Fy
ALBERT R. MANN LIBRARY AT CORNELL UNIVERSITY
i /
wi ry Ad
» 4 att y tari i aes
as asap i m Yh a . p sty ee iN y, on
t Ay 4 °) or at ee y TOS ae A) AG Ge ay ve ; Ata 4 : : bil ( Af i it) 7 i As Ay I ‘ uj fy 4 s i M . ie aw ee \' i iy a ba a ‘|
Ar Ge 7 Loe By ii Bi - ‘
MY COTAXON
AN INTERNATIONAL JOURNAL DESIGNED TO EXPEDITE PUBLICATION OF RESEARCH ON TAXONOMY & NOMENCLATURE OF FUNGI & LICHENS
V ODUIMUR XXX V IOs 92919
COMPLETE IN TWO QUARTERLY ISSUES,
CONSISTING OF iv + 512 PAGES INCLUDING FIGURES
CO-EDITORS
G. L. HENNEBERT French Language Editor & Book Review Editor
Laboratoire de Mycologie systématique et appliquée Université de Louvain, B-1348 Louvain-la-Neuve, Belgium
RICHARD P. KORF English Language Editor & Managing Editor
Mycotaxon, Ltd., P.O. Box 264 Ithaca, NY 14851, USA
SUSAN C. GRUFF Associate Editor & Index Editor
Plant Pathology Herbarium, Comell University Ithaca, NY 14853, USA
Published by MYCOTAXON, LTD., P.O. BOX 264 ITHACA, NY 14851, USA
Printed in the United States of America
TABLE OF CONTENTS, VOLUME THIRTY-SIX
No. 1. October-December, 1989
Amanita eburnea — a new species from Central America. Rodham E. Tulloss Lecanora sect. Petrasterion (lichenized Ascomycotina) in North America: Lecanora weberi Ryan, sp. nov. (subsect. Pseudocorticatae), ATOMS OLCTACO ee pee ee ee FN ol ge Bruce D. Ryan Two new species of the genus Ascochyta Lib. Simeon G, Vanev and Ganka G. Bakalova Type studies in marasmioid and collybioid fungi (Tricholomataceae) II. PAVATICUSERTONUNUNT hen? fe eee oe bt Vladimir Antonin Vaccinium fungi: Helicoma vaccinii sp. nov. ............ L. M. Carris Long-chain fatty acid composition as a tool for differentiating spoilage wine yeasts... M. Malfeito-Ferreira, A. St. Aubyn, and V. Loureiro Description of the anamorph of Valseutypella multicollis in culture.
Julia Checa and Angel T. Martinez Studies on J. B. Cleland’s fungal herbarium — 2: Cortinarius subgenus Myxacrum (Corunatiates)e eee. Cheryl A. Grgurinovic
Studies on Galeropsis and Gastrocybe (Bolbitiaceae, Agaricales). G. Moreno, M. Heykoop, and C. Ilana
Phytophthora erythroseptica ................. H. H. Ho and S. C. Jong
Deuteromycotina from Antarctica. New species of hyphomycetes from Danco, Coast, ‘Antarctic Peninsula’... 22 0 Marta N. Cabello
Studies on Pholiota in culture...................... Stig Jacobsson.
Scientific names in the Endogonales, Zygomycotina . . Rogério T, Almeida Contribution to the lichen flora of Brazil. XXIII. Lichens from Sao Paulo AG RAEI one Lit gms a ees ese whee Héctor S. Osorio A new species and first record of Gymnopaxillus (Hymenogastrales) from AATDEN tina apie ae x sates ire Susana Calvelo and Laura Lorenzo A taxonomic revision of the genus Cheilymenia — 1. Species close to Chey ymeniar brat 2h MNO US aN REMIND LS | Raa Jiri Moravec
R. Duran and P. M. Gray Meliolaceous fungi from the State of Kerala, India I. V. B. Hosagoudar and R. D. Goos Cultural, enzymatic and cytological studies in the genus Pholiota. Jaroslav Klan, Dana Baudisova, and Ivana Rulfova Two new Glomus species from arable land... J. P. Skou and I. Jakobsen The genus Pezizella I: nomenclature and history. Wolf-Riidiger Arendholz
Three new species in the lichen genus Parmelia (Parmeliaceae, Ascomycotina) from southern Africa, with further notes. Franklin A. Brusse
ill
305
No. 2. January-March, 1990
Entolomataceae in eastern North America I: new species of Claudopus and Rhodocybe from the southern Appalachian Mountains. Timothy J. Baroni 313 Contribution to the lichen flora of Brazil. XXIV. Lichens from Nova Petropolis, Rio Grande do Sul State. Héctor §. Osorio and Mariana Fleig 325 Nidulispora gen. nov., a hyphomycete genus with crateriform conidia. A. Nawawi and A. J. Kuthubutheen 329 A preliminary checklist of the Agaricales of Tulsa County, Oklahoma. Estelle Levetin, Nora Jones, and Kerry Owens 337 Teleomorph-anamorph connections and correlations in some Xylaria SPECles .auiuaeer tae ec eres Brenda E. Callan and Jack D. Rogers 343 First record of Catenaria auxiliaris in Illinois. C. M. Stiles, D. A. Glawe, and G. R. Noel 371 Halophytophthora, gen. nov., a new member of the family Pythiaceae. H. H. Ho and S.C. Jong 377 A monograph of the discomycete genus Strossmayeria (Leotiaceae), with comments on its anamorph, Pseudospiropes (Dematiaceae). Teresita Iturriaga and Richard P. Korf 383 Nomenclature and synonymy of Stereum spadiceum var. plicatum. B. De 455 Phoma proboscis sp. nov. pathogenic on Convolvulus arvensis.
Dana Kelly Heiny 457
Taxonomical studies on Ustilaginales. V. .......----- Kalman Vanky 473 BEG Re VICW SAARI oats gee tke aay ahha wl» aipnauany G.L. Hennebert 483
R.SINGER; S. T. MOSS; Tadashi ARAI; D. MOORE, L. A. CASSELTON, D. A. WOOD, & J. C. FRANKLAND; J. A. von ARX; M. E. NOORDELOOS; R. A. SAMSON, E. S. HOEKSTRA, & C. A. N. VAN OORSCHOT; sod, (CHANG -“@ 22H. QUIMTIO;) Ku" H. STEINKRAUS; Ingo NUSS;
E. J. H. CORNER; DEUTSCHEN GESELLSCHAFT FUR MYCOLOGIE;
J. B.iSMITHS oD. TC GIBSON; R. WATLING & N. M. GREGORY;
S. A. GUARRERA, I. GAMUND{ de AMOS, & D. RABINOVICH de HALPERIN; E. PARMASTO; Sheila BARRY, D. R. HOUGHTON,
G. COLELEWELLYN,, & C..5B. OCREAR, CG. is: HENNEBERT & AL.
NOTICES: Fourth International Mycological Congress: Second Circular ......-- 493 XI Congress, International Society for Human and Animal Mycology... 494 Subscription price increase ......----- esses eee r eet etsy 495 Important change in editorial policy requiring submission of reviewers CCT TRCLITS Poe te ine, ghee teers i sce le haat ra 496 Arathor INDEX 0c oe ele eeu ni te 9 Alam See tthe Nirah atin Oe 497 INDEX to Fungous and Lichen Taxa ......---- +--+ sere terres 499 Reviewers vine hee BE Oi aves Bla. xs Bogetn ar oat ane en ce Sea 511 Publication Dates, MYCOTAXON Volume 35(2), SG) OEE ee ee 511
Berri lk ee ee orn se at Sic ene le Votre oa PERRY fe 98 Ae SRS Ral Sn 512
Digitized by the Internet Archive in 2025 with funding trom Cornell University Library
https ://archive.org/details/mycotaxon36unse
A)
aaa
NS
ay ee M
he ub ae ' i al he : ‘ th, di ; Th, - 1 A } " Pip ato s Ay ee tf ‘ Me oy
7 i | cy ‘f vita Mit aa Wy jaa we ; 4 He ‘ ; ‘i ) se | 4, Mi . M P AGA P a Vy ia 9 ¥) at bats ‘ie AS ‘¥ 7 oP
Sage
«7 LYCOTAXON
AN INTERNATIONAL JOURNAL DESIGNED TO EXPEDITE PUBLICATION OF RESEARCH ON TAXONOMY & NOMENCLATURE OF FUNGI & LICHENS
Vol. XXXVI October-December 1989 No. 1
CONTENTS
Amanita eburnea — a new species from Central America. Rodham E. Tulloss | Lecanora sect. Petrasterion (lichenized Ascomycotina) in North America: Lecanora weberi Ryan, sp. nov. (subsect. Pidacoricate’ PM COOMA Sk ie ke iy eae oe es le Bruce D. Ryan 9 Two new species of the genus Ascochyta Lib. Simeon G. Vanev and Ganka G, Bakalova 15 Type studies in marasmioid and collybioid fungi (Tricholomataceae) II.
ASOCUS STOMOVEN 605 SS ee Vladimir Antonin 19 Vaccinium fungi: Helicoma vaccinii sp.nov. ............ L.M. Carris 29
Long-chain fatty acid composition as a tool for differentiating spoilage wine yeasts... M. Malfeito-Ferreira, A. St. Aubyn, and V. Loureiro 35 Description of the anamorph of Valseutypella multicollis in culture. Julia Checa and Angel T. Martinez 43 Studies on J. B. Cleland’s fungal herbarium — 2: Cortinarius subgenus Myxacium: (Cortnariales) oo 5 es Cheryl A. Grgurinovic 47 Studies on Galeropsis and Gastrocybe (Pelbiiscece. Agaricales). G. Moreno, M. Heykoop, and C. Ilana 63
Phytophthora erythroseptica ................. H.H. Hoand$.C.Jong 73 Deuteromycotina from Antarctica. New species of hyphomycetes from
Danco Coast, Antarctic Peninsula............. Marta N. Cabello 91 Stuties on Phokote:m cultare 2 a es os ce Stig Jacobsson 95
Scientific names in the Endogonales, Zygomycotina .. Rogério T. Almeida 147 Contribution to the lichen flora of Brazil. XXIII. Lichens from Sao Paulo
CE ec s s nis ore us nk Etec eae Héctor S. Osorio 161 A new species and first record of Gymnopaxillus (Hymenogastrales) from
Areentas 46 ee . Susana Calvelo and Laura Lorenzo 163 A taxonomic revision of the genus Cheilymenia — 1. Species close to
CHEM OTE Oh ees ey Jiti Moravec 169 Size and shape of urediniospores as influenced by ambient relative
Mrmndity 3) Ses os Larry J. Littlefield and Wanda K. Schimming 187
[CONTENTS continued overleaf]
ISSN 0093-4666 MYANAB (30. (1) “T-32L (1989)
Published quarterly by MYCOTAXON, LTD., P. O. Box 264, Ithaca, NY 14851. For subscription details, availability in microfilm and microfiche, and availability of articles as tear sheets, see back cover.
[CONTENTS continued from front cover]
Nuclear DNA, an adjunct to morphology in fungal taxonomy. R. Duran and P. M. Gray Meliolaceous fungi from the State of Kerala, India I. V. B. Hosagoudar and R. D. Goos Cultural, enzymatic and cytological studies in the genus Pholiota. Jaroslav Klan, Dana Baudisova, and Ivana Rulfova Two new Glomus species from arable land... J. P. Skou and I. Jakobsen The genus Pezizella 1: nomenclature and history. Wolf-Ridiger Arendholz Three new species in the lichen genus Parmelia (Parmeliaceae, Ascomycotina) from southern Africa, with further notes. Franklin A. Brusse
[(MYCOTAXON for July-September 1989 (35(2): 201-512) was issued August 28, 1989.]
205 221
249 273
283
305
MYCOTAXON
Vol. XXXVI, No. 1, pp. 1-7 October-December 1989
AMANITA EBURNEA—A NEW SPECIES FROM CENTRAL AMERICA AR. MANN LIBRARY
Rodham E. Tulloss FEB 0 2 1990
21 Lake Drive Roosevelt, New Jersey 08555
Summary
Amanita eburnea is described as new from pine forests of Honduras and Belize.
Amanita eburnea Tulloss sp. nov. Holotypus: HONDURAS, Siguatepeque, vi.1976 M. H. Ivory s/63 (K).!
Etymology: eburneus, ivory white.
