features reflecting a shared ancestry with Ctenopteris, features that mark it as abundantly distinct from Polypodium. The most conspicuous of these differences from Polypodium are: rhizomes short and non-branching, paleae basally attached, axes terete, and fronds pectinate. From Ctenopteris, diagnostic differences are: spores neither trilete nor green, hairs never acicular from a broad base and not dark maroon, and stipes articulate. These and other characters are detailed below. Pecluma Price, gen. nov. Polypodium subg. Pectinatum Lellinger, Amer. Fern J. 71:93. 1981. eve, non ramificans nec glaucescens, radices saepe prolificas amina angustaque, plerumque sursumdecurrentia, marginibus non cartilagineis pilis ascendentibus multiseptatis dispositis, infra pagina pilis multiseptatis glandulosis appressis praedita. Venae liberae vel fortuito anastomosantes. Stomata polocytica atque anomocytica vel copolocytica. Sori orbiculati paraphyses trichoideas continentes. Sporae monolaetae, non chlorophyllosae. “TYPE: Polypodium pectinatum L. ETYMOLOGY: Pecluma is a compound of the epithets pectinatum and plumula which have been used to exemplify the group. DISTRIBUTION: Tropics and subtropics of the New World, from Bermuda, Florida, and northern Mexico to southern Brazil and northern Argentina (Evans, 1969). Species about 28, generally epiphytic or epipetric. The only previous author to accord formal taxonomic rank to Pecluma was Lellinger (1981), who noted that it was “sharply distinct” from the rest of Polypodium. As a genus, Pecluma is immediately recognizable and clearly defined and circumscribed. Separation from Polypodium (sensu stricto or lato) is important for the portrayal of the evolutionary lines involved, since Pecluma appears to have Preserved characters that are possibly primitively simple among Polypods. Some of these character elements, especially of rhizomes, paleae, and axes, are fundamen- tally different from the usual condition in not only the genus Polypodium but also i Division of Biological Sciences and Herbarium, University of Michigan, Ann Arbor, MI 48109. 110 AMERICAN FERN JOURNAL: VOLUME 73 (1983) the entire subfamily Polypodioideae (the Polypods). The removal of Pecluma thus leaves Polypodium far more uniform, definable, recognizable, and phylogenetically meaningful. However, Pecluma does agree with Polypodium in a number of crucia features that require microscopy for detection (characters of the spores, sporangial stalks, and gametophytes), as well as the articulate stipes, which taken together are my basis for maintaining it in the Polypodioideae. In the other direction, Pecluma agrees with many members of subfamily Grammitidoideae in various characters by which it differs from the other genera of subfamily Polypodioideae, again characters of rhizomes, paleae, and axes, and also of frond-form and venation. Loxogramme exhibits a mosaic of characters otherwise associated with éither the Polypodioideae or the Grammitidoideae, and therefore is living evidence that those two are linked evolutionarily. A subfamily Loxogrammoideae may therefore be appropriate. I believe that Pecluma must be retained in subfamily Polypodioideae, rather than be placed in a subfamily of its own, but I am convinced that its numerous character states so similar to those widespread in the Grammitidoideae cannot be attributed to evolutionary convergence or reversal, but rather indicate shared ancestry. Pecluma, then, provides further evidence of the affinity between the Polypod and the Grammitid ferns and strengthens the case for ranking them as subfamilies of a greater Polypodiaceae, as apparently first done by Wagner (1961). n the following discussion of the characteristics of Pecluma, I will not attempt to repeat the morphological observations made in the excellent paper of Evans (1969), except to place emphasis on certain character states. A summary of major characters compared with Polypodium and Ctenopteris,' both confused with Pecluma in the past, is presented in Table 1. n Polypodium, the rhizome is wide-creeping, even rampant, regularly branching or producing short, dormant side branches, and is often glaucous beneath the paleae. Pecluma differs strongly in all these features, with the rhizome invariably short, whether ascending or creeping, and even when creeping having phyllopodia close together (exceptionally to 1 cm apart), and never regularly branching. The absence of rhizome branches is sometimes compensated for by the production of proliferations from the roots, already confirmed for seven species by Evans (1969, pp. 195, 196, and 203), and one more first noted here, P. funicula. The rhizome surface is never glaucous. All these rhizome characters, as they contrast with Polypodium, are the general condition of Grammitid ferns, which usually have short, non-branching, non-glaucous rhizomes, although the Malesian Ctenopters taxodioides and the American C. moniliformis, for example, have slender, long- creeping rhizomes. Root proliferations are known among Polypods only in the highly-specialized Platycerium, where they should not be construed as evidence of affinity to Pecluma. Among Grammitids, root prolifery has only been confirmed for Adenophorus subg. Oligadenus (Bishop, 1974), perhaps first shown in the careful illustration of A. pinnatifidus Gaud. by Brackenridge (1855, pl. 2, fig. 3.)- In the Poly- ne . —- . si here for expedience to refer to miscellaneous pinnate oe pepe rammitids, although its Malesian type is v ely related to the type © described genus Prosaptia. ee ype M. G. PRICE: PECLUMA, A NEW FERN GENUS Ill TABLE 1. COMPARISON OF MAJOR CHARACTERS OF Pecluma WITH Polypodium SENSU STRICTO AND THE PINNATE OR SUBPINNATE GRAMMITIDS (“Crenopteris”). Structure Rhizome Roots Paleae Stipes Axes Lamina segments Hairs Venation Stomates Paraphyses Sporangial Stalks Spores Chromosome numbers podiaceae as here circumscribed, root prolifery is also exhibite Polypodium peltately attached often clathrate sometimes with rhizoids often marginate often scattered on lamina articul ate variously channeled usually not dark usually relatively few often lanceolate : occasionally sursumcurrent various or glabrescent free to regularly osing usually polocytic + copolocytic and/or ocytic scales, hairs, or monolete not green c= SF a Close ally of Loxogramme. Basally attached paleae characterize the Gram tous condition is peltate attachment; even among attached paleae occur only in a few highly speci i.e., Platycerium, a few large Pyrrosia species, an has paleae basally attached both on the rhizomes and t frond they are situated only along axes, never scattere clathrate in whole or in part, and not strongly species of Pecluma as well as Polypodium and Cten Pecluma short unbranched not glaucous often proliferous sometimes with rhizoids margin not differentiated only along axes or absent from lamina articulate perfectly terete dark numerous usually sursumcurrent multiseptate, dense on rachis above, appressed on lamina below and glandular free or casually anastomosing polocytic + anomocytic or copolocytic hair-like 2 or 3 cells thick monolete not green x Ctenopteris rt not glaucous sometimes proliferous basally attached usually not clathrate sometimes with izoids margin usually not differentiated not on lamina usually not articulate usually terete often dark various, often acicular, broad-based, dark m free or casually nastomosing usually polocytic + hair-like or apparently none uniseriate at base trilete chlorophyllose x=37 d by Anarthropteris, mitids. In Polypodium, the ubiqui- the entire Polypodioideae, basally alized cases unrelated to Pecluma, d some drynarioid ferns. Pecluma he fronds, and if borne on the d on the laminar surface, not differentiated at the margins. Some opteris have rhizoids on the surfaces of the rhizome scales which appear to be homologous with root hairs, and SO presumably function for absorption. Marginal hairs on the paleae, when they ‘ rp arg wie occur in any of these three, are morphologically quite different. Surficial hairs on 112 AMERICAN FERN JOURNAL: VOLUME 73 (1983) the paleae are also found in Tricholepidium (Ching, 1978) of Pleopeltid affinity, where they resemble the slender, maroon setae of the Grammitids, in Leucostegia of the Davalliaceae, in Pleurosoriopsis, where they seem to be rhizoidal, and in the Thelypteridaceae (Holttum, 1971), where they are related to the hairs on the fronds. Pecluma agrees with the Polypodioideae rather than the Grammitidoideae in the important character of stipes articulate to the phyllopodia, but this is not an exclusive feature, as Ctenopteris celebica, for example, has articulate stipes. The stipe and rachis of Pecluma are dark and virtually perfectly terete; they do not shrink during drying to become grooved, channeled, or differentially dimensional. No Polypodium has this character, although it is widespread in the Grammitidoideae, and occurs in some species of Pleopeltis of the Polypodioideae. In checking for this character, one must be wary of several deceptive conditions. If immature, still- developing fronds are dried before the deposition and hardening of the sclerenchy- matous sheath responsible for the character, the axes will shrink along certain lines and appear to be channeled, or if unusually heavy or unequal mechanical pressure is used in drying, portions of the axes may appear slightly compressed. Laminar tissue is long-decurrent down the stipe, forming a very narrow strip on each side, but the actual shaft of the stipe is invariably terete. Laminar segments (pinnae) are very numerous (over 40) and long and narrow, the ratio of length to breadth ranging from 6:1 to 14:1, giving the fronds a marked pectinate appearance, and although nearly the same character state occurs in a few species of Polypodium, the situation is far more frequent in Ctenopteris. In most species, some or all of the segments are sursumcurrent, the bases extending upwards along the rachis more markedly than downwards. This character is more common in Polypodium than in Ctenopteris, but occurs in both. The margins are never cartilaginously reinforced as in many Polypods. Fronds are always hairy with multiseptate hairs, copious on the rachis adaxially, less so abaxially, and at intervals along the margins (except in Pecluma curvans and P. pectinatiformis); the lamina beneath almost always has appressed, pale, multiseptate hairs with glandular terminal cells (except P. hygrometrica, which has erect, non-glandular hairs). The morphology and distribution of these hairs are most similar to those of some species of Polypodium, and although a few Ctenopteris have hairs of similar appearance and distribution, the widespread condition in the Grammitidoideae of dark maroon, broad-based, slender, acicular hairs is never found in Pecluma. The combinations of stomatal types are similar to those widespread among the Grammitids (van Cotthem, 1970) and Polypods (Sen & Hennipman, 1981). Pecluma plumula has a combination of polocytic and copolocytic stomates, whereas the other species I have examined (P. pectinata, P. hygrometrica, P. ptilodon, and P. pectinatiformis) combine polocytic with anomocytic stomates. orl are orbicular, never elongate as in some Polypodium and Ctenopter's, surficial (slightly impressed in P. sursumcurrens), and always contain hair-like paraphyses. Setose sporangia are common in Pecluma and Ctenopteris, but are rare in Polypodium. Wilson (1959) pointed out that although no fundamental differences exist between the sporangial capsules of Polypods and Grammitids, P. pectinata and M. G. PRICE: PECLUMA, A NEW FERN GENUS 113 P. plumula both agree with the Polypodioideae in having two rows of cells at the base of the sporangial stalk (vs. a single row at the base in the Grammitidoideae). Spores are normally monolete and fabiform (globose in P. dispersa), finely tubercu- late, and achlorophyllous. Spore characters are probably most consistently useful for distinguishing between subfamilies Polypodioideae and Grammitidoideae, as excep- tions are very rare, and in only one case is a Grammitid known to have monolete spores (F. S. Wagner, in prep.). Gametophyte characters agree best with Polypodium as to early appearance of the first rhizoid, short duration of the filamentous stage, form of the mature thallus, and distribution and types of hairs (Stokey, 1959), although gametophytes of the apogamous Pecluma dispersa agree with the Grammitidoideae in several characters (Evans 1969, 07). Most similar to Pecluma is the group of species’ around Polypodium hartwegianum Hook., as pointed out by Evans (1969, p. 217) and Smith (1981, p. 187), but all these species have the usual differences in rhizomes, paleae, and axes. The Old World Thylacopteris, a segregate of Polypodium, is similarly pectinate, but has a very slender, long-creeping, branched rhizome, peltate paleae, grooved axes, and glabrescent laminae. I am indebted to Prof. W. H. Wagner, Jr. and David M. Johnson for critical comments and helpful discussion. ENUMERATION OF SPECIES a Pecluma absidata (Evans) Price, comb. nov. Polypodium absidatum Evans, Ann. Missouri Bot. Gard. 55:238, f. 20. 1969. Pecluma alfredii (Rosenst.) Price, comb. nov. Polypodium alfredii Rosenst. Fedde Repert. 22:15. 1925. Pecluma atra (Evans) Price, comb. n Polypodium atrum Evans, Ann. Missouri Bot. Cad 55:237, f. 18. 1969. Pecluma bermudiana (Evans) Price, comb. n Polypodium bermudianum Evans, Ann. Missouri Bot. pied: 55:228, f. 17. 1969. Pecluma boliviana (Rosenst.) Price, comb. nov. Polypodium bolivianum Rosenst. Fedde Repert. 5:236. 1908. Pecluma camptophyllaria (Fée) Price, comb. nov. Polypodium camptophyllarium Fée, Huit. Mém. 86. 1857. Pecluma chiapensis (Evans & Smith in Smith) Price, comb. nov Polypodium chiapense Evans & Smith in Smith, Amer. Fern J. 70:23, f. 16-17. Pecluma choquetangensis (Rosenst.) Price, comb. nov. Polypodium choquetangense Rosenst. Meded. Rijks Herb. 19:18. 1913. Pecluma consimilis (Mett.) Price, comb. n Polypodium consimile Mett. Ann. Sci. Nat. (Paris) Z A of -54° Prince ~ Albert F North Battleford ss Saskatoon a 52° » ra Swift Current, * * Regina 50° 49° -—_ MONT. = ! ' 1 eS DAK. 110 106° 102 FIGURE 1. Map of Saskatchewan, Canada, showing localities for populations of A. Mia ide fe sitchense (closed circles), of var. michauxii (open circles), and of those with intermediates a6 a ae! var. sitchense (half-open circles). Symbols for the regions are: A =Cypress Hills Provincia ge B=Clearwater River, C=South re of Lake Athabasca, D=Reindeer Lake—Wollaston ; E = Wapawekka Hills, and F = Pasquia Hills. V. L. HARMS: THE LADY FERN IN SASKATCHEWAN 119 DISTRIBUTION OF ar ee ATHYRIUM FILIX-FEMINA : | : s.iat. IN NORTH AMERICA QQ = va: michauxii i aati UT =— var, asplenioides S <“F wd Pa cama =- var. californicum 60 py > oS LZ {2 SY ( @ -A. filix-femina sg, lat. Gyn “gy disjunct isolates Zips R H Lie O -A. filix-femina sg. lat. Soe. present Saskatche- wan records x NSS ee ® thy LA Le, (40 fp x \ & sia, es Ss : 0 500 1000 km FIGURE 2. FIG. 2. Map of North America showing the main ranges of A. filixfemina giana (by gest location of previously reported disjunct isolates (by closed circles), me aes Fee stars Saskatchewan collections (by open circles), indicating the geographical aeons sd oO ie iiaied be to the former. Based mainly on Hultén (1964, pp. ee 155-156) Boivin information from McGregor et al. (1977, p. 5). Scoggan (1957, p. 63, 1978, -. JJ usr (1967, pp. 29-30), Gleason and Cronquist (1963, p. 17), and Hitchcock et al. (1907, pp. ®°—)"”- the usual Lady Fern habitat here and elsewhere in the province aiscggtta ape wooded or shrub-thicketed to more or less open stream sides, lake shores, an seepage areas. : : The Saskatchewan specimens of A. filix-femina are especially a nomen they occur in the mid-continental distributional gap between the _ asaeeneiote and American var. michauxii (Spreng.) Farw. [syn. subsp. angusturm ie ~yclosorum the western var. sitchense Rupr. [syn. var. cyclosorum Rupr. and subsp. ©) (Rupr.) C. Chr.] (Fig. 2). The distinguishing characteristics between these varieties, 120 AMERICAN FERN JOURNAL: VOLUME 73 (1983) as compiled from various published keys and descriptions including those of Butters (1917), Scoggan (1978, pp. 155-156), Hultén (1968, p. 48), Gleason (1952, p. 43), Fernald (1950, p. 40-41), Boivin (1967, pp. 29-30), and Hitchcock et al. (1969, pp. 63-64), are summarized in the following key: 1. Indusia more than | mm long, less than half as broad as long, straightish or crescent-shaped, not crossing the vein, short-ciliate; spores light brownish, smooth to sparingly papillate; pinnules more narrow, mostly 5 mm wide or less, slightly to only medium-toothed; laminae mostly less than 6 dm long, widest below the middle; stipes relatively longer, the laminae mostly less than 2(2.5) times as long as the stipes; stipe base scales 7 mm long or less, mostly dark-brown, often with blackish streaks; rhizomes horizontal to oblique-ascending, with the new frond growth appearing lateral to the tuft var. michauxii Indusia mostly less than | mm long, about half to as broad as long, horseshoe-shaped to suborbicular, often crossing the vein, conspicuously long-ciliate; spores yellowish, more densely papillate to appearing finely warty; pinnules broader, the best-developed ones over 5 mm wide, strongly toothed or lobed to pinnatifid; laminae often more than 6 dm long, widest at or somewhat above the middle; stipes relatively shorter, the laminae mostly more than 2.5 times as long as the stipes; stipe base scales usually pale brown, mostly over 10 mm long; rhizomes strongly ascending to erect, with the new frond growth appearing central in the tuft var. sitchense Most Saskatchewan Lady Fern collections clearly belong to the western var. sitchense, rather than to the eastern var. michauxii, despite the wider geographical gap apparently remaining to the west than to the east of the Saskatchewan populations. All the collections (including 70 individual plants) from the more western Saskatchewan regions of the Cypress Hills, Clearwater River and Lake Athabasca south shore, as well as those from the Wapawekka Hills, were consis- tently identifiable as var. sitchense (Fig. 1, A, B, C, and E). The collections (including 24 individual plants) from the more eastern Saskatchewan regions 0 Reindeer Lake—Wollaston Lake and the Pasquia Hills were also mostly determined as nearest to var. sitchense (Fig. 1, D and F), but at least some specimens showed various degrees of intermediacy to the eastern var. michauxii. The only two Saskatchewan collections to be determined as nearest to var. michauxii were from Courtenay Lake and from 20 km west of Numabin Bay of southern Reindeer Lake (Ternier & Jasieniuk 1420 and 2113, respectively). _The Lady Fern seems to display a relatively common boreal North American distributional pattern, with major eastern and western ranges of relatively high plant frequency, but an intervening central region where the plants are less common to rare and sporadic, or even absent. Our recent findings of A. filix-femina at widely spaced localities across Saskatchewan suggest it should be looked for as well across central and eastern Alberta and in western Manitoba. The formerly apparent mid-continental gap for the species, from western Alberta to central Manitoba, may actually be nonexistent. It is interesting that farther south across the Great Plains a similar series of “stepping stones” seems to be formed by disjunct isolates in north-central Nebraska, the Black Hills in southwestern South Dakota, and Sheridan County in northeastern Wyoming (Fig. 2). Most likely all of these apparently disjunct isolated populations represent relicts of a formerly continuous west-east distribution. The author expresses his sincere appreciation to Wayne C. Harris. and Sheila M. Lamont for their valuable contribution to this paper in first discovering and — V. L. HARMS: THE LADY FERN IN SASKATCHEWAN 121 collecting A. filix-femina in the Cypress Hills Provincial Park, and for subsequently measuring the varietally diagnostic characteristics on a standing population sample of 21 plants at the collection site. Thanks are also given to field associates, Robert A. Wright (along on all collecting expeditions), and to John H. Hudson, Donald F. Hooper, and Les Baker (each accompanying us on one field trip). LITERATURE CITED BOIVIN, B. 1967. Flora of the Prairie Provinces. Part I—Pteroids, Ferns, Conifers, and Woody Dicopsids. Provancheria, Université Laval, Quebec City. BUTTERS, F. K. 1917. The genus Athyrium and the North American ferns allied to Athyrium filix-femina. Rhodora 19:170—207. FERNALD, M. L. 1950. Gray’s Manual, 8th ed. American Book, New York. GLEASON, H. A. 1952. The New Britton and Brown Flora of the Northeastern United States and Adjacent Canada. Vol. 1-The Pteridophyta, Gymnospermae, and Monocotyledoneae. New York Botanical Garden, New York. —, and A. CRONQUIST. 1963. Manual of Vascular Plants of Northeastern United States and Adjacent Canada. Van Nostrand, New York. HARMS, V. L. 1978. Athyrium filix-femina new to Saskatchewan. Amer. Fern J. 68:119—120. HITCHCOCK, C. L., A. CRONQUIST, M. OWNBEY, and J. W. THOMPSON. 1969. Vascular Plants of the Pacific Northwest. Part I-Vascular Cryptogams, Gymnosperms, and Monocots. Univer- sity of Washington Press, Seattle. HULTEN, E. 1964. The Circumpolar Plants. |-Vascular Cryptogams, Conifers, Monocotyledons. Almqvist & Wiksell, Stockholm. . 1968. Flora of Alaska and Neighboring Territories. Stanford University Press, Stanford, California. McGREGOR, R. L., R. M. BARKLEY, et al. 1977. Atlas of the Flora of the Great Plains. Iowa State University Press, Ames. SCOGGAN, H. J. 1957. Flora of Manitoba. Bull. Natl. Mus. Canada. Ottawa, 140:i-v, 1-619. ————. 1978. The Flora of Canada. Part 2-Pteridophyta, Gymnospermae, Monocotyledoneae. National Museums of Canada, Ottawa. / REVIEW HYBRIDS IN EUROPEAN ASPLENIACEAE (PTERIDOPHYTA), by T. Reichstein, Botanica Helevtica 91:89-139. 1981.—Two-thirds of the more than 30 species of European Aspleniaceae are as fully promiscuous as those of the Appala- chian Asplenium complex in America, and the number of hybrids they produce iS even greater. The species and their hybrids have been studied intensively since the 1950’s, principally by Bouharmont, Lovis, Meyer, Reichstein, Sleep, and Vida. The present paper summarizes the accumulated knowledge of past research and presents it in the form of annotated checklists of the species and of the hydrids. Several new hybrids and new cytological results are described in two appendices, and many hybrids are illustrated with line drawings. Extensive introductory material is applica- ble to fern hybridization in general, as well as to European Asplenium—D.B.L. 122 AMERICAN FERN JOURNAL: VOLUME 73 (1983) AMERICAN FERN JOURNAL Manuscripts submitted to the JOURNAL are reviewed for scientific content by one more of the editors, and, often, by one or more outside reviewers as well. During the past year we have received the kind assistance of D. S. Barrington, L. E. M Evans, D. R. Farrar, F. R. em, J.D, Montgomery, Kee sion Taylor, to whom we are deeply indebted. We welcome suggestions of other i reviewers.—D. INDEX TO VOLUME 73 perencaee subg. Oligadenus, 110; pinnatifidus, 110 Adian 8, 95; furcatum, 28; pedatum, 86, 90 Adlets hal ii and Saceu a ae 28 Alston, ; A. @.- Jem . M. sie nkin. The genus Seapine in eae ge ‘men bey. ), 4 J Apaloplebia costata, 77 As 85 pac ce 7, 14, 28, 31, 42, 88, 93, 95, 100; Hanmi 28; and 5 nigram x subsp. trichoetises; 80: x trudellii, 3; X wherry x Asplenosorus, 31; boydsto: 28; Xshawneensis, 31 Athyrium, 12, 16, 42; fis denies, sas 14, 62, 117, 119-121, S osorum, 119, var. 8 aceon Nagel 118-12 Azolla as a green manure: use od management in crop mae (rev.), aishya, A. K. & R. R. Rao. Ferns and fern-allies of Meghalaya State, ning (rev.), 93 Ballard, H. E., Jr. A new combination for an Asplenosorus hybrid, Bisporangiate a sporophylls in oe from Rajasthan, 64 Blechnum “omg i, 65, 66, 71; viviparum, 63 Blotiella, 105- 7 Bohra, D. R. (se Ronseieria, £2; 16 hoe 12; hispida, 12; pedata, 12; subpaleacea Cha apman’s eget reconsidered, 39 0; subg. Botrychium, 53; boreale, 3; echo e as se, : + 57-59, subsp. hesperium oss . 53, 59; minganense, 55, ; paradoxum, 53, 61; 12 pinnatum, 53, 34, 57; simplex, 53; rownsey, P. J. Polyploidy and aneuploidy in Hypolepis and the evolution of the Dennstaedti Camptosorus, 28, 95; rhizophyllus. i, 3i Campyloneurum, 73 Cardiomanes, 42 Ceratopteris dhalictndides: 12 Ceterach, 95 — osylxanthones in diploid and tissue culture- induced auto- paenieidt nee llia fejeensis, 43 nsidered, Cheilanthes, 95; californica, 95; eatonii, 89-92, f. eatonii, = feei, 89; fendleri, 87, 89, 92; s of osa, 95: wootonii, 87, 89, 9 i ps Churchill, S P. e C. C. Freeman) alge eris Cra yi The ‘aibeus of Woodwardia areolata, 46 Crypto 5 eee 109-112: gene 112; funicula, 114, 115; monili- eerie 1 Vittaria gametophytes discovered in a new physio- ike re graphic aie 33 Cyatheaceae, 65, 104, 106, 107 ; pe Cyclophorus, 73; borneensis, 78; dispar, 78: interme MB; be macrocarpa, 78; sect. Nipho bolus, 73; sect. Niphopsis, /: winckler 12, 90; dickieana, 12; fragilis, Cystopteris, "16 bulbifera, : : _ 12; reevesiana, 12; oe ‘ow, De avallia ens 42, Davallia in i Death Notice: gpa * Correll (1908-1983), 94 = spnenciger 04-106; <— ae aoe punctilobula, F 105; ‘nies. a wilfor Dense ee subfam. Syinaee 105, subfam. Monachosoroideae The stb of “abst pe — 46 Donov: orrell (1908-19 i id in copodium staan and L, annotinum foun xa sans: 3; x benedict : Heer 3: celsa, 31; oe a x marginalis, 3: Dryopteris, 3, 77, 94, 95, 100 * margina as, a marginalis, 3; nee sates 3. Xleedsii, 3; "x ne0 aise a spinulosa, 3, 117, var. interme x triploidea, 3 Edgar T. Wherry and his contributions to pteridology, | AMERICAN FERN JOURNAL: VOLUME 73 (1983) Elaphoglossum, i furcatum, 28 62; sci 62 Fems and allied plants, with ectel ‘reference to tropical America (rev.), 94 a0 and fern-allies of opiate et India (rev Ferns and fern allies of t less Area a sg Iowa, Ginent and Wisconsin ri rev.), age The ferns bus esi Mountain, Arizona, 85 Field gui i ferns (rev.), 96 ora Ma alesiana, series I[-Pteridophyta, volume 2, part 5, Thely- Lee Freeman C. C. & S. P. Churchill. Noteworthy pteridophyte records raska, 29 Gastony, G. J. (see D. E. Soltis) The genus — in tropical South America (rev.), 45 roi enia “ . D. Adiantum furcatum and snare furcata, 28: Mii in the xylem of Blechnum viviparum, 63 Grammitis, 9: Greller, A. M. & D. .. ee Stone fort at Fort A oor Last Habitat for \ ree a Island, New York?, 6 Harm Lady Fern, Athyrium filix-femina in Saskatche- in, - mre C. H. (see D. E. Soltis) Henry, - Dz Spread of Marsilea quadrifolia in McDonough Coun- ty, Illinois, 30 Haters, 195-1 107 Holttu . Flora Ma — series II-Pteridophyta, volume 2, part 5 Th Thlyteidacese (re 42 Hybrids in Europea (Pteridophyta) (rev.), 121 Hypolepis, 104-107; altegracillima _punctata, 101 bivalvis, 98; bogotensis, 101; br , 99, 04, 10 istans, 98-100, 102, 104, 105; hispaniolica, 99; millefolium i ens, 99, :.. : 105; pa 99, 1 1, 102, rufobarbata, 1 inct 97; repens 10 was 97, 98, 101; na viscidum, 101; viscosa, 101 “Soe Isoétes 41 23: fa eS; ~ j » 41; ape , 12, 41; chapmanii, 39; coroman- delina, 64; hese i o a ccida, 39-41, var. chapmanil, 39-41, var. flaccida, aris r. rigida, 41; melanopoda, 41 Jermy, A. C. (see A. H. G. Alst es Key, J. S. Field Guide to Missouri ferns (rev.), 96 The Lady Fern, Athyrium ‘is tocnien, in percents 117 an 112 Llavea, 95 see A. M. Greller) , 106 Lumpkin, T. A. & D. L. Plu sie Azolla as a green manure: use : and management in crop production (rev.), 96 “asians 62; annotinum, 62; sah A , 62 si ape complanatum and L. annotinum found in the Black ill re tachys protostelicus, 64 i. , 85; japonicum, 12 ecg a, 42; quadrifolia, 30 atteuccia struthiopteris, 117 sgn . 107 i, in the xylem of Blechnum viviparum, 63 Microstaphyla, 28; bifurcata, 28 123 Mildella, 95 Monachosorum, 105-107; maximowiczii, 106 Nephrolepis, 71, 73; exaltata, 12 A new combination for an eager hybrid, 31 A new combination in Asplenium, 28 Sapa 73; mollis, 77 Ops Notes on ih ecology and development of Plagiogyria fialhoi, 79 Noteworthy pteridophyte records for Nebraska, 29 Observations on the structure and function of hydathodes in 6 echnum Onoclea sensibilis, Ophioglossum ron 12; vulgatum var. pseudopodum, 29 Osmunda os a, 28 Paesia, 94, eo 104-107; st 98, 104 Ita, H . ee see P. M. Ric Peck, J. H. Ferns and fern allies i the Driftless Area of Illinois, Ow: Ot Pecluma, 109-113, ; absidiata 3; alfred . 113; atra, ns di: chiapensis, 113, ¢ etange lep! : curvans, 112, ee ; no ybasis, 114; ferruginea, 114: filicula, 114; funicula, 110 114; hygrometrica, 112 iseae, 115 ata, 112 115 tinati s, 112, 115; plumula, 112, 115 ptilodon, 142,115; rec ita, H rod ee 1 currens, 112, 115; tru m, 115; venturii, Pecl W al American genu: oss Pellaea, 12, 92; andromedifolia, 12, 14, 26; atropurpurea 2: glabella, 12; longimucronata, 90; truncata, 87, 90; wrightiana, 87, Pereira-Noronha, M. (see we G. Windisch) Pityrogramma, 73; triangul s, 90 Nie! ed mic Se 79-82: glauca, 83; pycnophylla, 83; Plagiogyria semic watt 79, 83, 84; triquetra, Platycerium, 110, 111; andinum, 9: Pleopeltis, 112 Pleurosoriopsis, | Plucknett, D. L. A. Lumpkin) Polyploidy and aneuploidy in Hypolepis, and the evolution of the Dennstaedtial Polypodiaceae ey 110, subfam. Grammitidoideae, 110, 112, 113, subfam aaa, 110, subfam. Polypodioideae, 110-11 Polypodium, 71, 109-113; ee 113; alfredii, 113; angustatum, 77 , 113, aureum, 65; bermudianum, 113; pect ctinatum, 97. 102, 105, 106; aquilinum, 12, var. Pteris, 106, Pyrrosia, a s bor! aoe . 