Pileus albus vel fumeus, 70 - 90 mm in mensura diametrica, primo globosus, sed deinde qui fit late campanulatus vel plano-convexus, margine nonstriata, nonappendiculata; materies volvica absentes. Lamellae condensae, albae, liberae. Stipes 45 - 110 x 10 - 18 mm, albus. Volva membranacea, alba vel subflavida, vel 45 mm alta. Basidiae tetrasterigmaticae; fibulae absentes. Sporae (7.0-) 8.0 - 11.0 (-12.0) x (4.8-) 5.2 - 6.5 (-7.5) pum, ellipsoidae vel elongatae, amyloideae.
Amanita eburnea (Fig. 1) is a member of section Phalloideae. It is a medium-sized white to smoke gray mushroom with a smooth dry pileus surface and an annulate stipe. It has a membranous universal veil forming several limbs on the rounded to slightly pointed stipe base; the limbs are free for about one third of their height. Microscopic characters serving to distinguish the species are ellipsoid to elongate spores with an overall average Q of 1.58; a ramose subhymenium containing many uninflated, short hyphal elements; and rather plentiful inflated cells in the partial veil. This entity is known from pine forests in Belize and Honduras.
PILEUS: 70 - 90 mm diam, white to smoke gray, dry, globose at first, then broadly campanulate, then planoconvex to planar; margin nonappendiculate, nonstriate;
a DTI - private herbarium of Dr. D. T. Jenkins, University of Alabama, Birmingham, U.S.A. IVORY - private herbarium of Dr. M. H. Ivory, Oxford Forestry Institute, University of Oxford, U.K. K - Herbarium, Kew Botanic Gardens, U.K.
eee “SS SS Ga Cs SS Ss it
Fig. 1 Amanita eburnea. M. H. Ivory s/63 [x1.2].
pileipellis ‘‘papery’’ (Ivory); context white, firm to spongy. LAMELLAE: white, free, drying brown (6D6 or 7.5YR 5/8)’, up to 6 mm broad, margin at times buff® and remaining pallid in exsiccatae. STIPE: 45 - 110 x 10 - 18 mm, white, cylindric, decorated with longitudinal striations; context white, firm to spongy, stuffed to solid; annulus white to buff, superior to subapical, membranous to submembranous, persistent or left in patches over gills or collapsing and sliding down stipe; base swollen to slender bulb, rounded or pointed below, 25+ mm long; universal veil membranous, white to buff, appressed to base, limbate in several free lobes reaching to 45+ mm from base of bulb, with limb free for up to 15+ mm. One specimen (s/111) which was old enough to have lost its annulus, had a ‘‘moderately unpleasant smell’’ (Ivory).
PILEIPELLIS: filamentous undifferentiated branching hyphae 1.5 - 19.0 um diam, gelatinizing just at surface, interwoven, not radially arranged; oleiferous hyphae 3.0 - 13.3 um diam, with walls of the broadest thickened slightly up to 0.7 um. PILEUS CONTEXT: tangled undifferentiated filamentous branching hyphae 1.8 - 15.0 um diam; inflated cells ovoid, subpyriform, subglobose, plentiful, to 107 x 57 um; branching oleiferous hyphae 1.0 - 7.5 um diam. LAMELLA TRAMA: bilateral; frequently branching tangled filamentous undifferentiated hyphae 2.4 - 7.2 um diam, dominating, occasionally with slightly inflated terminal segment (e.g. 56 x 10 um) at 30°-40° to central stratum; inflated cells to 32 x 17 um, thin-walled, intercalary, clavate to ovoid to ellipsoid; branching oleiferous hyphae present, relatively common, 2.2 - 7.0 um diam. SUBHYMENIUM: branching chains of uninflated narrow short hyphal segments to slightly inflated short hyphal segments and ovoid to subglobose inflated cells (to 18 x 12.5 um); basidia arising from cells of all types. BASIDIA: 37.5 - 46 x 9.7 - 11.0 um, 4-spored, thin-walled; clamps not seen. UNIVERSAL VEIL: On the stipe base at the exterior surface: a thin layer of loosely interwoven undifferentiated filamentous branching hyphae largely without sublongitudinal orientation, 1.8 - 5.8 um diam, at times in fascicles; some inflated terminal cells subclavate to clavate to ventricose, to 166 <x 34 um, all below the surface. On the stipe base below the surface: filamentous undifferentiated branching hyphae plentiful, 1.8 - 11.5 tum; inflated cells plentiful, ellipsoid to ovoid to subglobose (to 85 x 61 tum) and clavate (to 95 x 31 um), occasionally with walls thickened up to 1.0 um; oleiferous hyphae present 4.5 - 4.8 um. On the stipe base, inner surface: extensively gelatinized, all structures similar to interior; in some regions there is a thin layer of sublongitudinally oriented hyphae. STIPE CONTEXT: acrophysalidic; branching filamentous undifferentiated hyphae 4.2 - 10.5 um diam; oleiferous hyphae 2.8 - 7.7 um diam; acrophysalides dominant to 315 x 64.5 um; clamps not seen. PARTIAL VEIL: dominated by terminal inflated cells, difficult to reinflate, subglobose to ovoid (to 45 x 43 tum), subclavate to pyriform (to 82.5 x 25 um, most less than SO tm long) with walls infrequently thickened to 1.0 um;
2: Color codes of the form ‘*6D6’’ are from (Komerup & Wanscher, 1978). Color codes of the form **7. SYR 5/8’’ are from (Munsell Color, 1975).
3. The color *‘buff’’ is mentioned for several parts of the fruiting body in the collector’s notes. Dr. Ivory informs me (pers. corresp.) that this name was taken from Rayner (1970) in which it is noted as equivalent to the Munsell notation 1.2 Y/8.1/4.1 and the ISCC-NBS name ‘'‘pale yellow’’.
filamentous undifferentiated branching hyphae 1.5 - 7.8 um diam, mostly quite narrow, collapsing; oleiferous hyphae present, 4.0 - 10.0 tum diam. All tissues pale yellow in 2% and 5% KOH and 10% NH,OH.
BASIDIOSPORES: [108 from 5 specimens] (7.0-) 8.0 - 11.0 (-12.0) x (4.8-) 5.2 - 6.5 (-7.5) um, (average length (per specimen) = 9.0 - 9.8 1m; overall average length = 9.3 um; average width (per specimen) = 5.7 - 6.0 um; overall average width = 5.9 um; Q = (1.29-) 1.34 - 1.84 (-2.08); average Q (per specimen) = 1.50 - 1.66; overall average Q = 1.58), amyloid, thin-walled, hyaline, ellipsoid to elongate, rarely broadly ellipsoid or cylindrical, sometimes swollen at one end, often slightly adaxially flattened; contents guttulate; apiculus sublateral, cylindrical; white in deposit.
Distribution and habitat: Solitary to gregarious in troops under Pinus oocarpa Schiede & Deppe in grass on well-drained loamy soil and P. caribaea Mor. in bushy forest over poorly drained sandy soil, May to October, at elevations between 500 and 1100 m, in Honduras and Belize.
Figs. 2-4 Amanita eburnea. 2. Elements of hymenium and subhymenium (M. H. Ivory s/63). 3. Elements of partial veil (M. H. Ivory s/46b). 4. Elements of universal veil from free portion of limb near exterior surface (M. H. Ivory s/63). The bars in Figs. 2-4 represent 20 um.
Collections examined: BELIZE: Augustine Forest Station, vi.1976 M. H. Ivory s/85 (K), vi.1976 M. H. Ivory s/93 (K, portion in IVORY), vi.1976 M. H. Ivory s/111 (K, portion in IVORY). HONDURAS: Siguatepeque, v.1976 M. H. Ivory s/46b (K, portion in IVORY), vi.1976 M. H. Ivory s/63 (holotype K, isotype with faded color photograph in IVORY).
DISCUSSION
The amyloid spores, nonappendiculate pileus margin and membranous volva on an expanded stipe base require A. eburnea to be placed in section Phalloideae (Bas, 1969). Five white taxa of section Phalloideae described from North America have spores of approximately the size or shape of those from A. eburnea (Jenkins, 1986). The group with similar values of overall average Q include the following (values in parentheses obtained from (Jenkins, 1986)):
— Amanita parviformis (Murrill) Murrill (1.40) - a small mushroom with pileus about 25 mm diam described from Florida (U.S.A.), differing from A. eburnea by having few inflated cells in its universal veil tissue, having an annulus composed exclusively of hyphae, and having spores 7.8 - 9.4 x 5.5 - 6.3 um (based on study of the type by Jenkins (1979));
— Amanita gwyniana Coker (1.45) - a small mushroom with pileus up to 40 mm diam described from North Carolina (U.S.A.). differing from A. eburnea by having markedly rounded ends to the lamellae at the pileus margin, having a distinct odor of chloride of lime, and having spores 9.2 - 11.0 x 6.5 - 7.4 um (based on the protologue (Coker, 1927));
— Amanita hygroscopica Coker (1.47) - described from North Carolina (U.S.A.), differing from A. eburnea by having lamellae that become pinkish with maturity, having a subhygrophanous pileus, having an annulus that is subsuperior to submedian (Coker, 1917: pl. 17 and 18), and having spores 9.0 - 12.2 x 6.2 - 8.1 um (Jenkins (1986) states that the characters of lamellae and pileus here cited for A. hygroscopica are quite distinctive in Amanita);
— Amanita magnivelaris Peck (1.47) - described from Long Island, New York (U.S.A.), differing from A. eburnea by having no inflated cells in its partial veil, having few inflated cells in the universal veil at the stipe base, and having spores 8.6 - 10.9 x 5.5 - 7.8 um (based on study of the type by Jenkins (1978));
— Amanita elliptosperma Atkinson (1.49) - described from North Carolina (U.S.A.), differing from A. eburnea by its having few inflated cells in its partial veil, having sparse inflated cells in its universal veil, and having spores 9.37 - 10.15 x 6.25 - 8.59 tm (based on study of the type by Jenkins (1982)).
Amanita magnivelaris has been reported from Mexico by Singer (1958). White species of section Phalloideae having globose to broadly ellipsoid spores which are also broader than those of A. eburnea (A. bisporigera Atkinson, A. verna (Bull. : Fr.) Roques, and A. virosa ss. auct. amer.) have also been reported from that country by several authors including Pérez-Silva et al. (1970). Pegler (1983) reports no records of any species in section Phalloideae in the Lesser Antilles. There is no white species of section Phalloideae reported for any locality in South America (Garrido & Bresinsky,
1985 and Raithelhuber, 1986).
The whitish pileus, habit, microscopic structure of the annulus and the overall average Q of the spores of A. eburnea are suggestive of the Mediterranean taxa A. ovoidea (Bull. : Fr.) Link, A. ovoidea var. proxima (Dumée) Bon & Courtecin, and A. aminoalifatica Filippi.* These entities are considered to form a closely related group in section Amidella (Gilbert, 1940 & 1941; Bas, 1969; Kiihner & Romagnesi, 1974; Filippi, 1985). A distinguishing character of section Amidella is an appendiculate pileus margin which is absent in A. eburnea.
The collection M. H. Ivory s/244 (HONDURAS, Siguatepeque, x.1976) is believed by Dr. Ivory (pers. corresp.) to be conspecific with the collections of A. eburnea herein cited. Portions of Ivory s/244 were deposited in both K and DTJ. Neither portion can be located at present despite repeated searches.
ACKNOWLEDGMENTS
I am very grateful to all of the following: Dr. C. Bas, Rijksherbarium, Leiden, The Netherlands, for his comments on this article and for providing working space in his laboratory for the study of collections of A. ovoidea; Sig. Ilario Filippi, Florence, Italy, who provided the loan of co-type and paratype material of A. aminoalifatica and allowed me to retain some parts of these collections as well as a collection of A. ovoidea and several color transparencies; Dr. M. H. Ivory, Oxford Forestry Institute, University of Oxford, U. K., for his informative correspondence, for a gift of photographs, for providing me with a copy of his field notes and a loan of specimens, and for reviewing drafts of this article; Dr. David T. Jenkins, Department of Biology, University of Alabama in Birmingham, for his reviewing this article; Ms. Mary A. King, Roosevelt, New Jersey, for assistance in final preparation of the manuscript for publication; Mr. Neal Macdonald, Princeton, New Jersey, for preparing the illustrations; Dr. David N. Pegler,, Herbarium, Royal Botanic Gardens, Kew, U. K., who originally put me in contact with Dr. Ivory, hosted me at K, located a number of collections, and provided work space and facilities for examination of material at K.
LITERATURE CITED
Bas, C. 1969. Morphology and subdivision of Amanita and a monograph of its section Lepidella. Persoonia 5(4): 285-579.
Beeli, M. 1930. Notes mycologiques. Champignons nouveaux pour la flore Belge II. Bulletin de la Société Royale de Botanique Belgique 62(2): 127-132.
Coker, W. C. 1917. The amanitas of the eastern United States. Journal of the Elisha Mitchell Scientific Society 33(1-2): 1-88 + plates.
An additional taxon possibly belonging to this group is known to me only from its protologue: Amanita ovoidea var. ammophila Beeli (1930: 129). The spore dimensions given by Beeli suggest a much thinner spore than is found in A. ovoidea: 7 - 10x 4-5 um. It appears average Q = 1.9+. No type is designated; but a collection made in September in Genck, Belgium is cited.
. 1927. New or noteworthy basidiomycetes. Journal of the Elisha Mitchell Scientific Society 43(1-2): 129-145.
Filippi, I. 1985. Una nuova Amanita dell’area mediterranea. Micologia Italiana 3: 15- 19.
Garrido, N. and A. Bresinksy. 1985. Amanita merxmuelleri (Agaricales) eine neue Art aus Nothofagus-Waldern Chiles. Botanische Jahrbiicher fiir Systematik, Pflanzengeschichte und Pflanzengeographie 107(1-4): 521-540.
Gilbert, J.-E. 1940 & 1941. Amanitaceae. Iconographia Mycologica. 27. xx + 427 pp. + pls.
Jenkins, D. T. 1978. A study of Amanita types I. Taxa described by C. H. Peck. Mycotaxon 7(1): 23-44.
. 1979. A study of Amanita types III. Taxa described by W. A. Murrill. Mycotaxon 10: 175-200.
. 1982. A study of Amanita types IV. Taxa described by G. F. Atkinson. Mycotaxon 14(1): 237-246.
. 1986. Amanita of North America. Mad River, Eureka. vi + 198 pp.
Kornerup, A. and J. H. Wanscher. 1978. Methuen handbook of colour. Methuen, London. 252 pp.
Kiihner, R. and H. Romagnesi. 1974. Flore analytique des champignons supérior. Masson et Cie., Paris. xiv + 557 pp.
Marchand, A. 1973. Champignons du nord et du midi. Hachette. 2: 273 pp.
Merlo, E. G. and M. Traverso. 1983. Le amanite. Sagep Editrice, Genoa. 151 pp.
Munsell Color. 1975. Munsell soil color charts. Baltimore.
Pegler, D. N. 1983. Agaric Flora of the Lesser Antilles. Kew Bulletin Additional Series IX. vi + 668 pp.
Pérez-Silva, E., T. Herrera and G. Guzman. 1970. Introduccién al estudio de los macromicetos téxicos de México. Boletin de la Sociedad Mexicana de Micologia 4: 49-53.
Raithelhuber, J. 1986. Amanitaceae in Sudamerika. Metrodiana 14(1): 3-21.
Rayner, R. W. 1970. A Mycological Colour Chart. Kew, Commonwealth Mycological Institute.
Singer, R. 1958. Fungi mexicani, series prima—Agaricales. Sydowia 11(1-6): 354-374.
MYCOTAXON
Vol. XXXVI, No. 1, pp. 9-14 October-December 1989
LECANORA SECT. PETRASTERION (LICHENIZED ASCOMYCOTINA) IN NORTH AMERICA: LECANORA WEBERI RYAN, SP. NOV. (SUBSECT. PSEUDOCORTICATAE) , FROM COLORADO
BRUCE D. RYAN
Department of Botany, Arizona State University, Tempe, Arizona 85287 U.S.A.
ABSTRACT A new crustose-squamulose lichen, Lecanora_ weberi Ryan, sp. nov., is described from rocks in the vicinity of Boulder, Colorado. It is treated here
under sect. Petrasterion subsect. Pseudocorticatae Poelt, and is probably closely related to L. novomexicana Magnusson, but is distinguished especially by its short, rather flattened and squamule-like lobes, which are grayish tinged and have a very thin upper cortex. It is also similar to forms of Rhizoplaca melanophthalma (DC.) Leuck. & Poelt sensu lato, but lacks a distinct umbilicus and lower cortex, and has adnate apothecia with orangish to yellowish brown discs and scarcely prominent margins.
This paper describes a new crustose-squamulose species of Lecanora, from Colorado. Although the new species resembles forms of both Lecanora novomexicana Magnusson sensu lato and Rhizoplaca _ melanophthalma (DC.) Leuck. & Poelt sensu lato, it does not fit well within either, and retains its identity when growing side by side with then. The type material has already been distributed (as "Lecanora sp. nov. Ryan") in the exsiccati series from the University of Colorado (COLO), and a diagnosis and description of the new species are given now. Until the relationship between the genera lLecanora Ach. emend. Massal. and Rhizoplaca Zopf can be clarified by further investigations, the supraspecific classification of the several apparently intermediate taxa (including L. nigromarginata Magnusson and L. opiniconensis Brodo, as well as the new species described below) is uncertain. For the present, taxa such as these, which lack a distinctly umbilicate-foliose thallus and well-developed lower cortex, appear to be best placed within Lecanora subg. Placodium (Pers. emend. Poelt) Poelt. The new species described here 1S; treated: under ect. Petrasterion subsect. Pseudocorticatae Poelt.
Unless noted otherwise, the methods and terminology used in this paper are as described by Ryan (1989a,b).
10
Colors (followed by numbers in parentheses), as viewed through a dissecting scope with fiber optic lighting, refer to the system of Kelly (1965). Chemical analyses were made by the standard thin-layer chromatographic (TLC) method of Culberson (1972), modified as described by Ryan (1989a).
Lecanora weberi Ryan, sp. nov. (Fig. 1, 2)
In rubipus siliceis crescens, Colorado. Thallus areolatus ad squamosus, squamulis marginalibus eis centri subsimilibus vel nonnihil elongatis et lobiformibus, olivaceo-griseis, marginibus pallide flavovirescentibus instructis, non vel leviter pruinosis. Apothecia adnata ad sessilia, discis aurantiacis ad flavovirescentibus pruina flavoalbida tectis, margines leviter prominentes, continui ad crenati. Cortex superior valde tenuis, cellulas algarum emortuas includens; fasciculi hypharum subcorticales desunt vel non evoluti; cortex granulis flavescentibus inspersus; cortex inferior deest vel indistincte evolutus. Paraphyses subconglutinatae, apicibus clavatis; sporeae octonae, ellipsoidae, ca. 8-11 x 4-6 pm. Spermatia filiformia, + curvata. Continet acidum usnicum in cortice, acida pingua (acida allo-pertusaricum, et al.) in medulla.
TYPE: Ua (iSciAwe COLORADO: Boulder Co.: Boulder Mountain Parks, just S of city of Boulder, 1850 m, on scattered low fine-grained sandstone or quartzite boulders at base of Flatiron Mountains, in open stands of Pinus ponderosa, 10 Aug. 1986, W. Weber, COLO Exs. 685, Holotype (COLO!), Isotype (ASU!).
THALLUS areolate-squamulose, indistinctly radiating, closely appressed, not easily removed intact; areoles scattered to contiguous or weakly imbricated (fertile thalli) or becoming densely imbricated (sterile thalli); lobes similar to central areoles or somewhat larger and more lobe-like or squamule-like, isodiametric to moderately elongated, 1-2 mm long, 0.5-1.5 mm wide, simple or slightly branched, often strongly crenate, partly overlapping each other, flat, to concave or) ,undulate 0.20). 3) nm cnack. appressed to slightly ascending, broadly adnate at least on side toward thallus center, often becoming free on side toward thallus margin; thallus center areolate-verrucose or partly areolate-squamulose in fertile thalli; areoles flat to slightly convex or undulating, with more-or-less
Figures 1-4. Lecanora weberi Ryan and similar taxa. Scale = mm. -1. Part of the holotype of L. weberi (COLO Exs. 6855). (COLO): 2). Part of a collection of L. weberi (Shushan and Weber S-3218, M), showing fertile thalli with verrucose areoles (upper right) and sterile thalli with
imbricate squamules (below). -3. A specimen of Lecanora novomexicana Magnusson from the type locality of L. weberi (Weber, S.n.,)' (COLO). A, Subcrustose form of Rhizoplaca
melanophthalma (DC.) Leuck. & Poelt sensu lato from the type locality of L. weberi (Weber, s.n., COLO).
11
PULCAVAL TEAL DT ETE
|
i
titi
|
L
PELLEL
1
|
iit
i
|
WH
{
Hii
i
biitt
i
|
Plaid
I
I
12
downward-turning margins, irregular in outline, to 1(-2) mm wide, sometimes rimosely divided into secondary areoles, 0.2-0.5(1.5) mm thick, thinner toward edge, becoming crenate, usually narrowed below, broadly attached to substrate centrally or more towards one side, by about one
fourth to one half the lower side, the margins often becoming free from substrate (especially in sterile thalli); color above in fertile thalli when fresh usually with distinctly grayish green tone, light grayish olive (109) to light olive gray (112), paler and more grayish towards thallus margin, with the squamules edges mostly pale yellowish green (121) to pale greenish yellow (104), in older herbarium specimens fertile thalli grayish yellowish green (122) to grayish greenish yellow (105), partly with orangish yellow tinges, and sterile thalli grayish yellow (90) to moderate yellow (87); upper surface mostly epruinose, more-or-less matt; squamule edges often somewhat roughened, with patches of whitish pruina; color below pale, more-or-less moderate yellowish brown (77), darker brown near attachment area; upper cortex with few to many dead algal cells, very thin, 5-10(-15)pm, even, without conelike hyphal bundles, mostly inspersed with fine yellowish granules (soluble in kK); algae trebouxioid; algal layer 30-50(-75) pm thick, continuous to interrupted; medulla moderately loose, partly filled with clumps of gray granules; hyphae 3-5 pm diam.; lower cortex absent, or (on ascending squamules) 12-15 pm thick, often poorly developed, broken and penetrated by medullary hyphae; hyphae ca. 5 pm diam., lumina 2 pm.
APOTHECIA usually numerous and becoming crowded (but sometimes absent or rare), to three per areole, 1-1.3 mm diam., borne submarginally to laminally on the squamules, adnate to broadly sessile; disc often slightly concave when young, becoming plane to undulate when old, usually densely whitish-yellowish pruinose, but often epruinose when young, grayish yellow (90) to yellowish gray (93) or sometimes almost pale greenish yellow (104) with pruina, light to deep orange (51-52) to moderate yellowish brown (77) under pruina, often darker and more olivaceous when young; margin about 0.1-0.2 mm thick, soon irregularly with irregularly foveolate surface, entire or somewhat crenate, becoming flexuous or distorted in age, sometimes distinct and more- or-less raised from the start, persistent, often somewhat paler than thallus and weakly pruinose; hymenium hyaline, 50-65 pm high; epihymenium brownish, weakly inspersed with fine granules (partly soluble in K) and covered by 15-20 pm thick superficial layer of coarse granules (soluble in K); paraphyses distinct and more-or-less free in water, about 1.5-2 pm thick below, the tips 2-2.5 pm thick, colorless; subhymenium grayish, with oil droplets; excipulum hyphae gelatinized but somewhat distinct in water, densely packed, irregularly oriented, ca. 3-5(-7) pm diam., the lumina more-or-less short, 1 wm wide; parathecium hyphae parallel; hypothecium distinct from subhymenium, hyaline to slightly yellowish-brownish, to 300 pm thick in center; amphithecium similar in structure to thallus but cortex to 30-50 pm
13
thick in lower part; algal layer about 50 pm thick below hypothecium, continuous to interrupted; algae filling most of the margin; asci 35-40 x 10-12 pm, Lecanora-type; spores ellipsoid to oblong-ellipsoid (L:W = 1.8-2.4:1), about 8- 11(-13) x (3-)4-6(-7) pm, with one or two oil droplets, 8 per ascus, the wall about 0.5 pm thick.
SPERMOGONIA immersed, the ostiole pale, the cavity ca. 100-150 um diam.; spermatia curved, (15-)20-25 pm long.
SPOT TESTS AND CHEMISTRY: Thallus and apothecia C-, Pd-; cortex K- or + yellowish; KC+ yellowish; with usnic acid (often only traces); medulla K-, KC-, I5jKI-; with fattyviacidas Mof, thei cprotolichesterinic’) group! (allo-
pertusariciwiiand) dihydropertusaric;, constipatic, protoconstipatic and dehydroconstipatic acids), and sometimes traces of unknowns; discs K-, KC-. A specimen
was analyzed by C. Leuckert (Shushan & Weber S-3218, COLO).