74; a? 74, 76-78, 74, The . 74; costat r. Costatae ~ Deion Sar 2 fs sect. ome ptiree snes 77; set. Moles, 7. eg took. "Th do ii, 74; dispar, 78: tichocarpa, 78; drakean el eberhardtii, 74, 77; heteractis, 73, 77; intermedia, 78; can: ae? 4, 78; mollis, 124 TT, subg. oT 77: nummularifolia, 77; polydactilis, 74; sect. Pyrrosia, 76; ser. Pyrrosia, 76; subg. Pyrrosia, 76; 78: samarensi 7; strigosa, 77; winckleri, 78 i A reclassification of the fern genus Pyrrosia. ferns new to Saag ag — aA Reichstein, T. Hybrids in iso Aspleniaceae (rev.), ; Flora Mal 5. anger , 42; ge’ h America, 45; Hybrids in European pia eas (Pteridophyta), 121 Richardson, P. M. & H. K. Palta. C-glycosylxanthones in diploid 4% Stare air eign ploid Davallia fejeensis, 43 Schizaea, 94 Selaginella, 8g SS; an Sharma, B Singh & D. R. Bohra. So anomalous eee in mes st Rajasthan Pees H. Are classificat ation of the fe P ia, 73 Asplenium 28 — = ines B. D. Sharma) Sone, » E., a H. Heuties, D.C, Depron. & O. J. Gertony. ~— and electrode buffers, and aining — Sperry, J. S. Observations on the ture and Joni of hyda- thodes in ae aaa Sphaerostep Spread of a pal ap in McDonough County, Illinois, 30 AMERICAN FERN JOURNAL: VOLUME 73 (1983) Starch gel electrophoresis of ferns: A compilation of grinding Ts Stone Fort at Fort Totten: Last Habitat for Woodsia o eu platyneuron in Queens County, Long Island, New Teens, 105 Tectaria, 95 Thence e, 42, 106, 112 fags seeatip i simulata, 52 on) Tryon, R. M. & A. F. ‘i . Ferns and allied plants, with special reference to tropical Amerie (rev.), 9 wo moonworts of the Roc untains: << hesperium and a new species ebnes pear with Vittaria gametophytes discovered in a new prin province, Vittaria, 32, 35-37 Wagner, F. S. (see W. lan Wagner, Jr.) gar T. Wherry and his contributions to agner, W. in Jr. pteridolo; oad ah H., . & F. S. Wagner. Two moonworts of the Rocky M shim hespesions and a new species formerly ria wit Windham, M. ” gc ferns | of Elden Mountain, Arizona Windisch, p ra-Noronha. Notes on the — and development of Paeioeves fialhoi, 79 ers toM d Delaware, 31 sia, 6, 7, 92; mexicana, 87, 91, 92: oregana, 12, 87, 91; 9 ardia areolata, 46-51; subg. Lorinseria, 46 ERRATA FOR 1982 Page 85, line 11 should read: “(Rousseau, 1974; Boivin, 1966; Hultén, 1958). Abbe (1936) reporting on the northern range of this species in. . . ” Page 85, line 29: For “marina” read “mariana” BRITISH PTERIDOLOGICAL SOCIETY Open to all who are interested in growing and studying ferns and fern-allies. Full members receive THE FERN GAZETTE and the BULLETIN OF THE BRITISH PTERIDOLOGICAL SOCIETY. Membership subscriptions are £7 for full members, £ 5 for ordinary members (not receiving the GAZETTE), and £5 for student members (under 25 years of age). For particulars, U. S. residents should apply to Dr. J. Skog, Biology Department, George Mason University, Fairfax, VA 22030. Non-U. S. residents should apply to Lt. Col. P. G. Coke, Robin Hill, Stinchcombe, Dursley, Gloucestershire, England. Ce eee eee re 4 Sy” yf _.. from TRIARCH We are grateful for the purchase orders and collec- tions of material received during Triarch’s 56 years of service. We try to say thank you by maintaining quality of product, variety of listings, and by supporting biolog- ical societies with memberships, donations, and advertising. Having completed 25 years in the management of Triarch, | add this written word of thanks for your support, without which Triarch would not exist! Paul L. Conant, President TRIARCH PREPARED MICROSCOPE SLIDES P.O Ripon, Wisconsin 54971 AMERICAN FERN JOURNAL Volume 74 Number 1 January-March, 1984 QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY Promotion of Apogamy in Matteuccia struthiopteris, the Ostrich Fern A New Filmy Fern from Puerto Rico Chromosome Numbers of Neotropical Isoétes Two New Phenolic Glycosides in Asplenium septentrionale A Remarkable Cyathea Hybrid The Ligule of Isoétes Shorter Notes: Two Species of Adiantum Newly caped in Florida; A New Station for Trichomanes petersii in Alabama; Trichomanes Gametophytes at Bartholomew’s Cobble Reviews ‘8 . VON ADERKAS GEORGE R. PROCTOR R. JAMES HICKEY FILIPPO IMPERATO R. E. HOLTTUM B. D. SHARMA and R. SINGH oth NICAL gourt BO" I 7 The American Fern Society Council for 1984 TERRY R. WEBSTER, Biological Sciences Group, University of Connecticut, Storrs, CT 06268. President FLORENCE S. WAGNER, Dept. of Botany, University of Michigan, Ann Arbor, MI 48109. Vice-President MICHAEL I. COUSENS, Faculty of Biology, University of West Florida, Pensacola, FL 32504 Secretary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Treasurer LESLIE G. HICKOK, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Records Treasurer DAVID B. LELLINGER, Smithsonian Institution, Washington, DC 20560. Journal Editor . SMITH, Dept. of Botany, University of California, Berkeley, CA 94720. Memoir Editor DENNIS Wm. STEVENSON, Barnard College, Columbia University, New York, NY 10027. Fiddlehead Forum Editor — Aba Journal DAVID B. LELLINGER U = Se 1 Herbarium NHB-166, berger meer ashington, DC 2 ASSOCIATE EDITORS DAVID W. BIERHORST Rt. 3, Box 188, Picayune, MS 39466. GERALD J. GASTONY Dept. of Biology, Indiana University, Bloomington, IN 47401. JOHN T. MICKEL New York Botanical Garden, Bronx, NY 10458. TERRY R. WEBSTER ....... Biological Sciences Group, University of Connecticut, Storrs, CT 06268. “American Fern Journal” (ISSN 0002-8444) is an illustrated quarterly devoted to the general study of ferns. It is owned by the American Fern Society, and a. at the Smithsonian Institution, Washington, DC 20560. Second-class postage paid at Washington. Claims for missing issues, made 6 months (domestic) to o. aoe (foreign) after the date of issue, and the matters for publication should be addressed to the Edito Changes of address, dues, and applications for membership saad be sent to Dr. Leslie G. Hickok, Department of Botany, University of Tennessee, Knoxville, TN 37916. Orders for back issues should be addressed to Dr. James D. Montgomery, Ichthyological Associates, R.D. 1, Berwick, PA 18603. eneral inquiries concerning ferns should be addressed to the Secretary. Sahecsigeins $9.00 gross, $8.50 net if paid through an agency (agency fee $0.50); sent free to members of the American Fern Society on dues, $8.00 + $4.00 mailing surcharge beyond U. Canada, and Mexico; life membership, $160 Back volumes 1910-1978 $5.00 to $6.25 ak ‘single back numbers of 64 pages or ist $1.25; 65-80 pages, $2.00 each; over 80 pages, $2.50 each, plus shipping. Back volumes 1979 et seq. $8. 00 each; single back numbers $2.00 each, plus shipping. Ten percent discount on orders of six an or more. Fiddlehead Forum Prof. Dennis Wm. Stevenson, Dept. of Biological Science, Barnard College, Columbia University, New York, NY 10027, is editor of the newsletter “Fiddlehead Forum.” The editor welcomes contribu- tions from members and non-members, including miscellaneous notes, offers to exchange oF purchase materials, personalia, horticultural notes, and reviews of non-technical books on ferns. Spore Exchange Mr. Neill D. Hall, 1230 Northeast ek Street, Seattle, WA 98115, 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 its services to members and to others interested in ferns. Botanical books, back issues of the Journal, and cash or other gifts are always welcomed, and are tax-deductible. Inquiries should be addressed to the Secretary. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) l Promotion of Apogamy in Matteuccia struthiopteris, the Ostrich Fern P. VON ADERKAS The facultative development of sporophytes from vegetative gametophyte cells has been recorded for many species (Steil, 1939). There are also cases in which individual sporophyte organs such as sporangia develop directly on gametophytes (Lang, 1898; Lawton, 1932). The appearance of isolated organs or tissues is considered to be evidence of apomixis if the gametophytes asexually produce sporophytes (Dépp, 1967). Tracheary tissue in gametophytes of Matteuccia struthiopteris (L.) Todaro (Ostrich Fern) cultured over a prolonged period was not interpreted as an apogamous phenomenon, since the gametophytes did not produce buds or shoots. Induction and promotion of apogamy in ferns has been investigated by Whittier and co-workers with an emphasis on factors such as carbohydrates (Whittier & Steeves, 1960; 1962), ethylene (Elmore & Whittier, 1975a, b), and osmotica (Whittier, 1975). These studies of Preridium aquilinum have been concerned with apogamous development of buds. Attempts to induce similar development in Matteuccia are described in this note. MATERIALS AND METHODS Spores used in this study originated from a single fertile frond of a plant collected by the author (no. 821) from the Five Mile River, near South Maitland, Hants County, Nova Scotia (voucher in ACAD). Following separation from the sporangia, spores were stored in glass vials at — 20°C until required. Spores were sterilized for 2 min in a solution of 3% “Clorox,” a commercial bleach, to which a drop of Tween 80 surfactant had been added. Spores were trapped on a Millipore membrane filter (pore size = 8 xm) and then were washed twice with sterile, deionized water. They were plated onto Petri plates containing Knudson’s medium supplemented with Nitsch’s modified trace elements (Whittier, 1964), FeEDTA (1 »M), and 1% agar. These plates were placed in growth chambers set at 23°C with light level of 67 WE-m~?-s~! and a light/dark cycle of 16:8 h. Plants were subcultured monthly, and these cultures served as stock for the experiments. oo Gametophytes were incubated in Erlenmeyer flasks stoppered with silicon bungs each penetrated by a long intake and a short outlet tube. Each flask had 100 ml of Knudson’s medium supplemented with varied amounts of either sucrose or sorbitol. Sorbitol is very poorly metabolized by ferns, but is preferable to mannitol which may be inhibitory to development (Hirsch, 1975). The osmolarity of the media was increased by 0, 0.06, 0.12, 0.19 osmolal, (as measured on a Fisons Osmometer Model 3W), which corresponded to concentrations of 0, 2, 4, and 6% sucrose, or 0, 1.1, 2.3, and 3.4% sorbitol. * Atlantic Research Laboratory, National Research Council of Canada, 1411 Oxford St., Halifax, Nova Scotia B3H 3Z1, Canada. N.R.C.C. no. 22710. olume 71, number 4, of the Journal was issued December 29, 1983. 0 AMERICAN FERN JOURNAL: VOLUME 74 (1984) A\ FIGS. 1-4. Sporophytic features on Matteuccia struthiopteris gametophytes raised in vitro. All bars = mm. FIG. 1. Awl-like structure (A) arising from the anterior end of the central cushion (CC). FIG. < Awl-like projection from meristematic tissue and two leaves, L1 from the marginal meristem and L2 from the central cushion (CC). FIG. 3. Apogamous bud (B) surrounded by numerous glandular scales (GS). FIG. 4. Dissected juvenile leaves of apogamous sporophyte (L). P. VON ADERKAS: PROMOTION OF APOGAMY IN MATTEUCCIA 3 FIGS. 5-6. Sporophytic features on Matteuccia struthiopteris gametophytes raised in vitro. All bars=| mm. FIG. 5. Juvenile leaf (L) of sporophyte of sexual origin. FIG. 6. Cluster of sporangia (S) at base of awl-like structure (A). The flasks were connected by sterilized plastic tubing (Tygon) to a glass manifold, and this assembly was linked to a 2] Erlenmeyer flask containing distilled water that served as a humidifier. One set of flasks was supplied from the central air supply, and the other from a cylinder containing | ppm ethylene in air (CanOx). Flow rates were maintained at 10 ml-min~! with calibrated flowmeters. Gametophytes were transferred to the flasks in a laminar flow hood. The flask assembly was transferred to a growth chamber and raised under the same conditions as the stock cultures. The total weight of all plants in a replicate was recorded at the beginning of the experiment, and individual plant weights, as well as total replicate weights, were recorded at the end of a 35-day period. There were 20 plants per treatment. The experiment was run three times. RESULTS Structures that developed on gametophytes fell into three categories: The most frequent outgrowths were awl-like projections (91.5%) from either the marginal meristem or sub-meristematic tissue (Figs. / and 2). Less frequent (8.3%) were reduced leaves (Fig. 2), and observed only once was an apogamous bud ( Fig. 3). The sporophyte which developed from this bud had dissected leaves (Fig. 4) with an abundance of ramenta along the stipe, unlike sporophytes of sexual origin (Fig. 5). ere were noticeable intergradations between awl-like projections and reduced leaves, but the latter were distinguished by the obvious development of a leat surface Fig. 2). These leaves developed from either the meristem or sub-meristematic tissue. The most extreme sporophyte reduction was a cluster of sporangia at the base of an awl (Fig. 6). No spores were produced, the sporogenous tissue having aborted Prior to meiosis as revealed by aceto-carmine squash of fixed material. Sporangia removed from the plant and raised on nutrient media did not develop further. Sporophytic organs were not restricted to plants of a particular size, as the smallest Plants of either experiment had awl outgrowths. 4 AMERICAN FERN JOURNAL: VOLUME 74 (1984) TABLE 1. NUMBER OF GAMETOPHYTES (n=60) RAISED IN DIFFERENT CONCENTRATIONS OF CARBOHYDRATE AND ETHYLENE, WHICH DEVELOPED SPOROPHYTIC STRUCTURES. Carbohydrate Ethylene (1 ppm) 4 0% sucrose 6 6 2% sucrose 37 35 4% sucrose 41 24 6% sucrose 38 35 1.1% sorbitol 30 19 2.2% sorbitol 26 20 3.3% sorbitol aT. 13 TABLE 2. MEAN RELATIVE INCREASE (W,/W,) IN FRESH WEIGHT OF PLANTS RAISED IN DIFFERENT CONCENTRATIONS OF CARBOHYDRATE AND ETHYLENE. Carbohydrate Ethylene (1 ppm) + ae 0% sucrose 2.3 23 2% sucrose 5.6 6.5 4% sucrose 53 4.5 6% sucrose 2.8 4.8 1.1% sorbitol 2.6 3.6 2.2% sorbitol 3.0 3.0 3.3% sorbitol 2.3 mo Apogamous development occurred most frequently on gametophytes raised on sucrose and | ppm ethylene (Table ]). Increasing the concentration of sucrose above 2% had no effect on apogamous development. The effect of ethylene was only marginally promotive, the more pronounced effect being attributable to sucrose. Sorbitol promoted apogamous growth compared to controls, but at a level much lower than that of parallel sucrose treatments. Sorbitol- and ethylene-treated plants had a higher frequency of apogamy than those without. Increases in plant weight were greatest overall on media supplemented with sucrose (Table 2), and these were also the treatments which elicited the highest apogamous response. However, treatment with sorbitol and ethylene elicited an apogamous response equal to that of sucrose alone, even though the former had only half the weight increase of the latter. That increase in weight of gametophyte and apogamous response are not related is made clear from the 4% and 6% sucrose treatments. DISCUSSION apogamy, in spite of their tracheary tissue. Dopp (1967) stated that the mere presence of a sporophytic organ or tissue on gametophytes which never asexually developed whole sporophytes was insufficient evidence of apomixis. The appearance of tracheids in gametophytes of the otherwise non-apomictic fern Todea barbara \s a P. VON ADERKAS: PROMOTION OF APOGAMY IN MATTEUCCIA 5 similar case (DeMaggio, 1972). However, apogamous buds of M. struthiopteris do occur, and have also been isolated, albeit very infrequently, from maintained stock cultures. Apomictic sporophyte development is much less frequent in this species than in Matteuccia orientalis (Lloyd, 1973). Unlike some other ferns in which development of apogamous structures was preceded by formation of either a special cylindrical process (Lang, 1898) or a prothallial cushion (Whittier, 1962), in M. struthiopteris the reduced leaves and awls developed either directly from the marginal meristem or from tissue of the central cushion. Promotion of apogamy has long been known to occur when a carbohydrate is added to the substrate (Whittier & Steeves, 1962). Initially, only the nutritional effects of carbohydrate were considered important (Whittier, 1964), but recently the role of osmotic potential of the media in promotion of apogamy in Pteridium was shown by Whittier (1975). For the Ostrich Fern there appears to be a threshold response to sucrose, but much of this sucrose effect is certainly of an osmotic nature. Similar to experiments with Pteridium in which an osmoticum replaced some of the requirement for sucrose (Whittier, 1975), organ development on the Ostrich Fern gametophytes occurred on up to 33% of the plants raised on sorbitol. Nevertheless, the effects of sucrose and ethylene on apogamy differ markedly from those of a similar study on Pteridium aquilinum (Elmore & Whittier, 1975a). In Bracken, induction of apogamy resulted in whole plant formation, but in the Ostrich Fern, with a notable exception, only reduced sporophytic organs developed. The general failure to promote bud development in Matteuccia struthiopteris by the addition of compounds promotive of apogamy in Pteridium does not imply that the mechanisms for the two species are necessarily different. The short duration of each experiment, when compared with the 3-5 months required to induce apogamy in Ampelopteris prolifera (Mehra & Sulklyan, 1969) or 15 weeks for Osmunda cinnamomea (Whittier & Steeves, 1960), may account for the low level of sporo- phyte formation observed. LITERATURE CITED DeMAGGIO, A. E. 1972. Induced vascular tissue differentiation in fern i 133:311-317. ; DOPP, W. 1967. Apomixis bei Archegoniaten. Pp. 531-549 in W. Ruhland, ed. Encyclopedia of Plant Physiology, vol. 18. Springer, Berlin, Heidelberg, New York. ELMORE, H. W. and D. P. WHITTIER. 1975a. The involvement of ethylene and sucrose in the inductive and developmental phases of apogamous bud formation in Pteridium gametophytes. Canad. J. Bot. 53:375-381. : , and D. P. WHITTIER. 1975b. Ethylene and carbohydate requirements fo’ induction in Pteridium gametophytes. Canad. J. Bot. 52:2089-2096. : HIRSCH, A. 1975. The effect of sucrose on the differentiation of excised leaf tissue into either metoph _ Pl. Physiol. 56:390-393. LANG, W. H. 1398. On wa se acai of sporangia upon fern prothalli. Royal Soc., B, 187-239. LAWTON, re Re aa induced polyploidy in ferns. Amer. J : Bot. 19:303-333. LLOYD, R. M. 1973. Facultative apomixis and polyploidy in Matteuccia orientalis. Amer. Fern. J. gametophytes. Bot. Gaz. r apogamous bud Phil. Trans. s on apogamy, apospory, and controlled ueues HRA, P.N. and D. S. SULKLYAN. 1969. In vitro studie y lopteris prolifera (Retz.) Copel. Bot. J. differentiation of rhizome segments of the fern Ampe Linn. Soc. 62:431—443 / 6 AMERICAN FERN JOURNAL: VOLUME 74 (1984) MOTTIER, D. M. 1927. Behaviour of certain fern prothallia under prolonged cultivation. Bot. Gaz. 1244-266. STEIL, W. N. 1939. Apogamy, apospory, and parthenogenesis in the pteridophytes. Bot. Rev. 53 WHITTIER, D. P. 1962. The origin and development of apogamous structures in the gametophyte of Pteridium in sterile culture. Phytomorphology 12:11—20. : . The influence of cultural conditions on the induction of apogamy in Pteridium gametophytes. Amer. J. Bot. 51:730-—736. 1975. The influence of osmotic conditions on induced apogamy in Pteridium aquilinum. Phytomorphology 25:246-249. ——_——., and T. A. STEEVES. 1960. The induction of apogamy in the bracken fern. Canad. J. Bot. 38:925-930. ————, and T. A. STEEVES. 1962. Further studies on induced apogamy in ferns. Canad. J. Bot. 40:1525-1531. REVIEW ILLUSTRATIONS OF PTERIDOPHYTES OF JAPAN, VOLUME 3, S. Kurata and T. Nakaike, eds. x + 730 pp. + folding map. 1983. University of Tokyo Press, yen 12,000.—This is the third in a continuing series of magnificent volumes with detailed treatments of 100 Japanese ferns. The third closely follows the format of the first two volumes which were reviewed in this JOURNAL 72:11. 1982 and 72:48. 1982. The unusually sturdy volumes contain line drawings, photos, and maps for each species, plus descriptions and lists of localities in Japanese. The nomenclature is as presented by T. Nakaike (Enum. Pterid. Japon. Filices, 1975) where synonymies are listed. Volume 3 is mainly devoted to Diplazium, Polystichum (continued from volume 1) and the thelypterids. Regrettably, the nomenclature of the thelypterids was not updated to incorporate the latest publications of Prof. R. E. Holttum. Two other minor points are that Diplazium petri Tard. was reduced to D. latifrons v.A.v.R. by Iwatsuki & Price (South East Asian Studies 14:565. 1977), and that Salisbury should not be the parenthetic author for Thelypteris palustris because his name was illegitimate. Furthermore, the nomenclature of the Marsh Fern in Japan, as well as in the United States, cannot be settled without a new taxonomic study. Holub (Taxon 21:331-332. 1972) believes it should be called T. thelypteroides (Michx.) Holub on the basis of earlier work by Morton, but Tryon and Tryon (Amer. Fern J. 63:67. 1973) are skeptical. Fern people in North America will be especially pleased with this third volume, since it includes several other ferns besides the Marsh Fern, with which they are familiar. These are: Polystichum braunii, P. lonchitis, P. tsus-simense (cultivated here), Phegopteris connectilis, Thelypteris torresiana, Cyclosorus dentatus, and Diplazium esculentum, the most delicious of all ferns, with a photo showing a whole field of it—M. G. Price, Herbarium, North University Building, University of Michigan, Ann Arbor, MI 48109. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) 7 A New Filmy Fern from Puerto Rico GEORGE R. PROCTOR* The writer is presently engaged in field work sponsored by the Puerto Rican Dept. of Natural Resources, which is intended to lead to the production of an up-to-date volume on the ferns and allied plants of this island. Already, after five months of collecting and collating other information, it appears that 45 taxa have been added to the 320 listed by Liogier & Martorell (1982), several of them new to science. At the end of one year, I intend to publish a complete checklist of Puerto Rican ferns in order to summarize new additions and describe any new species or varieties, as a preliminary to final preparation of the book. The purpose of the present paper is partly to publicize the work being done, and partly to provide a name for a species of Trichomanes of which fragmentary material has already been seen from other parts of the Antilles, and which has been confused in the past with another species known only from Jamaica. Trichomanes (Subg. Achomanes) padronii Proctor, sp. nov. Fig. 1. A Trichomanes pinnatifido rhizomatibus crassioribus, stipitibus longioribus, laminis 6-15 cm longis pinnato-pinnatifidis, pilis ramosis vel stellatis effusioribus differt. Rhizome wide-creeping, finely cordlike, 0.5-0.8 mm in diameter, densely clothed toward apex with flexuous, reddish, hairlike rhizoids ca. | mm long. Fronds distant, 9-18 cm long, long-stipitate; stipes 3—4.5 cm long, shorter than the blades, nonalate except at the extreme apex, loosely and deciduously clothed with branched or stellate, pluricellular hairs up to 1.3 mm long; similar hairs also on vascular parts and margins of the blade. Blades oblong-lanceolate, 6-15 cm long, 2.5-3.5(5) cm broad, pinnate-pinnatifid, acuminate at the apex, subtruncate at the base; rhachis alate (wings 0.3-0.7 mm wide on either side), pinnae ascending, alternate, 7-13 on a side below the pinnatifid apex, 1.5—2.5 cm long, short-stalked (the stalks winged like the rhachis), the expanded portion rhombic-acuminate, 1-1.5 cm broad below apex subtruncate to slightly flaring; receptacle becoming long-exserted. TYPE: South of Road 120 ca. 0.5 km ESE of Observation Tower, Maricao State Forest, Municipio San German, Puerto Rico, elev. 850-860 m, 23 November 1983, G. R. Proctor 39833 (US). Scandent on trunk of a tree-fern in subtropical wet forest, serpentine substrate. Collected with, and named for, Sr. Rubén Padron, diligent and well-informed Biologist in Charge of the Maricao State Forest. This species has been recorded from Hispaniola under the name Trichomanes pinnatifidum v. d. Bosch (Christensen, 1937, P.- 10), but that species as now understood is endemic to Jamaica. Trichomanes padronii clearly differs from T. Pinnatifidum in its thicker, more robust rhizome, larger and more divided fronds with much longer stipes, longer and more loosely-spreading hairs, and differently *Dept. of Natural Resources, P.O. Box 5887, Puerta de Tierra, PR 00906. 8 AMERICAN FERN JOURNAL: VOLUME 74 (1984) ie fi bp = co FIG. 1. Portion of holotype of Trichomanes padronii (Proctor 39833, US). located sori. A peculiar feature exists in the branching pattern of 7. padronii, which is neither clearly anadromous (as in subg. Trichomanes and Pachychaetum) ies catadromous (as is usually the case in subg. Achomanes). In fact, the basal divisions of the pinnae are almost exactly opposite, as are many of the ultimate veins arising from the costules of the segments. The species is assigned to subg. Achomanes because of the branched and stellate marginal hairs. LITERATURE CITED CHRISTENSEN, C. 1936. The collection of Pteridophyta made in Hispaniola by E. L. Ekman 1917 and 1924-1930. Kungl. Svenska Vetens. Akad. Handl. III, 16(2):1-93, 1. 1-20. i. LIOGIER, H. A. and MARTORELL, L. F. 1982. Flora of Puerto Rico and Adjacent Islands: Systematic Synopsis. Universidad de Puerto Rico, Rio Piedras. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) 4 Chromosome Numbers of Neotropical Isoétes R. JAMES HICKEY* Chromosome counts have been made for a number of /soétes found in eastern North America, Europe, and India. Typically the haploid complement of n= 11, or a multiple thereof, has been reported (Love et al., 1977; Kott & Britton, 1980). Polyploid series range from 2n= 22 to 2n= 110. The aneuploid /. durieui is notable in having the lowest chromosome number in the genus, with 2n=20. Several taxa from India, including J. coromandelina Linn. f., 7. panchananii Pant & Srivastava, I. indica L. Rao, and J. pantii Goswami & Arya, are reported to possess one or two supernumerary chromosomes (Goswami, 1975; Pant & Srivastava, 1965). Only Stylites gemmifera Rauh [=Isoétes andicola (Amstutz) L. D. Gémez] shows a strong deviation from the presumed base number of x=11. According to Rauh and Falk (1959), the diploid number for this species is ca. 58. Mexico, Central America, the Antilles, and tropical South America, with over fifty taxa, are exceptionally rich in /soétes. In addition, the morphological diversity in these taxa is perhaps greater than in any other part of the world. Among the neotropical species, only the count for J. andicola has been reported (Rauh & Falk, 1959). As part of an overall systematic revision of the Neotropical /soétes, the of these counts have been previously mentioned in Tryon & Tryon (1982), at which time I ascribed a count of 2n=44 in J. ticlioénsis H. P. Fuchs nom. nud. to I. lechleri Mett. 7, METHODS AND MATERIALS The specimens of neotropical /soétes examined in this study were grown in the alpine greenhouse of the Biological Sciences Group at The University of Connecti- cut. All taxa, with the exception of J. andicola, were cultivated fully submerged in glass aquaria in the Alpine Room under ambient day length. Specimens of I. andicola were potted in rich humus constantly saturated from beneath and kept at 4°C in a cold room under a 16 hour day length. Voucher specimens are deposited in the Gray Herbarium, Harvard University (GH), and duplicates are in the author's herbarium. To obtain roots for cytological study, corms were rinsed in water and all roots removed; the plants were then grown as floating aquatics until new roots emerged from the fossae. The roots were harvested when 1-2 cm long and were soaked for 4—6 hours in a saturated solution of paradichlorobenzene at room temperature. Roots were harvested in the morning, as suggested by Kott and Britton (1980). Following Pretreatment, the roots were blotted dry and fixed in Farmer’s Solution for a minimum of 24 hours. The roots were hydrolyzed in hot IN HCl for 30 minutes, deacidified in 95% ethanol, and stained with Whittman’s hematoxylin. The root tips rp then destained in glacial acetic acid and squashed in Hoyer’s medium (Willey, 1). *Box U-43, Biological Sciences Group, The University of Connecticut, Storrs, CT 06268. 10 AMERICAN FERN JOURNAL: VOLUME 74 (1984) ys Ys vad ¥ ; ™ . ‘= p : d Diploid and dodecaploid species of Isoétes, x 1500. a. Isoétes storkii, 2n=22. b. Isoétes Meslastils 2n= 22. c. Composite photo of I. novo-granadensis, 2n= 13 R. J. HICKEY: CHROMOSOME NUMBERS OF ISOETES ll TABLE 1. NEW CHROMOSOME COUNTS OF NEOTROPICAL ISOETES Isoétes alcalophila Halloy 2n=22: Argentina: Prov. Tucuman, Laguna Muerta, October 1980, Halloy s.n. (Fig. 1b). Isoétes andicola (Amstutz) L. D. G6mez 2n=44: Peru; Dept. Junin, Lago Junin, April 1979, Karrfalt s.n. (Fig. 2c). 2n=44: Bolivia; Prov. Bautista Saavedra, Feuerer 59 Isoétes boliviensis U. Weber 2n=22: Bolivia; Prov. Omasuyos, Hickey 753 & Eshbaugh Isoétes glacialis Asplund 2n=44: Bolivia; Prov. Murillo, Hickey 839 & Eshbaugh (Fig. 2b). Isoétes herzogii U. Weber 2n=44: Bolivia; Dept. Cochabamba, Hickey 819 & Eshbaugh (Fig. 2a). 2n=44: Bolivia; Dept. Cochabamba, Hickey 821 & Eshbaugh Isoétes novo-granadensis H. P. Fuchs 2n= 132: Ecuador; Prov. Napo, Sperling, Allgard & Roth 5189 (Fig. Ic). Isoétes storkii T. C. Palmer 2n=22: Costa Rica; Prov. Alajuela, Volcan Pods, Churchill 3284 (Fig. 1a). 2n=22: Costa Rica; Cerro de La Muerte, Anderson 1439 Isoétes ticlioénsis H. P. Fuchs, nom. nud. 2n=44: Peru; Dept. Lima, Ticlio Pass, Hickey 840 & Eshbaugh 2n=44: Peru; Dept. Lima, Ticlio Pass, Hickey 842 & Eshbaugh (Fig. 2d). Slides were examined with a Zeiss research microscope under bright field and phase contrast. Photographs were taken using Kodak High-Contrast film. RESULTS AND DISCUSSION The recently described J. alcalophila Halloy (Halloy, 1979) from Argentina, /. boliviensis U. Weber from Bolivia, and the Costa Rican /. storkii T. C. Palmer are diploids, 2n=22 (Fig. la, b, Table 1). Isoétes glacialis Asplund, I. herzogii U. Weber, and the undescribed /. ticlioénsis of H. P. Fuchs (Fuchs-Eckert, 1982), are ) _ In 1980, L. D. Gomez transferred Stylites andicola Amstutz, the type of Stylites, into Isoétes, and recognized the gemmiferous plants as /. andicola var. gemmifera h G6 production is facultative, and so J. andicola should be considered a monotypic species. Eight chromosome counts from both gemmiferous plantlets (Karrfalt mate- tial) and young, non-gemmiferous plants (Feuerer material) yielded a diploid number of 44. The discrepancy between these counts and the previous report of 2n=ca. 58 (Rauh & Falk, 1959) may well be the result of chromosome fragmenta- tion, which I have found to be a common occurrence in many neotropical species. However, the possibility of a cytologically distinct population cannot be ruled out. My counts provide additional support for the inclusion of Stylites in Isoétes. Two cells of J. novo-granadensis have yielded counts of 132 (Fig. Ic), and several others have yielded counts of 126, 128, and 129. The dodecaploid J. novo- Sranadensis has the highest chromosome count yet recorded for the genus. The 12 AMERICAN FERN JOURNAL: VOLUME 74 (1984) FIG. 2. Tetraploid species of Isoétes, x 1500. a. Isoétes herzogii, 2n=44. b. Isoétes glacialis, 2n= 44. c. Isoétes andicola, 2n=44. d. Isoétes ticlioensis, 2n= 44. R J. HICKEY: CHROMOSOME NUMBERS OF ISOETES 3 The use of chromosome numbers and other cytological data promises to be of considerable significance in understanding the evolution of Isoétes species. The large size and variable morphology of the chromosomes (e.g., secondary constric- tions, satellite DNA) in the neotropical species suggest that karyotype analysis might lead to a better understanding of the evolutionary history of the genus. For example, karyotype analysis has proven useful in the elucidation of triploid derived /soétes aneuploids (2n = 33+ 1) in India (Kuriachan & Ninan, 1974). Such investigations are particularly needed in /soétes, where morphological characters are notoriously plastic. I thank Gregory J. Anderson, The University of Connecticut, Hugh Churchill, University of Vermont, Tassillo Feuerer of Munich, Stephan Halloy of Fundacion Miguel Lillo, Argentina, and Calvin Sperling, Harvard University, for providing plant material used in this study. I also thank Carl Taylor, Milwaukee Public Museum, and Gregory J. Anderson for suggestions on cytological techniques. Special thanks are due to W. Hardy Eshbaugh, Miami University, for assisting in the collection of several taxa studied and for making this research possible. This study was supported in part by NSF grants BSR 78-23389 to W. Hardy Eshbaugh and BSR 82-07125 to G. J. Anderson. LITERATURE CITED FUCHS-ECKERT, H. P. 1982. Zur heutigen Kenntnis von Vorkommen und Verbreitung der ; sudamerikanischen Isoétes-Arten. Proc. Ned. Akad. Wet. C85:205-260 GOMEZ P., L. D. 1980. Vegetative reproduction in a Central American Isoétes (Isoétaceac). Its morphological, systematic and taxonomical significance. Brenesia 18:1—14. GOSWAMI, H. K. 1975. Chromosome studies in natural populations of Isoetes pantii, with heterosporous sporangia. Cytologia 40:543-551. ALLOY, S. 1979. Dos nuevos Isoétes (Lycopsida) de alta montafa, con datos ecolégicos de las lagunas Muerta y Escondida (Cumbres Calchaquies, Tucuman, Argentina). Lilloa 35:65-95. KOT. L. & and D. M. BRITTON, 1980, Chromosome numbers for Isoétes in northeastern North America. Canad. J. Bot. 58:980-984. : KURIACHAN, P. I. and C. A. NINAN. 1974. Karyomorphology of a triploid Isoetes coromendeline A with a note on karyotype evolution in the species. New Botanist 1:160-167. ave. A DLO R. E. G. PICHI SERMOLLI. 1977. Cytotaxonomical Atlas of the Pteridophyta. J. Cramer, Vaduz. BREED. D cua GK. SRIVASTAVA. 1965, Cytology ‘and reproduction oF Some Indian species of Isoetes. Cytologia 30:239-251. RAUH, W. and H. FALK. 1959. Stylites E. Amstutz, eine neue Isoétaceae aus den Hochanden Perus. der Vegetationsorgane. Sitzungsber. Heidelberger Akad. Wiss. Math.-Naturwiss. KI. 1959:3-83. TRYON, R. M. and A. F. TRYON. 1982. Ferns and Allied Plants. Springer-Verlag, New York. 14 AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) Two New Phenolic Glycosides in Asplenium septentrionale FILIPPO IMPERATO* On the basis of data from morphological and cytological analyses, Wagner (1954) suggested the concept of reticulate evolution in the Appalachian Asplenium com- plex, which was subsequently confirmed by Smith and Levin (1963) using chemical analysis. It is interesting to note in this regard that the morphological characteristics of the hybrids are generally intermediate between their parents, but that the chemical constituents of the hybrids show a total addition of parental attributes; Appalachian Asplenium plants clearly demonstrate additive inheritance of chemical characters. Recently it has been shown that the chemistry of European Asplenium adiantum- nigrum complex is analogous to that of Appalachian Asplenium complex (Richard- son & Lorenz-Liburnau, 1982). In spite of the foregoing studies, the chemistry of most species of Asplenium is not well known. In the present investigation, a new kaempferol tetraglucoside (I) and a new sulphate ester of 1-p-coumarylglucose (II) have been isolated from the fronds of A. septentrionale; moreover two other sulphate esters of hydroxycinnamic acid-sugar derivatives (III and IV) have been found in this plant material. Previous work on the chemical constituents of A. septentrionale has led to the identification of amino-acids (4-hydroxy-4-methylglutamic acid, 2-amino-4-hydroxy- pimelic acid and the corresponding lactone, N-acetylornithine) by Berti and Bottari (1968), keto acids (pyruvic, hydroxypyruvic, 2-oxoglutaric, 2-oxopimelic, and 4-hydroxy-2-oxo-pimelic acids) by Virtanen and Alfthan (1954), and cyanogenic glucosides by Hegnauer (1961). Very recently Imperato (1983) reported isoquercitrin and a new flavonoid (quercetin 3-0-(3’-sulphate) glucoside) from A. septentrionale. MATERIALS AND METHODS An A. septentrionale plant was collected on Mt. Etna, Italy. Fresh fronds were homogenized and extracted with hot 95% ethanol. The extracts were concentrated to a small volume in a vacuum and were filtered. Phenolic glycosides were isolated by preparative paper chromatography on Whatman 3MM paper in n-butanol-acetic acid-water (4:1:5, upper phase). The bands, revealed under ultraviolet light with ammonia vapor, were cut off, eluted with 70% ethanol, concentrated, and rechro- matographed in 15% acetic acid and in n-butanol-ethanol-water (4:1:2,2). Total acid hydrolysis of isolated phenolic glycosides was carried out with 2N HCl (1 hr at 100°C); controlled acid hydrolysis was carried out with 10% acetic acid (3.5 hr under reflux). Enzyme hydrolysis with B-glucosidase was carried out in citrate- Phosphate buffer (pH 4.5 at 37°C for 20 hr). The aglycone:sugar ratio was determined according to Nordstrém and Swain (1953). Aglycones were identified by ultraviolet spectral analysis with the customary shift reagents (Mabry et al., 1970), paper co-chromatography with authentic samples (four solvent mixtures) and poly- amide TLC. Sugars were identified by paper co-chromatography (three solvent mixtures) and SiO, TLC. Sulphate was identified using barium chloride. Flavonol * Institute of Chemistry and Industrial Chemistry, University of Catania, V. le A. Doria 8, Catania 1-95125, Italy. F. IMPERATO: NEW GLYCOSIDES IN ASPLENIUM 15 SPECTRAL PROPERTIES AND R; VALUES OF KAEMPFEROL 3-SOPHOROTRIOSIDE-7-GLUCOSIDE Spectral properties yoy Ry values Shift reagent Nae Solvent mixture Ry (x 100) 265,352 BAW oF NaOAc 263,375 BEW 48 NaOAc/H;BO; 260,3 H,0 90 AICI, 274,297 343,385 15% HOAc 74 AICI,/HCl 272 300,339,385 PhOH 51 273,385 (inc) ZrOCl,/citric acid 263 332 BAW =n-butanol-acetic acid-water (4:1:5, upper phase), BEW =n-butanol-ethanol-water (4:1:2,2), PhOH = phenol saturated with water, inc = shift accompanied by increase in intensity. glycosides obtained by controlled acid hydrolysis and by enzyme hydrolysis were identified by paper co-chromatography with authentic samples (four solvent mix- tures), by ultraviolet spectral analysis with the customary shift reagents (Mabry et al, 1970) and by acid hydrolysis. Phenolic glycosides were methylated with methyl iodide in dimethylformamide in the presence of silver oxide (18 hr in the dark at room temperature under stirring), and the permethylated products were hydrolysed by refluxing for 4 hr with 0.3 N HCl. Methylated sugars were identified by paper co-chromatography with authentic samples (Petek, 1965) and SiO, TLC; the partially methylated aglycones were identified by paper co-chromatography and UV Spectroscopy. RESULTS AND DISCUSSION Color reactions (brown to yellow in ultraviolet + NH;), Ry values and ultraviolet spectral characteristics (see Table 1) of the isolated flavonoid (I) were consistent (Mabry et al., 1970) with those of a 3,7-disubstituted flavonol glycoside with free Phenolic hydroxyl groups at positions 5 and A’. Total acid hydrolysis gave kaempferol and D-glucose; quantitative examination gave an aglycone:sugar ratio of 1:4.1. Controlled acid hydrolysis gave kaempferol, kaempferol 7-O-glucoside, D-glucose, sophorose (2-O-B-glucosylglucose) in small quantities, and a sugar which On paper chromatography behaved as a trisaccharide. This sugar, isolated by Preparative scale chromatograms, gave glucose by total acid hydrolysis, whereas by controlled acid hydrolysis it gave glucose and sophorose; these data are consistent with this trisaccharide being sophorotriose (O-B-glucosyl-(1 2)-O-B-glucosyl- (1. 2)-glucose). Moreover, since this sugar was obtained from flavonoid (I) by H:O> oxidation (Chandler & Harper, 1961), it must be linked to the 3-hydroxyl group of kaempferol. On enzyme hydrolysis with B-glucosidase, (1) gave D-glucose, kaempferol 3-O-sophorotrioside, and kaempferol in very small quantities. Methyla- tion of (I) followed by acid hydrolysis gave 2,3,4,6-tetra-O-methyl-D-glucose, 3,4,6-tri-O-methyl-D-glucose, and kaempferol 5,4'-dimethyl ether. These results show that the isolated flavonol tetraglucoside (1) must be kaempferol-3-O-sophorotri- oside-7-O-glucoside, which is a new natural product (Fig. /). Flavonol 3,7-diglucosides have already been found in two Asplenium species, A. rhizophyllum (Harborne et al., 1973) and A. trichomanes (Imperato, 1979). From the 16 AMERICAN FERN JOURNAL: VOLUME 74 (1984) CH,OH G. 1. Structural formula of compound (I), kaempferol-3-O-sophorotrioside-7-O-glucoside. FIG. 2. Structural formula of compound (II), 1-p-coumarylglucose 3”-sulphate. Systematic viewpoint, this is of interest because it has been suggested (Harborne, 1965) that flavonol 3,7-diglycosides are of restricted distribution. Of the three hydroxycinnamic acid-sugar derivatives isolated from A. sep- tentrionale, compound (II) has been proved to be a new natural product in the following way. The compound (color reactions: colorless to blue in ultraviolet + NH) was electrophoretically highly mobile (toward the anode) and strongly sugges- tive of a sulphate derivative. On both total acid hydrolysis and controlled acid hydrolysis, p-coumaric acid, D-glucose, and sulphate were obtained. Since the ultraviolet spectrum (ANsC" 312 nm) showed a large bathochromic shift (50 nm) in the presence of NaOMe, the phenolic hydroxyl group of p-coumaric acid must be free (Harborne & Corner, 1961). Methylation followed by acid hydrolysis gave p-methoxycinnamic acid and 2,4,6-tri-O-methyl-D-glucose. These results show that compound (II) must be 1-p-coumarylglucose 3”-sulphate, which is a new natural product (Fig. 2). The two remaining phenolic glycosides (III and IV) isolated from A. septentrionale have been characterized as sulphate esters of 1-p-coumarylglucose by the above methods, but the position of sulphation on the glucose has not been determined because these products could not be obtained in sufficient amount for their characterization to be completed. F. IMPERATO: NEW GLYCOSIDES IN ASPLENIUM 17 Sulphate esters of hydroxycinnamic acid-sugar derivatives were reported for the first time by Cooper-Driver and Swain (1975), who found sulphate esters of p-coumaryl- and caffeylglucose in Pteridium aquilinum and in ten Adiantum species. Subsequently, the following compounds have been isolated from ferns: 2-O-p-coumarylglucose 6-sulphate from Asplenium fontanum var. obovatum (Imperato, 1981), 2- and 3-sulphates of 1-caffeylglucose from Ceterach officinarum (Imperato, 1981), and 1-p-coumarylglucose 2-sulphate and 1-caffeylgalactose 6-sulphate from Adiantum capillus-veneris (Imperato, 1982). Although sulphate esters of hydroxycinnamic acid-sugar derivatives have been found mainly in ferns, they have been isolated also from higher plants, €.g., from the grass Paspalum convexum (Harborne, 1977). The author thanks the Consiglio Nazionale delle Ricerche (Rome) for financial support (grant CT 810164803) and Prof. H. Wagner (Institut fir Pharmazeutische Biologie der Universitit Miinchen) for a sample of kaempferol 3,7-diglucoside. Thanks are also due to Mr. A. D’Urso (Botanic Institute, University of Catania) for identification of plant material. LITERATURE CITED BERTI, G. and F. BOTTARI. 1968. Constituents of Ferns. Jn L. Reinhold and Y. Liwschitz (eds). Progress in Phytochemistry, vol. 1. Interscience Publishers, London, New York, Sydney. CHANDLER, B. V. and K. A. HARPER. 1961. Identification of saccharides in anthocyanins and other flavonoids, Aust. J. Chem. 14:586—595. COOPER-DRIVER, G. C. and T. SWAIN. 1975. Sulphate esters of caffeyl- and p-coumarylglucose in ferns. Phytochemistry 14:2506—2507. HARBORNE, J. B. 1965. Characterization of flavonoid glycosides by acidic and enzymic hydrolyses. Phytochemistry 4:107—120. . 1977. Flavonoid sulphates. /n Reinhold, L., J. B. Harborne, a Phytochemistry, vol. 4. Pergamon Press, Oxford. ,and J. J. CORNER. 1961. Hydroxycinnamic acid-sugar derivatives. Biochem. J. 81 242-249. "'C. A. WILLIAMS. and D. M. SMITH. 1973. Species-specific kaempferol derivatives in ferns of the Appalachian asplenium complex. Biochem. Syst. 1:51-54. HEGNAUER, R. 1961. Distribution of hydrocyanic acid in cormophytes. IV. Distribution oo sis. Pharm. Weekblad 96:577-596. IMPERATO, F. 1979. Two new kaempferol 3,7-diglycosides and trichomanes. Experientia 35:1134—1135 . 1981. New sulphate esters of hydroxycinnamic aci (London) 19:691—692. "~~: 1982. Sulphate esters of hydroxycinnamic acid-suga veneris. Phytochemistry. 21:2717-2718. . 1983. A new flavonol glucoside (London) 9:390-391. MABRY, T. J., K. R. MARKHAM, and M. B. THOMAS. 1970. The syste flavonoids. Springer-Verlag, Berlin, Heidelberg, New York. ; see NORDSTROM, CG. and T. SWAIN. 1953. The flavonoid glycosides of Dahlia variabih oh General Introduction. Cyanidin, Apigenin and Luteolin glycosides from the variety “Dandy”. bce J. Chem. Soc. 1953:2764-2773. K, F. 1965. Chromatographies des sucres methyles. Bul ICHARDSON, P. M. and E. LORENZ-LIBURNAU. 1982. C-glycosylxanthones adiantum-nigrum _ Fern J. 72:103-106. a SMITH, D. M. sary a ee A chromatographic study of reticulate evolution in the Appalachian Asplenium complex. Amer. J. Bot. 50:952-958. nd T. Swain (eds). Progress in kaempferitrin in the fern Asplenium d-sugar derivatives in ferns. Chem. Ind. r derivatives from Adiantum capillus- in the fern Asplenium septentrionale. Chem. Ind. matic identification of |. Soc. Chim. Fr. 1965:263-266. in the Asplenium 18 AMERICAN FERN JOURNAL: VOLUME 74 (1984) VIRTANEN, A. I. and M. ALFTHAN. 1954. New a-keto acids in Green Plants. Acta Chem. Scand. 8:1720-1721 WAGNER, W.H.. Jr. 1954. Reticulate evolution in the Appalachian Aspleniums. Evolution 8:103-118. REVIEW L A MONOGRAPH OF THE FERN GENUS PLATYCERIUM (POLY- PODIACEAE), by E. Hennipman and M. C. Roos. 1982. North Holland Publish- ing Company, Amsterdam and Oxford, New York. $63.00.—This little 124-page book on the staghorn ferns, Platycerium, is attractively presented on glossy paper with numerous figures and plates, some of the latter in color. No cultivated ferns are more spectacular than these showy plants, so familiar in conservatories the world around. The book is divided into two sections: general and taxonomic. The former contains the taxonomic history, habitats, morphology, phylogeny, generic relation- ships, species concepts, and geography. Notable is the detailed character analysis for primitive and divergent states, which emphasizes leaf structure and trichomes, including scales, hairs, and paraphyses. The phylogeny is constructed according to the sister group method, but the resulting graph, unfortunately, is so complicated, the actual lines partially obscured by character bars that are too wide, that it resembles a damaged Venetian blind. Character 18 (frond structure) is practically chaotic, as are some of the other divergent, multistate characters in which the Separate trends have not been sifted out. On the other hand, the geographical analysis is especially interesting and well done. The taxonomic section contains the generic description, keys, descriptions, and range maps. The authors base their taxonomic conclusions on studies of dried materials from 22 herbaria and living plants in seven botanical gardens. They reduce the number of species from the 18 recognized by Barbara Joe Hoshizaki to 15 by treating several of her species as infraspecific taxa. All characters considered, Platycerium stemaria appears to be the most primitive of the present-day species, being closest to the groundplan by having the least number of divergences. In the taxonomic treatment, I was disappointed to see the taxa arranged alphabetically rather than according to clusters of relationship worked out in the construction of monophyletic groups. I agree with the authors that it is not necessary to name the groups formally. The end of this fine book contains not only an index of collections, but also valuable notes on collecting and preparing specimens of these ferns. All in all this is a splendid contribution to pteridology, and I strongly recommend it to all students of ferns, including fern horticulturists. The authors are to be congratulated. aes H. Wagner, Jr., Department of Botany, University of Michigan, Ann Arbor, MI 8109. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) 19 A Remarkable Cyathea Hybrid R. E. HOLTTUM* I recently discovered, in the Herbarium at Kew, specimens of a hybrid Cyathea plant cultivated at the Birmingham, England Botanical Gardens in 1890 and of the two plants stated to have been parents of the hybrid. The specimens were sent to Kew by W. B. Latham, who was Curator of the Birmingham Gardens from 1868 to 1903. The only published reference to the hybrid known to me is in an account of his career by Latham (1901), in which he stated that he raised the hybrid about 1870 and that the parent species were “the Mexican Cyathea insignis and the Norfolk Island Alsophila excelsa”; he did not give a distinctive name to the hybrid. He also referred to a Dicksonia hybrid which he raised in 1862 and which was named D. lathamii by Thomas Moore (1885).! Thus Latham was a pioneer in fern hybridizing, although he is not mentioned in the book by E. J. Lowe on the subject. Lowe (1895, p. 11) recorded that when he began experiments in 1851, Thomas Moore and others told him that hybridization of ferns was impossible; for comments on Lowe’s work as recorded in his book, see Lovis (1967). Although Latham accepted the nomenclature which placed in different genera the parent plants of his Cyathea hybrid, he evidently recognized their close relationship. They both belong to Cyathea subg. Sphaeropteris in my arrangement of Cyatheaceae of the Old World (Holttum, 1963, 1964), where about 100 species of the subgenus are recognized. — In 1970, R. M. Tryon recognized Sphaeropteris as a distinct genus, including in it not only the American species mentioned by me (1963, Pp. 124), but also several others which in my judgement are quite unrelated. The six American species which I regard as true Sphaeropteris were described by Tryon (1971) as “the group of Sphaeropteris horrida.” In this group, two species are closely allied; they are named by Tryon S. insignis (D. C. Eaton) Tryon (from the Greater Antilles) and S. horrida (Liebm.) Tryon (from southern Mexico). Baker (1874, p. 17) united the two as Cyathea insignis D. C. Eaton, but in cultivation the Mexican species became known as C. princeps E. Mayer, and Latham so named the specimen of the Mexican parent of his hybrid; see Tryon (1971) for a full synonymy. Latham’s specimen Is sterile, but its scales indicate that it belongs either to S. insignis or to S. horrida, as named ‘Latham claimed that this was a hybrid between D. arborescens L antarctica Labill. of Australia. The habit of the plant as described by ee him to Kew, indicate that it is intermediate between the alleged parents piace : oie wad sae plant, now 120 years old, is still alive (its height was lowered by air-layering #0 ©xperimental work on it is now in progress; | hope a repo Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, England. 20 AMERICAN FERN JOURNAL: VOLUME 74 (1984) Latham’s specimen of the other parent shows clearly that it belongs to the species from Norfolk Island originally named Alsophila excelsa R. Brown. When transfer- red to Cyathea, the name must be changed to C. brownii Domin because there is a different species named C. excelsa. Thus there is no doubt that Latham’s two parent plants came from countries about 10,000 km apart. His hybrid is certainly intermediate in its characters of indusia and scales; I therefore regard his claim as justified, and name his hybrid as follows: Cyathea (Sphaeropteris) x lathamii Holttum, hybr. nov. Int . princepem E. Mayer (Gartenflora 17:10. 1868) et C. brownii Domin (Pteridophyta 262. 1929); a C. principe differt indusiis rudimentariis ambitu irregulari, squamulis costarum pinnularum brevioribus; a C. brownii differt soris indusiis rudimentariis, non squamulis discretis, cinctis, squamulis costarum pinnularum majoribus, setis multis castaneis instructis. TYPE: Cultivated in the Birmingham Botanical Gardens, England, 9 July 1890, W. B. Latham (K). In C. princeps the indusia are quite large, opaque, and of a firmer texture than those of all other species of the subgenus except C. insignis. They completely cover the young sori, but at maturity develop an apical aperture to form cups with contracted rims; these cups soon develop two or three splits to the base (Tryon, 1971, figs. 7, 8). In C. brownii there are no indusia; each sorus is surrounded by several narrow scales, as in most exindusiate species of the subgenus. In the hybrid the base of the receptacle is surrounded by a continuous disc of irregular size and very uneven outline. The costal scales in C. princeps are commonly 5 mm long and nearly all have firm, brown, marginal setae; the larger scales are wholly brown, whereas some others are colorless but firm. In C. brownii the costal scales rarely exceed | mm long and nearly all are thin and pallid with short, pallid, marginal hairs. The scales of the hybrid are about 1.5 mm long and nearly all have firm, brown setae. Schneider (1892, p. 354; 1893, p. 83) gave a contemporary report on the cultivation of the parent species in Britain. His comment on C. brownii is that young plants grow very quickly and are “every year sacrificed in enormous quantities for indoor decoration.” His comment on C. insignis indicates that plants so named were then well known in cultivation in English greenhouses; he did not distinguish between C. insignis and C. princeps. Among other specimens at Kew made from cultivated plants of Cyathea is one labelled “Madeira (cult.), comm. C. L. Power, 25 April 1904.” All sori are old and all show indusia comparable with those of Latham’s hybrid, although most are somewhat larger. The costal scales are small, mostly pallid but with rather long, marginal setae, colored or not. The pinnule lobes are very firm and rather deeply lobulate, not evidently glaucous. I suggest that this represents another hybrid within subg. Sphaeropteris, one parent being possibly C. cooperi (F. v. Muell.) Domin of Queensland, which has become widely distributed in cultivation (some years ago it was a weed in the fern houses at Kew). I know of no other records of fern hybrids the parents of which are so widely Separated geographically as those of C. x /athamii, but it is notable that such have R. E. HOLTTUM: A REMARKABLE CYATHEA HYBRID 1 been produced in angiosperms (e.g., Eucryphia X intermedia Bausch, the parents of which are E. glutinosa from Chile and E. lucida from Tasmania). So far as I know, no other hybrids, either wild or cultivated, have been reported between species of Cyathea subg. Sphaeropteris sect. Sphaeropteris Holttum (1963). However, in sect. Schizocaena there is a probable wild hybrid between C. moluccana R. Br. in Desv. (simply pinnate and indusiate) and C. squamulata (Blume) Copel.; it is named C. alternans (Wall. ex Hook.) Pres] (Holttum, 1963, p. 145). The existence of Latham’s hybrid is one more piece of evidence that in Cyathea sensu Holttum (1963) species with indusia and species lacking indusia may be related genetically. In his account of the genus Cyathea, as restricted by him, Tryon (1976) described five putative hybrids between indusiate species which he includes in Cyathea and exindusiate ones which he includes in Trichopteris. Tryon (1970, p. 15) separated these genera solely on the presence or absence of indusia; this separation seems to me unnatural. In Holttum and Edwards (1983, pp. 163-164) I suggested that Cyathea, Trichopteris, and part of Sphaeropteris sensu Tryon (1970) need to be united and resubdivided in a new way; such a subdivision presents very complex problems. Experimental hybridization might offer clues to their solution. As Tryon remarked (1970, p. 42), hybrids between two indusiate, or two exindusiate, species are difficult to detect from herbarium specimens and may be more abundant than is at present apparent. The problem of the origin of the American species of subg. Sphaeropteris remains uncertain. As the subgenus is very much more diverse in the Old World (ca. 100 species, with rather distinct divisions named Schizocaena and Fourniera) than in the New World, an origin in southeastern Asia is surely indicated. Tryon (1971, Pp. 7) wrote “Sphaeropteris insignis and S. horrida [Cyathea princeps] may be considered as the most primitive species in the [tropical American] group on the basis of their open indusia and their crozier scales which bear wholly antrorse teeth.” But as no Old World species has such indusia, I suggest that the condition 1s more likely to be a local development, also the antrorse setae on the stipe scales of the other species. LITERATURE CITED BAKER, J. G. 1874. Synopsis Filicum, ed. 2. Hardwicke, London. HOLTTUM, R. E. 1963. Cyatheaceae. Flora Malesiana, II Pteridophyta, 1:65-176. . 1964. The tree-ferns of the genus Cyathea in Australasia and the Pacific. Blumea 12:241-274. | , and P. J. EDWARDS. 1983. The tree-ferns of Mt. Roraima and neighbouring areas of the Guayana Highlands with comments on the family Cyatheaceae. Kew Bull. 38:155—188. LATHAM, W. B. 1901. An account of his career. Gard. Chron. III, 29:296. : LOVIS, J. D. 1967. Fern hybridists and fern hybridizing, I. The work of E. J. Lowe. British Fern Gaz. 9:301-307. LOWE, E. J. 1895. Fern Growing. Nimmo, London. MOORE, T. 1885. Dicksonia lathamii, n. hyb. Gard. Chron., 1s. 24: 584, 689, f. 154. SCHNEIDER, G. 1892. The Book of Choice Ferns, vol. 1.Upeott Se London. ————. 1893. The Book of Choice Ferns, vol. 2. Upcott Gill, London. TRYON, R. M., Jr. 1970. The classification of the Cyatheaceae. Contr. Gray Herb. 200:3-53. ——. 1971. The American tree ferns allied to Sphaeropteris horrida. Rhodora 73:1-19. ———.. 1976. A revision of the genus Cyathea. Contr. Gray Herb. 206:19-98. 