DISTRIBUTION AND ECOLOGY: Montane areas of western U. S204. 4 1(Col orado, to) idaho! (andi S. (Dakota); on. siliceous rocks (sandstone, conglomerate, quartzite, arkosite), usually in moderately shaded areas, on horizontal to sloping faces near tops of boulders or outcrops, in open forest, sometimes in canyons, 1050-1875 m; often very abundant, covering whole level surfaces of boulders; associated organisms include the vascular plants Pinus ponderosa or P. edulis, or Cercocarpus montanus, and the lichens Lecanora muralis, L. novomexicana, L. sp. (L. polytropa group?), Rhizoplaca melanophthalma sensu lato R. chrysoleuca sensu lato (" subdiscrepans" morphotype), Acarospora fuscata, Aspicilia caesiocinerea, Candelariella spp., Dimelaena oreina, Lecidea atrobrunnea, Melanelia spp., and Xanthoparmelia spp.
OTHER REPRESENTATIVE SPECIMENS EXAMINED: Ust Swe A. COLORADO: Boulder Co.: near Boulder, Shushan & Weber S- 3218 (COLO, M, WIS); near Lyons, Weber L-72985 (COLO); Garfield Co.: 7 km N of Rifle, Ryan 20678 (ASU); Larimer Co.: km SW of Ft. Collins, Anderson S-20272 (COLO); Moffat
CO. 2b oa kmuN of CAxXial. Ryan: 320637 (ASU). IDAHO : Bonneville Co.: Palisades Reservoir, Ryan 19590 (ASU). SOUTH DAKOTA: Pennington Co.: 6.5 km NW of Hermosa,
Wetmore 10285 (MIN, UPS).
DISCUSSION: The new species is named after the collector of the type material, William Weber of the University of Colorado at Boulder, in honor of his work on lobate species of Lecanora in North America.
The two taxa with which L. weberi is most likely to be confused are L. novomexicana sensu lato and Rhizoplaca melanophthalma sensu lato (Figs. 3 and 4, respectively). All three taxa have pruinose apothecia and an upper cortex containing dead algal cells, and contain usnic acid and fatty acids of the pertusaric/constipatic group. At the type locality of L. weberi and other locations, the three taxa grow side by side, yet each remains distinct, with a different overall appearance produced by a combination of features. Lecanora weberi differs from the other two
14
species especially by its very thin cortex and lack of psoromic acid, and by its more grayish, pale-edged,
frequently imbricated squamule-like lobes. Its marginal lobes are shorter, flatter and more loosely attached than those of L. novomexicana. Lecanora weberi differs from R.
melanophthalma in that the new species lacks a distinct umbilicus and lower cortex and has apothecia that are adnate, with orangish to yellowish brown discs and scarcely prominent margins. Many of the characteristics of L. novomexicana and R. melanophthalma (e.g., chemistry, growth form, and colors of the thallus and apothecia) that are useful for distinguishing those species from L. weberi at the type! locality (of the) latter ‘do’ not! apply (at other localities. Especially in the case of R. melanophthalma, the material from the type locality of L. weberi is not very typical. In fact, both of these other species (as presently delimited by most authors) are extremely variable and plastic, can also be very difficult to distinguish from each other, and may well consist of several taxa.
The morphology and color of the thallus of L. weberi itself are also somewhat variable. Especially interesting is the difference between the more common, predominantly fertile forms (areolate-verrucose thallus center, squamulose-lobed margin) and the occasional, mostly sterile forms (more-or-less imbricate squamulose throughout) that occur on or next to the parent thalli. The origin of such dimorphism, which also occurs in various other lobate taxa, is unknown.
ACKNOWLEDGEMENTS This study was supported by awards from the National
Science Foundation (#BSR-8511556), Smithsonian Institution, Arizona Federation of Garden Clubs, and Arizona State
University Graduate Student Association. Thanks go to the curators of the following herbaria: ASU, COLO, M, MIN, UPS, WIS. Special thanks also go to the Arizona State
University Media Development center for photography, C. Leuckert for chemical analyses, J. Flock for help with fieldwork, and I. M. Brodo, T. H. Nash III, J. Poelt and W. Weber for comments on the manuscript.
LITERATURE CITED
Culberson, C. 1972. Improved conditions and new data for the, identification’ of Lichen, products by “a standardized thin-layer chromatographic method. J.
Chromatogr) 72 (bs -125¢
Kelly, K. L. 1965. ISCC-NBS Color-Name Charts Illustrated with Centroid Colors. Supplement to NBS Circular 553. Washington, DC.
Ryan! By 1989a. The genus Cladidium (LIchenized Ascomycotina). Mycotaxon 34(2): 697-712.
Ryan, B. 1989b. A monograph of Lecanora sect. Endochloris (Lichenized Ascomycotina). The Bryologist (accepted
for publication).
MY COTAXON
Vol. XXXVI, No. 1, pp. 15-18 October-December 1989
TWO NEW SPECIES OF THE GENUS ASCOCHYTA LIB.
SIMEON G. VANEV AND GANKA G. BAKALOVA Institute of Botany, 1113 Sofia, Bulgaria
ABSTRACT. Two new species of genus Ascochyta Lib. - A.urticicola Vanev et Bakalova, sp.nov. and A.digitalina Vanev et Bakalova, spenov., from Bulgaria, are described and illustrated.
During inventory investigations of genus Ascochy- ta Lib. (order Sphaeropsidales, class Deuteromycetes) in Bulgaria, two new species belonging to this genus were es- tablished.
ASCOCHYTA URTICICOLA Vanev et Bakalova sp.nov.
Maculae 0,1-0,8 cm in diam., orbiculares vel an- gulatae, solitariae interdum comfluentes, atro-brunneae vel nigrae, saepe concentric zonatae. Pycnides epiphyllae, Ssparsae, solitariae, globoSae vel depresso-globosae, palli- de brunneae, 110-1804¢m in diam., poro orbiculari 25-35 Mmm in diam., cellulis parvis obscurioribus cincto. Conidia cylindrica vel elliptica utrinque rotundata, uni-vel bisep- tata, saepe ad septum leniter constricta, hyalina, (10) 195-411. 5 21) 2 4-5(6)wm (Fig. 1A).
Im foliis Urticae dioicae L., Bulgaria, mons Pi- rin, FPredela, 26.09.1975, S.G.Vanev, G.G.Bakalova, SOM 18962 M, holotypus.
Leaf spots 0,1-0,8 cm in diam., numerous, roun- ded or angular, single or often confluent, dark brown or black, more pale in the centre, occasionally concentric. Pycnidia spheric or flattened, single, scattered, pale- brown, ae ee With icipentar ostiol, ae ah in vole.
16
surrounded by zone of darker cells. Conidia cylindric with rounded both ends or ellipsoidal, straight or slightly cur- ved, hyaline, 1-2(rarely 3)septate, occasionaly constric- ted at the septa, (10)13,5-17 x 4-5(6) pm
Specimen examined: on living leaves of Urtica di- oica 1l., Bulgaria, Pirin mount., Predela, 26.09.1975, S.G. Vanev and G.G.Bakalova, SOM 18962 M (holotype).
There is only one Ascochyta species registered on plants belonging to genus Urtica- A.urticee A.L.Sm.et Ramsb.
Melnik(1977) delimited 2 subgenera of Ascochyta on the basis of the number and disposition of conidial sep- ta. According to this author A.urticae belongs to subgenus Libertia Melnik since the conidia have only 1 central sep- tum dividing the conidium into 2 nearly equal cells.
Ascochyta urticicola clearly differs from A.urti- cae in the size of conidia and the number of the conidial Septasiil Or 2, Tarely oo) (lao len a),
Table 7. Comparative morphologic data of Ascochyta urticicola and A.urticae
Dimensions of Number of Subgenus Species conidia in ane septa in conidia
Ascochyta | A.urtici4 (10)13,5- cola 17 5X21)
Libertia A.urticae| 7-12
(by Mel- nik,1977)
For this reason it is necessary to place A.urti- cicola in subgenus Ascochyta Melnik.
There are 2 other Ascochyta species parasitic on plants of the same family (Urticaceae)- A.parietariae
17
Roum.et Fautr.(on Parietaria officinalis L.) and A.boehme- riae Woronichin (on Boehmeria ssp.) but they differ from A.urticicola not only in the host-plants but morphologica- lily also.
ASCOCHYTA DIGITALINA Vanev et Bakalova, sp.nov.
Maculae 0,3-1,5 cm in diam., orbiculares, solita- riae, globosae vel confluentes, ochraceae vel pallide bru- nneae, indistincte atropurpureo-marginatae. Pycnides epi- phyllae sparsae, solitariae, globosae vel depresso-globo- sae, pallide brunneae immersae, 110-175am in diam. poro orbiculari 20-35 em in diam., cellulis atrofuscis cincto. Conidia cylindrica utrinque rotundata, interdum oblongo- elliptica vel: clavata, recta vel leniter curvata, distin- cte uni-vel biseptata, ad septum haud vel vix constricta interdum inaequaliter bicellulata, hyalina, (12,5)15-17,5 x (4)4,5-5(6) mom (Fig. 1B).
in toliis vivas Dveltalidis vivid. tiorge Lindr., Bulgaria, mons Rodopi, Beglica, 2.08.1981, S.G.Vanev, SOM 18964 M, holotypus.
Leaf spots 0,3-1,5 cm in diam. rounded, single or confluent, ochraceous or palebrown with dark- brown or dark-violaceous wide border. Pycnidia more or less spheric, separate, scattered thin-walled, pale-brown, 110-175 in diam., with a single, circular ostiol 20-354. in diam., surrounded by darker cells. Conidia cylindric with rounded both ends, or ellipsoidal, rarely clavate, straight or sli- ghtly curved, hyaline, 1-2-septate, occasionally constric- ted at the septa (sometimes with 2 unequal cells), (12,5 15-17,5 x (4)4,5-5(6) mom
Specimen examined; on living leaves of Digitalis viridiflora Lindr., Bulgaria, Rodopi mount., Beglica, 2.08.1981, S.G.Vanev, SOM 18964 M, (holotype).
Two different Ascochyta species parasitic on plants of the genus Digitalis are described in the litera- ture- A.digitalis Fuckel and A.moelleriana Wint. Melnik (1977) has studied the type specimens of both species and has rejected A.digitalis as a misdetermined species. Acc-
18
ording to this author A.moelleriana is a later synonym of A.euphrasiae Oud. He has placed A.euphrasiae in subgenus Libertia because of the t-septate conidia.
The new species A.digitalina belongs to subgenus Ascochyta and clearly differs from A.euphrasiae in size of
conidia and the number and disposition of the septa (Ta- ble i2)..
Table 2.
Comparative morphologic data of Ascochyta digitalina and A.euphrasiae
Dimensions of Subgenus Species conidia in jem
Gi) 5) 15 17,5(20)
Libertia | A.euphrasiae (Melnik, 1977)
Fig.1 Conidia of: A. Ascochyta urticicola
B. A. digitalina
LITERATURE CITED
Melnik V.A. 1977. Opredelitel gribov roda Ascochy- ta Lib. Nauka, Leningrad, 245 p.
MY COTAXON
Vol. XXXVI, No. 1, pp. 19-27 October-December 1989
TYPE STUDIES IN MARASMIOID AND COLLYBIOID FUNGI (TRICHOLOMATACEAE ) II. AGARICUS GRAMINUM
Vladimir Antonin Moravian Museum, Dept. of Botany Alam. 25 Uno Laul6, 1659) 137) (Bono Czechoslovakia
ABSTRACT
The results of the revision of the type specimens of Agaricus graminum Libert /= Marasmius graminum (Libert)Berk. et Br./ are published. Agaricus graminum in the original Ssenaemor lLibert. differs from: Manasmius gramigum) in sense of contemporary authors. It is proposed to use the name Maras- PMUISeCULLeY VDELK (Ot OE a TOnV the Latiers
During preparation of a monograph of Central-European species of the genus Marasmius and type studies in these fun- gi, I was very surprised to find that the type specimen of Agaricus ghaminum Libert distributed in ithercollection of exsiccata "Plantae Cryptogamae Arduennae" differs from Ma- rasmius graminum in sense of contemporary authors. I have found the same fungus in the Favre s herbarium in Geneve, which was published by Favre (1960) as a bisporic form of M. graminum. Marasmius graminum in the sense of contemporary Authors isa cosmopolite species and all modern mycologists have this taxon in the same conception. Therefore, this fact mentioned above is very interesting. In the present paper, I Give. the descriptions of both species and also the reasons POL va, Proposal to)iselect jasq correct’ mame .Marasmius curreyi Berk. et Br. for the distinguished second species.