22 AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) The Ligule In Isoétes B. D. SHARMA and R. SINGH* The ligule was first illustrated in Lepidodendron by Solms-Laubach (1892). Williamson (1894) described its structure under the name “adenoid organ.” Maslen (1898) found the ligule to be a small triangular point in Lepidostrobus, while Seward (1910) described the occurrence of “ligule pits” in the leaf bases of Lepidodendron and Lepidophlois. Scott (1920) suggested size variation in the ligule, from a small, pointed body as in Lepidostrobus to an elaborate structure, e.g. in Miadesmia. Recently Grierson and Bonamo (1979) described the presence of a ligule in the Middle Devonian species Leclercquia complexa. Among the living representatives of lycopods, this structure exists in Selaginella, Isoétes, and Stylites; on this basis these genera are sometimes grouped together as the “Ligulatae” (Sporne, 1975). A number of functions have been assigned to the ligule. It is believed to be a secretory or nutritive organ, or both, which by exuding water and mucilage serves to keep the young leaves and sporangia moist (Foster & Gifford, 1959; Bierhorst, 1971; Sporne, 1976). Michaux (1973) studied the ultrastructure of the cells of the ligule and found that the mucilagenous secretion possessed polysaccharides. Horner et al. (1975) believed the ligule to be a physiologically active organ which was involved in the movement of selective substances. Goswami (1976) considered the basal portion of the ligule (i.e., the glossopodium) to be a haustorial structure; Sporne (1976) stated that no one knows its true function. Recently Kristen and Biedermann (1981) and Kristen et al. (1982) studied the ultrastructure of the ligule of J. lacustris and considered it to be a vestigial organ, which, in extinct genera, might have functioned as a digestive organ. The ligule is a comparatively well developed structure in Jsoétes which occurs above the sporangium on the adaxial surface of the leaf. It has a large, embedded portion called the glossopodium (Bierhorst, 1971; Goswami, 1976). The cells of the glossopodium are isodiametric and distinct from the surrounding cells of the leaf and the labium or velum. Sometimes the glossopodium is surrounded by transfusion cells (Sharma et al., 1980). The free portion of the ligule is a thin, flat, triangular, upward-facing structure parallel to the adaxial surface of the leaf. MATERIALS AND METHODS Three species of Isoétes, large I. coromandelina L. (JAC 2025), medium-sized I. rajasthanensis Gena & Bhardwaja (JAC 2044), and small J. reticulata Gena & Bhardwaja (JAC 2052) were studied (Gena & Bhardwaja, in press). The type Specimens of the latter species are deposited at Government College, Ajmer, India. Isoétes coromandelina was collected from Daosa, Jhalawar, and Atru, while /. rajasthanensis came from Mt. Abu and J. reticulata from Atru in Rajasthan. The material was fixed in FAA, and after passing through dehydration alcohol series was embedded in wax. Microtome sections were cut at 8-12 jm (Johanson, 1940) and stained with safranin and Fast green or safranin and haematoxylin. The latter combination was better for studying the ligule. * Department of Botany, University of Jodhpur, Jodhpur 342001, India. SHARMA & SINGH: THE LIGULE IN ISOETES 23 FIG 2 FIG 3 GS. ~3. Reconstructions of Jsoétes ligules, X55. FIG. 1. I. coromandelina. FIG. 2. 1. rajasthanensis. FIG. 3. I. reticulata. The abbreviations are: G = glossopodium, L=ligule. DESCRIPTION To study the ontogeny of the ligule, serial longitudinal and transverse microtome sections were cut through the rhizomorphs of the three species of Jsoétes. In all three species the ligule is fundamentally similar. In none of the species is there a single apical cell, nor is there any plate of apical cells produced (Bhambie, 1957; Paolillo, 1963). The superficial cells divide anticlinally, while the inner cells divide at random. Leaf primordia appear on either side of the apex. The ligule develops from a single initial cell and is differentiated adaxial to the leaf primordium. It divides periclinally, and so a short filamentous structure is produced. In comparison to the leaf, the ligule develops faster; a very young leaf contains an almost fully developed ligule (Fig. 4). In I. coromandelina the glossopodium starts to form as two anchor-shaped patches of parenchyma, one on either side of the leaf bundle (Fig. 5). These are then connected by a transverse bar that is 5-7 cells thick (Fig. 6). The anchor-shaped patches also develop a proboscis-shaped structure from the lower side of the bar (Figs. 7 and 8). The bar increases in thickness and forms a large, pad-like structure towards the interior side (Fig. 9), while on the outer side a large flap or whip-like portion of the ligule is produced. The pad remains embedded in the tissue of the leaf. The glossopodium in /. coromandelina thus becomes a Complicated structure with anchor shaped, curved side arms and a central pad-like rtion _ The free, flap-like portion of the ligule and the glossopodium are made up of Similar, small, isodiametric, thin-walled cells (Fig. 10). However, the outer 3 or 4 Peripheral layers of cells of the glossopodium differ in being comparatively smaller AMERICAN FERN JOURNAL: VOLUME 74 (1984) a A *, i oy a. roy O1 " mys. s - i ee Pane, Ke aie, re es a ® * ad 1 Naa uP ha oS < 11 FIGS. 4-12. Anatomical details of /. ake nagcseme FIG. 4. Longisection of apical portion of a x 47. FIG. 5. Cross-section of a leaf showing origin of the 13 IG. rhizomorph with young leaves and ligules, glossopodium as anchor-shaped patches of parenchyma, x 13.5. Cross-section of a leaf Showing anchor-shaped patches connected _ iG. . Cross- section of a sporophyll showing glossopodium shape, x 13.5. FIG Shape, x 13.5. FIG. 9. Longisection of a sporophyll showing the ligule w upper labium is also seen, x 13.5. FIG. 10. Same, showing ligule ae different from those of the labium, which have transfusion cells; *76,5. me, ri the non-vascularized ligule, IG. 12. Basal portion of a labium with transfusion cells, x SHARMA & SINGH: THE LIGULE IN ISOETES 5 (Fig. 10). The glossopodium does not receive any vascular supply (Fig. //). However, in J. coromandelina it is surrounded by transfusion cells (Fig. 12) (Sharma et al., 1980). In this species, the free portion of the ligule is 4.5-6 mm long. Its epidermis is non-stomatiferous, but possesses unicellular and multicellular glandular hairs (Fig. 13). In J. coromandelina an upper labium covers the proximal portion of the ligule. In the medium and small-sized species of /soétes, a distinct velum is present which protects the sporangium (Fig. 74). In the latter species, the ligule is comparatively smaller and the basal portion is less complicated (Figs. 14 and 1/5). The glossopodium starts to differentiate either as a triangular structure, as in /. reticulata, one on either side of the leaf bundle (Fig. /5) or as two circular patches of parenchyma, as in J. rajasthenensis (Fig. 16). These structures are then connected by a transverse bar (Figs. 17 and 18); as in I. coromandelina, an interior, pad-like structure is produced (Fig. 19). However, the glossopodium is not surrounded by transfusion cells. On the basis of the present study, an attempt has been made to reconstruct the structure of the ligule in the three species of /soétes studied (Figs. /-3). In J. coromandelina the glossopodium is a complex, branched structure and the ligule is quite large. With reduction in plant size, the complexity of the glossopodium and the size of the ligule decrease and the transfusion cells found in association with the glossopodium disappear. DISCUSSION Heterospory in pteridophytes originated in the lycopods sometime during the Devonian Period (Pettitt, 1970). The ligule also appeared during the same geological time (Grierson & Bonamo, 1979) and is known to occur in the extinct as well as extant heterosporous lycopods. Whether its occurrence was just a coincidence or Whether some relationship really exists between the ligule and heterospory needs further investigation. Recent discovery of a ligule in the presumably homosporous species Leclercquia complexa Banks (Grierson & Bonamo, 1972) increases further the importance of this relationship. The structure of the ligule is not well preserved in L. complexa, nor can its position on the leaf be related to any of the known ligulate fossil or living genera. Could it be a reduced, sterile telome associated with the fertile telome (sporangium)? In Lepidostrobus a number of megaspores were Produced per sporangium and the ligule was a small structure, whereas Miadesmia had a single megaspore in the sporangium but the ligule was quite elaborated (Scott, 1923; Eames, 1936; Bierhorst, 1971). In /soétes a large number of megaspores are Produced per sporangium and the ligule is also a well developed structure. Thus hardly any parallel can be derived between the evolution of heterospory and the Sabon of the ligule. _ In addition to its peculiar type of tracheary el 1S unique in having ; velum sae sporangium. In large plants the velum is absent, but the upper labium is present (e.g., /. coromandelina, I. indica, and I. pantii), Whereas in comparatively smaller species (e.g., /. rajasthanensis and I. reticulata) a ements (Singh et al., 1982), [soétes AMERICAN FERN JOURNAL: VOLUME 74 (1984) FIGS. 13— 19. Maa ian of Isoétes species. FIG. 13. Epidermis of J. i cnsatelieg ligule with a glandular haris, x 180. FIG. 14. Longisection of an J. reticulata sporophyll showing ligule and vel x27. FIG. 15. Same, _ Showing glossopodium, x54. FIGS. 16-18. Cross-sections of /. siaschinan sporophylls Showing different stages of glossopodium development, 19. ng 54. “ohaonton ; n of an J. rajasthanensis sporophyll showing pad-like glossopodium and free portion of igule, x 54. SHARMA & SINGH: THE LIGULE IN ISOETES 7 distinct velum is present. The elaborated size of the ligule and the complexity of the glossopodium of the larger species may be related to the presence of an upper labium, while reduction in size of the ligule and simplification of the glossopodium in smaller species are probably associated with the presence of a velum. However, this conclusion needs further investigation. The presence of glandular hairs on both the margins of the ligule favors its secretory function, whereas the occurrence of transfusion cells surrounding the glossopodium (Sharma et al., 1980) and mucilage with polysaccharides (Michaux, 1973) suggest its purpose is a water reservoir to protect the young leaves and sporangia from desiccation. The present study does not favor the haustorial function of the glossopodium, as suggested by Goswami (1976): because the glossopodium cells are small, isodiametric, and uniform, and especially the 2 or 3 layers of cells which occur in the interior portion of the glossopodium are smaller than the surrounding leaf cells. Haustorium cells are generally large and irregular in shape (Maheshwari, 1950). e absence of a vascular supply and a cuticle in the ligule suggests that it can neither be related to leaf nor to stem. Harvey-Gibson (1896) considered the ligule as a specialized ramentum commonly found in pteridophytes. If this is true then the phylogeny of such a specialized ramentum is yet to be discovered. The authors are thankful to Prof. H. C. Arya, the Head, Department of Botany, for providing laboratory facilities and to the U.G.C. and C.S.I.R. for financial assistance. LITERATURE CITED BHAMBIE, S. 1957. The shoot apex of Isoetes coromandelina L. J. Indian Bot. Soc. 36:491-502. BIERHORST, D. W. 1971. Morphology of Vascular Plants. Macmillan, New York. EAMES, A. J. 1936. Morphology of Vascular Plants (Lower Groups). McGraw Hill, New York. FOSTER, A. S. and E. M. GIFFORD, Jr, 1959. Comparative Morphology of Vascular Plants. Freeman, San Francisco. Reprinted by Allied Pacific Private Ltd., Bomb GENA, C. B. and T. N. BHARDWAJA. In press. Three new species India. J. Bombay Nat. Hist. Soc. GOSWAMI, H. K. 1976. A revision of ligule and labium in Isoétes. Acta. Soc. Bot. ay. of Isoetes L. from Rajasthan, Poloniae 5:69-—76. wie ye GRIERSON, J. D. and BONAMO, P. M. 1979. Leclerequia complexa: earliest ligulate lycopod (Middle Devonian). Amer. J. Bot 66:474—476. : HORNER, H. T. Jr., C. K. BELTZ, R. JAGELS and R. E. BOUDREAU, 1975. Ligule ge aga and fine structure in two heterophyllous species of Selaginella Canad. J. Bot. 53:127-143. JOHANSEN, D. A. 1940. Plant microtechnique. McGraw-Hill, New York. ie 4 os KRISTEN, U. and M. BIEDERMANN. 1981. Ultrastructure, origin, and composition of the protein bodies in the ligule of Isoetes lacustris L. Ann. Bot. 48:655-663. oe KRISTEN, U., G. LIEBEZEIT, and M. BIEDERMANN. 1982. The ligule of Tsoctes aa Ultrastructure, mucilage composition, and a possible pathway of secretion. Ann. Bot. aang McGraw-Hill, New MAHESHWARI, P. 1950. An Introduction to the Embryology of Angiosperms. S'c raw-Hill, ork. MASLEN, A. J. 1898. The ligule in Lepidostrobus. Ann. Bot. 12:256-259._ MICHAUX, N. 1973. La ligule chez rpostdt setacea Lam. C. R. Acad. Sci. Paris, D, engage PAOLILLO, D. J ., Jr. 1963. The developmental anatomy of Isoetes. Illinois Biol. Monogr. 31:1-130. PETTITT, J. 1970. Heterospory and origin of the seed habit. Biol. Rev. 45:401-415. SCOTT, D. H. 1920. Studies in Fossil Botany, Part I, ed. 3. A. & C- Black, Ltd., London. 28 AMERICAN FERN JOURNAL: VOLUME 74 (1984) SEWARD, A. C. 1910. Fossil Plants, II. Cambridge Biological Series, London. SHARMA, B. D., D. R. BOHRA, and R. SINGH. 1980. Transfusion cells in Isoetes coromandelina L. Curr. Sci. 49:872-874. SINGH, R., D. R. BOHRA, and B. D. SHARMA. 1982. Tracheary elements in Isoetes coromandelina L. Bionature 2:23-25. SOLMS-LAUBACH, H. 1892. Ueber die in den Kalksteinen des Kulm von Glatzisch-Falkenberg in Schlesien erhaltenen structurbientenden Pflanzenreste. I. Bot. Zeit. 50:49-56, 105-113. pl. Il. SPORNE, K. R. 1975. The Morphology of Pteridophytes, 4th ed. Hutchinson, London. Reprinted 1976 by B.I. Publications, Delhi. WILLIAMSON, W. C. 1894. On the organization of the fossil plants of the Coal Measures, XIX. Phil. Trans. R. Soc. London., B, 184:1-38. REVIEW PTERIDOPHYTIC FLORA OF GARHWAL HIMALAYA, by S. S. Bir et al. Jugal Kishore & Co., viii + 83 pp. illustr. 1982. Rs. 95.—This is an account of the pteridophytes in the Garhwal district of Uttar Pradesh, India (78-79° E Long, 30-31° N Lat), especially of the area within 50 km of the town of Mussoorie, an old and ever popular collecting locality. The region mostly lies at 600-2400 m elevation and has a tropical or subtropical monsoon climate. A total of 157 pteridophytes, = of them widespread species, are enumerated, and others whose presence in the region is problematical are mentioned. No new taxa are described. The generic name Microsorum Link (1833), rather than Microsorium, is adopted following Sledge (1960) and Pichi Sermolli (1977), although Link (1841) did correct the original spelling to Microsorium. A map of the region, photographs of some of the species, and an index conclude the volume, which is available from Jugal Kishore & > Co., 23-C Rajpur Road, Dehra Dun, India.—D.B.L AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) 29 SHORTER NOTES TWO SPECIES OF ADIANTUM NEWLY ESCAPED IN FLORIDA.—Two previously unrecorded species of Adiantum have recently been discovered near Homestead in southern Dade County, Florida. Both are commonly cultivated tropical species known from single sites, so the plants almost certainly are escapes from cultivation. Adiantum trapeziforme L. was discovered by the late George N. Avery in February 1980. He found a single plant growing at the top of a low spoil bank in an area kept humid by runoff from a nearby water-pumping station, and he collected a single fertile frond (Avery 2207, FTG). The fern grows in an area shaded by the exotic weed trees Schinus terebenthifolius and Albizzia lebbeck. Avery took us to see the plant in late 1982; we visited the site again in January 1984. Both times we looked for evidence of young Adiantum plants. No sporelings were ever found, although the original plant appears to be growing well and produces spores regularly. The Adiantum might represent a persistent plant from cuttings dumped at the site, but, as no evidence for dumping was found, we concluded that the plant is a true escape. The future of this species in South Florida is problematical. It grows in a relatively protected area and is not likely to be destroyed by human activities in the near future, but it apparently has not spread in the four years since its discovery. Whether it is capable of becoming truly naturalized is not clear. In March 1983 the senior author found approximately seven plants of Adiantum anceps Maxon & Morton growing on the vertical walls of trenches cut into rocky ground, along with Ctenitis sloanei (Poeppig ex Sprengel) Morton. The plants were growing in the shade of Schinus terebenthifolius. Undoubtedly water collecting at the bottom of the trenches helped maintain a humid environment. Two voucher specimens have been collected (Herndon 702 and Avery 2501, FTG). This species has spread within its rather specialized habitat, suggesting that the climate in South Florida is conducive to its growth. The colony is in an area of active commercial development and will likely be destroyed before the plants spread to more protected areas. Still, the habitat in which the plants grow is quite similar to the “pinnacle rock” areas found in some Dade County parks. Given the continuous supply of Spores from cultivated plants and the availability of apparently suitable habitat in Protected areas, it is likely that this species will sooner or later become an established member of our naturalized flora.—Alan and Rhonda Herndon, 15301 S.W. 306th St., Homestead, FL 33033. 30 AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) A NEW STATION FOR TRICHOMANES PETERSII IN ALABAMA.—The filmy fern Trichomanes petersii A. Gray (Hymenophyllaceae) was first described from Winston County, northern Alabama, and the fern occurs most abundantly in the northern part of that state. Nevertheless, relatively few stations are known, and the plant is considered rare and endangered by Dean (Ferns of Alabama and Fern Allies, 1964, p. 31) and by Short (Distribution of Alabama Pteridophytes, M.S. Thesis, Auburn University, 1978, p. 105). The entire range of 7. petersii in the United States includes North and South Carolina, Georgia, Florida, Tennessee, Illinois, Arkansas, Mississippi, and Louisiana. Only a single station is known in many states. While doing fieldwork in the vicinity of Tuscaloosa on 12 November 1983, I discovered a dense colony of T. petersii covering a standstone boulder in a small, north-facing ravine ca. 0.1 km south of Holt Lock and Dam on the Black Warrior River at an elevation of ca. 65 m, in Tuscaloosa Co., west-central Alabama (Wyatt 1622, AUA, GA, NCU, UNA). The new collection extends the range of 7. petersii approximately 100 km south of the areas of its greatest abundance on the Sipsey Fork of the Black Warrior River. Its occurrence in the Fall Line Hills of Tuscaloosa County on the southern edge of the Cumberland Plateau parallels its restriction to ravines near the western (Franklin, Lawrence, Winston, and Lamar Cos.) and eastern (DeKalb and Etowah Cos.) edges of the Plateau. The only exception in Alabama is the collection by Correll (Amer. Fern. J. 29:141. 1939) from the Valley and Ridge Province in Cleburne County. All collections appear to be from sandstone of the Pennsylvanian Upper Pottsville Formation. The steep, protected ravines of the Holt Lake area north of Tuscaloosa harbor a number of rare or disjunct angiosperms, including Neviusia alabamensis A. Gray, Croton alabamensis E. A. Smith ex Chapman, Cladrastis kentuckea (Dum.-Cours.) Rudd, and Aesculus parviflora Walter (Brooks, A Checklist of the Vascular Plants of the Holt Lock and Dam Area, Tuscaloosa County, Alabama. M. S. Thesis, Univ. of Alabama, 1969, p. 46). Fortunately, construction of the Holt Dam did not flood the ravine in which T. petersii occurs, as happened at the type locality for this fern (Dean, Ferns of Alabama and Fern Allies, 1964, p. 31) and for Leptogramma pilosa var. alabamensis (Crawford) Wherry (Short & Freeman, Amer. Fern J. 68:1-2. 1978) on Sipsey Fork in Winston County. It is possible that 7. petersii occurs at additional sites on the Black Warrior River north of Holt Lock and Dam and that this very diminutive fern has simply been overlooked. In addition to the seven counties for which herbarium records are known, attempts should be made to document literature reports for Marion Co. by Mohr (Contr. U. S. Natl. Herb. 6:319. 1901), Marshall Co. by Graves (Amer. Fern J. 10:78. 1920), and Walker Co. by Dean (Ferns of Alabama and Fern Allies, 1964, p. 174). I thank Ann Stoneburner and Ireneusz J. Odrzykoski for field assistance and Robert R. Haynes for advice. A grant from the Whitehall Foundation helped make this research possible.—Robert Wyatt, Department of Botany, University of Georgia, Athens, GA 30602. , AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 1 (1984) 31 TRICHOMANES GAMETOPHYTES AT BARTHOLOMEW’S COBBLE. —Distribution of the little known asporophytic fern gametophytes of Vittaria and Trichomanes in the northeastern United States was recently described by Farrar, Parks, and McAlpin (Rhodora 85:83-91. 1983). Their study was prompted by the discovery in 1976 by McAlpin and Farrar of Trichomanes gametophytes in Franklin County, Massachusetts, more than 400 miles above the previously known northern- most station (Amer. Fern J. 68:4. 1978). Both Trichomanes and Vittaria reproduce asexually by gemmae, which develop on the gametophyte, and are not known to develop sporophytes throughout their northeastern distribution. They are typically found on heavily shaded, moist outcroppings of non-calcareous rock, often deep in crevices or small caves. During the summer of 1982, I discovered a second Massachusetts station for Trichomanes at Bartholomew’s Cobble in southern Berk- shire County on rocks which are recognized as largely calcareous. This discovery is of interest because of the historic nature of the site and because of the apparent problem of the gametophytes growing on the “wrong substrate.” Bartholomew’s Cobble was aptly described by Weatherby (Amer. Fern 12 STA. 1947) as a picturesque natural preserve located in the southwestern corner of Massachusetts in the Housatonic River valley, several miles from Ashley Falls. Like other cobbles in the valley, it is formed as a rocky hill projecting from the underlying limestone. It is a National Natural Landmark protected and administered by The Trustees of Reservations of Massachusetts and is recognized by field naturalists for its impressive natural concentration of plants. Weatherby (Amer. Fern J. 37:1. 1947) cited a total of 276 species of flowering plants and ferns within the 25 acre preserve; however, literature distributed at the Cobble boasts almost 500 wildflowers, 100 kinds of trees, and 49 species of ferns and fern-allies, including somewhat elusive specimens of Scott’s Spleenwort. During the summer of 1982, while hiking the picturesque Ledges Trail overlook- ing the Housatonic River, I was attracted to investigate the little pockets and caves scattered around the outcrops, which are covered with various ferns and other plants. Having recently accompanied Dr. Donald Farrar in the field in Lancaster County, Pennsylvania, where he was studying the distribution of Trichomanes and Vittaria, | developed some skill in their field identification. To my surprise, I discovered the typical little fuzzy green tufts of Trichomanes growing on the sidewalls of several of these little “limestone” caves. These tiny plants are superficially very moss-like and may be easily overlooked; however, with a good hand lens and a little experience, they can be readily recognized. Farrar (pers. comm.) has indicated that neither Trichomanes nor Vittaria have ever been reported on a calcareous substrate. _ [was pleasantly surprised to discover that Wherry reported a somewhat similar incongruity when he discovered acidophilic Spike-moss ( Selaginella rupestris) growing on the same ledges of Bartholomew’s Cobble (Amer. Fern J. 41:1. 1951). The explanation, he found, was that acid soils accumulate in the pockets formed in the veins of milky-white quartz mingled with the limestone. It was In these very pockets that I discovered the Trichomanes. Dr. Wherry warned against erroneous inferences as to the soil preference of individual species on the basis of superficial 32 AMERICAN FERN JOURNAL: VOLUME 74 (1984) observations. I would like to think he would be pleased that another acid-loving plant was discovered growing on this predominantly limestone cobble. Bartholomew’s Cobble is surely an atypical site for Trichomanes. And yet, as field naturalists become more familiar with these interesting little ferns and involved in the search for new sites in the interest of extending the distributions recently mapped by Farrar, et al. (Rhodora 85:83-91. 1983), we do well to note Wherry’s warning and not overlook habitats on the assumption that they are calcareous and therefore unfavorable to Trichomanes.—Kenneth G. Miller, Roddy Science Center, Millersville University, Millersville, PA 17551. STATEMENT OF OWNERSHIP. MANAGEMENT AND CIRCULATION Required by 39 U.S.C. 3685) 1A, TITLE OF PUBLICATION B. PUBLICATION NO. 2. DATE OF FILING AMERICAN FERN JOURNAL @)/0/0/2|/81/4/4/4] 1 Oct 1983 3. FREQUENCY OF ISSUE ANNUALLY PRICE 4 $8 mem; $9 subscr U. S. Nat'l. Herbarium, 10th & Constitution Ave. N.W., Washington, DC 20560 U. S. Nat'l. Herbarium, 10th & Constitution Ave. N.W., Washington, DC 20560 6. FULL NAMES AND COMPLETE MAILING ADDRESS OF PUBLISHER, EDITOR, AND MANAGING EDITOR /This item MUST NOT be blank) PUBLISHER (Name and Complete Mailing Address) AMERICAN FERN SOCIETY, INC. (see 5.) EDITOR (Name and Complete Mailing Address) Dr. David B. Lellinger (see 5.) 7. OWNER (If owned by a corporation, its address must be stated and also immediately names and addresses of stockholders posbiethas Iveta y diam peas ir iemsetenep ie If not owned by a corporation, ac manos dee onvester OF te individual owners must onan ai rth hse ynchnlt cel girs sired aspepseckry po swe aha of each mda mute given. If the publica- by @ nonprofit organizatic just be stated.) (Item must be completed. FULL NAME ICAN FERN SOCIETY, INC. 8. KNOWN BONOHOLDERS, MORTGAGEES, AND OTHER SECURITY HOLDERS OWNING OR HOLDING 1 PERCENT OR MORE OF TOTAL AMOUNT OF BONDS, MORTGAGES OR OTHER SECURITIES (Uf there are none, 30 state) " . ULL NAME COMPLETE MAILING ADDRESS none 9. Has NOT CHANGED DURING HAS CHANGED DURING kk) PRECEDING 12 MONTHS PRECEDING 12 MONTHS. change with this a ——$$ $$ 10 AVERAGE NO. COPIES EACH ACTUAL NO. COPIES OF SINGLE ISSUE DURING PRECEDING ISSUE PUBLISHED NEAREST TO 12 MONTHS ame TES es A. TOTAL NO. COPIES (Net Press Run) 1300 1300 8. PAID CIRCULATION 1. Sales through dealers and carriers, street vendors and counter sales 0 Ww 2. Mail Subseri iption 1202 1212 © TOTALP, 1202 1212 . FREE DISTRIBUTION BY MAIL, CARRIER OR OTHER MEANS SAMPLES, COMPLIMENTARY, AND OTHER FREE COPIES 1 1 Or E. TOTAL DISTRIBUTION (Sum of C and D} 1203 1213 F. COPIES NOT DISTRIBUTED |. Office use, left over, unaccounted, spoited atter printing oF 87 sae Ae | certify that the statements made by me above are correct and complete Secleliecaen pile cee 1300 1300 AN kik Sen America and treated it as P. lemmonii Underw. The variety of P. munitum known es — Est imbricans D. C. Eaton he considered to be a distinct species. P. imbricans (D. C. Eaton) D. Wagner. He thus give the hybrid formula as P. imbricans x lemmonit. Herbarium and Department of Botany, University of Michigan, Ann Arbor, MI ge H1X 2B2 eect Marie-Victorin, Institut Botanique, 4101 Est, Rue Sherbrooke, Montreal, Que. 2B2, ada, including new record from Newfoundla ons wetwarin : > Pq \ hr a 7 Polystichum Xscopulinum, southerly slopes of dry serpentine ridge, near ee head of North Arm, Newfoundland, Rouleau own range of Polystichum & scopulinum, S pen “wl: oe te 8 ow an 884 (MT). FIG. (p861) 2 SWMOA ‘TWNYNOL NY34 NVOINSWY WAGNER & ROULEAU: WESTERN HOLLY FERN IN NEWFOUNDLAND 35 distance, the plant may be confused with P. lonchitis (L.) Roth, a common northern species, and thus may be overlooked in routine collecting. It may also superficially resemble exposed or dwarfed forms of P. braunii (Spenner) Fée, a fern with bipinnate, thinner-textured, and scalier fronds. These three species are contrasted in Fernald’s key (1950, p. 37). Rouleau found P. Xscopulinum for the first time in Newfoundland on 18 July 1950 while exploring the southerly slopes of a dry serpentine ridge near the head of North Arm in the Humber District (Rouleau 884, MT). The plant resembled a small, fertile P. braunii (Fig. 1), and its true identity was not recognized until March 1978. Rouleau has found both P. braunii and P. lonchitis to be common and widespread in Newfoundland, but thus far he has collected P. x scopulinum only at this single site. This discovery extends the known range of P. X scopulinum nearly 400 miles (560 km) northeast of the colony on Mt. Albert in Quebec. Although fully fertile, the herbarium specimen is smaller than average for this species. The stipes are unusually long, nearly equal in length to the blades, presumably as a result of growing in a deep pocket. The pinnae of the lower half of the blade are strongly auricled at the anterior base, and some of the auricles are contracted basally to a stalk. A distribution map (Fig. 2) of P. x scopulinum based on records from W. Wagner (1973), D. Wagner (1979), Taylor (1970), and the Newfoundland specimen shows not only the vast disjunction between west and east and the far greater number of localities in the west, but also the extension of the taxon in the west to much lower latitudes. Damman (1965), in his review of distribution patterns in the flora of Newfound- land, recognized a category of species that are restricted to serpentine areas, to which P. x scopulinum must now be added. He described the habitats as bleak and exposed with sparse vegetation. He believed that serpentine species occur on these sites because of a “lack of competition, the basic soils, and their ability to tolerate frost churning.” Damman discussed Fernald’s “Nunatak Theory” and concluded from reports by a number of workers that Fernald’s nunatak areas had been covered with ice at some time or other during the glacial period. : Rather than assume that P. Xscopulinum persists as a relict ona Newfoundland nunatak, it may be more reasonable to attribute the occurrence of P. x scopulinum to long distance dispersal by spores. Bouchard et al. (1977) recently gave another striking example of probable long distance dispersal in Newfoundland: Thelypterts limbosperma (Allioni) H. P. Fuchs [syn. Oreopteris limbosperma (Allion!) Holub] was unknown in eastern North America until 1973, when a large colony was found in Gros Morne National Park. The species is known to occur otherwise in North America only from Alaska to central Washington. In view of the number of examples now known 0 Canada and the United States and the evidence that single spores have the potentiality for establishing new colonies (Wagner, 1972), the presence of P. *scopulinum in the east seems no longer as exceptional as It was once thought he . The existence of this fern in Newfoundland several hundred miles from wes nearest other colony could have resulted from a wind-blown spore from as close as the Gaspé Peninsula or as far as the west coast of North America. f west to east disjunctions in 36 AMERICAN FERN JOURNAL: VOLUME 74 (1984) We thank André Bouchard, A. W. H. Damman, David H. Wagner, and Florence S. Wagner for help in various ways. LITERATURE CITED BOUCHARD, A., D. BARABE, and S. HAY. 1977. An isolated colony of Oreopteris limbosperma (All.) Holub in Gros Morne National Park, Newfoundland, Canada. Naturaliste Can. 104:239-244. DAMMAN, A. W. H. 1965. The distribution patterns of northern and southern elements in the flora of Newfoundland. Rhodora 67:363-392. FERNALD, M. L. 1924. Polystichum mohrioides and some other subantarctic or Andean plants in the northern hemisphere. Rhodora 26:89-95. ————.. 