Marasmius graminum (Libert) Berkeley et Broome Agaricus ograminum\ibert,, Pl. Crypt. Arduennae, (Fasc. I, Now 2ho. eae: Marasmius graminum (Libert) Berk. et Br., Outl. Brit. bung wei ondony Com i22 26 heed).
Macrofeatures (partly according to Favre s notes and partly from the type-specimens): PILEUS up to 4.5 mm broad but usually smaller, convex, opaque, with or without papilla, plicate, red-brown with the darker centre. LAMELLAE distant, L = (4-)6-8, 1 = 0, collariate, collarium very fine but prominent, not pure white but cream-whitish. STEM up to 25 mm long, filiform, smooth, lustrous, whitish above, black- brown to almost black below.
20
Figs. 1-3. Marasmius graminum (type-specimen): 1. Pileipel- Lisi. 2. Cheilocystidia. 3. Basidiospores. Scale bar =
10 um.
ra
Microfeatures (figs. 1-3): BASIDIOSPORES ellipso- id, broadly ellipsoid to slightly amygdaliform, indextrinoid, hyaline, thin-walled, smooth, of; twor sizes: 7=12.-x%.2.5-4 um and (9-)11-16(-18) x (5-)6.2-8(-10) um; the second one is mo- re common. BASIDIA clavate, clamped, 2- and 4-spored, e.g. 24 x 8 um. BASIDIOLES clavate, broadly clavate, fusiform, so- metimes with a broad and obtuse top, thin-walled, clamped, 13-29(-32) x (4.5-)6-10(-11) um. CHEILOCYSTIDIA of the simi- lar form as the broom cells of the pileus surface, clamped, thin-walled, 16-27 x (5-)6-10 um, projections up to 2-4(-5) um long. PLEUROCYSTIDIA none. SUBHYMENIUM of thin-walled, branched, cylindric, slightly dextrinoid hyphae, 2-4 um broad. TRAMA of the lamellae of cylindric, thin-walled, dextrinoid, less branched, clamped, 3-8 um broad hyphae; flesh of the pi- leus of interwoven, thin-walled, clamped, more or less cylin- dric, branched, slightly dextrinoid hyphae, sometimes finely incrusted,.2.2-9 um broad; flesh of) the istem in cortex of pa- rallel, slightly dextrinoid, cylindric, clamped, slightly thick-walled (up to 0.8-1.2 um), 3-4 um broad hyphae, on the surface smooth, in medulla of parallel, dextrinoid, cylind- ric, thin-walled (up to 0.5 um), clamped, 2-8 um broad hy- phae. PILEIPELLIS hymeniform, of cylindric-clavate, clavate, broadly clavate to almost globose cells, clamped, thin-wal- led, 13-28 x 8-24 um, with irregular, simple to corraloid branched projections above, 1-3(-6) um long, these broom cells mostly of the Rotalis-type but sometimes of a type transient to Siccus-type, the upper part of broom cells and the projections brown pigmented; between these cells are so- metimes almost smooth to smooth cells and rare thin-walled hyphae with 4-6 um long projections.
Material revised:
BEoLUN- EAs worypt. Arduennae,! Fasc ig Dis); No.l Libert,, feoZour key 107065). (Lectotype) ,BP I, BP) belss.,) BRe Ky CutiAga- ricus graminum).
SULSSOELo Entre: le God Trid ets ie, bod Purchers vers! 1900: m; val Trupschun pr. S-chauf, Hm Engadine, dans une aunaie sur graminées pourissantes, 9.1X.1955, J.Favre, G 14465 (ut Ma- rasmius graminum forme bisporique).
This species is characterized macroscopically especial- ly by the low number of lamellae and microscopically by the size of spores, character of broom cells of the pileus epi- cutis and especially of their projections.
The type-specimens from Libert s collection "Plantae Cryptogamae Arduennae" are preserved in many herbaria. The specimen from BPI was designated as an "isotype" (Gilliam, 1976). However, all the specimens of these exsiccata must be considered syntypes. The "isotype" (BPI) consists of two parts of the stem, with the absence of pileus, and thus this specimen has not any significance as a type. I revised these type-specimens from 5 important herbaria (BP, BPI, BR, K, PRM); the best specimen of them was that from the herbarium of the National Museum in Prague. Therefore, I propose it (PRM 707065) as a lectotype. However, the best revised ma-
22
terial of this species was the Favre s collection from Suisse (G 14465)
The oldest synonym for Marasmius graminum treated in va- rious papers and monographs of marasmioid fungi is Marasmius pruinatus Berk. et Curt. 1859 (not M. pruinatus Rea 1916).
In monographs of Singer (1958, 1965, 1976), this name is men- tioned as a synonym while Gilliam CL9OTG6) after the type stu- dy of the holotype-specimen (K) and authentic material of Berkeley et Curtis (FH) considers Marasmius pruinatus an in- dependent species. According to this author, this species differs from M. graminum especially by the finely divided projections on the cuticular cells of M. pruinatus which con-= trast with the broader, discrete projections of M. graminum.
I have revised the specimen of M. pruinatus from Far- low Herbarium and my results agree with Gilliam s solution. The microfeatures were the following (figs. 4-5): BASIDIO- SPORES clavate to drop-shaped, indextrinoid, hyaline, thin- walled, smooth, 10.1-14.2 x 3.2-4.2 um. BASIDIA clavate, 4- spored, clamped, 21-24 x 6.2-7.7 um. BASIDIOLES clavate, fu- siform, clamped, often with prominent obtuse top, thin-walled, 14-26 x 4.7-8 um. CHEILOCYSTIDIA of the similar form as the broom cells of the pileus epicutis, clavate, with more or less nodulose projections, thin-walled, e.g. 16 x 7.5-8 um. PLEUROCYSTIDIA none. SUBHYMENIUM of dextrinoid, thin-walled, clamped, 2-3.5 um broad, hyaline hyphae. TRAMA of the lamel- lae of dextrinoid, clamped, thin-walled, sometimes branched, 2.5-7 um broad hyphae; flesh of the pileus of dextrinoid, thin-walled, branched, clamped, hyaline,3-6 um broad hyphae; flesh of the stipe of parallel, clamped, dextrinoid, 3-7 um broad, thin-walled (in medulla) and thick-walled (up to 1-1.5 um in cortex) hyphae; surface of the stipe smooth. PILEIPELLIS hymeniform, of broom cells of the Siccus-type, cells clavate, sometimes clamped, thin-walled to thick-walled in the upper part, withyup to’ 5.5 umiilong, more or less) nodulose, ‘obtuse projections, 9-14 x 5.2-8 um; some cells coralloid branched with some thick-walled projections.
These microfeatures show unambiguously that this species belongs to the section Sicci. These results agree with con- clusions of D.E.Desjardin on the revision label dated 3.I1. 19868) and; LO SUPE ooo.
Therefore, I have studied the type-specimen of the second older known synonym - Marasmius curreyi Berk. et Br. 1879. In my opinion, features of this specimen as well as Cooke s ta- ble 1130B (Cooke, 1890), agree well with descriptions of the common "M. graminum" in the sense of contemporary authors and both taxa are conspecific. According to Dennis Clo Maras- mius exustus Berk. et Curt. 1879 could be identical with "M, graminum”, too. The type-specimen is preserved in FH (not in K). However, according to Singer (1976) who revised this ty- pe, M. exustus represents a species from the section Hygrome- trict). Therefore, I propose to use the name Marasmius cur- reyi a6 the| correct’ one: for the species’ called: “Miaiinaminun: now. A full description of this species follows now.
23
Marasmius curreyii) Berkeley et Broome, Ann. Mag, Nat. AUS SV Pa ZO9K, OMB SY
Marasmius graminum (Libert) Berk. et Br. ss. auct., e.g. Gilliam (1976), Singer (1976). Marasmius tritici Young, Phytopathology, 25:116, 1925.
Macrofeatures: PILEUS 4-6 mm broad, conical-hemispheri- cal, then convex to expanded, sometimes with slightly revolu- te margin, with a small papilla, especially when young, then often with small umbilicus, smooth, under lens slightly to- mentose, slightly plicate, young red-brown to rusty-brown to pinkish-brown. LAMELLAE distant, L = 9-13, 1 = 0, collaria- te, very rarely without distinct collarium, with concolorous edge, white. STEM 18-26 mm long, filiform, lustrous, smooth, whitish to white above, through brown to dark red-brown to black-brown toward the base.
yiiil FG ao
Figs. 4-5. Marasmius Tuinatus (auth i i - : MEE SLE Oy entic mater : ripe leipellis. 2. Basidiospores. Scale bar = 5 a ba i
24
Figs. 6-8. Merasmius, Curreya: 6, Pileipellis. 7) /Ened locyve. tidia. 8. Basidiospores. Scale bar = 10 um.
Microfeatures (figs. 6-8): BASIDIOSPORES ellipsoid, cylindric-ellipsoid to slightly amygdaliform, smooth, thin-wal- led, hyaline, (7.7-)8-11(-13) x 4-5.7(-6.5) um. BASIDIA clava- te, clamped, 4- rarely 2-spored, 15-30 x 4-7 um. BASIDIOLES clavate, cylindric-clavate, slightly fusiform, clamped, 18-28 x 4-8 um. CHEILOCYSTIDIA of the similar form as the broom cells of the pileipellis, more or less clavate, hyaline,
25
10-16(-20) x 6-12 um, with 1-7(-8) um long, hyaline to slightly yellowish projections above. PLEUROCYSTIDIA none. SUBHYMENIUM of thin-walled, cylindric, clamped, branched, hyaline, 2-4 um broad hyphae. TRAMA of the lamellae of dex- trinoid, thin-walled, clamped, hyaline, 3-8 um broad hyphae; flesh of the pileus of dextrinoid, branched, interwoven, thin-walled, clamped, hyaline, 3-7(-10) um broad hyphae; flesh of the stem in cortex of parallel, clamped, slightly thick-walled (up to 1 um), yellow-brown to brown pigmented, in medulla dextrinoid, parallel, thin-walled, clamped, hya- line, 2.2-8 um broad hyphae. PILEIPELLIS hymeniform, of broom cells of the Siccus-type, clavate or short cylindric-clava- te, some of them thin-walled and hyaline, other (usually a great part of them) slightly thick-walled above, 9-22 x (6-) 8-14(-18) um, with 2-5(-7) um long, obtuse projections, cells hyaline below, yellow-brown obove and in the projections.
Habitat: on remnants of Poaceae, Juncaceae, Cypera- Cceae and rarely of some other plants.
Beet Cr bad Ce.viics.e d : BELGIUM: Westerloo (prov. Antwerpen), 1941, Tuymans (BR). - Bruxelles (Brabant), 1891, Delogne (BR). CZECHOSLOVAKIA: Dolany near Unho&St, 1939, Herink (PRM 139045) - Repy mear Praha, 1952, Pouzar (PRM 707068). - Praha-7iz- kov, 1968, Svréek (PRM 671895). - Praha, Kinského sady, 1963, Wichansky (PRM 600982 and 624000). - Praha, Stromovka, 1952, Pouzar (PRM 707057); 1948, Vacek (PRM 707075). - Mnichovice, 1918, Velenovsky (PRC and PRM 707061). - Mnichovice, MySlin, 1939, Velenovsky (PRM 153986). - Mnichovice, Hubaéov, 1939, Velenovsky (PRM 154305). - Libochoviéky, 1916, Velenovsky (PRC and PRM 707062). - KroGehlavy near Kladno, 1941, Herink (PRM 707064). - Cernolice near Dobfichovice, 1950, Pouzar (PRM 707069). - Mofinka near Dobfrichovice, 1947, Svréek (PRM 707080). - Pyskoéely near Stfibrnd Skalice, 1951, Pouzar (PRM 707059, 707067, 707070, 707071). - Ruda near Nové Stra- Semi loo, terink CPRM 159124). = Poriéko, (1950, *Pouzar CPEvo 7070/2). —) Kosot, 1950, Vacek ‘CPRM.707073).)'-. Libfice, 1941, Herink (PRM 707060). - KarlStejn, 1946, Vacek (PRM 707081); Svréek (PRM 707078). - Draheléice, 1947, Vacek (PRM 514995). - Hornfi Slavénice near Lomnice n. Luz., 1962, Svr- Gek (PRM 567936). - Smrzov near Lomnice n. Luz., 1961, Ku- biéka (PRM 616353); 1962, Svréek (PRM 567935). - Hamr-Kosky, 1980, Kubiéka (CB 2344). -_ Zdbofi near Blatnd, Skalicky CPRE). -iLutova pear Trebon, 1945,' Svréek (PRM 707126)%)- Tfebon, 1979, Kubiékova et Kubiéka (CB 2161). - Vodnany, 1937, 1938 and 1943, Herink (PRM 490571, 490566, 499642 and 707077). = Cimelice near Pisek, 1961, Svréek (PRM 616352). - LazisSté near Pisek, 1963, Svréek (PRM 612757). - Vrdabsko near Cimelice, 1966, Svréek (PRM 626072 and 626073); 1963, Pouzar (PRM 612756). - Zvikovské Povltavi, 1954, Pouzar (PRM 617489). - Golétv Jenikov, 1940, Herink (PRM 707063 and 707066). - NemySl near Tabor, 1943, Svréek (PRM 707058). - Vidnava (Wei- denau), 1912 and 1919, Hruby (BRNM 07311/39 and 07310/39). - Bojkovice-Bzova, 1985, Antonin (BRNM). - Kufim, 1942, Smar- da (BRNM 313942). - Zdravda Voda near ZaroSice, 1947, Vacek
26
(PRM 707076). - Prenéov, 1898 and 1901, Kmet (BRA). - Tur- Ciansky Martin, 1951 Smarda (BRNM 313941). “RaCa ibook; Krippelova et émarda (BRA). - Zemianské Podhradie (Ns.Podhra- gy), 1884, Baumler (BP 19704).
ENGLAND: on Rye, Cooke (K). - Kew, Royal Bot. Gardens, 1966 (K). - Coughton (Warwicks), Clark (K). - Canseway, Slapton (Dorset), 1981, Clark (K). - Arington Court (N.Devon), 1978, Clark (K). - West Nolesey (Surrey), 1979, Spooner (K). - Ha- le Barnes Playing Fields (Cheshire), 1980, Newton (K). - Kew, 1912 (K). - Arbrook Common, Esher (Surrey), 1981, Spooner (K). - Holm Fen (Huntongdonshire), 1965, Houtton (K). - Cran- bourne Park, Windsor Park (Berkshire), 1967, Dennis and Reid (K). - Lickey Hills (Worcestershire), 1968, Price (K). - Maidenhead (Berkshire), 1970, Verdcourt (K).
FIJI: Viti Levu, Savura Creek, 1976, Maddison and Kirby (K). FRANCE: Santes, Courtecuisse (Herb. Courtecuisse).
GERMAN DEMOCRATIC REPUBLIC: Dienstadt (Ostthuringen), 1975, Hirsch (JE B 591/50). - Halle/Saale, Dolauer Heide (Sachsen- Anhalt), 1974, Hirsch and Braun (JE B 475/42). - Halle-Neu- stadt (Sachsen-Anhalt, 1971, Hirsch (JE B 48/4). - Brauns- dorf (Sachsen), 1970, Zschieschang and Ebert (JE). - Dubener Heide (Sachsen-Anhalt), 1977, Dorfelt (HAL); 1970, Hirsch (JE B 74/4). - Eisleben (Sachsen-Anhalt), 1972, Hirsch (JE
BY 2017 LS),
HUNGARY: Tiszacsege, Hortobagy (Hajdu-Bihar), 1974, Babos CBPOSLS92 00 Tah a Mts Pn lig i) o oot Bonus anu Banos «Oe 30833). - Cstcshégy, Mts. Budai, 1957, Bohus and Babos (BP tet - Tokhegy, Mts. Budai, 1954, Bohus and Babos (BP 39250).
THE NETHERLANDS: Osgstgeest (Zuid-Holland), 1955, Bas (BRNM 96794 andi kK)’,
POLAND: Wroclaw (Breslau), Schroeter (BRA).
ROMANIA: Cluj, 1959,Silaghi (PRM 533819).
SWEDEN: Goteborg, Stora "Anggarden" (Vastergotland), 1937, Nathorst-Windahl (PRM 103546 and K).
WEST PAKISTAN: Lahore and Sialkot, 1959, Sultan Ahmad (K).
Marasmius curreyi differs from the above mentioned M. graminum ss.str. especially by the broom cells of the pilei- pellis solely of the Siccus-type and the smaller, rather el- lipsoid to cylindric-ellipsoid spores. The number of the la- mellae is usually lower in the first species.
According to these results, seven species of Marasmius section Marasmius occur in Europe - Marasmius alniphilus Favre). My buiandias Quelice mM WeurrevirBerk wet (Om hn Mes iar minum ' (Cibert) Berk. et Br., M. limosus Quél., M. rotula
(Scop, Fr. Fr. amdy M, wettsteinii Sacc. et Syd.
ACKNOWLEDGEMENT
I wish to thank to the curators of herbaria BP, BPI, BR, BRA /BRNMYUBRNU SY CBs) Gs PHOS HAL JEO Ky OLP.) PROCS PRM anda ee R. Courtecuisse (Wattignies, France) for making it possible
70)
to study of the type and other herbarium specimens and to Dr. Z. Pouzar (National Museum, Prague) for valuable notes to my manuscript.
REFERENCES
Cooker. Co uLez 0 Ll lustpati onset british spungi is isvondoni,
Dennis, R.W.G. 1951. Some Agaricaceae of Trinidad and Vene- zuela. Leucosporae: Part I. Trans. Br. Mycol. Soc. 54: 411-482.
Favre, J. 1960. Catalogue descriptif des champignons supé- rieurs de la zone subalpine du Parc National Suisse. ai Wiss. Unters. Schweiz. Nationalparks, Aarau, 682) b
Gilliam, M.S. 1976. The genus Marasmius in the Northeas- tern United States and adjacent Canada. Mycotaxon 4: 1-144,
Singer, R. 1958. Studies toward a monograph of the South American species of Marasmius. Sydowia, Ann. Mycol., pers lit) 2s) S4a148.
Singer, R. 1965. Monographic studies on South American Ba- Sidiomycetes, especially those of the East Slope of the Andes and Brazil. 2. The genus Marasmius in South Ameri- Paw, SoyHOWla, WANN. UMYCOL oer Olt Lon lO06— 550.
Singer, R. 1976. Marasmieae (Basidiomycetes - Tricholoma- taceae). Flora Neotropica 17: 1-347.
Wy
; 1 i" iN “ d ty, ae
i La . 4 Wilby \ 4% 4 ' / £ | Ta ah, "i; + : Wy i] yf j he ee ap ¢ yn) . a ay ea ¥ ; iM ha \ Ae MM %
i nae 1 an Wee af i es
Nae
gus
tie Lap ate ah hey rq
tae be
Wye
ant fr ren + sy Vs vy
nae
ai ng, 7 Lig ‘AS 4% 3) a“ ee i aes aie
” ’ | Bee st ee Doe
a
ih ‘ai
. NET) 1 e. ; an it a ; us ? cy <, ‘ Mate Ae) Lk 34 Bi: va any hy , Asie ete TOW a Wry cv Be OY) sy hee Ugur oT Mh { ci, } ¥ | a dn! he 5 | ch : ‘ Ny > yt be tigi : , 4 Tha ~s ‘ , 1 "sd : o) i Ay aN ie , LyADEA it ; RAW ae a st “_ A Li 2 \ D ; he ’ } ‘ 1 ? : pikeni ate fa ft, ' : Me ty. uy 7 . ihe Rit, aT: in| a > 7 ¥ it pt ah AY ‘fe 2 : rit 4) i pea! Ay ry, Mi, eet | if ty nee f i on Yaa & f A) fy) AY ee hee ty Ata Oe . eee te ae? oe ‘ e pas a 5 F . ae : Arte Ar ‘ a ne ¢ ” ae f i ie vi Ven UR MeL Cunt eel i iM a RAN of ee Walt ati , ; « Pa j a Aa ee ae Kai 2 a “haa y ei ; i bd vate ee , } + /% vl Ll ie Yee oo ee | it ie ve r i hit : a Dea Lt yA ny z Tih. \ i} iy 4 is p > s. iw eae i ; Bia ih rr tere vant BRO Be Gave te : LGN ea ‘ oA P| 4 nr a: j Ms oN i . 4) qi ‘ A rahe i { Ne ow ‘ J j s ve » ; va ' 5 if ; wh pet) “ Lei x a aA a J | a be ve ; i ' i tt Ty Ate ate : ie em ; "| My Ndi uy: es Vids i 7 AL an iar Yi ‘7 ment ae 71 s a * 1 , ; ay ahh \ rye is are 4 id ae &* 7 a + | Lt" aed ol a NAW the Wes hes i fi J pe ie a is os i ol Be it Weg Aba a ‘ ‘ : 2 ‘ i eis A bes! Hes Cr Ri up “ih : { ng é i} y* 0 RU aii} ; i, Wig.e at at: t ht ay WAN : 44 “ v; i oe ; ut } a} ay , : } AQ\ th) eee y \s F Py Pan : he 1h 0 ' dy oy Ta PN: Ph ye wis 4 ive ph ny pe ’ ft fied b < { : Hel “ib A (oa a a } \ i 4 a nis ' ; + ih rit ‘ Wy) : iat’ { hy 12 L's ‘ e. Al ¢ ‘ ae oe ’ } ' . “ay ‘ } : t f it ’ an Ah Ment Po ath Cog A nh AN Poe eee Ye Leeann on ees ris Boy oe ’ "er - et 4 del ee ty | ‘ ba ‘ NG see } Th r ‘ ; . +) ; \ / te i¢ ad J 2 : ’ p LAY t y a Se ‘ aan F, ay “| rele ul eve. I, / a > t a ir , n , f p ‘ iy ¢ Ay } Wy tus + ay ETRY, ook A‘ a ve HAT ea ; Lady d 4 ie ¢ i ‘ ; i ee : “ay | Ny abu 6 MGM tia Deel ‘iy 5A Salil # iu ay . yet any ey an ‘s Bie af t : eM vad | vite } io Tr bayia Aa L pi ba D te ya >. a" Fe. ia! ; ' ras ha Pe mie St 1 9 ‘ Wh it 4
fq d (own
as ; vhs ‘ol vy A hI 5 Bc ny Lo
MYCOTAXON
Vol. XXXVI, No. 1, pp. 29-34 October-December 1989
VACCINIUM FUNGI: HELICOMA VACCINII SP. NOV. La Mod Cares
Department of Plant Pathology Washington State University Pullman, WA 99164-6430
An apparently undescribed species of Helicoma Corda was found on Vaccinium elliotii Chapman stems during a survey of fungi on Vaccinium spp. in the eastern U.S. The fungus is described as a new species and illustrated from culture and host tissue. It represents the first report of a Helicoma from Vaccinium.
The Helicoma sp. was colonizing a vertical scar on a stem section of V. elliotii collected near the Satilla River, Ware Co., Georgia. Conidia were streaked onto Difco Bacto-Agar and single germinating conidia trans- ferred to the following types of media: Difco malt extract agar (MEA), Difco corn meal agar (CMA), full and half-strength Difco potato dextrose agar (PDA) and V8 agar (Stevens, 1974). Cultures were maintained under labora- tory bench conditions of fluctuating light and temperature (18-20; C) ;
Helicoma vaccinii L. M. Carris sp. nov. Bnes.*/ bay
Coloniae in agaro cum Zeae farina composito cultae aetate quinque hebdomadum diametrum 0.6-1.0 cm attingentes; mycelio atro-brunneo, lanato vel caespitoso. Coloniae in caulibus effusae, hirsutae, atro-brunneae. Mycelium immersum vel superficiale, ex hyphis septatis, brunneis, 1.7-3.4 wm crassis. Conidiophora plerumque simplicia vel raro ramosa, flexuosa, atro-brunnea, cellulis apicalibus pallide brunneis, interdum rugosa, 4-10 septata, 64-145 pm LONG 94s 2- DOU uN Pate age basem, /25-515 pm Lat. ad apicem, proliferationibus percurrentibus pluribus (1-3). Cellulae conidiogenae mono- vel polyblasticae, denticulatae; denticula 1.3-2.7 pm long., 1.0-1.3 pm lat. Filamenta conidica in 1.5-1.75 spiris convoluta, 4-8 septata, apice rotundato, basi truncata, 2.0-4.0 pm crassa. Conidia hyalina vel pallide brunnea, laevia, 8.0- 13°20) pm. Tata.
30
Habitat in caulibus Vaccinii elliotii, Satilla River, Ware Co., Georgia. N. Vorsa, 9.vii.1988 (herb. BPI) HOLOTYPUS.