1950. Gray’s Manual of Botany, 8th ed. American Book, New York. TAYLOR, T. M. C. 1970. Pacific Northwest Ferns and their Allies. Univ. of Toronto Press, Toronto and Buffalo. WAGNER, D. H. 1979. Systematics of Polystichum in western North America north of Mexico. Pteridologia 1:1—64. WAGNER, W. H., Jr. 1965. Pellaea wrightiana in North Carolina and the question of its origin. J. Elisha Mitchell Sci. Soc. 81:95-103. . 1972. Disjunctions in homosporous vascular plants. Ann. Missouri Bot. Gard. 59:203-217. . 1973. Reticulation of holly ferns (Polystichum) in the western United States and adjacent Canada. Amer. Fern J. 63:99-115 REVIEW FERNS AND FERN ALLIES OF GUATEMALA, PART III. MARSILEACEAE, SALVINIACEAE, AND THE FERN ALLIES (INCLUDING A COMPRE- HENSIVE INDEX TO PARTS I, II, AND IID, by Robert G. Stolze. Fieldiana: Botany, NS, 12. 91 pp. 1983.—This is the final part of the Ferns and Fern Allies of Guatemala, and includes the heterosporous ferns and fern allies—eight genera in all—plus an index to all parts. Lycopodium is treated by B. @llgaard and Isoétes with the help of R. J. Hickey. The third part follows the first two in format (Part I reviewed in Amer. Fern J. 67:94, Part II in 72:14), and the clear, concise style is maintained here also. I found no errors of consequence. The keys seem well constructed with contrasting statements in each pair of leads. I tried several species of Lycopodium and found no problems in use. A nice feature is the discussion within the generic treatments of excluded species In Marsilea, Azolla, and Salvinia which have not been found but could be expected in Guatemala, and how these could be distinguished from included taxa. Stolze also points out the need for monographic work in these genera and the difficulties in taxonomy and identification when much (most in Azolla) material is vegetative only. The author is to be commended on the completion of this important work on tropical American pteridophytes. Stolze’s Ferns and Fern Allies of Guatemala and Man R. Smith’s Flora of Chiapas—Pteridophytes have made an excellent start on filling in our knowledge of the ferns on this region.—James D. Montgomery, Ichthyological Associates, Inc., Berwick, PA 18603. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 2 (1984) 37 Drymoglossum Under Water Stress C. §, Hew? Drymoglossum is an epiphytic fern found commonly on the trunks of some local trees. It exhibits typical CAM features (Hew & Wong, 1974; Wong & Hew, 1976). It has fleshy fronds for storing water, but it is not uncommon to observe shrivelling of its fronds during dry spells. Different CAM plants seem to respond to drought differently. Under water stress, CAM plants such as Tillandsia increase their CAM activities, but plants such as Opuntia reduce their CAM activity instead (Kluge & Ting, 1978). We have reported recently that tropical succulent orchids behave just like Opuntia when placed under water stress (Fu & Hew, 1982). 240Fr = 200 D a, D g = 160 - se, e 3) Pink A Control © 120 A * -10bars 3 % ® -i5bars S 2 =e Oo -18bars e 40 x 0 0800 3200 1600 2000 « e..0400 0200 1200 Time (hours ) FIG. 1. Effect of water stress on diurnal fluctuation of titratable acidity in Drymoglossum fronds after imposing two days of water stress. s in amino acid metabolism are aspects Inhibition of protein synthesis and change olist been extensively studied in relation to of metabolic changes in plants which have : water stress (Hsiao, 1973). Accumulation of proline concentration was the main reason for the increase of free amino acids (Singh et a 1973a, b). Proline accumulation is a result of a stimulation of its synthesis and an inhibition of its utilization (Stewart, 1980). Works on proline accumulation under water stress have * as 5 OADOES. > ic of Singapore. Department of Botany, National University of Singapore, Singapore 0511, Republic of Singapore 38 AMERICAN FERN JOURNAL: VOLUME 74 (1984) > 80 < = 160} 2 ad x 2 560 f a A ret 3 120+ Cg Control g so -10 bars c 3 - nee < 80F . -15bars o x £ £ 3 me S w Q20F- = 40+ ~ —18 bars i i J f- 1 Ll i. 2 0 -10 -12 6 =18 No.of Days under Stress PEG concentration (bars ) FIG. 2. Effect of length of time of water stress on titratable acidity in Drymoglossum fronds. FIG. 3. Proline accumulation in Drymoglossum fronds under various PEG concentrations 56 hrs after imposition of water stress. been carried out with Chlorella (Greenway & Setter, 1979), barley (Hanson et al., 1977; Singh et al., 1973a, b; Tully et al., 1979), Bermuda grass (Barnett & Naylor, 1966), halophytes (Cavalieri & Huang, 1979), spinach (Huang & Cavalieri, 1979), and tobacco (Boggess & Stewart, 1980). This paper reports the effect of water stress on CAM activity and proline accumulation in Drymoglossum. Sterile, mature fronds of Drymoglossum piloselloides (L.) Presl were collected from Acacia tree trunks. Eight uniform-sized fronds were placed in each petri dish with filter paper serving as substratum. These were kept in a temperature-controlled room (26° C) with a 12 hr photoperiod and at a light intensity of 1200 ft-c. The light source was a 700-watt HPLR mercury lamp. Water stress was imposed by placing the fronds in various concentrations of polyethylene glycol (PEG) 1000, giving a range of osmotic potential from about ~ 10 bars to — 18 bars. The control was kept in distilled water. All treatments were done in triplicate. Titratable acidity was measured as described by Wong and Hew (1976). The extraction and determination of proline were done according to the method de- scribed by Singh et al. (1973a) and by Troll and Lindsley (1955). Unless stated otherwise, measurement of titratable acidity was done at 0800 hr. Both controls and stressed plants displayed pronounced diurnal fluctuation of titratable acidity (Fig. /). Pronounced decrease in titratable acidity was observed 2 days after water stress. Titratable acidity decreased with a corresponding increase un PEG concentration and with days of water stress (Figs. | & 2). The decrease In titratable acidity in Drymoglossum under stress, therefore, resembled that of tropical orchids (Fu & Hew, 1982). The drop in titratable acidity in water-stressed C. §. HEW: DRYMOGLOSSUM UNDER WATER STRESS 39 Drymoglossum could also be attributed to a decrease in CO) fixation, as has been observed in orchids (Fu & Hew, 1982) and in other plants (Osmond, 1978 Proline accumulation increased in fronds of Drymoglossum following water stress (Fig. 3). This is in agreement with previous reports obtained for barley, grass, wheat, spinach, and tobacco. Our findings support the generalization that most if not all plants have the ability to accumulate proline, provided that the imposed water stress is severe enough (Hsiao, 1973). Proline accumulation in succulent halophytes under stress has been reported (Cavalieri & Huang, 1979), but it is yet to be shown that the succulent halophytes exhibit CAM features (Webbs & Burley, 1965). We now have evidence to indicate that under PEG stress, CAM plants such as Drymoglossum start to accumulate proline around — 10 to —15 bars. The same has also been observed for succulent tropical orchids, also CAM plants (Hew, unpublished data). LITERATURE CITED BARNETT, N. M. and A. W. NAYLOR. 1966. Amino acid and protein metabolism in Bermuda grass during water stress. Pl. Physiol. 41:1222-1230. BOGGESS, S. F. and C. R. STEWART. 1980. The relationship between water stress induced proline accumulation and inhibition of protein synthesis in tobacco leaves. Pl. Sci. Letter. 17:245-252. CAVALIERI, A. H. and A. H. C. HUANG. 1979. Evaluation of proline accumulation in the adaptation of diverse species of marsh halophytes to the saline environment. Amer. J. Bot. 66:307-312. FU, C. F. and C. S. HEW. 1982. Crassulacean acid metabolism in orchids under water stress. Bot. Gaz. 143:294—297. GREENWAY, H. and T. L. SETTER. 1979. Accumulation of proline and sucrose during the first hour after transfer of Chlorella emersonii to high NaCl. Austral. J. Pl. Physiol. 6:69-79. HANSON, A. D., C. E. NELSON and E. H. EVERSON. 1977. Evaluation of free proline accumula- tion as an index of drought resistance using two contrasting barley cultivars. Crop ac 17720-7236, HEW, C. S. and Y. S. WONG. 1974. Photosynthesis and respiration of ferns in re Amer. Fern J. 64:40—48. HSIAO, T. C. 1973. Plant responses to water stress. Ann. Rev. Pl. Physiol. 24:519-570. HUANG, H. C. A. and A. J. CAVALIERI. 1979. Proline oxidase and water stress-induced proline accumulation in Spinach leaves. PI. Physiol. 63:531-535. ae oe KLUGE, M. and L. P. TING. 1978. Crassulacean Acid Metabolism. Analysis of an Ecological Adaptation, Springer. Berlin. OSMOND, C. B. 1978. Crassulacean acid metabolis 29:379-414. SINGH, T. N., L. G. PALEG, and D. ASPINALL. 1973a. Stress metabolism I. and growth in the barley plant during water stress. Austral. J. Biol. Sci. 26a =e SINGH, T. N., D. ASPINALL. L. G. PALEG and S. F. BOGGESS. 1973b. Stress = ; Changes in proline concentration in excised plant tissues. Austral. J. Biol. Sct. 26:57-63. s STEWART, C. R. 1980. Proline accumulation: Biochemical aspects. /” Paleg. site ms “as 7 (eds). Physiology and Biochemistry of Drought Resistance 10 Plants. Harcourt brace Jovanovich, New York. TROLL, W. and J. LINDSLEY. 1955. A photometric metho Chem. 215:655— i ies e a ene and C. E, NELSON. 1979. Proline accumulation In sere oot barley leaves in relation to translocation and the nitrogen budget. PI. one =) Sas It WEBBS, K. L. and J. W. A. BURLEY. 1965. Dark fixation of CO, by obligate and facultative sa marsh halophytes. Canad. J. Bot. 43:281-285. WONG, S. C. and C. S. HEW. 1976. Diffusive resistance, tit tropical epiphytic ferns. Amer. Fern. J. 66:121-124. — lation to their habitat. m: a curiosity in context. Ann. Rev. Pl. Physiol. Nitrogen metabolism d for the determination of proline. J. Biol. ratable acidity. and CO, fixation in two 40 AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 2 (1984) Problems in Asplenium, with Some New Species from Ecuador ROBERT G. STOLZE* While studying the genus Asplenium for the “Flora of Ecuador,” I encountered great difficulties with circumscription of A. myriophyllum (Swartz) Presl. This is a highly dissected species of wet forests, reported from Florida, the Greater Antilles, Mexico, much of Central America, Colombia, Venezuela, Ecuador, and Peru. It occurs rather frequently throughout much of its range, has several very close relatives, and is itself highly variable. Among its most diagnostic characters are: tripinnate or subtripinnate (often only bipinnate in Florida) laminae rather abruptly reduced at the base; dull grayish or reddish brown petioles up to 15 cm long; castaneous to blackish, rather shiny, scarcely clathrate rhizome scales; and pinnae about 20-25 pairs, mostly ascending, sessile, with the basal pinnules overlapping wae rachis. splenium myriophyllum has been found in herbaria, often identified as A. Poise Fée, A. cristatum Lam. (syn. A. cicutarium Swartz), A. divaricatum Kunze, and A. rutaceum (Willd.) Mett. (syn. A. conquisitum Underw. & Maxon ex Christ). It is easy to understand the confusion, for these species share many similar features, and whereas each seems to be readily identifiable in the typical habit, there can always be found several collections in which characters tend to merge with those of other species in the group. Asplenium rutaceum (common in the neotropics) is the most distinctive, as it is the only species which has highly lustrous, castaneous to blackish petioles and lower rachises. Furthermore the lamina is strongly and gradually reduced to a naked and flagelliform apex, this often with a proliferous tip. In the other species of the group, the petiole is dull and lighter in color, and the lamina tapers to a pinnatifid apex—this sometimes attenuate, but never flagelliform and radicant. Asplenium cristatum (also with wide distribution in the neotropics) is the next most distinctive of the species listed above, as the lamina is scarcely, if ever. reduced at the base; i.e., the basal pair of pinnae are nearly or quite as long as the central ones. The lamina in all the other species is strongly (either gradually or abruptly) reduced at base, with two to many proximal pairs of pinnae conspicuously shorter than the central ones. Nevertheless, I have found several specimens of A. cristatum, from Guatemala to Ecuador, in which one or two proximal pairs of pinnae are significantly reduced. Asplenium cladolepton and A. divaricatum are species which more properly might be considered variants of A. myriophyllum. The former, known from Colombia, Venezuela, and rarely in Ecuador, differs from A. myriophyllum primarily in its smaller size and narrower lamina. The epipetric Peruvian species, A. divaricatum. appears to be a depauperate A. myriophyllum with leaves only 3-10 cm long and tiny ultimate segments. It may be merely an ecological variant. *Department of Botany, Field Museum of Natural History, Chicago, IL 60605. R. G. STOLZE: PROBLEMS IN ASPLENIUM 4] Asplenium myriophyllum varies strongly throughout its range. Many collections from Florida and Guatemala are much smaller and often only bipinnate, whereas leaves in South American plants are typically luxuriant, up to 40 cm long and fully tripinnate. Petioles are commonly 8-15 cm long, yet those of some plants from Florida and the Greater Antilles measure only 2-3 cm. It is no wonder then that ferns in this group are universally misunderstood. Obviously detailed research is needed, in which comparisons are made of specimens throughout the neotropics. Furthermore, it is probable that much hybridization is occurring here—a situation not uncommon in the genus. Mickel (“How to Know the Ferns and Fern Allies,” p. 56. 1979; Wm. C. Brown, Dubuque, Iowa) delineates several known hybrids in North America which involve A. myriophyllum. But no such studies have been undertaken yet throughout the broad range of its distribution. One should be cautious in naming a new species within a group already so confused. However, in studying the genus for “Flora of Ecuador,” I came upon a large number of Ecuadorean specimens which appeared to be clearly distinct in lamina architecture from the other components of the species complex. These were variously determined as A. cicutarium, A. conquisitum, A. cristatum, A. myriophyllum, A. rhizophyllum, and A. rutaceum. As herbarium specimens were separated on the basis of obvious morphological features, other, less obvious, characters came to light, which convinced me that this was a species much more distinct than most others already recognized in the group. Oddly enough, although it has been collected frequently in Ecuador, I can find no record of its occurring IM neighboring countries. Asplenium ecuadorense Stolze, sp. nov. Figs. 96 Rhizoma erectum, paleaceum; paleae 2-4 mm _ longae, deltato-lanceolatae ve -ovatae, brunneae vel fuscae, conspicue clathratae: folium 20—45 cm longum, 0— ipl , fascic plerumque brunneus, numquam_ nitidus, lamina gradatim: pinnatifidam vel flagelliformem versus, saepe apice prolificanti, ‘ Ee arater angustata; pinnae 34—60-jugae, lanceolato-oblongae, confertae vel poten i patentes, vel pinnae proximales deflexae, sessiles, pinnulis basalibus a i. Superpositis supra rachin; venae segmentorum ultimorum simplices; sori in quog Segmento solitarii; indusium delicatum, subintegrum Chimborazzo [Prov. Bolivar, Ecuador], 500 m, ornans,” Rimbach 16 [Rosenstock exsicc. no. isotypes GH, S, US). ae In wet forests, commonly on trunks of trees, very rarely on the forest floor, 100-1400 m, thus far found only in Ecuador. Plants epiphytic strial; rhi stout, erec _ rarely terrestrial; rhizome * Pole: - lanceolate aa. : n or gray-brown scales, these 2-4 mm long, conspicu 10] (US, 2 sheets: 1197005-6: > tripinnate or subtri asciculate; petiole 1-3 ¢ , terete, ¢ reddish brown oe “ mang glabrous OF with ee tek Nute, filiform scales; lamina glabrous, thin-herbaceous to membranaceous, AMERICAN FERN JOURNAL: VOLUME 74 (1984) FIGS. adaxial side, x 6. FIG. 3. Rhizome seale,: <1 1-3. Asplenium spipgorneb as FIG. 1. Habit, x 0.5 12. _ FIG. 2. Portions of rachis and two pinnae. R. G. STOLZE: PROBLEMS IN ASPLENIUM 43 green, elliptic-attenuate, strongly and gradually reduced to a pinnatifid or flagelli- ex and often with a proliferous tip, strongly and gradually narrowed at base, with 8-18 slightly to greatly reduced pinnae; rachis dark or reddish brown, marginate or narrow-alate; pinnae (30)34—-60 pairs, crowded to imbricate, mostly ome imal ones commonly deflexed, oblong-lanceolate, their of their length, sessile, the basal pinnules strongly overlapping the rachis; pinnules 10-20 pairs, cut nearly or quite to the costa into of segments, these elliptic or ovate-elliptic, entire (or often the basal SELECTED SPECIMENS EXAMINED: ECUADOR: Bolivar: Western Cordillera, western slope of the river Suquibi, 500 m., Rimbach 29 (topotypes GH, US). Cotopaxi: Epiphyte, produces new crowns from a terminal bud. On road from Quevedo to Quito, ca. 500 m, Haught 2917 (GH, S. UC, US). Guayas: Crescit in silv. trop. prop. Puente de Chimbo, 8/1891, Sodiro 20/47 (P). Los Rios: Epiphytic fern, lightly disturbed forest on hills: Centinela Ridge area, 12.5 km E of Patricia Pilar, 1400 ft, Hansen et al. 7746 (AAU, MO). Manabi: La Morena, ca. 15 km NNE of Flavio Alfaro, remnants of seasonal rain forest, ca. 100 m, Harling & Andersson 18909 (GB). Pichincha: Seasonal rain forest, 20 km W of Santo Domingo de los Colorados, 1000 ft, Cazalet & Pennington 5264 (NY, UC, US). Tungurahua: Western Cordillera, W of town of Ambato, 500 m, moist forest region, Rimbach 640 (F). Zamora-Chinchipe: Fallen trees on hilside, on tree trunk, road from Loja to Zamora, km 42, 1400 m, Dodson & Thien 843 (MO). , The general aspect of this new species 1s that of A. rutaceum in that both have the lamina strongly reduced at the apex and base, with the tip of the rachis often flagelliform and radicant. The pinnae of both are commonly borne at right angles to the rachis and are sessile, with the basal pinnules strongly overlapping the rachis, and the pinna margins parallel for most of their length. In addition, the rhizome scales are gray-brown and conspicuously clathrate. However, A. ecuadorense ditters obviously from A. rutaceum in that the pinnae are much more crowded (often strongly imbricate) and numerous (ca. 3550 pairs vs. 15-30 pairs), and the petioles and lower rachises are dull brown or reddish brown, VS- castaneous to atropurpureous and highly lustrous in A. ruftaceum. Sem Asplenium myriophyllum is the other species with which A. ecuadorense mig A e easily confused. Both are similar in gross morphological characteristics. anal’ or subtripinnate lamina, strongly reduced to both apex and base; dull scsi ae usually reddish brown petiole; pinnae sessile, with basal pinnules oan a rachis. But the new species has gray-brown rhizome scales that are fe ks clathrate, whereas in A. myriophyllum these are castancous to blackish and only obscurely clathrate (i.e., the lumina are greatly constricted). Also, OD AQT? Text P8 =~ a" ZI a Cy —_ as a Beceag Om ae ek cn NS SS = Sy Sa EES t and bequests to the Society enable it to expand its services to members and to others interested in ferns. Botanical books, back issues of the Journal, and cash or other gifts are always welcomed, and are tax-deductible. Inquiries should be addressed to the Secretary. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 3 (1984) 65 The Habitat Characteristics and Abundance of Equisetum ferrissii and its Parent Species, Equisetum hyemale and Equisetum laevigatum, in lowa LORENZ M. RUTZ and DONALD R. FARRAR* Equisetum Xferrissii Clute is the naturally occurring sterile hybrid of E. laevigatum A. Br. and E. hyemale var. affine (Engelm.) A.A. Eaton (Hauke, 1958). Although it is morphologically distinct and relatively easy to identify, it does not consistently appear on floristic lists of Iowa’s prairies, nor in broader discussions of lowa or Midwest floras. Only recently has the commonness of E. X ferrissii become recognized. Peck (1976) documented the presence of E. x ferrissii in 74 of lowa’s 99 counties. He felt (pers. comm.) that a thorough search would turn up E. x ferrissii in the remaining counties as well, making it as widespread in Iowa as its parent species. E. hyemale has been recorded in all counties, and E. laevigatum in 91 of 99 counties. The current study was designed to quantify the presence of E. x ferrissii in one county considered representative of much of Iowa and to address ecological questions regarding the dispersal and maintenance of the sterile hybrid and its parental species. Ecological information on these three Equisetum taxa is sparse. Hartman (1958) recognized what are now known as E. x ferrissii and E. laevigatum as plants of ditches and embankments, noting also that E. x ferrissii grew in moister sites than E. laevigatum and that E. hyemale preferred a moister habitat than either E. Xferrissii or E. laevigatum. Because they are agricultural weeds and toxic to livestock, E. palustre and E. arvense have received considerable attention. Borg (1971) did an extensive study on E. palustre as a weed in Finland. He recorded rhizome growth of 30 cm to several meters per year, reaching depths of 180 cm below the soil surface. In regard to vegetative dispersal, he found that rhizome fragments of E. palustre were not good propagules and did not support the idea that E. palustre was spread by road building activity. a As field weeds, E. palustre and E. arvense have been tested for perbientc responses. E. arvense topgrowth was found to be sensitive to most chemicals, and thizome growth could be retarded to various degrees (Hoyt & Corder, 1962). Equisetum palustre, on the other hand, was found to be quite resistant to the chemicals tested. Topgrowth could be damaged, but rhizomes remained virtually unaffected (Holly, 1953). - The ease of vegetative propagation in Equisetum has been noted severa ue (Hauke, 1963: Praeger, 1934; Schaffner, 1931; Wagner and Hammitt, 1970). "he reports concentrate on the proliferation of aerial culm fragments floating on water or placed in or on saturated soil. Hauke (1963) suggested that E. X ferrissit culms = transported outside the range of its parent species by human activity. He note ; however, that preventing desiccation during translocation was critical for pub ge Dosdall (1919) also found that culms rapidly lost viability upon eye ts “Department of Botany, Iowa State University, Ames. IA 50011. Volume 74, number 2, of the JOURNAL was issued July 18, 1984 66 AMERICAN FERN JOURNAL: VOLUME 74 (1984) contrast to Borg’s findings (1971), that rhizome fragments were the most suitable propagules for starting Equisetum in drained soil. Sexual reproduction in Equisetum has been studied mostly in laboratory cultures, but Duckett and Duckett (1980) described the ecology of in situ gametophytes of E. arvense, E. fluviatile, and E. palustre in Great Britain. The sites described were previously flooded muds, bare of vegetation. They found that spores and gameto- phytes had a very low range of moisture tolerance and proposed this as an explanation for the frequent absence of gametophytes in apparently suitable sites. Gametophytes in the American Midwest were discovered by Walker (1921, 1931) on stream banks in Iowa, Nebraska, and Kansas. All gametophytes were found on the narrow mudbelt along the water, typically associated with Riccia. Unfortunately, Walker’s identifications must be treated with caution for two reasons. She reported gametyophytes of “E. kansanum” and E. “laevigatum,” which are now known respectively as E. laevigatum and the sterile (!) E. Xferrissii, and, according to Walker (1931), the identifications depended “entirely upon the species growing in the vicinity.” fey 4 oe @ 92° ae £24 ; ‘ = Field o> 6 S < S a ie 9° o Fes ay oy = Depth @ Width Fence- row Lisanti pels ar oe er eis FIG. 1. Schematic cross section of a typical lowa road showing ditch features; not drawn to scale. METHODS Natural areas in central and northwestern Iowa were visited to gain familiarity with the three taxa in their native habitats and in roadside ditches. Hauke’s (1963) monograph was used to identify the plants. To quantify the relative frequencies of the three taxa, and to obtain ecological information on ditch populations, a survey of 64 miles (103 km) of roadside in Story County, central lowa, was made on foot in the summer of 1981. Story County 1s composed of 16 townships of 36 sections each. Sections are one mile (1.6 km) square and usually are delimited on four sides by roads. The roads are bordered on each side by ditches 4-10 m wide and 1-1.5 m deep, depending on the road type (Fig. 1). Information regarding the design, construction, and maintenance of Iowa’s roads and ditches was obtained from the Iowa Department of Transportation (IDOT), RUTZ & FARRAR: HABITAT CHARACTERISTICS AND ABUNDANCE OF EQUISETUM 67 TABLE 1. PRESENCE OF Equisetum SUBGENUS Hippochaete IN IOWA PRESERVES OF NATIVE VEGETATION. Vegetation Equisetum Equisetum Equisetum Preserve type laevigatum x ferrissii hyemale Bixby State Park Forest - = 4 layton Co. Black’s Prairie Prairie + + - tory Co. Cayler Prairie Prairie + + _ Dickinson Co. Crossman Prairie Prairie + i oward Co. Dolliver State Park Forest = = x ster Co Doolittle Prairie Prairie aie 5 mE ory Co. Freda Haffner Kettlehole Prairie rk aE a kinson Co. Gitchie Manitou State Preserve Prairie & + + + Lyon Co. forest Hayden Prairie Prairie ae + . Howard Co. ee gee * Kalsow Prairie Prairie - = Pocahontas Co Ledges State Park Forest oe ng. = Boone Co. Woodman Hollow State Forest as ne i eserve, Webster Co. + = present, — = not observed, * = present only as invasion from bordering roadside ditch the Story County Engineer, and the Story County Weed Commissioner. The survey unit was the four ditches bordering a section. If any of the ~— were missing, the equivalent ditch of the adjacent section was surveyed. One primary section in each of eight alternating townships was selected randomly. Along primary sections, the following ditch and Equisetum stand characeristics were measured: drainage, topographical position, ditch depth, apparent soil drift, species aang (number of species present), road type, ditch position relative to field (N, E, 2 : estimate of snow drift, *type and width of fencerow, *adjacent field use, . . material, *slope of ditch bottom, kind of Equisetum, overall density of the -~- : maximum density found in the stand, height of Equisetum, stand length, stan Width, *percent of culms with cones, and *health of plants. Asterisked characteris- tics were found to be closely linked to others or essentially non-varying. For clarity of presentation, these are not included in the results. : ee To obtain a larger statistical base for abundance data, two “secondary — Were likewise selected from each of the eight townships. Along these sections, only the taxon present, stand length, and overall density were recorded. ae ar Secondary sections is included only in Table 2. Equisetum stands typically eae the entire ditch from fence row to road bed. Thus stand length, eed aa e the road, was directly proportional to the total physical extent of the stand. 