Five-wk-old colonies on CMA 0.6-1.0 cm diam, dark brown, woolly to floccose, sporulation abundant, border even, slightly crenate, reverse dark brown. Colonies on stem effuse, hairy, dark brown. Mycelium immersed and superficial, hyphae septate, brown, 1./7-3.4 ym diam. Conidiophores (Figs. 1B, 1E, 3, 9) simple or rarely branched, cylindrical, flexuous, dark brown with pale apical cells, occasionally roughened, 4-10 septate, 64-145 pm long, 4.2-5.0 um wide at base, 2.5-3.3 wm wide at apex, usually with 1-2 percurrent proliferations (Fig. 6). Conidiogenous cells (Figs. 4, 5) mono- or polyblastic, denticulate, denticles 1.3-2.7 pm in length, 1.0-1.3 pm wide. Conidial filament coiled 1.5-1.75 times, 4-8 septate, rounded at apex and tapering to a truncate base, 2.074 e0" am dlam, i Conmara terres). al yh Die 107. Orn al Oem) hyaline to pale brown, smooth, 8.0-13.0 mum diam.
HABITAT*:".On Stem’scars’ of Vaccinium’ elliotii: DISTRIBUTION: "Ware (Coy Georgia, Ul S i Ay
MATERIAL EXAMINED: LMC 0043, HOLOTYPE, deposited as stem section, dried culture, and prepared slides in BPI. ISOTYPE specimen deposited at WSP, living culture deposited as ATCC 66068.
OTHER PERTINENT MATERIAL EXAMINED: Helicoma muelleri Corda, IMI 78575, FH-Linder No. 1341; Tubeufia pezizula (Berk. & Curt.) Barr (H. muelleri anamorph), IMI 73306; H. ambiens Morgan, FH-co-type.
Only minor differences were noted between colonies on host tissue (Fig. 2) and in culture. Conidiophores formed in culture frequently had roughened walls and enlarged cells with a dark, vesicle-like outer wall layer (Fig. 1E) that were not evident in colonies on host material.
Fig. 1. Helicoma vaccinii. A. Conidiophore developing from cell of conidium in culture on CMA. B. Conidio- phores on Vaccinium elliotii. Note proliferations. C. Immature conidia. D. Mature conidia. E. Conidiophores formed in culture on CMA. Arrows indicate vesicle-like outer wall.
31
32
Colonies were restricted on all media used, and did not exceed 6-7 mm diameter after two months’ growth.
Goos (1986) monographed the genus Helicoma, recognizing 32 species and four sections. The sections are delimited primarily on mode of conidial development and conidium attachment to the conidiogenous cell. Helicoma vaccinii fits well in Sect. Helicoma, a group comprised of species with conidia borne acrogenously on distinct denticles. Goos (1986) noted the morphological similarities between the ten species in Sect. Helicoma. There is considerable overlap in characters, including: conidiophore length; number of septa; and number of coils in the conidium. The diagnostic characters most useful in differentiating species include diameter of conidia and width of conidial filaments. Nine of the species in Sect. Helicoma produce conidia >13 wm in diameter. Only H. taiwanensis Matsushima has relatively small conidia (7-15 pm in diameter) in the size range of the conidia of H. vaccinii. The conidia of H. taiwanensis are otherwise different from the conidia of H. vaccinii in having wider conidial filaments (4.0-6.0 wm versus 2.5-4.5 wm for H. vaccinii) ‘and’ blunt’ basalvcebie”” ‘Invaddition. iH: taiwanensis produces a second, phragmosporous type of conidium and conidiophores with multiple, inflated conidiogenous nodes (Matsushima, 1983).
Goos (1986), Linder (1931), and Pirozynski (1972) noted the shape of the conidium basal cell as a useful character for differentiating H. muelleri and H. ambiens, two morphologically similar species in Sect. Helicoma. Likewise, the long, tapering basal cells of H. vaccinii (5.4-14.7 pm in length) distinguish this fungus from other species in Sect. Helicoma. For example, the basal cells of H. muelleri conidia (IMI 78575) are 4.7-6.7 pm long.
The conidiogenous cells of H. vaccinii are predomi- nately monoblastic, both on host tissue and in culture (Figs. 1B, 1E, 4-6, 9). Occasionally, conidiogenous cells with 2-3 denticles are found. More frequently, the
Figs. (2-13. Helicoma vaecinii.. 2. \Gonidiophores.on Vaccinium elliotii. X 90. 3. Conidiophores formed; in culture on CMA. X 600. 4, 5. Conidiophore apices. Note denticles. X 1400. 6. Proliferation of conidiophore apex with ruptured outer wall. X 1400. 7, 8. Immature conidia. X 2000. 9. Conidiophore formed on CMA. X 800. 10,411; . Mature conidia. ~X 10002) “1204137 . Contdiopho ne. developing from cells of conidia. X 600.
33
34
conidiogenous cells proliferate percurrently, with a new conidiogenous cell emerging through the ruptured darkened outer wall of the conidiophore apex (Figs. 1B, 6). This type of proliferation was illustrated by Pirozynski (1972) for the H. muelleri anamorph of Tubeufia pezizula [cited as Thaxteriella pezizula (Berk. & Curt.) Petrak]. An illustration of a conidiophore apex of H. ambiens (Goos, 1980; Fig. 29) also resembles the conidiogenous cell proliferation in H. vaccinii shown in Fig. 6.
Germination of H. vaccinii conidia frequently occurs from the basal cell, in a way similar to H. muelleri as reported by Linder (1929), but germ tubes may develop from any cell in the conidium of the former. In colonies on CMA and MEA, conidia often become swollen and melanized, giving: rise directly to eoniditophores (Pigs. (1A 12 js).
ACKNOWLEDGEMENTS
PPNS 0046, College of Agriculture and Home Economics Research Center, Project 0836, Washington State University, Pullman, WA 99164. I thank the curators at IMI and FH for the loan of specimens examined in this study, Dr. D. P. Rogers for kindly correcting the Latin diagnosis’, and )DrsiV Ri Dy; Coos, Jd.) D. Rogers ands Ci yk, Shearer for reviewing the manuscript. Special thanks to Dr. N. Vorsa for collecting plant material in Georgia, and Rutgers Blueberry and Cranberry Research Center for use of facilities.
LITERATURE CITED
Goos, R. D. 1980. Some helicosporous fungi from Hawaii. Mycologia 72:595-610.
Goos, R. D. 1986. A review of the anamorph genus Helicoma. Mycologia 78:744-761.
Linder, D. H. 1929. A monograph of the helicosporous Fungi Imperfecti. Ann. Mo. Bot. Gard. 16:227-388.
Linder, D. H. 1931. Brief notes on the Helicosporeae with descriptions of four new species. Ann. Mo. Bot. Gard. 18:9-16.
Matsushima, T. 1983. Matsushima Mycological Memoirs. No. 3. Published by the Author, Kobe.
Pirozynski Ko Any ALO. Mierorung lof Ranzaniann wits Miscellaneous fungi on oil palm. Mycol. Pap. 129:1- 64.
Stevens, R. B., Ed. 1974. Mycology Guide Book. Univ. of Washington Press, Seattle.
MYCOTAXON
Vol. XXXVI, No. 1, pp. 35-42 October-December 1989
LONG-CHAIN FATTY ACID COMPOSITION AS A TOOL FOR DIFFERENTIATING SPOILAGE WINE YEASTS
M. MALFEITO-FERREIRA
Centro de Microbiologia e Industrias Agricolas Instituto Superior de Agronomia 1399 Lisboa Codex - Portugal
A. ST. AUBYN
Centro de Informatica do Instituto Superior de Agronomia Instituto Superior de Agronomia 1899 Lisboa Codex - Portugal
AND
V. LOUREIRO
Centro de Microbiologia e Industrias Agrtcolas Instituto Superior de Agronomia 1399 Lisboa Coder - Portugal
SUMMARY
The use of conventional methods for yeast identification is not pratical enough for industrial application due to the complexity and morosity of the techniques. Fur- thermore, misinterpretation of the results is frequent. The use of chemotaxonomic methods for pratical uses enables a faster and easier identification. In this work the analysis of long-chain fatty acids was used to differentiate several yeast strains belonging to the genera Saccharomyces, Zygosaccharomyces, Torulaspora, Candida and Pichia.
INTRODUCTION
The identification of yeasts according to morphological, sexual, physiological and biochemical characteristics has been traditionally used by Lodder (1970), Barnett et al. (1983) and Kreger-van Rij (1984). The work of Lodder (1970) and Kreger- van Rij (1984) results from a revision of the book edited formerly by both authors (Lodder and Kreger-van Rij, 1952). Unfortunately for those who deal with practical aspects of yeasts the frequent changes of nomenclature make the identification more difficult, due to an increase in the synonymy among the species classified by these authors in their different editions, which led van der Walt (1987) to propose that the fermentation industry should develop a specific taxonomy.
36
Nowadays the more significant developments on taxonomy are due to molecu- lar methods (for a review see Kurtzman and Phaff, 1987). The chemical constituition of each microorganism is regarded as a result of natural evolution and so, the species with similar composition are seen as being closely related. The fact that the above mentioned methodologies are time consuming and/or of difficult industrial use, lead to the development of more rapid techniques, such as analysis of compounds and metabolites. These, by revealing also a phylogenetic relatedness, enable an easier practical delimitation of species.
Among other compounds analysed are the long-chain fatty acids, which have been used with several microorganisms since the work of Abel et al. (1963) using bacteria. In our work we tried to differentiate several spoilage wine yeasts based on the cellular long-chain fatty acid composition following, primarily, the methodology proposed by Lategan and his co-workers (Kock et al. 1985, 1986; Cottrell et al. 1985, 1986; Viljoen et al. 1986, 1987, 1988; Tredoux et al. 1987a, 1987b). The spoilage wine yeasts can be divided into three main groups: the film-forming (e. g. Candida vint, Pichia membranaefaciens and Brettanomyces spp.), the fermenting (Saccharomyces cerevisiae), and those of more difficult control, the “sensu stricto” spoilage yeasts (e.g. Zygosaccharomyces bailit). Our aim was to obtain a test which could rapidly determine whether or not the isolated yeasts belonged to the species more dangerous to wine stability. Z. bazlzt is difficult to differentiate from Torulaspora delbruecki: when the sexual conjugation is not evident. Therefore several strains of these species, isolated from other origins, were also analysed.
MATERIAL AND METHODS
The yeast strains were either obtained from the yeast culture collection of the Instituto Gulbenkian de Ciéncia (IGC), Oeiras, Portugal, or isolated from Portuguese bottled dry white wines with “sandy” sediments eee 1). Stock cultures of all strains were maintained on Wickerham agar slopes and the isolation was made by plating, using the same medium. All isolated strains were identified according to conventional methods (Barnett et al. 1983, and Kreger-van Rij, 1984) in the IGC laboratory.
Growth on liquid medium followed the methodology proposed by Kock e¢ al. (1985). Cells were grown in 250 mL Erlenmeyer flasks with 80 mL of medium com- posed by 80 g.L~! glucose (Merck) and 6.7g.L~! Yeast Nitrogen Base (YNB) (Difco) at 30+0.1 C for 16 h, in a water bath with magnetic stirring. Then, 10 mL of the suspension were transferred to 1 L Erlenmeyer flasks with 400 mL of the previous medium, in the same environmental conditions, for 48 h. The cells were harvested during the stationary phase by centrifugation and washed twice with cold demineral- ized water. The biomass obtained was either freeze dried or used immediately. For growth on solid medium the cells, from stock cultures, were suspended in Ringer solution and a drop was placed in plates of Wickerham agar. Incubation was carried at 25+0.1 C for 48 h, after which the biomass was taken from the agar and used immediately.