68 AMERICAN FERN JOURNAL: VOLUME 74 (1984) TABLE 2. RELATIVE ABUNDANCE OF Equisetum laevigatum, E. * ferrissii, and E. hyemale IN ROADSIDE DITCHES IN STORY COUNTY, IOWA. Sum of Average No. of stands! stand stan No. of length length Low Medium High Taxon Sections stands (m) (m) density ensity density E. laevigatum imary 166 22,370 135 0 5 By Secondary” 198 22,445 113 93 74 34 Total 364 44,723 123 173 133 87 E. X ferrissii i 3,868 64 a3 7 Secondary” 2,923 59 40 10 1 Total 110 6,791 62 93 17 I E. hyemale im 8 611 l 3 4 Secondary” 9 1,363 152 2 1 6 Total 17 1,974 116 3 4 10 All Grand total 49] 53,291 109 268 154 98 'Low = 0-50 culms per square meter, med = 50-200, high = 200-400. Some extensive stands were measured at several points along their length. *Time constraints allowed the survey of only half of the secondary sections initially selected, con- sequently their distribution was biased towards the north portion of the county. OBSERVATIONS AND RESULTS IN NATURAL HABITATS Prior to European settlement, Iowa lands formed a network of upland prairies, prairie potholes, and prairie and woodland streams, providing an abundance of habitats suitable to E. x ferrissii and its parent species. The distribution of the three taxa, as observed in natural areas of a number of Iowa’s parks and preserves, reflects this earlier distribution (Table 1). Equisetum hyemale is common on the banks and terraces of Iowa’s creeks and rivers. It may become a dominant part of the understory of the gallery forest, attaining densities, at Ledges State Park for example, of 500 culms per square meter over large areas. Stands continue along tributary branches to various degrees, sometimes well into steep gullies with only intermittently flowing water. Low density stands occasionally appear on old terraces well above the present floodplain. An example of this occurs at Woodman Hollow State Preserve. Equisetum hyemale grows on or very near the water table. Subsurface soils of this habitat are unlikely to ever become dry. Stands of E. laevigatum and E. x ferrissii in native Iowa habitats always occur in full sunlight on prairie uplands. In undisturbed prairie, both may occur conspicu- ously or in very low densities. The presence of E. laevigatum at Crossman Prairie was substantiated by only two individual culms. In disturbed areas of native prairies, densities become substantially higher. Along an old drainage ditch cut through the northwest corner of Kalsow Prairie, for example, E. laevigatum attains densities of 60 culms per square meter. E. ferrissii is less frequent than E. laevigatum, but it also 18 not uncommon in native prairie habitats. Soils in Iowa’s prairies range from dry to very wet, reflecting the poor drainage conditions of much of northern and central Iowa prior to artificial drainage by European settlers. As with soils of the native riparian habitat of E. hyemale, it is unlikely that the more poorly drained prairie soils ever become dry. Equisetum laevigatum, however, grows on higher, well-drained sites, along the rim of the Freda RUTZ & FARRAR: HABITAT CHARACTERISTICS AND ABUNDANCE OF EQUISETUM 69 Haffner Kettlehole in northwestern Iowa, for example. The substrate there is a coarse glacial till, which becomes very dry in summers of sparse rainfall. E. x ferrissii grows in the same area, but is confined to the lower slopes of the kettle im Two sites with Equisetum gametophytes were located. Sporophytes from these gametophytes were transplanted, allowed to mature in the greenhouse, and identified as E. hyemale in both cases. In the summers of 1980 and 1981, gametophytes of E. hyemale were common on the vertical cut bank of the Des Moines River at Ledges State Park. A few gametophytes were also found on the silts and muds of the level floodplain a few meters back from the bank. The second site was along the cut banks of Kegley’s Creek in Story County (section 36, Lafayette Township). The cut banks at both sites were 1 —2 m high, kept moist by constant seepage, faced north to west, and received only late afternoon sun, if any at all. New sporophytes were common in late summer, but their fate was uncertain, as frost wedging cleaves most of the bank faces, burying the sporelings in mud. Associated plants included Ricciocarpus natans and Riccia frostii, several unidentified mosses, and a variety of angiosperm seedlings and annuals. OBSERVATIONS AND RESULTS IN ROADSIDE DITCHES As the prairies were converted to farmland, roads were cut into the landscape in the middle 1800's. Equisetum x ferrissii and E. laevigatum were displaced from their native prairie but found a suitable habitat along the roadsides where they are now found in abundance. Today, with the virtual elimination of prairies, roadside ditches have become the primary habitat for E. x ferrissii and E. laevigatum, and often provides suitable habitats for E. hyemale as well. ae Characteristics of ditch stands of Equisetum.—The number, size, and distribu- tion of stands among three density classes of each taxon was surveyed (Table 2). Equisetum laevigatum was by far the most common, having the largest number of stands and the greatest sum of stand length. Because of the difficulty in delimiting individual stands, the latter figure probably expresses the dominance of E. laevigatum more meaningfully than does the number of stands. The total length of ditch occupied by E. laevigatum was over six times that of E. x ferrisstl, which in turn was over three times that of E. hyemale. The total stand length of E. laevigatum alone represents 43% of the 103 kilometers surveyed. Discounting overlaps In stands, 52% of the extent of central lowa’s roadside ditches contains one of these three Eguisetum taxa. The Sianbation of stands in three density categories is different for each of the taxa (Table 2). Equisetum laevigatum stands regularly occur In all categories, re a more frequently of low or medium density. Equisetum x ferrissil occurs oop exclusively as low density stands. Equisetum hyemale shows the opposite, ces mostly as high density stands. This relationship 1S repeated in the mean be ‘ ao shown in Table 3. The maximum density also indicated that E . X ferrissit ( : = . m*) had by far the lowest density of the three taxa. Equisteum hyemale = s highest mean maximum density (156 culms m2), but did not greatly exceed ©. laevigatum (105 culms m7”). 70 AMERICAN FERN JOURNAL: VOLUME 74 (1984) BS E. laevigatum 60-- E. X ferrissii = 50 E.hyemale w a z Q. o Rhizoctonia solani 50 50 100 100 > 1000 »1000! 100 Botrytis cinerea 5 500 >1000 >1000 1000! >1000! 500 Pythium debaryanum 100 5 5 500 500 1000 50 P. middletonii 50 50 50 50 50 50 P. ultimum 50 50 100 100 500 >1000! 50 Bacteria Corynebacterium 100 500 1000 1000 500 100 50 poinsettiae C. fasciens 500 500 100 500 500 1000 50 Erwinia amylovora 50 50 100 50 50 50 100 E. carotovora 100 50 100 50 100 100 50 ‘stimulatory at low concentrations local necroses, indicated as leaf spots or brown necrotic tissue especially around damaged areas, and ‘Curl-tip’, which was shown by curling and darkening at the apex of the pinnae. The amount of infected tissue was estimated. In order to isolate micro-organisms from infected plant tissue, the tissue was surface sterilized with 1 ppm mercuric chloride and washed in 70% ethanol followed by sterilized distilled water. The tissue was dried and placed on suitable growth medium, PDA for isolation of fungi and nutrient agar (NA) for bacteria. Plates were incubated at 30° C. Seventeen different bacteria and eight fungi were isolated from infected areas and obtained in pure culture. Attempts were made to demonstrate the pathogenicity of these fungi, but it was difficult to demonstrate re-infection of Bracken plants in the field (cf. Hutchinson, 1976), either by inoculating the leaf surface by scraping or by injecting the stem at the junction of the petiole and the stem. RESULTS Bracken gametophyte tissue was surprisingly completely devoid of flavonoids. The major phenolic compounds present were p-coumaric, caffeic, ferulic acids, and p-hydroxybenzoic acid; vanillic, protocatechuic, and o-coumaric acids, which were present in the sporophyte, could not be detected. There were few qualitative or quantitative changes in these acids in the _—_* from one week up to ten weeks. The acids were present mainly as gluc or quinic acid esters, and electrophoresis at pH 2.2 and treatment with Sa confirmed the presence of SAN FRANCISCO & COOPER-DRIVER: ANTI-MICROBIAL ACIDS IN PTERIDIUM 9] sulfated derivatives of caffeic and p-coumaric acids (Cooper-Driver & Swain, 1975). Phenolic acids in the gametophytes were present at concentrations of 0.15—0.20%/gm dry weight. Base hydrolysis of the alcohol-insoluble fraction gave no indication that hydroxy-cinnamic acids were bound to the cell walls (Hartley & Jones, 1977), whereas p-coumaric and ferulic acids were isolated from the base hydrolysate of the methanol insoluble residue of the sporophyte. No flavonoids were detected in sporophytes growing from the gametophytes, whereas vegetatively produced sporophyte plants on emergence synthesized flavonoid glycosides. The effect of these acids on the growth of Botrytis cinerea, Rhizoctonia solani, and Pythium species, together with Corynebacterium poinsettiae, C. fasciens, and Erwinia species is shown in Table | and is expressed as minimum concentrations of acids causing 50% inhibition of fungal or bacterial growth. Overall, cinnamic acids, especially ferulic and p-coumaric acids, were the most effective inhibitors of fungal growth (the growth of Rhizoctonia solani was inhibited at concentrations as low as 50ppm). On the other hand, protocatechuic acid was stimulatory to growth of R. solani at all concentrations tested, and vanillic acid was only slightly inhibitory (18%) at concentrations as high as 1000ppm. In the case of Botrytis cinerea, the hydroxybenzoic acids together with caffeic acid had little effect on growth, and p-coumaric and ferulic only inhibited growth at concentrations of 500ppm or above, and then only by 55%. In combination, the acids were inhibitory to growth of R. solani at 100ppm (50%) and were completely inhibitory to growth at higher concentrations. For B. cinerea, 500ppm was inhibitory (74%), and no growth occurred at higher concentrations. Overall, however, R. solani was much more affected by phenolics than was B. cinerea. ae All Pythium species also showed much greater susceptibility to phenolic acids than did Botrytis. Growth of P. debaryanum was uniformly inhibited by phenolic acids at concentrations above 500ppm, and 100ppm ferulic acid and p-coumaric acid inhibited growth 13% and 50% respectively. Protocatechuic acid was the least inhibitory. The growth of P. middletonii was inhibited at all concentrations of the acids tested, whereas the growth of P. ultimum was stimulated by protocatechuic acid. In all Pythium species, no growth occurred when a combination of acids was added to the medium. The bacterial species tested responded to the phenolic acids in a more uniform manner. At 50ppm, the acids were mildly stimulatory or caused only slight inhibition of growth (<25%). As the acid concentration increased, there was a corresponding increase in inhibition. There were no differences in the inhibitory action of hydroxybenzoic acids or cinnamic acids. Combinations of the ants inhibited bacterial growth in all cases. No growth was observed at concentrations above 100ppm and, with the exception of E. amylovora, 98% inhibition was Observed at concentrations as low as 5Oppm. Seasonal changes in the concentration of ph Fig. 2. There was a slight increase during ' followed by a slight drop and a gradual increase reachin of 0.75% phenolic acids/gm dry weight of leaf material enolic acids in Bracken are shown in May, the period of leat expansion, g a maximum concentration (0.25%/gm fresh weight) in 92 AMERICAN FERN JOURNAL: VOLUME 74 (1984) 0.8 May | June | July | August |September ! FIG. 2 Seasonal changes in the concentration of phenolic acids (in % per gram of dry weight, @, and fresh weight, ©) in Pteridium aquilinum leaves. late July. There was then a drop in phenolic acid concentrations during August following a period of heavy precipitation (1.2 in rain) that caused leaching of the acids from the leaves. The concentration of eke acids in the leaf tissue dropped markedly as senescence set in during September and October. The number of fungal and bacterial species isolated from sterile leaf tissue showed a general increase as the fronds matured, with an especially sharp increase in the amount of infected tissue in early August. DISCUSSION AND CONCLUSION Phenolic acids, when added to a microbial growth medium, inhibit the growth of both fungi and bacteria. They have been shown to react with fungal enzymes and other proteins. Phenolic compounds in infected tissues are often oxidized and polymerize to form reactive oligomeric products that may have increased anti- microbial activity often associated with increased inhibition of the cell wall degrada- tion caused by extracellular enzymes produced by pathogens (Friend, 1979). This may account for the high inhibition of Rhizoctonia solani by phenolic acids, as SAN FRANCISCO & COOPER-DRIVER: ANTI-MICROBIAL ACIDS IN PTERIDIUM 93 oxidation products were seen as brown rings around the edges of the fungal mycelium. Hunter (1974) has demonstrated that the resistance of cotton plants to R. solani is due to oxidized catechin, which inhibits the polygalacturonase activity of the pathogen. Phenolic acids also act on membrane permeability and electric potentials, causing an enhanced ratio of oxygen consumption by uncoupling electron transfer (Glass & Dunlop, 1974). The variation in response of the fungi, Botrytis and Pythium to phenolic acids may be a reflection of the chemical composition of their cell walls. Chitin is known to be far more resistant to microbial decomposition than is cellulose (LeJohn, 1971). This may explain the relative resistance of Botrytis over Pythium species to the phenolic acids, as Rhizoctonia and Botrytis have cell walls of chitin, whereas those of Pythium are composed primarily of cellulose (Bartnicki-Garcia, 1968). Cell membranes of Pythium also lack sterols, which are present in both species of Deuteromycetes (Hall, 1979). Differences in cell wall composition between the two genera of bacteria (Stanier et al., 1976, p. 871) are not reflected in their response to the individual acids. The high resistance of B. cinerea to phenolic acids is interesting in light of the fact that this fungus has often been found associated with Bracken. Godfrey (1974) reported B. cinerea on the leaf surface (phylloplane); we also found this species to be present in leaf isolates. In addition, B. cinerea has been recorded on decomposing Bracken petioles (Frankland, 1976), and prothalli are also very susceptible to attack by this organism (Hutchinson, 1976). It is also interesting that the cinnamic acids, ferulic and p-coumaric, are generally the most inhibitory acids to the growth of fungi and bacteria. The cinnamic acids are of prime importance in lignin formation, and polysaccharide esters of p-coumaric and ferulic acids have been shown to be released from the cell walls of grasses by cellulase action (Hartley & Jones, 1977). It has been suggested that such acetylation of the cell wall polysaccharides by hydroxycinnamic acids may be a primitive ; i (Swain, 1979). Although gametophytes do not have bound cinnamic acids i shown by the lack of detectable acids in the hydrolysed residue, nevertheless these compounds were present in the soluble fraction and were the most effective anti-microbial compounds against the pathogenic organisms tested. Mixtures of simple phenolic acids have a synergistic effect and are better inhibitors of microbial growth when present in combination. These same acids have also been shown by Adams and Bernays (1978) to be more effective deterrents to feeding by Locusta migratoria in combination. Increased synergistic inhibitory effects of acids on radish and sorghum germination have also been shown ( Rasmus- sen & Einhellig, 1977; Einhellig & Rasmussen, 1978). From the field and laboratory studies, it was evident t Present in high enough concentrations in Bracken sporophytes 4500-7500ppm, phenolic acids/gm dry weight) and gametophytes 1500-2000ppm, phenolic acids/gm dry weight) 7 antimicrobial compounds. Damage to the tissues Dy : releases the acid vi as free ids (Woodhead & Cooper-Driver, 1979). mmnee Phenolic acids are effective inhibitors of microbial growth at concentrations of the hat phenolic acids are (0.25-0.75%, (0.15-0.20%, 94 AMERICAN FERN JOURNAL: VOLUME 74 (1984) order of 100ppm w/v, they obviously play an important anti-microbial role in Bracken. The concentration of phenolic acids present in gametophyte tissue was much lower, but was still high enough to inhibit microbial growth. Concentrations of phenolic acids in the leaves of Pteridium aquilinum changed over the season depending not only on the age of the plant, but also on abiotic and biotic factors. The sharp increase in infected tissue and the numbers of isolated microorganisms detectable in early August may be due to a preceding period of heavy precipitation followed by warm, humid weather around 40° C. The heavy rainfall would have resulted in leaching of the phenolic acids from the leaves, and thus a decrease in chemical protection. In July there was a heavy infestation of aphids on the Bracken fronds, and the presence of these insects could well have increased the points of entry for the fungal hyphae, as well as promoting greater cree of plant defense chemicals from the leaves. comparison of phenolic compounds in sporophytes and gametophytes of Polypodium aureum, Thelypteris patens, T. normalis, Asplenium aethiopicum, Cyrtomium falcatum, Gymnopteris rufa, and Pteris straminea (Cooper-Driver, unpubl.) has shown that there are two distinct patterns: (a) ferns in which the phenolic complement in gametophytes and sporophytes is identical with often a wide range of flavonoids being present in both, and (b) those in which the phenolics in the gametophyte are restricted to simple phenolic acids, as in the present case of Bracken. It is interesting to speculate whether this difference in gametophyte chemistry reflects the dependence of the species on the eae cs for reproduc- tion. As is well known, Bracken reproduces mainly vegetatively. Thus it has been shown that phenolic acids differentially enseon the growth of fern pathogens and other bacteria in vitro and are present for most of the growing season in high enough concentrations in vivo in the plant tissues to inhibit the growth of pathogenic microbes. We would like to thank Bruce Lund, Director of the Massachusetts Audubon Bird Sanctuary, Natick, MA, for his help and cooperation during the field work, Matthew Zavitovsky for his expert field advice, and Tony Swain for his evaluation of the manuscript. LITERATURE CITED ADAMS, C. M. and E. A. BERNAYS. 1978. The effects e combinations of deterrents on the feeding behaviour of Locusta migratoria. Entomol. Ex I. 23:101—109. BANERJEE, R. D. and S. P. SEN. 1980. Antibiotic =e of pteridophytes. Econ. Bot. 34:284-298. BARTNICKI-GARCIA, S. 1968. Cell wall chemistry, morphogenesis and taxonomy of fungi. Ann. Rev. Microbiol. 22:87-105. BOHM, B. A. 1968. Phenolic sep ers in ferns III. An examination of some ferns for caffeic acid derivatives. Soyer 7:1825- , and R. M. TRYON te aes ona in ferns I. A survey of some ferns for caffeic and benzoic acid peared Canad. J. Bot. 45:585-593. BRAID, K. W. 1940. The eradication of bracken. Scott. J. Agr. 33:31-36. CHOWDHURY, A., N. MUKHERJEE and N. ADITYACHAUDHURY. 1974. Sensitivity of some plant pathogenic fungi towards plant metabolites: antifungal activity of some chalcones, dihydrochalcones and flavanones. Experientia 30:1022—1023. SAN FRANCISCO & COOPER-DRIVER: ANTI-MICROBIAL ACIDS IN PTERIDIUM 95 COOPER-DRIVER, G. A. 1976. Chemotaxonomy and phytochemical ecology of bracken. Bot. J. Linn. Soc. 73:35—46. ——, and T. SWAIN. 1975. Sulphate esters of caffeyl- and p-coumaryl glucose in ferns. Phytochem. 14:2506— ———., J. J. CORN ER-ZAMODITS, and T. SWAIN. 1972. The metabolic fate of hydroxybenzoic acids in plants. Z. Naturforschung. 27B: te 946. , S. FINCH, T. SWAIN, and E. A. BERNAYS. 1977. Seasonal variation in secondary anpounds in relation to the ens of Pteridium aquilinum. Biochem. Syst. Eco 5:177-183. EINHELLIG, F. A. and J. A. RASMUSSEN. 1978. Synergistic inhibitory — of vanillic and p-hydroxybenzoic acids on radish and grain sorghum. J. Chem. Ecol. 4:425-436. FRANKLAND, J. C. 1976. Decomposition of bracken litter. Bot. J. Linn. Soc. 5 145-150. FRIEND, J. 1979. Phenolic substances and plant disease. Jn T. Swain, J. Harborne and C. F. Van mere, eds. Biochemistry of Plant Phenolics. Rec. Adv. ana 12:557-588. GLASS, a. D. M. 1976. The allelopathic potential of phenolic acids associated with the rhizosphere of Pteridium aquilinum. Canad. J. Bot. 54:2440—2444 , and B. A. BOHM. 1969. The accumulation of cinnamic and benzoic acid derivatives in Pteridium aquilinum and Athyrium filix-femina. Phytochem. 8:371-377. , and J. DUNLOP. 1974. Influence of phenolic acids on ion uptake IV. Depolarization of membrane potentials. Plant Physiol. 54:855-858. GLEISMANN, S. R., and C. H. MULLER. 1978. Allelopathic mechanisms of oe in bracken (Pteridium aquilinum) in Southern California. J. Chem. Ecol. 4:289- GODFREY, B. E. S. 1974. Phylloplane mycoflora of bracken, Pteridium ie Trans. Brit. Mycol. Soc. 62:305-311. GREGOR, M. J. F. 1938. Associations with fungi and lower plants. /n F. Verdoon, ed. Manual of Pteridology. M. Nijhoff, The H HALL, R. 1979. PS iaiswane and ser oe Bot. Rev. 47:1— HARBORNE, J. B. 1982. Introduction to Ecological Biochemistry, bi ed. Academic Press, New York. ad R. D. and E. C. JONES. 1977. Phenolic — and degradability of cell walls of s and legume species. Phytochem. 16:1531 HUNTER, “ E. 1974. Inactivation of pectic enzymes be polyphenolics in cotton seedlings of different HUTCHINSON. S. A. 1976. The effects of fungi on bracken. Bot. J. Linn. Soc. 73:145-150. —, and M. M. FAHIM. 1958. The effects of fungi on the gametophytes of Pteridium aquilinum L. Kuhn. Ann. Bot., , 22:117-1 ae JONES, C. G. 1983. iaisciburtcad variation, colonization and insect communities in the case of bracken fern, Pteridium aquilinum (L.) Kuhn. /n R. F. Denno and M. S. McClure, eds. Variable Plants and Herbivores in Natural aa Aone Systems. Academic Press, New York. ilinum and their involvement , and R. D. FIRN. 1979. Some allelochemicals of Pteridium aquil in resistance to Pieris brassicae. Biochem. Syst. Ecol. 7:187-192. s KHRISAGAR, M. K. and A. R. MEHTA. 1972. Antibacterial substances in ferns. Pl. Med. 22:386-390. KLEKOWSKI, E. J. 1969. Reproductive biology of the Pteridophyta III. A study of the Blechnaceze. Bot. J. Linn. Soc. 62:361-377. LeJOHN, H. B. 1971. Enzyme regulation, lysine r lines of evolution in fungi. Nature 231:164-168. icin LYNCH- pele WAITE, B. A., E. J. DUNCAN and C. E. SEAFORTH. 1975. A survey of ferns To antibacterial activity. Pl. Med. 27:173—200. MARUZZELLA, J. C. 1961. Antimicrobial substances from ferns. Nature 191: 518. PETERSEN, R. L. and D. E. FAIRBROTHERS. 1980 Flavonoid synthesis and antheridium initiation in Dryopteris gametophytes. Amer. Fern J. 70: 93-95. pathways and cell wall structures as indicators of 96 AMERICAN FERN JOURNAL: VOLUME 74 (1984) QUAMME, H. A., T. VAN DER ZWET and V. DIRKS. 1976. Relationship of fireblight resistance to young pear seedlings inoculated in the greenhouse to mature seedling trees naturally infected in the field. Pl. Dis. Reporter 60:660-—664. RASMUSSEN, J. A. and F. A. EINHELLIG. 1977. Synergistic inhibitory effects of p-coumaric and ferulic acids on germination and growth of grain sorghum. J. Chem. Ecol. 3:197-205. STANIER, R. Y., E. A. ADELBERG, and J. INGRAHAM. 1976. The Microbial World. Prentice Hall, New York. STEWART, R. E. 1975. Allelopathic potential of western bracken. J. Chem. Ecol. 1:161-169. SWAIN, T. 1979. Phenolics in the environment. /n T. Swain, J. B. Harborne, and C. F. Van Sumere, eds. Biochemistry of Plant Phenolics. Rec. Adv. Phytochem. 12:617—640. WHEELER, B. E. J. 1976. An Introduction to Plant Diseases. Wiley, New York. WOLLENWEBER, E. 1978. The distribution and chemical constituents of the farinose exudates in gymnogrammoid ferns. Amer. Fern J. 68:13-28. WOODHEAD, S. and G. COOPER-DRIVER. 1979. Phenolic acids and resistance to insect attack in Sorghum bicolor. Biochem. Syst. Ecol. 7:309-310. REVIEW The Phylogeny and Classification of the Ferns, A. C. Jermy, J. A. Crabbe, and B. A. Thomas, eds. Koeltz Scientific Books. 284 pp. Reprint 1984. DM150 (ca. $60.00).—This authorized reprint makes available again the results of the Sympo- sium on the Phylogeny and Classification of the Filicopsida, which was held in London in April 1972. The original volume was reviewed by C. Page (Fern Gaz. 11:47-48. 1974). The reprint is on slightly thinner and less opaque paper than is the original and there is a slight but inevitable loss in definition of the photographic illustrations, but the volume is sturdily bound and fully serviceable, and will make an excellent addition to any fern student’s library.—D.B.L. BRITISH PTERIDOLOGICAL SOCIETY Open to all who are interested in growing and studying ferns and fern-allies. Full members receive THE FERN GAZETTE and the BULLETIN OF THE BRITISH PTERIDOLOGICAL SOCIETY. Membership subscriptions are £ 7 for full members, £ 5 for ordinary members (not receiving the GAZETTE), and £5 for student members (under 25 years of age). For particulars, U. S. residents should apply to Dr. J. Skog, Biology Department, George Mason University, Fairfax, VA 22030. Non-U. S. residents should apply to Leo. P. G. Coke, Robin Hill, Stinchcombe, Dursley, Gloucestershire, England. en ne Em WANTED DEAD OR ALIVE! High quality, heart-shaped fern prothallia, live or FAA-preserved, for slide making. We need monoecious and dioecious prothallia as well as stages showing sporophyte develop- ment. Please send samples for viewing. We pay for high quality specimens only! ed microscope We ly high quality prepar é also supply high quailty best for their slides to those who need the very teaching or research. TRIARCH, INC. - P.O. Box 98 - RIPON, WI 54971 (414)-748-5125 AMERICAN — FERN 2 ee JOURNAL QUARTERLY JOURNAL OF THE AMERICAN FERN SOCIETY The Identification of Hawaiian Tree Ferns of the Genus Cibotium RICH BECKER 97 Two New Tree Ferns from Panama _ ROBERT G. STOLZE 101 Trunk Length and Frond Size in a Population of Nephelea tryoniana from El Salvador RALPH L. SEILER 105 An Unusual New Elaphoglossum from Peru ROLLA TRYON = 108 New Tropical American Ferns JOHN T. MICKEL 111 Shorter Note: An Urban Locality for Asplenium platyneuron 1985 A.I.B.S. Meeting—Call for Papers American Fern Journal Index to Volume 74 Errata MISSOURI BOTANICAL The American Fern Society Council for 1984 TERRY R. WEBSTER, Biological Sciences Group, University of Connecticut, Storrs, CT 06268. President FLORENCE S. WAGNER, Dept. of Botany, University of Michigan, Ann Arbor, MI 48109. Vice-President MICHAEL I. COUSENS, Faculty of Biology, University of West Florida, Pensacola, FL 32504, Secretary JAMES D. CAPONETTI, Dept. of Botany, University of Tennessee, Knoxville, TN 37916. Treasurer DAVID S. BARRINGTON, Department of Botany, University of Vermont, Burlington, VT 05401. ‘ecords Treasurer DAVID B. LELLINGER, Smithsonian Institution, Washington, DC 20560. Journal Editor ALAN R. SMITH, Dept. of Botany, University of California, Berkeley, CA 94720. Memoir Editor DENNIS Wm. STEVENSON, Barnard College, Columbia University, New York, NY 10027. Fiddlehead Forum Editor American Fern Journal EDITOR DAVID B. 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Subscriptions $9.00 gross, $8.50 net if paid through an agency (agency fee $0.50); sent free to members of the American Fern Society (annual dues, $8.00 + $4.00 mailing surcharge beyond U.S.A., Canada, and Mexico; life membership, $160.00). Back volumes 1910-1978 $5.00 to $6.25 each; single back numbers of 64 pages or less, $1.25; 65-80 pages, $2.00 each; over 80 pages, $2.50 each, plus shipping. Back volumes 1979 et seq. $8.00 each; single back numbers $2.00 each, plus shipping. Ten percent discount on orders of six volumes or more. Fiddlehead Forum Prof. Dennis Wm. Stevenson, Dept. of Biological Science, Barnard College, Columbia University, New York, NY 10027, is editor of the newsletter “Fiddlehead Forum.” The editor welcomes contribu- tions from members and non-members, including miscellaneous notes, offers to exchange or purchase materials, personalia, horticultural notes. and reviews of non-technical books on ferns. Spore Exchange Mr. Neill D. Hall, 1230 Northeast 88th Street, Seattle, WA 981 15. is Director. Spores exchanged and lists of available Spores sent on request. Gifts and Bequests Gifts and bequests to the Society enable it to expand its services to members and to others interested in ferns. Botanical books, back issues of the Journal, and cash or other gifts are always welcomed, and are tax-deductible. Inquiries should be addressed to the Secretary. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 4 (1984) 97 The Identification of Hawaiian Tree Ferns of the Genus Cibotium RICH BECKER™ The identification of Hawaiian tree ferns is difficult for most laymen and taxonomists because of past nomenclatural confusion, misapplication of the specific names, and the absence of a complete key to all of the species. Although Sadleria, a Hawaiian endemic genus of the Blechnaceae, has some arborescent species which are referred to as tree ferns, these are not treated in this paper. The correct nomenclature of the six Cibotium species found in Hawaii is presented. A simple key is given for the species. This is supplemented with descriptions of their characteris- tics as seen in their native habitats. I have deposited in the herbarium of the University of Hawaii my reference collection of entire fronds for each of the Hawaiian species described. The range of Cibotium extends from Assam through China, Malaya, Indonesia, the Philippine Islands, and Hawaii to southern Mexico and Guatemala. There are three Asiatic species (Holttum, 1963, pp. 164-166).: C. barometz (L.) J. Smith, C. cumingii Kunze, and C. arachnoideum (C. Chr.) Holttum. The Central American species are: C. schiedei Schlecht. & Cham. (Mexico), C. regale Versch. & Lem. (Mexico to Honduras), and C. wendlandii Mett. ex Kuhn (Mexico and Guatemala), according to Smith (1981, pp. 75-76) and Stolze (1976, pp. 94-97). The six endemic Hawaiian Cibotium species are: C. glaucum (J.E. Smith) Hook. & Arn., C. chamissoi Kaulf., C. splendens (Gaud.) Krajina ex Skottsb., C. nealiae Degener in Degener & Hatheway, C. hawaiense Nakai & Ogura in Ogura, and C. st.- johnii Krajina. Cibotium glaucum and C. chamissoi are found in the rain forest areas of Kauai, Oahu, Molokai, Lanai, Maui, and Hawaii. Cibotium splendens is common on Oahu; forms of it may also be on Molokai and Maui, but this has not been confirmed. Cibotium nealiae and C. st.-johnii are apparently restricted to Kauai. Cibotium hawaiense is endemic to the island of Hawaii; it is locally abundant on that island. I have seen it close to Wright Road in the Olaa Forest, at Puu Makaala near the Stainback Highway, at the end of the Olaa Back Road, above Laupahoehoe, and in the Kohala Mountains. The most common species on the island of Hawaii is C. glaucum. Cibotium chamissoi is the next most abundant. - The key to resolving the nomenclatural confusion associated with the Hawaiian tree ferns lies in understanding Hooker's treatment of the genus in the ny Filicum (1844). He separated C. chamissoi Kaulf. and C. glaucum (J.E. Smith) Hook. & Arn. and described a new species, c. menziesil Hook. Krajina in Skottsberg (1942, pp. 30-41) showed the latter to be identical to C. . Kaulf. and the name C. chamissoi sensu Hooker to be C. splendens (Gaud.) Krajina ex Skottsb. *P.O. Box 1358, Station A, Vancouver, B.C. V6CT2T, Canada. Volume 74, number 3, of the JOURNAL, was issued December 6, 1984. 98 AMERICAN FERN JOURNAL: VOLUME 74 (1984) Hillebrand may have used the name C. chamissoi Kaulf. in its original sense on some of his first collection labels (Skottsberg, 1942, p. 40). However, in the Flora of the Hawaiian Islands, Hillebrand (1888, pp. 545-548) used the incorrect nomencla- ture from Hooker’s Species Filicum, as did Rock (1913), in his The Indigenous Trees of the Hawaiian Islands. Both are standard works on the Hawaiian flora that were for a long time assumed to be correct, causing many later authors to unwittingly use an incorrect nomenclature. The credit for uncovering the synonymy belongs to Krajina, who has carefully studied the genus. The key given below is based on relatively constant characteristics that are easily observable in the field on mature plants. Other characteristics, such as trunk height, frond length, pinnule width, and number of sori per pinnule, vary greatly with the age and vigor of the plants. They are not reliable diagnostic features and have not been used in the key. KEY TO THE HAWAIIAN SPECIES OF CIBOTIUM 1. Stipes with reddish-brown or stiff purplish-black hairs along most of their length. 2. Fronds distinctly glaucous beneath; costae villosulous 2. Fronds not glaucous beneath; costae glabrous 2. C. chamissoi 1. Stipes naked, or with dull brown or golden brown hairs mainly at their base. 2. One or both of the basal pinnule segments auriculate, fronds glabrous and distinctly glaucous. 3. C. glaucum 1. C. st.-johnii 2. None of the pinnule segments auriculate; fronds not or only slightly glaucous beneath. 3. Stipes very dark brown to blackish with a conspicuous, white line along each side when fresh; pin- nule segments covered with tufts of hairs (as seen under 10 X lens) beneath ...... 4. C. nealiae 3. Stipes brown or greenish when fresh; pinnule segments covered with cobweb-like hairs (as seen under 10 x lens) beneath. 4. Trunk slender and smooth, stipes covered with inconspicuous, soft, matted, dull, brown hairs. 5. C. hawaiense 4. Trunks rough with stubs of dead fronds; stipe bases with golden brown hairs 6. C. splendens 1. Cibotium st.-johnii Krajina, Stud. Bot. Cechosl. 1(2):94. 1938. The most characteristic feature of C. st.-johnii is the copious amount of matted hairs on the full length of the stipes, and on the costae and costules. The hairs at the base of the stipes are, when fresh, reddish-brown. Further up the stipes and on the costae they are grayish-brown to whitish. Cibotium st.-johnii closely resembles C. glaucum, but has much smaller fronds. In 1966, Dr. Krajina annotated Heller 2818 (BISH 01703, 01703a 01703b) as “Cibotium glaucum (Sm.) H. & A. subsp. St.-Johnii (Krajina) c.n. var. fallax Krajina.” The description of Hillebrand’s (1888, Pp. 547). C. chamissoi var. B can be used to confirm the identity of C. st.-johnii. Distribution: Alakai swamp, Kauai. 2. Cibotium chamissoi Kaulf. Berl. Jahrb. Pharm. 21:53. 1820. C. menziesii Hook. Sp. Fil. 1:84, 1. 29C. 1844. This is the most robust species. The stipe bases are covered with long, reddish- brown hairs; further up the stipe, the hairs are still and blackish. The rachis and the costae are often straw-colored. The fronds are not at all glaucous beneath and usually are glabrous, but sometimes have minute, scattered tufts of hairs visible under 40x magnification. Distribution: Kauai, Oahu, Molokai, Lanai, Maui, an Hawaii. Hawaiian name: hapu’u’i’i. R. BECKER: IDENTIFICATION OF HAWAIIAN CIBOTIUM 99 On the islands of Molokai, Lanai, and Hawaii, I have seen several tree ferns that may be hybrids between C. glaucum and C. chamissoi. Their fronds were distinctly glaucous beneath and their basal pinnule segments auriculate. These are characteris- tics of C. glaucum; however, their stipe hairs were bristly and either dark purplish- brown or dark orangy-brown. 3. Cibotium glaucum (J. E. Smith) Hook & Arn. Bot. Beech. Voy. 3:108. 1832. Dicksonia glauca J.E. Smith, Rees’ Cyclop. 11(22). 1808. Cibotium glaucum is the most abundant species on the Island of Hawaii. The stipe bases are covered with a mass of glossy, golden-yellow hairs and are lustrous when dry. The distal part of the stipes and the rachis are essentially naked. The fronds are glabrous and bright glaucous beneath (slightly less so in the wetter parts of the range of the species). The pinnule segments closest to the costae are auriculate and overlap the costae. The auricles are largest on plants from the island of Hawaii; they are present but much smaller on plant forms from the other islands. One plant I examined on Molokai had one frond exauriculate. In this species, the stipe bases remain attached to the trunks for some time after dead fronds drop off. Distribution: Kauai, Oahu, Molokai, Lanai, Maui, and Hawaii. Hawaiian name: hapu’u pulu. 4. Cibotium nealiae Degener in Degener & Hatheway, Fl. Haw. 5: Family 12. 1951. The stipe bases of this species are very dark brown, almost black, and are covered with a thick mass of matted, yellowish hairs. Distally, the stipes are greener and naked. A conspicuous, long, white line (pneumatothode) extends along each side of the fresh stipes. I found the pneumatothodes to be very obvious when I saw the species in the field. Although the lines are a useful field identification, they did not remain visible on my herbarium specimens. The costules and veins are yellow and prominent. The frond underside is green, but not glaucous beneath. The costules are covered with very small tufts of hairs beneath. Distribution: Hanalei, Kauai. 5. Cibotium hawaiense Nakai & Ogura in Ogura, Bot. Mag. Tokyo 44:468. 1930 e The spelling of the epithet hawaiense follows Nakai and Ogura’s bala publication. The stipes of C. hawaiense may appear naked, but actually are aes with thick, flaky mats of dull brown hairs. When a stipe is pulled back eae the trunk, the basal part is seen to be covered with golden, glossy, and slightly sae hairs. The laminae are thin and bright green, the underside is slightly paler an somewhat glaucescent. Fine, cobwebby hairs can be seen, with a 10 x a see the undersides of the pinnules. The trunk is slender, and the fronds, alter arc ri upwards, are held almost horizontally. The dead fronds hang down close to the trunk, similar to those of the Date Palm. Eventually they break away cleanly, leaving a smooth trunk. Distribution: Olaa, Laupahoehoe, Kohala Mountains, Hawaii. Hawaiian name: meu. 6. Cibotium splendens (Gaud.) Krajina ex Skottsb. Acta Horti Gothob. 15:40. 1942. Uranie 96, 369, f. 21. 1827. Pinonia splendens Gaud. in Freyc. Voy. 1:83. 1844. Cibotium chamissoi sensu Hook. Sp. Fil. 100 AMERICAN FERN JOURNAL: VOLUME 74 (1984) This is the most delicate appearing species. It has been confused with C. hawaiense, although their ranges do not overlap. A glaucescent form from Molokai (Fosberg 9600, BISH 01741) was annotated with the unpublished name C. splendens var. glaucescens by Krajina. This specimen has pinnule segments that are somewhat glaucous and with arachnoid hairs on their undersides. Distribution: Oahu, and perhaps also on Molokai, and Maui, but not seen by me on these islands, or on Lanai or Hawaii. I wish to thank Sandra Becker for her assistance in mounting my reference collection of herbarium specimens. The review of the preliminary draft and discus- sion on the nomenclature of C. nealiae by Dr. V. J. Krajina were most helpful. The continued encouragement and support of Drs. D. Mueller-Dombois, K. W. Bridges, and C. Lamoureux, are much appreciated. LITERATURE CITED CHRISTENSEN, C. 1905-1906. Index Filicum. H. Hagerup, Copenhagen. HILLEBRAND, W. F. 1888. Flora of the Hawaiian Islands. Reprinted 1965, Hafner, New York. HOLTTUM, R. E. 1963. Cyatheaceae. Flora eee. I, 1:66-176. HOOKER, W. J. 1844. Species Filicum, vol. 1. ndon soe ce % F. 1913. The Indigenous Trees of ee Islands. Published under patronage, Honolulu, Haw: SKOTTSBERG, C. 1942. Vascular plants from the Hawaiian Islands. Part 3. Pteridophytes collected during the Hawaiian bog survey 1938. Acta Horti Gothob. 15:35-14 SMITH, A. R. 1981. Flora of Chiapas, part 2. Pteridophytes. Calif. Ac ad. Sci., San Francisco. STOLZE, R. G. 1976. Ferns and Fern Allies of Guatemala, part I. Fieldiana, Bot. 39: 1-130. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 4 (1984) 101 Two New Tree Ferns from Panama ROBERT G. STOLZE* Recently I was sent two specimens of tree ferns to identify, which were thought to be Cnemidaria: one by Henk van der Werff at the Missouri Botanical Garden, and another by David Lellinger at the U.S. National Herbarium, which was a duplicate he had received from Missouri. One proved to be new species of Cnemidaria, the other a new, simply pinnate Trichipteris. 1 am always hesitant to describe new species based only upon single collections, as is the case with these, but each is highly distinctive from others in their respective genera. It is curious that both of these new Panamanian species collected between 1975 and 1982 are very small (for tree ferns) and are simply pinnate. Cnemidaria glandulosa Stolze, sp. nov. Fig. 1. Folium pinnatum, tenuiter herbaceum, ca. 130 cm longum, | 25 cm latum, segmento apicali non conformi; petiolus haud spinosus, petioli paleis abundantibus, bicoloribus: lamina in pagina abaxiali dense glandulosa; glandes minutae, bacilliformes; rhachis plus minusve nuda; pinnae subse ine integro et revoluto; venae 2- vel 3-furcatae, liberae, vel raro venae basales anastomosantes et areolas costales efformantes, indusium atrobrunneum, plus minusve semicirculare; receptaculum ovideum, sine paraphysibus. VTYPE: North of San Felix at Chiriqui-Bocas del Toro border, on Cerro Colorado copper mine road along continental divide. Lower montane rain forest, 5,000—5,500 ft, Province of Chiriqui, Panama, Mori & Kallunki 5908 (US; isotype MO). Thus far known only from the type collection. Caudex not seen; leaf pinnate, thin-herbaceous, to 130 cm long and 25 cm broad, abruptly terminating in a nonconform apical section (7.e. broader at base than the i ntially naked, or with a few scattered scales or i ; pinnae about 12 bas we only abaxially, the glands dark brown, minute, appressed, bacilliform; costae naked adaxially, sparsely provided with broad, flaccid, amorphous, whitis abaxially; veins 2—3-forked, essentially = hg spaced He ge ee Be thon aie ae oe ay pe pitts indusia dark brown, more Or less id. lacking paraphyses, spores pale yellow to srl white, globose-tetrahedral, trilete, with one large aperture near the center of each face near the equator The he: species bears closest resemblance to C. decurrens of peas om northern Central America, especially in that some forms of the latter have cou entire pinnae. However, in C. decurrens the lamina is eglandular and coe - sen reduced to the apex, veins commonly (not rarely) merge to form costa pe eae the indusia are pale or yellowish (not dark) brown. The occurrence 0 *Department of Botany, Field Museum of Natural History, Chicago, IL 60605. 102 AMERICAN FERN JOURNAL: VOLUME 74 (1984) FIG. 1. Holotype of Cnemidaria glandulosa, Mori & Kallunki 5908 (US). glands on the abaxial surface of pinnae is unique in the genus. In Cc glandulosa the glands may be observed even at low magnification, and by using higher power (25 x) one can easily discern their rod-like shape. R. G. STOLZE: TREE FERNS FROM PANAMA FIG. 2. Holotype of Trichipteris oi S3 pinnata, Knapp 5881 (MO). Spanitade pinnata Stolze, sp. nov. m pinnatum, tenuite incor anicnll conformi, r vel firme herbaceum, la rca cecal es x :e i a sia eee: ere | 58822 > bhi PANAMA Fa ae 180 N terrestrial. Sori browne MISSOURI BOTANICAL GARDEN HERBARIUM ( Fig. 2. circa 50 cm longum, 20 cm | segmentum apicale ad basim articulatum; petiolus haud 103 104 AMERICAN FERN JOURNAL: VOLUME 74 (1984) spinosus vel interdum sparse muricatus, squamis ovatis vel lanceolatis, nitidis, brunneis vel interdum bicoloribus; pinnae nonnullae, breviter petiolulatae, ad rhachim articulatae, /TYPE: Santa Rita Ridge road, 21-26 km from Transisthmian Highway. Tropical wet forest, 500-550 m, Province of Colén, Panama, Knapp 588] (MO; isotype F). Thus far known only from the type collection. Caudex erect, ca. 6 cm long, | cm broad; leaf pinnate, thin- to firm-herbaceous, to 50 cm long and 20 cm broad, abruptly terminating in a conform apical segment which is articulate at the base; petiole 18-22 cm long, yellowish or grayish brown, lacking spines, but sometimes slightly muricate, amply scaly at least near the base, the scales 5-7 cm long, ovate to lanceolate, lustrous brown or some of them bicolorous (with narrow, short-stalked, articulate to the rachis, the margins crenate, the apex acuminate, the base subtruncate to broadly rounded, the tissue between the veins rather densely gland-dotted on the abaxial side; costae glabrous abaxially, densely provided with appressed tri- chomes as on the rachis adaxially; veins 4—6 pairs along the costule, only the distal ones reaching the pinna margin; sori borne on the proximal 1/3 or 1/4 of each vein branch; indusia lacking; receptacle globose, with short paraphyses among the sporangia; spores pale yellow to whitish, trilete, rather sharply tetrahedral. Compared to most tree ferns, this is a very diminutive one—the fronds are only half a meter long and the caudex is small, not trunk-like. Of course, it must be Pp 1 that more collections are made of this species, much larger plants also may be found. Trichipteris pinnata is most likely to be confused with T. williamsii Maxon of Panama, Colombia, and Venezuela, one of two species in the genus with entire or subentire pinnae (Trichipteris typically has huge leaves, 2—3-pinnate or more). Both species also have the lamina abruptly terminating in a conform apical segment which is articulate to the rachis, and the lateral pinnae are nodose-articulate as well. In 7. williamsii, however, the pinnae are subcoriaceous, with their margins entire, or at best slightly undulate, and the basal veins often or occasionally merge to form costal areoles. Pinnae of T. pinnata are thin- to firm-herbaceous, with their margins rather deeply crenate and veins are all free. Leaf tissue in T. williamsii is eglandular, whereas the abaxial surface of the pinnae in T. pinnata is copiously dotted with minute, castaneous glands. There is some disagreement about the valid spelling of the name Trichipteris, some authors preferring “Trichopteris.” Tryon (1970, p. 41) adhered to Presl’s original spelling, as did Barrington (1978, p. 19) in his revision of the genus. Holttum and Edwards (1983, p. 161) and others prefer to adopt Schott’s alteration to “Trichopteris” for orthographic reasons. I do not intend here to belabor either argument, as the rules according to the I. C. B. N. are rather ambiguous and/or flexible in cases of this kind. I prefer to follow Tryon, who separated the many species of Trichipteris from Alsophila, and Barrington, who did the revision. LITERATURE CITED TRYON, R. M., 1970. The classification of the Cyatheaceae. Contr. Gray Herb. 200:1-53. BARRINGTON, D. S. 1978. A revision of the genus Trichipteris, Contr. Gray Herb. 208:1-93. HOLTTUM, R. E., and P. J. EDWARDS. 1983. The tree-ferns of Mt. Roraima and neighbouring areas of the Guayana Highlands with comments on the family Cyatheaceae. Kew Bull. 38(2):155-188. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 4 (1984) 105 Trunk Length and Frond Size in a Population of Nephelea tryoniana from El Salvador Ralph L. Seiler* Field observations of tree fern fronds suggested that small tree ferns have longer stipes relative to lamina length than tall tree ferns of the same species. This study was done to determine how total frond length, lamina length and stipe length vary within a population. In April 1979, measurements were made of 146 fronds on 45 individuals of Nephelea tryoniana Gastony from a dense cloud forest at 2250 m in Bosque Montecristo, a wildlife preserve in northwest El Salvador. The forest is described more completely in Seiler (1981). The trunk length of each tree was measured, as well as the stipe and lamina length of all undamaged fronds. Usually three fronds per tree were measured, but occasionally four or five or as few as two per tree were measured. The majority of the fronds were sterile, but the presence of fertile fronds should have no effect on the results, for fertile and sterile leaves are practically monomorphic in Nephelea (Gastony, 1973). RESULTS The length of the fronds of a tree is fairly constant. The standard deviation within a tree averaged 4.55% (s=2.96, n=45). Analysis of 139 fronds on 43 trees indicates frond length is weakly related to trunk length (r=0.39, P<0.01). Two of the 45 trees were not included in the statistical analysis for reasons that will be discussed later. In Figure 1, frond length is plotted against trunk length. Each point represents the mean frond length from one tree. The ratio of lamina length to stipe length within a tree is also fairly constant, the standard deviation within a tree averaged 5.72% (s=3.01, n=45). Analysis of 139 fronds on 43 trees indicates that shorter tree ferns do have longer stipes relative to lamina length and that the lamina/stipe ratio is strongly correlated to trunk length (r=0.90, P<0.01). In Figure 2 the ratio of the lamina length divided by the stipe length is plotted against the trunk length; each point represents the mean lamina/stipe ratio for one tree. Two trees, numbers 17 and 22 (Figs. 1 and 2), were found that had been knocked over and, although the main trunks were covered with humus and litter, the — had subsequently turned and grown upright. Usually the trunk length is more or = equal to the height of the crown above the ground, but in both of 8 a = actual height of the crown was more than a meter less than the length 0 e wea ; The data gathered from these two trees were not included in the statistical ana yses because these two trees were different from the rest in that the effective height en much less than the trunk length and being forced to change the direction of growt may have had an unknown effect on the apical meristem. cacaanel The effective height of the crown is better predicted by the measured a ies ratio than is the total trunk length on the two fallen trees. In tree 22 the lamina/stipe *3977 S. 775 W., Bountiful, UT 84010. 106 AMERICAN FERN JOURNAL: VOLUME 74 (1984) Mean Frond Length, in meters Mean Lamind/Stipe Ratio ' T r T “Tr Gi T 2 3 Trunk Length, in meters ms. I, Relationship between mean frond length and trunk length in Nephelea tryoniana. See text for explanation of numbered points. FIG. 2. Relationship between lamina length divided by stipe length (lamina/stipe) and trunk length in Nephelea tryoniana. See text for explanation of numbered points. RL. SEILER: TRUNK LENGTH AND FROND SIZE IN NEPHELEA 107 ratio is 1.23, the trunk length is 160 cm, and the effective height is 16 cm. The predicted trunk length is 10 cm for this tree. Similarly, tree 17 has a lamina/stipe ratio of 1.42, a trunk length of 137 cm, and an effective height of 27 cm. The predicted trunk length is 41 cm. DISCUSSION More measurements of trees taller than two meters would have been desirable, but would have required cutting the trees down. Since all tree ferns are very rare in El Salvador and are protected in Bosque Montecristo, only the 5 m tall tree was cut down. The sampling bias towards small trees could cause. undue weight to be put on the values for the two tallest trees, suggesting a correlation between frond and trunk length that may not really exist. Omission of the data from the two tallest trees reduces the correlation coefficient; however, the correlation is still significant =0.22, P<0.05). The lamina/stipe ratio is strongly correlated with the trunk length. The effective height of the crown seems to be the determining factor for the lamina/stipe ratio. The relative length of the stipe in the two trees that had been knocked over is much better predicted by the effective height of the crown, rather than by the total length of the trunk. Although leaf angle was not measured, it was observed that the fronds of the smaller tree ferns are held more erect than those of larger ferns. Functionally, it may be that the more erect leaf angle and the relatively longer stipes on small tree ferns combine to lift the lamina further toward the light. For mechanical reasons, the stipe may have to be shorter to support the more massive, more horizontal lamina of taller tree ferns. Further investigation is needed to determine what combination of environmental and internal developmental factors determine the relative stipe length and its func- tional significance. The developmental age of the apical meristem of the fern may be important in determining the lamina/stipe ratio, while the most probable environ- mental factors would be light intensity and quality. LITERATURE CITED GASTONY, G.. 1973. A revision of the fern genus Nephelea. Contr. Gray Herb. 203:81-148. SEILER, RL. 1981. Leaf turnover rates and natural history of the central American tree fern Alsophila salvinii. Amer. Fern J. 71:75-81. 108 AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 4 (1984) An Unusual New Elaphoglossum from Peru ROLLA TRYON* A distinctive species of Elaphoglossum has been recognized among specimens received for identification from the Missouri Botanical Garden.This new species has a combination of characters that adds to the diversity of its large genus. It also emphasizes the need for more intensive collecting in the species-rich eastern Andes of Peru. Elaphoglossum pascoense R. Tryon, sp. nov. Figs. 1, 2. Caulis terrestris dorsiventralis valde repens, petioli adaxialiter sulcati fasciculis vascularis sex, phyllopodiis nullis, lamina sterilis oblongo-ovata ca. 12—24 cm longa .5—12 cm lata profunde cordata subcuspidata venis liberis longis, lamina fertilis oblonga ca. 12 cm longa 2 cm lata, sporae reticulato-echinatae. Say ¢ = 4 FIG. 1. Spore of Elaphoglossum pascoense, X 5555. = res vith dark, appressed scales, these few except at the apex, stele dorsiventral; eaves 28-54 cm long; petioles gradually merging into (long-decurrent on) the stem spaced, as long as to usually longer than the lamina, more or less covered by scales; fertile lamina densely covered beneath by sporangia, these intermixed with es *Harvard University Herbaria, Harvard University, Cambridge. MA 02138. R. TRYON: A NEW ELAPHOGLOSSUM FROM PERU 109 x 0.56. FIG. 2. Holotype of Elaphoglossum pascoense, with fertile leaf in center, 110 AMERICAN FERN JOURNAL: VOLUME 74 (1984) TYPE: Road between Oxapampa and Villa Rica, Dept. Pasco, Peru, 75°20”W, 10°73”S, on flatter parts of steep ridge, 11 October 1982, Robin B. Foster 9127 (GH; isotypes MO, others to be distributed). The most unusual features of Elaphoglossum pascoense are the relatively broad, deeply cordate sterile lamina with wholly free venation. The sterile lamina can only be compared to that of E. crinitum, which has fully areolate venation. This new species agrees with Elaphoglossum sect. Setosa (Mickel & Atehortua, 1980) in the lack of phyllopodia, the long-creeping stem, the presence of hyda- thodes, and the reticulate-echinate spores. It has the long-creeping stem and dark stem scales of subsect. Alpestria. However, it is quite different from all of the enumerated species of that subsection, which have the sterile lamina rather narrow and more or less cuneate at the base. I am indebted to Dr. Alice F. Tryon for SEM preparations of the spores. This research was partially supported by National Science Foundation Grant DEB 81-05726 to Rolla Tryon and Alice Tryon. LITERATURE CITED MICKEL, J. T. and L. ATEHORTUA, G. 1980. Subdivision of the genus Elaphoglossum. Amer. Fern . 10:47-68. AMERICAN FERN JOURNAL: VOLUME 74 NUMBER 4 (1984) 111 New Tropical American Ferns JOHN T. MICKEL* In the course of identification work for various Latin American field programs at the New York Botanical Garden, several new species of ferns have come to light. Three of the new taxa are in Anemia, a genus of which there is yet much to be learned. trols Anemia elaphoglossoides Mickel, sp. nov. Fig. 3B Frondes dimorphae, lamina sterilis indivisa integra glabra coriacea, lamina fertilis bi- ad tripinnata. Rhizome horizontal, compact, ca. 4 mm diam., clothed with red-orange hairs, 5-8 mm long; fronds clustered, 10-20 cm long, dimorphic, the stipe /2-¥s of the frond length, stramineous, glabrous, slender, 0.5 mm diam., 7-13 cm long; sterile blade simple, elliptic, glabrous, coriaceous, 3.5—4.5 cm long, 1.0-1.2 cm wide, broadly cuneate at the base, the apex obtuse; veins free, dichotomously forking, leaving the midvein at an acute angle; margin thickened; fertile frond about equalling the longest sterile frond in length, bi- to tripinnate; spores unknown. UTYPE: Brazil: Est. Goids: ca. 15 km S of Niquelandia; gravelly hillside and sandy cerrado, rooted at base of overhanging rock; elev. 1000 m; 21 Jan 1972, H. S. Irwin et al. 34664 (UB; fragment NY). Known only from the holotype, two plants, one sterile and one fertile. This is the only known species of Anemia with totally undivided sterile blades. Its relationship within the genus probably in subg. Anemia, but the precise relationship is not known. The spores are too young to see the ornamentation; the stomates are floating. 10 Anemia marginata Mickel, sp. nov. Fig. ~ Frondes dimorphae, lamina sterilis ternata pinnata, segmenta lanceolata coriacea glabra viridi-grisea ad marginem incrassata flava, frondes fertiles bi- ad tripinnatae quam steriles multo longiores. ; Rhizome horizontal, compact, short-creeping, Ca. 3 mm diam., hairs on colored near the apex, maturing to orange-brown; fronds dimorphic, sterile see erect, 3.5-7 cm tall; stipe /2—%4 of the frond length, stramineous, glabrous; - e ca. 1 cm broad, ternate; lateral pinnae small, ca. 7 X 3 mm, entire, ssi ation ps ascending, with an acute apex, the base broadly attached; terminal sere arger, ¢ “a 14x4 mm, lanceolate, entire to slightly , Oe atc glabrous, Veins free; ish- with distinct thickened yellow margin, ©. | a a 53, 15 cm tall, much surpassing the sterile fronds in length, 2-3 pinnate. TYPE: Brazil: Est. Goids: Alto Paraiso, Chapada dos Veadeiros, 5 km E of od Paraiso, elev. 1600 m; locally common in patches of humus and sand os ees as spots near boulders in high sandy flat exposed place; 15 Feb pr, eres Estabrook 206 (UB; isotype NY). *New York Botanical Garden, Bronx, NY 10458. 112 AMERICAN FERN JOURNAL: VOLUME 74 (1984) FIG. 1. SEM photomicrographs of _— spores. A, B. Proximal and distal faces of A. salvadorensis ale 947), 510. C. Proximal face of A. sanctae-martae (Steyermark & Rabe 96462), x510. Spr face of A. hirsuta (Mickel 62 41), X510. E. Distal face of A. oblongifolia (Irwin 34329), F. Proximal face of A. clinata (Williams 2584), x 510 J. T. MICKEL: NEW TROPICAL AMERICAN FERNS 113 ois Tt Anemia salvadorensis Mickel & Seiler,' sp. nov. Fig. 2C. Rhizomata adscendentia; frondes usque 17 cm longae, usque 3.5 cm latae; stipes quam lamina sterilis quintuplo brevior;, lamina sterilis anguste obovato-oblonga vel oblonga pinnata, sparse pilosa; pinnae steriles 3-9 jugae oblique oblongae obtusae dimidiatae margine denticulatae; nervi liberi; frondium fertilium stipites laminas subaequantes; pinnae fertiles patentes vel oblique adscendentes, a pinnis sterilibus remotae. Rhizome ascending, clothed with reddish-brown hairs; fronds to 17 cm long, 3.5 cm wide; stipe of sterile frond ca. 1/6 of the frond length, deeply grooved adaxially and laterally, stramineous, wiry, moderately hirsute with reddish, tortuous hairs, glabrate; blade narrowly obovate-oblong or oblong, terminating in a flabelliform or obcordate apical segment, pinnate, thin, dull, sparsely pilose on both surfaces; sterile pinnae 3-9 pairs, subsessile, slightly ascending, to 1.9 cm long, .8 cm broad, obliquely oblong, obtuse, strongly inaequilateral at the base (dimidiate), truncate above and cuneate below, erose denticulate, the margin slightly cartilaginous, a distinct midrib lacking; veins free, evident, slightly raised on the adaxial surface; stomates floating; fertile fronds similar to the sterile but longer-stiped, stipes up to half the total frond length, the fertile pinnae 0.7—1.7 cm below the sterile blade; fertile pinnae to 3.2 cm long, spreading to slightly ascending, short-petiolulate, tripinnate, the ultimate divisions slender and virtually lacking lamina; spores tetrahedral-globose, striate, the ridges slender, few, with noticeable space between 6-7 «um, the space being about the same width as the ridges), 76-94 xm diam. TYPE: El Salvador: Depto. Ahuachapan: Montana de Agua Prieta, near San Benito, among grass on steep, disturbed, sunny slope, elev. 700 m, 20 Feb 1979, Seiler 947 (NY; isotypes F, MHNES). Known only from the type collection. This species is distinct in the habit of the fertile pinnae and pinnate blade. In the habit of the fertile pinnae, the ascending rhizome, and floating stomates it resembles members of subg. Coptophyllum sect. Adetostoma (A. brandegeea, A. intermedia, and A. clinata), which are found in western Mexico (the first two) and Panama to Bolivia (A. clinata), according to Mickel (1962, 1966). However, the pinna form is more like that of A. sanctae-martae of Colombia, and in form of the pinnae except for their undivided nature like that of the more ubiquitous A. hirsuta, which 1s closely related to A. sanctae-martae (subg. Anemia sect. Hirsutae). The very narrow form of the blade and the very short stipe suggest a relationship to A. oblongifolia (subg. Anemia sect. Oblongifoliae). The spores of A. salvadorensis most closely resemble those of the A. hirsuta complex (incl. A. sanctae-martae) (Figs. 1A-E). t clear, it is our conclusion that the the evolution of the genus, lying The section Adetostoma was placed in subg. its spores with many closely set ridges, as In “. ; The floating stomates were thought to be an anomaly or ete pone ane subgenus. On the other hand, they are known in some species of SUD. = a and in all species of subg. Anemia. On the basis of the ascending rhizome, (Mr. Seiler served for two years in the Peace Corps in Fl Salvador. ye a uae much of the country collecting plants and preparing specimens ee Historia Natural de El Salvador (MHNES). 114 AMERICAN FERN JOURNAL: VOLUME 74 (1984) . 2. A. Habit of Saccoloma membranaceum. B. Pinna of S. membranaceum. C. Habit of Anemia salvadorensis. D. Habit of Anemia marginata. J. T. MICKEL: NEW TROPICAL AMERICAN FERNS 115 floating stomates, and the distinctive, obliquely held fertile pinnae, we would place the new species in sect. Adetostoma, but with the similarities to sect. Hirsutae and sect. Oblongifoliae of subg. Anemia, we would also remove sect. Adetostoma from subg. Coptophyllum and place it in subg. Anemia. Patent fertile pinnae are known from several groups within the genus Anemia: A. aspera and A. perrieriana (subg. Coptophyllum sect. Anemiaebotrys), A. elegans and A. eximia (subg. Coptophyllum sect. Trochopteris), A. colimensis (subg. Anemiorrhiza) and all of sect. Adetostoma (subg. Anemia). These four groups are probably not closely related, and whether any of them actually represents the ancestral type in the genus is not known. It is clear that the patent fertile pinnae represent the primitive condition in the genus (Mickel, 1967), being derived from fronds with sporangia distributed over the abaxial surface and then limited to the basal pair-of pinnae, but it is not known whether the four groups within the genus today that have patent fertile pinnae are remnants of this condition or whether they are reversions to this condition. There are very likely other species of Anemia to be found in Latin America that hopefully will shed light on the phylogenetic relation- ships within the genus. fol en acutiserratum (Hieron.) Mickel, comb. nov. me 10'S 2 Asplenium cirrhatum L. C. Rich. var. acutiserratum Hieron., Hedwigia 60:259. 1919, as “acutiserrata.’~ LECTOTYPE (selected here): Trinidad, Fendler 140 (presumably B; isolectotype NY). | i ticaiam longicaudatum Mickel & Stolze, sp. nov. Fig. 3A. Ab. A. cirrhato Rich. ex Willd. frondibus grandibus et apice pinnarum acuto differt. Plants terrestrial; rhizome stout, erect, provided with lanceolate to ovate, acute scales, these 2-4 mm long, 0.6-1.0 mm wid k brown or (more commonly) bicolorous, with narrow, lighter brown margins, subclathrate, the lumina isodiamet- ric or slightly longer than broad; fronds 70-100 cm long, 12-18 cm pir cet iole 10-30 cm long, terete, dart brown to atropurpureous, lustrous or sublustrous, ith e iv crenulate; veins indistinct or obscure, commonly eto: ot se ee pees inéar to narrowly oblong, slightly arcuate, indu- sium linear, yellowish, firm-textured, subentire. oe Ri “TYPE: Ecuador: Pcia. Morona-Santiago: Cordillera Cutucu, ridge between K10s Itzintza and Chupiasa, 1200-1350 m, Camp E-1283 (NY: esate oe. This species belongs to the A. radicans complex. Bis. somewnat fe bide is widespread A. cirrhatum in that they are beg, ones pee vie She more considerably smaller with obtuse to subacute pinna apices, It resem a. ‘ata closely A. acutiserratum (Hieron.) Mickel of Trinidad, aan ee pig ee nee apices but with serrate pinna margins, auriculate pinna bases, an normally reach 35—45 cm in length. AMERICAN FERN JOURNAL: VOLUME 74 (1984) 116 Sal + se 1 | Mi, M 4 = WA | Wy, WIR tA CPI V4 gicaudatum. B. Habit of Anemia elaphoglossoides. C. Habit of FIG. 3. A. Habit of Asplenium lon Grammitis kirkbridei. J. T. MICKEL: NEW TROPICAL AMERICAN FERNS 117 Grammitis kirkbridei Mickel, sp. nov. Fig. 3C. . knightii (Copel.) Seymour frondibus parvioribus, lobis approximatioribus, stipitibus brevioribus differt. Haec species Joseph H. Kirkbride, Jr. botanico dicatur. Plants epiphytic; rhizome erect, ca. 0.6 mm diam, clothed with yellow-brown, lanceolate scales ca. 1 mm long; fronds fasciculate, 3-5 cm long, 5—9 mm wide; stipes 3-9 mm long, clothed with stiff, spreading, reddish-brown hairs 1-—1.5 mm long; blade pinnatifid, cut /2-% to the midvein, the lobes obtuse, slightly ascending, 10-13 pairs, with abundant, stiff, erect, reddish-brown hairs ca. 0.4 mm long; veins obscure; sori mostly 3 or 4 pairs per lobe, ca. | mm iam., medial between the midvein and the margin, which is often slightly recurved. TYPE: Panama: Pcia. Darién: Tres Bocas on the Rio Coasi, 1 May 1968, J. H. Kirkbride Jr. & J. A. Duke 1381 (NY). Grammitis kirkbridei belongs to the group of G. basiattenuata (Jenm.) Proctor of the West Indies and G. truncicola (Klotzsch) Morton of South America, but has fewer lobes and shorter laminar hairs. It most nearly resembles G. knightii (Copel.) Seymour, from which it differs in its smaller, less deeply cut fronds, pinnae closer together, and shorter stipe. Polystichum hottense C. Chr. Figs. 4A, B. For many years this species was known only from the type collection from the Massif de la Hotte in southwesternmost Haiti (Ekman H-10130) and part of a mixed collection from Ma Blanche, Haiti (Ekman H-5/3). It has recently been collected at the mining site of Las Abejas, Prov. Pedernales, in southwestern Dominican Republic (Liogier 14173, 26784; Mickel 8168; Zanoni 16718). This striking plant with bipinnate fronds and abundant, spreading, often black-centered rachis scales has never been well illustrated and is thus presented here. / O l ¢ L Pteris striphnophylla Mickel, sp. nov. Figs. 4C, D. b P. longifolia L. frondibus parvis, lamina sclerotica glabra, apice pinnarum obtuso differt. ‘ ; compact, the scales re guage light brown, 1.5-2.5 mm long, in a tangle of wiry roots and old stipe bases; sti 2-4 cm son with hairs eine to narrow scales at the base; fronds approximate, 7-11 cm long, 1.5-2.4 cm wide, pinnate, with a conform terminal Sag pinnae 6-9 pairs, mostly 10-12 mm long and 4-5 mm wide, entire, pe 0 sag ie obtuse, the base rounded to cordate, extremely coriaceous, pinna er es tops rachis swollen, the pinnae tending to be deciduous; veins obscure, = = =P fo l-forked; blade glabrous except for a few short hairs at pinna pores meals continuous along the margin, interrupted only at the pinna apex; 1n us coriaceous. : TYPE: Dominican Republic: Pcia. Pedernales: Sierra de Baoruco, a i Las Mercedes to Aceitillar, 1000 m, growing in a ravine, the Sieg a rock cliff, 11 Feb 1969, Bro. Alain Liogier Spe (NY; — JBSD). This is clearly related to P. longifolia, judging trom Its spores, vine bases, swollen pinna attachments, and arent te conceal easily distinguished by its small size, glabrous blade, and heavily texture. 118 “esa AA Sx 4 => "ag “thi x SS LAY LAG SOD) aoe HL SY AMERICAN FERN JOURNAL: VOLUME 74 (1984) N 4 . UC / eZ WS, eZ LUTE? Saad (WISI ‘AW Wag (Pkéic, Wiki. kes WAOe az, Stam NI arg... a : ANNA és i wa prnttitce. Liz SSK. DUBBO: . -) NQ) ThKiz, Wii ED ES AM Sane A. Habit of Polystichum — pin FIG. 4. Pteris striphnophylla. D. Same, 5mm (Liogier 14173). B. Same, rachis and pinna base. C. Habit of J. T. MICKEL: NEW TROPICAL AMERICAN FERNS 119 Saccoloma membranaceum Mickel, sp. nov. Figs. 2A, B. Ab aliis speciebus generis frondibus parvibus et ne membranacea differt. Rhizome horizontal, short-creeping, 2-3 mm dia with coarse, black, roots 0.40.6 mm diam.: rhizome clothed with Sadie reddish-brown, hea indurated, linear-deltate scales 1-3 mm long; fronds many, approximate, 10-1 ong, 2.0-3.4 mm wide; stipe 2-3 cm long, castaneous to light brown, bina 0.3-0.4 mm diam aa ade n arrowly lanceolate, 1-pinnate to a pinnatifid apex, the t pinnae cut less than y e midvein, the basal acroscopic lobe slightly hip accept veins free, Me simple or dichotomously orking, more divided in larger lobes; lamina membranous, al on veins, ca. | labrou mm from the margin, indusia cup-shaped; pried se with low, dichoto- mously branching ridges. VTYPE: Brazil: Est. Acre: Cruzeiro do Sul, Rio Jurua and Rio Moa, Serra do Moa, Rio Moa 6 km above school; forest on mountain; 25 Apr 1971, Prance et al. 12432 (INPA; isotype NY). The label states that the type specimen is epiphytic, but the roots hold consider- able sand and all the relatives are strictly terrestrial. This species is strikingly different from other members of the genus in size and texture. The spores are similar to those illustrated by Tryon and Tryon (1982, p. 386) in their low, dichotomously forking ridges. The rhizome scales and sorus structure and location also support its placement in Saccoloma. ACKNOWLEDGMENTS These studies were supported in part by a grant from the National Science Foundation (DEB 77-25582). Thanks also to Dr. Rupert Barneby and Joann Ward for their help with the Latin diagnoses, Mr. Joel Huang for the SEM photomicrographs, and Bobbi Angell and Charles Clare for the drawings. Furthermore, Mr. Seiler expresses his appreciation to the Museo de Historia Natural de El Salvador and to the Unidad de Parques Nacionales y Vida Silvestre for their support during his stay in El Salvador. LITERATURE CITED MICKEL, I T. 1962. A monographic study of the fern genus A 22. ce 36:349-482. "1966. A new species of Anemia from South —_———. 1967. The phylogenetic position of Anemia co TRYON, R. M. and A. F. TRYON. 1982. Ferns and Allie nemia, subgenus Coptophyllum. lowa America. Amer. Fern J. 56: 58-60. limen sis. Amer. J. Bot. 54: 432-437. d Plants. Springer-Verlag, New York. 120 AMERICAN FERN JOURNAL: VOLUME 74 (1984) SHORTER NOTE An Urban Locality For Asplenium platyneuron—The ebony spleenwort, Asplenium platyneuron (L.) B.S.P., was recently discovered at a site in Brooklyn (Kings County), New York. Vouchers consisting of only sterile fronds are deposited at BKL (W. Sax-Cedar & J. Goldbaum, 15 Feb 1984; T.J. Delendick, 22 Feb 1984). The population consists of fourteen individuals growing in a vertical stone wall at a well-developed and heavily trafficked intersection in the Park Slope district, approx- ‘imately midway between Prospect Park and the Gowanus Canal. The wall is part of an access well to a below-ground basement entrance to a commercial building. The well is approximately 12 feet deep, 5 feet wide and 40 feet long, with an east-west orientation. The ferns occur in the uppermost seven feet of the wall, among clumps of moss in the mortar between the granite stones; they are restricted to the east end of the well, thus having a western exposure. None of the plants examined had fertile fronds. This Brooklyn station represents the second verified locality for A. platyneuron known to exist on western Long Island at the present time, and the only one which is current for Kings County. The other site for the species in the western portion of the island is at Fort Totten on the north shore of Queens County, reported by Greller and Locke (Amer. Fern J. 73:6-8. 1983). This situation contrasts strongly with the distribution of the species on Long Island in the 19th and early 20th Century, when it was noted as “frequent throughout the island” by Jelliffe (The Flora of Long Island, 1899), including at least three locales in Kings County: East New York, G. B. Brainerd in 1866; Rockaway Beach, G.D. Hulst in 1889; and Central Park [now designated as Prospect Park], J. McCallum in 1902 (all BKL). It is, or at any rate was, fairly common in the eastern counties of Nassau and Suffolk. Stalter, (Bull. So. Carolina Acad. Sci. 41:72. 1979) cited it as occurring on Long Island, and the late Stanley Smith of the New York State Museum, Albany, noted it in both Nassau and Suffolk counties (manuscript checklist). This find also represents the only current site for A. platyneuron on Long Island south of the terminal moraine that forms the backbone of the island. There is a specimen of the species at BKL from Rockaway Beach (Kings County, 1889) and at NY from Valley Stream, Nassau County (Bicknell, 8 Aug 1903). The latter includes the collector’s note, “Bank of Aqueduct causeway. The only plants I have found south of the terminal moraine.” The outwash plains from the glacial moraine which form the southern portion of Long Island provide few habitats suitable for Asplenium species. The occurrence of A. platyneuron there seems, on the basis of available data, to be restricted entirely to man-made sites. It is remarkable that A. platyneuron has been able to extend its range into an area which has been so long and so heavily used as to lack virtually any trace of the original flora, and that its extension was made possible through commercial development. I am grateful for the assistance of Wynne Sax-Cedar, who made the first collections from the newly discovered locale, and to Andrew Greller and Richard Stalter for their comments and review of the text.—Thomas J. Delendick, Brooklyn Botanic Garden, 1000 Washington Ave.., Brooklyn, NY 11225. AMERICAN FERN JOURNAL: VOLUME 74 (1984) 121 AMERICAN FERN JOURNAL Manuscripts submitted to the JOURNAL are reviewed for scientific content by one or more of the editors and, often, by one or more outside reviewers as well. During the past year we have received the kind assistance of J .D. Caponetti, W. D. Hitz, H. T. Horner, Jr., J. D. Montgomery, M. G. Price, H. E. Robinson, A. R. Smith, R. G. Stolze, W. C. Taylor, D. P. Whittier, E. Wollenweber, and J. J. Wurdack, to whom we are deeply indebted. We welcome suggestions of other reviewers. Dr. Alan R. Smith, Department of Botany, University of California, Berkeley, CA 94720, will become editor of the American Fern Journal effective 1 Jan 1985. Dr. David B. Lellinger, U. S. Nat’l. Herbarium NHB-166, Smithsonian Institution, Washington, DC 20560, will become editor of Pteridologia on the same date. —D.B.L 1985 A.1.B.S. MEETING-CALL FOR PAPERS The American Fern Society and the Botanical Society of America will meet with the A.I.B.S. Annual Meeting at the University of Florida, Gainesville, on 11—I5 August 1985. Those members of the American Fern Society wishing to present a paper or poster and who have not received abstract forms may obtain forms from the program chairman of the Pteridological Section of the Botanical Society: Dr. Christopher Haufler, Dept. of Botany, University of Kansas, Lawrence, KS 66045. INDEX FOR 1984 divaricatum, 40: ecuadorense, 41-43: fontanum var. obovatum, jongicaudatum, 115, 116: Adiantum, 29; anceps, 29; capillus-veneris, 17; pedatum at : 40, 41, 43: oellgaardii, 44. 45. aleuticum, 62. subsp. calderi, 62, subsp. pedatum, 62, ilum, 62, ee subpumilum, 62: trapeziforme. 9 , 19, an 0 Ampelopteris prolifera, 5 ; ; A . TL, 112, 115; sect. Adetostoma, 113, | subg tum, 50: sect. Sphenopteris, 49 . : pees = Anemia, 111, 113, 115; sect. Anemiaebotrys, 115: subg 47, 49, trichomanes 4 var. herbace Anemiorrhiza, 113, 115: aspera, 115: brandegeea, 113: clinata x trudellii. 63 plier 12, 113; colimensis, 115: subg ee 113. 115: Athyrium subquadripinnatum. 57; tenuifolium, elaphoglossoid 11, 116; elegans, 115; exima, 115: hirsuta, Azolla, 112, 113: sect rsutae, 113, ae Ses ae i Becker. R. es identification of Hawaiian tree ferns of the genus 111; 114: oblongifolia, 112, 113: s ig salvadorensis, | 12-114: : oF OP 12, 113; s t. Trochopteris, 115 Anti- saleechlal activity of phenolic acids in Pteridium aqilinum, 8 as gaa cristatum X marginale, 62: cubense. 60: dicksonioides. Coke 14. 15. 40, 49, 50, 120; acutiserratum. adiantum-nigrum, 14: aethiop S| a conquisivem, 40, 41: cristatum, ect. Oblongifoliae. 113. sanctae-martae. icum, 94: cegriscee oe cirrhatum, Cibotium, 9 ah S.S. etal. era flora of Garhwal Himalaya (rev.). Botrychium, 79_85: dissectum, 78. 79. 83-85, f: ‘ikem. na Sctgeetaieel ——e oe of cma Isoétes. 9 Cibotium, 97. 98: sist 97: barometz, 97: ae 97-99: cumingii, 97: glaucum 97-99, var. fallax. 98: - menziesii, 97. ‘98: nealiae, 97-100: regale _ var. glaucescens, 100: 97, d st.-johnii, 97, 98: we Cnemidaria. 101: glandulosa. oa so decurrens. 101 122 Cooper-Driver, G. (see M. San Francisco) Crabbe, J.A. (see A.C. Je — ae 56: haitiensis, 56; protensa var. dicksonioides, 56; sloanei, 29 Cees Biggs 58; chaseae, 58; congesta, 58; cryptosora, 8 56; fen . 58; flexuosa, 58: longipinnata, 5 Pie roman eae, 56 usick, A.W. Graves’ > ort in Ohio, 63 pris 19-21: alternans, 21; atahuallpa, 56; bradei, 57: nii, 2 staal 0: excelsa, 20: insignis, 19, 20: intramarginalis, 57; lathamii, 20: oe ots luccana, 21; parianensis, rs princeps, 19-21; sect Oe ar 21: sipapoensis. 57: sect. Spinco oI; g. Sphaeropteris, 19-21: squamulata, 2 Pr eae, Cyrtomium falcatum, 94 Cyclosorus supine 6 Davallia novae-guineae, 56 Death notice: Gu alts Looser (1898-1982), 60 Delendick, T.J. An urban locality for Ae pa platyneuron, 120 Dicksonia, 19; antarctica, 19: arborescens, 19; glauca, 99, lathamii, 19 Dicranopteris rent 57; peruviana, 57 Dictymia mc ah Bor ot ‘sce, 6; ee 6; petri, 6; subquadri- 7: tenuifolium Pescporceend 37-39; ites 38 Drymoglossum under $s, 37 Drynaria acuminat. “_ Dryopteris aripensis, 56; cristata, 62: cristata x marginalis, 62: ——: var. Paranensi 56: ireneae, 60: marginalis, 62: , x slossonae, 62; x slossoniae, 62; subincisa var. ensis, 56 ———— 108: subsect. Alpestria. 110: crinitum, 110: ee 110; sect. Setosa, 110 opine inte; Equiset 61, GL: 69-73, 75, 76; arvense, 65: 66; x ferrissii, 61 67 . 73-76; fluviatile, 66: subg. dopasanee . affine, 65, 67-71, 6l, 65-75; palustre, 65, 66, 75; ramosissimum, 61 ramosissimum in Louisiana, 61 Exonotins flexuosus, Farrar, D.R. (see L.M. Rut Ferns and fern allies of Sau temala, part [II (rev.), 36 Flora of Ecuador 14(4): eR Se TARE (rev.), Frequency of cyanogensis in Bracken in relation to shading and winter severity, 51 Gleichenia angusta, 57: ponderous insidiosum, 57; integrum boliviensis, 57; peruviana. 57 m, 57: nesioticum, 57 Gra saiaagrcnie in Ohio, 63 sont roides, 58; angus' _ 58: basalis, 58: besiattenuata, 7 uesii, 58: chase , 58: ciliolepis. 58: congesta, 58: oe 61: fendle: eure feta 58; humilis, 58: Kegelian SDs, killipii, 58: cick idei. 116, 117; knightii 117; Seana . 58; maxoniana ayoris, 59: mela- notricha, 59: ene 5: perp la. 59: shaferi. 59: truncicola, 117: williamsii, Gualterio Looser (1898-1982) (death notice), 60 Gymnogramma a . 61 Gymaoptens rufa, pee ITissii S parent species, Equisetum hyemale and Preah rae in Iowa, 65 aki Hag isetum ramosissimum in Louisia Hensipean. E. .C. Roos. A monograph Py sy genus AMERICAN FERN JOURNAL: VOLUME 74 (1984) Platycerium Sync (rev.), Herndon, A. & R mdon. Two sah of Adiantum newly m under water stress. 37 R.J. Chro of neotropical Isoétes, 9 The ‘ion of Faveiies tree apie of the genus Cibotium, fiscal of pteridophytes of Japan, volume 3 ( ,6 new phenolic shail i in aia 11; andicola, 9, Isoétes Ho, 22 , 25, 36; alcalophila, 10, 11, 12; boliviensis, 11; coromandelina 26; durieui glacialis, 11, 12; herzogii, 11, 12; indic 9 25; lacustris, 11. 2; lechleri, 9; macrospora, 11; anadensis, 10, 11; anchananii, 9; pantii, 9, 25; rajasthanensis, 22, 23 3120; retic slat 22, 23, 25, 26; storkii, | 1; ticlioensis, 9, 11, 12 Jermy, A. C., J. A. Crabbe, and B. A. Thomas (eds.). The 6 ~ tag and — tio a) Kur. . & T. Nakaike. hataliéod of saison of Japan, pay ata x are 62 aa; 22) ew Sena. and some new names in $,.96; rns on North American ferns, ee rev. Lastrea cristat: Leclercquia aes Lellin _ D. Leptogram ar ee var. alabamensis, 30 iol e ligule of — , 61; cuspidata, 61; wallichiana, 61 ‘ scopodiolin x brue ei, 63 Lycopodium, 36; appressum, 63; X brucei, 63; prostratum, 63 Marsilea, 36 Matte euccia, 1; struthiopteris, 1-5; orientalis, 5 Mickel, J. vee w tropical American ferns, 111 Microgramma acuminata, 59; latevagans, 59; recreensis, 59: tuberosa, 5 Microsorium, 28 Microsorum, 28 Miller, K. G. Trichomanes gametophytes at Bartholomew's cob- bie, 31 A monograph of the fern genus Platycerium (Polypodiaceae) 18 . T. Usos de los helechos en Suramérica con especial relists a Colombia (rev.), Nakaike, T. (see S. Ki slossonae, 62 names in ferns, 56 A new station for ‘aerate eer in Alabama, 30 New — American ferns, 111 Not rican ferns, II, 62 rma, 35 organic nutrition of Botrychium gametophytes, 77 Osmunda, 84: cinnamomea, 5 Parris, B. S. PY taxonomic revision of the genus Grammitis . . . in New Guinea (rev.), 86 Pecluma camptophyllaria var. lachnifera, 59; eurybasis var. AMERICAN FERN JOURNAL: VOLUME 74 (1984) glabrescens, 59. var. villosa, 59; ptildon var. robusta, 60 Pellaea < wrightiana, 33: longimucronata. 33: ternifolia, 33 Phegopteris conne ee 6 The ee and classification of the ferns (rev.), 96 Pinonia splen ship Platycerium, iar semana. 18 “Shai pleolepis, 60 lypodium alike 58: angestipalcani, 56: aureum. 94: asale, 58; buesii, 58: ciliol 8. cryptosorum, 56: decipiens urybasis var. villosum, 59; flexuosum, 58 umile, 58: insidiosum, 57: keg um, 58: lachniferum, 59 var. glabrescens, 59: latevagans, 59; loretense, 59; mayoris, 5 mckeei, 60; melanotricha, 59: ticum, 57: perpusillum 59; cape 60: sans 58: recreense, 59: robustum ; : sha feri : . 59; tubero: Po fae cubensis, 60 Polystichum 6; "none 33; aculeatum, 33: braunii, 6, 35: hottense, 117, 118: imbricans, 33, mbricans. 33: imbricans X lemmonii, 33: lonchitis, = eh aa BA 6 osum, 59: williamsii, 59 e, 61 es from Ecuador. 40 m Pue rto pe p Promotion ok apogamy in Matteuccia struthiopteris, the Ostrich =k . 5; aquilinum, 1, 5. 17, 51. 87, a softs flora of Garhwal Himalaya (r Pteris longifolia, 117; peas 94. pence 117, 118 A remarkable Cyathea hybrid, Revi Ferns and lies i. roe part III, 36; Flora of — = 94 : ge sine Guinea, 86; Usos de los helechos en | referencia a Reins ia ~ MC. (see EB. ‘Heonipmad nape E. (see W. H. Wagner, Jr. utz, L. M. & D. R. Farr ee habitat characteristics and bundance of Equise ‘cial and its parent species uisetum hyemale ne Equisetum laevigatum, in Lowa, 65 Saccoloma, 119: membranaceum, 114, 119 Sadleria Salvinia, 36 : . Cooper-Driver. Anti- microbial activity of phenol acid in Pteridium aquilinum, Schre _ Nafus & D. Pimentel. Frequency of cyanogensts in Baas in relation to diading and winter severity, 51 Seiler, R. L. Trunk length and frond size in a population of 123 Nephelea mares from El oo 105 Selaginella, 22, 8 a Selliguea sem sitg harma, B. D. & ee The ligule of Isoétes, 22 e B. D. Sharma) . R. Flora of Ecuador 14(4): Polypodiaceae—Thelyp- ih otis Wi} 30 sia Ne oe atahuallpa, 56: bradei, 57; horrida. 19. = intramarginalis, 57: nsis, ri ® (se lock woodiana. 36: Problems in Asplenium gon some new y tpecied Sonn ; a, 101 gemmifera, 9 A taxonomic revision of the genus Grammitis . . . in New Guinea . 86 Breer 55: subg. Amauropelta, 55: ekmanii, 60; subg. Glaphyropteris. 55: ireneae ; limbo: ; subg. Cyclosorus, 55: Tmy) Three new cabiioieicie in » bicsecaise, 6l, odea ara, cee Fess; ies chaetum, 7: padronii Trichomanes, 7 subg. Achomanes, 7: . Pachy- nii, 7, 8; petersii, 30: sien 7: subg. tophytes at scoieisirenmial s cobble. 31 . 1 i wo new p lic g r ov icgieeiinne. 4 Ti rs Dp. ma 1 ] rwo new 0 Two species of Adiantum ig escaped in Florida, 29 Lane Se 104: pinnata, 103. 104; williamsii, 104 richopteri aes ae os i + was ee ae | — El — = usu. Elap! gone um a thon Peru An urban ie . de platyneu Usos de los helechos en Suramérica con cde referencia a Colombia (rev.) ee Vittaria, 31 on Aderkas, P. Promotion of apogamy in Matteuccia struthiop- teris. the Ostrich Fern, H.. Jr. & E. Rouleau. A sara Holly Fern. “aE x scopulinum, in N riccinconil n Whittier, P. The organic nutrition of ae gameto- ytes Wyatt, R. A new Station for Teichomanes petersii in Alabama, 30 Xiphopteris killipii, 58: morton ERRATA eae ih Page 59, line 27: For “recreense’” read “recree bb Page 60, lines 23 and 24: For “Gaulterio” ae mGualterio.” 124 AMERICAN FERN JOURNAL: VOLUME 74 (1984) STATEMENT OF OWNERSHIP, MANAGEMENT AND CIRCULATION Required by 39 U.S.C. 3685) 2. DATE OF FILING 1A. TITLE OF PUBLICATION AMERICAN FERN JOURNAL 28 Sept 1984 ANNUALLY PRICE quarterly 4 $8 mem; $9 subscr 4, COMPLETE MAILING ADDRESS 0! U. S. Nat'l. Herbarium, 10th & Constitution Ave. N.W., Washington, DC 20560 U. S. Nat'l. Herbarium, 10th & Constitution Ave. N.W., Washington, DC 20560 6. FULL epee MAILING ADC SoRUSntRs (Name and Complete ddress) CAN FERN ae INC. (see 5.) Dr. David B. Lellinger (see 5.) none ” OWNER (if owned by a corporation, its name and address must be stated and also immediately thereunder the names and addresses of stockholders owning or holding I percent or more of total amount of stock. 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Each entry is annotated with world distribu- tion, bibliographical references to descriptions and illustrations, ecolog- ical and horticultural requirements, growth habit, and variations. A comprehensive introduction and discussion on categories of hardiness facilitates maximum use of this book. 70 pages, softbound. Distributed in the U.S.A., Canada, and Mexico by Trainpower, Inc., Publications Division, P.O. Box 1255, Vienna, VA 22180 ($8.00 postpaid). Elsewhere: The British Pteridological Society Booksales, 46 Sedley Rise, Loughton, Essex 1G10 1LT, England (£4.50 postpaid). WANTED DEAD OR ALIVE! High quality, heart-shaped fern prothallia, live or FAA-preserved, for slide making. We need monoecious and dioecious prothallia as well as stages showing sporophyte develop- ment. Please send samples for viewing. We pay for high quality specimens only! We also supply high quality prepared microscope slides to those who need the very best for their teaching or research. TRIARCH, INC. - P.O. Box 98 - RIPON, WI 54971 (414)-748-5125 ee BRITISH PTERIDOLOGICAL SOCIETY Oe Open to all who are interested in growing and studying fens ane “OF THE Full members receive THE FERN GAZETTE and the BULLETIN ake BRITISH PTERIDOLOGICAL SOCIETY. Membership OF eerie and for full members, £ 5 for ordinary members (not receiving the san : : £5 for student members (under Fe years of age). For sguoteelt pene residents should apply to Dr. J. Skog. Biology Songhai iy Se University, Fairfax, VA 22030. Non-U. S. residents ca ag uF nd 5 Coke Robin Hill, Sanchcomie, OMe Oe Ferns of Jamaica A guide to Pteridophytes G. R. Proctor amaica has one of the richest fern floras in the world, yet hitherto there has been no publication dealing authoritatively with the Jamaican ferns i in their entirety. Ferns of amaica will rectify this situation, by p of all the knowz ferns and fern allies of the island. The aim has been to present a scheme of classification with broad validity, reconciling as far as possible older views which empha- sized resemblances with more recent attitudes that emphasize differenc This work names and describes the 579 ‘Species and 30 varieties of ferns recorded ir in Jamaica. Keys to the subfamilies of P eae ie ails relevance and potential for wide use in the whole ‘American tropics. the min distinguishing features of closely related plants. ni stily mp the eighty- -three genera are illustrated, many with splendid nineteenth century drawings taken from rare and often inaccessible sources. Where suitable illustrations could not be found, new drawings have been specially commissioned for this work. Each taxon is succinctly but fully described, and data on ga local and general geographical distribution are given. Habitat and altitudinal range are stated for most of the species. The author has _ — waioaiae all previous literature dealing with Jamaican ferns in the course of his research, a An extensive bibliography is seid ae care has been taken to present all information in a standardized format for ease of consultation Ferns of Jamaica is an immensely practical flora, Sapctially in the information it provides on habitat and ecology. The work’s definitive and authoritative nature will ensure its recognition as the standard work on the subject for many years to come. sabe bleh oo Acknowledgements, — ion Pteridophyta, Conspectus of Major Taxa, Systematics and Descriptions of Species: Class I. Psilopsida, Class II pe sats Class IIL Lycopsida, Class IV. Nera ag Bibliography, Glossary, Index. The Author One time Senior Botanist in agence of the ephyge of the Science Museum, Kingston, Jamaica, Dr. G. R. Proctor is botanist with thirty years of experience in the West Indian islands, including Tanciaize. and the Central American countries. 610pp, 135 line illustrations, 22 maps. Hardback. 0 565 00895 1 Publication due 3rd January 1985. ORDERS TO: Rudolph Wm. Sabbot, Natural History Books, 5239 Tendilla Avenue, Woodland Hills, Calif, 91364. Tel: (818) 346 7164. [Authorised agent of the British Museum (Natural History)]. Price $75.00 plus 10% postage and packing. British Museum (Natural History)