The cellular long-chain fatty acids were extracted and methylated according to Moss et al. (1974), using a solution of 5% (w/v) NaOH in 50% (v/v) aqueous methanol and borontrifluoride methanol sees The final methyl ester mixture was evaporated under a nitrogen stream and redissolved in hexane. The samples were analysed in a gas chromatograph (Perkin Elmer 8410) with a FID detector, using a stainless steel column (1/8 in x 3.5m) packed with 8% Carbowax 20M w-Aw (100-120 mesh). The column temperature was 230 C at an outlet N>2 flow of 40 mL. min~?. The methyl ester identification was done by retention times of myristic (C14:0), myristoleic (C14:1), palmitic (C16:0), palmitoleic (C16:1), stearic (C18:0), oleic (C18:1), linoleic (C18:2) and linolenic (C18:3) acid esters, relative to the palmitic acid ester. The peak areas were determined by automatic integration and the results, for each ester,
of
Table 1. List of analysed yeast strains
Species Designation Origin”
Saccharomyces cerevisiae Meyen ex
Hansen A000 and A063 Commercial wine starters S. cerevisiae (syn.”) S. bayanus Sac-
cardo) A065 Commercial wine starter S. cerevisiae (syn. S. bayanus Sac-
cardo) A028, A026 and A029 Isolated from bottled wines Zygosaccharomyces batlit (Lindner)
Guilliermond A006 Type strain, IGC 2470,
CBS 680
Z. batlit (Lindner) Guilliermond G227 IGC 4227, NRRL S9 144 Z. bailtt (Lindner) Guilliermond A022, A023, A024, Isolated from bottled wines
A025, A027 and A031 Z. batlit (Lindner) Guilliermond (syn. Saccharomyces elegans
Lodder & Kreger-van Rij) G899 IGC 2899 Pichia membranaefactens
(Hansen) Hansen A030 Isolated from bottled wines Candida vint (Desmaziéres) van Uden
& Buckley A007 IGC 2597, CBS 639 Torulaspora delbrueckit (Lindner)
Lindner G166 Type strain, IGC 4166,
CBS 1146, NRRL Y-886 T. delbrueckit (Lindner) Lindner (syn.
Saccharomyces delbrueckst Lindner) G477 IGC 2477 G500 IGC 2500. kefyr grains G501 IGC 2501, kefyr grains G502 IGC 2502, bottled kefyr
T. delbruecktt (Lindner) Lindner (syn. Saccharomyces roset (Gui-
liermond)Lodder & Kreger-van Rij) G209 IGC 3209, CBS 865, potato starch G713 IGC 2713, CBS 817
T. delbrueckst (Lindner) Lindner
(syn. S. roset and Schwannto-
myces hominis Batista et al.) G844 IGC 2844, human skin T. delbrueckii (Lindner) Lindner (syn.
S. roset and Torulopsts stellata
(Kroemer & Krumbholz) Lodder var.
cambrestert Lodder & Kreger-van Rij) G907 IGC 2907 G911 IGC 2911, CBS 158, sugar G998 IGC 2998, man G999 IGC 2999, seawater
T. delbrueckii (Lindner) Lindner (syn. Torulopsis colliculosa
(Hartmann) Saccardo) G916 IGC 2916, CBS 133, javanensi ragi Unindentified strains A032, A033, A067, A068, Isolated from bottled wines
A069, A070, A071. A072. A074, A075, A076, A077. A078, A079, A080, A081 and A082
a) Abbreviations: IGC, Instituto Gulbenkian de Ciéncia, Oeiras, Portugal; CBS, Centraalbureau voor Schimmelcultures, Delft. The Netherlands and NRRL. Northern Regional Research Center. US Department of Agriculture, Peoria, Illinois. USA.
b) Synonyms of the species described by Barnett et al. (1983) and Kreger-van Rij (1984).
38
expressed as a percentage by weight of the total area.
Principal Component Analysis (PCA) enables us to analyse observed values of a set of continous variables (in our case, percentages of fatty acids) for a set of experimental units (in our case, yeast strains), in order to build new variables, called principal components, which represent the directions of the axes of greatest variability. By projecting the initial variables on the planes defined by the first few principal components, we obtain graphical representations whose appropriate analysis enables the identification of the principal contrasts among the initial continuous variables. The coordinates of the strains in the first four axes were used to perform an ascending hierarchical classification according to the reciprocal neighbours method. Next. we apply a cut-off in the dendogram obtained from the hierarchical classification, this obtaining three classes which were characterized by the initial variables. All these analyses were carried out by the SPAD (Systéme Portable pour I’Analyse des Données) software (Lebart et al. 1985).
RESULTS AND DISCUSSION
The results from growth in liquid medium (Fig. 1) showed a clear distinction between the yeast groups previously established, enabling their separation in agree- ment with their practical spoilage importance. Other authors (Tredoux et al. 1987a, 1987b) have characterized a larger number of yeasts but the groups proposed have not a consistent enological meaning.
The methods in wine microbiological control should give rapid results and be simple to operate, therefore, in order to reduce time of analysis and to simplify the procedures, the yeasts were grown on solid medium. The fatty acid profiles (Fig. 2) were similar to those obtained on liquid medium with the same basic differences among the three groups. The major difference is on the percentage of C18:2 in Z. baili: strains, but this does not affect the distinction among the three groups. The influence of growth conditions on the fatty acid composition is largely referred in literature (for a review see Ingram and Buttke, 1984, and van Uden, 1985). This similarity of the profiles in different conditions was, therefore, somewhat unexpected although, probably, the media composition and incubation conditions are not distinct enough to produce large differences. For example, Hunter and Rose (1972) had already noticed that there was little change in the fatty acid composition of S. cerevisiae cells grown at 15 C and 30 C. These results are of particular importance to industry because the procedures are simplified and, principally, because it is possible to make the analysis directly from microbiological control plates, if the biomass grown is enough for the determination.
The statistical treatment by PCA establishes a correlation matrix, comprising the long-chain fatty acid compositions for each strain. This allows distribution of the strains in a projection plan according to their fatty acid similarities, confirming the differences between the three spoilage yeast groups (Fig. 3). The unindentifyed strains A068, A070, A074, A076, A077, A079, A081 and A082 were clustered on the S. cerevisiae group, strains A032, A067, A075, A078, and A080 were in the Z. batli group, and strains A033, A069, A071 and A072 were in the P. membranaefaciens group (Fig. 3). Further tests, on a larger number of strains and species, are being made in order to improve the reliability of this analysis. for that it will be necessary to ensure the stability of the defined clusters. If this analysis proves suitable it will be possible to create a data bank allowing a direct yeast identification according to its phylogenetic proximity.
The PCA of the thirteen studied strains of T. delbrueckit showed that their fatty acid profiles are clearly different from those of the Z. bailli strains (Fig. 4). In fact, whereas all strains of Z. batlli appear in one single cluster, except for one strain, G899 formerly classified as Saccharomyces elegans, strains of T. delbruecki are
39
Mean fatty acid proportion (%) Ww je)
a a
C14:0 C141 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3
Fig. 1 - Long-chain fatty acids profiles from strains grown on Wickerham liquid medium, at 30 C for 48h. [[]]-Fermenting yeasts, S. cerevisiae A000, A063, A065, A028 and A029. [_]-“Sensu stricto” spoilage yeasts, Z. bailii A006, A024, A027 and A031. KQ-Film-forming yeast, P. membranaefaciens A030. Results are the mean of two or three replicates. Standard deviations indicated by small vertical bars.
70 60 << <= 50 S 42 5 40 Qa o a 30 =) = 20 = Av in Sol da= ea_ ii) CNRS
C140 9 C14:1 C16:0 C16:1 C180 C181 C18:2 C183
Fig. 2 - Long-chain fatty acids profiles from strains grown on Wickerham solid medium, at 25 C for 48h. [)-Fermenting yeasts, S. cerevisiae A000, A063, A065, A028, A026 and A029. [_]-“Sensu stricto” spoilage yeasts, Z. baila: A006, G227, A022, A023, A024, A025 and A031. [Q-Film-forming yeast, P. membranaefaciens A030. Standard deviations indicated by small vertical bars.
-0.
40
°o HH > o a a
if
|
1
|
|
1 gate
I A067 ae : ; A077 A07 (v2 2 I I f A069 I 3 + (020) . I I I I A081 6 I I AO74 . A071 I I A070 I -6 + I I I ps I se bd _ HH tH at 0 3 3-4 A072 -0.9 -0.6 -0.3 0.0 0.3 0.6 0.9 Te2
Fig. 3 - Projection plan of the yeasts, isolated from wines and from commercial wine starters, on the axes 1 (horizontal) and 2 (vertical). S. cerevisiaeQ ; Z. bailii—:; and P. membranaefaciens . Unidentified strains do not have symbols.
a a ea Ch
I | 622724025 I
I A023 I
I A031A022 I -6 + I I I
I I
I : A024 I
I A082 : A080 I it A000 I I A063 I
x A075 te I
I ; I \e50/ A076A065 : I : Mast Me acl bt eb 7 so) é A067 I
I A028 Gd. I
I a077 A079A029 : I
I A069 I ye I I A068 I
I ; r
I A081 : A030 I
I A070 ; I
6 + Ls I I ros I
:
I I Jo et A033—-+ A072
=i -0.9 016 Ons 0.0 0.3 0.6 0.9 de
Fig. 4 - Projection plan of the yeasts, isolated from wines, from commercial wine starters and from other origins, on the axes 1 (horizontal) and 2 (vertical). S$. rose and S. hominis©; S. delbrueckiV;: T. stellata var. cambresiers>; T. colliculosaQ); T. delbrueckii (type strain) A ; S. elegans: Z. batlit( ; and C. vint YX. Strains isolated from wines and from commercial wine starters do not have symbols, the yeasts isolated from other origins are considered illustratives, not influencing the definition of the axes. Strains A026, A078 and G477 are covered by strains G209, G227 and G501,
respectively.
41
scattered on the projection plan. Nevertheless it was possible to distinguish several subgroups among strains of the species now considered as T. delbruecki (Barnett et al. 1983, and Yarrow, 1984), approximately related to the former classification for the species S. delbrueckii, S. roser and T. stellata var. cambrestert.
From all this a question arises: are the former classifications more consistent, in phylogenetic terms, than the present ones (Barnett et al. 1983, and Yarrow, 1984)?
Although the number of studied strains is too small to enable a definite con- clusion, our results showed that the long chain fatty-acid composition might be of great convenience in the distinction between Z. batli: and T. delbrueckii.
REFERENCES
Abel, K.; de Schmertzing, H. and Peterson, J.1. — Classification of microorganisms by analysis of chemical composition. J. Bacteriol. 85, 1963, 1039-1044.
Barnett, J., Payne, R. and Yarrow, D. - Yeasts: characteristics and identification. Cambridge University Press, Cambridge, 1983.
Cottrell, M.; J.L.F.; Lategan, P.M.; Botes, P.J. and Britz, T.J.— The long-chain fatty acid compositions of species representing the genus Kluyveromyces. FEMS Microb. Lett. 90, 1985, 373- 376.
Cottrell, M.; Viljoen, B.C.; Kock, J.L.F. and Lategan, P.M. — The long-chain fatty acid composition of species representing the genera Saccharomyces, Schwanniomyces and Lipomyces. J. Gen. Microb. 132, 1986, 2401-2403.
Hunter, K. and Rose, A. — Lipid composition of Saccharomyces cerevisiae as influenced by growth temperature. Bioch. et Bioph. Acta, 260, 1972, 639-653.
Ingram, L.O. and Buttke, T. — Effects of alcohols on microorganisms. Adv. Microb. Physol. 25, 1984,, 253-300.
Kock, J.L.F.; Botes, P.J.; Erasmus, S.C. and Lategan, P.M. — A rapid method to differentiate between four species of the Endomycetaceae. J. Gen. Microb. 181, 1985, 3393-3396.
Kock, J.L.F.; Cottrell, M. and Lategan, P.M. — A rapid method to differentiate between five species of the genus Saccharomyces. App. Microb. Biotech. 28, 1986, 499-501.
Kreger-van Rij, N. (ed.) — The yeasts, a taxonomic study. Elsevier Science Publishers, B.V., Amster- dam, 1984.
Kurtzman, C. and Phaff, H. — Molecular taxonomy. In “The Yeasts”, vol. 1 (Rose, H. and Harrison, |. ed.). Academic Press, London, 1987, 63-94.
Lebart et al. — Systéme Portable pour I’Analyse des Données ( SPAD). Version 1985. Paris, 1985.
Lodder, J. (ed.) - The yeasts, a taxonomic study. North-Holland Publishing Company, Amsterdam, 1970.
Lodder, J. and Kreger-van Rij, N. - The yeasts, a taxonomic study. North-Holland Publ. Co., Ams- terdam, 1952.
Moss, C.W.; Lambert, M. and Merwin, W. —- Comparison of rapid methods for analysis of bacterial fatty acids. Appl. Microb., 28 (1), 1974, 80-85.
Tredoux, H.; Kock, J.; Lategan, P. - Identification and characterization of yeast strains related to the wine industry. Proc. 8th Int. Oenol. Symp., Cape Town, Rep. South Africa, 1987a, 23-37.
Tredoux, H.G.; Kock, J.L.F.; Lategan, P.M. and Muller, H.B. — A rapid identification technique to differentiate between Saccharomyces cerevisiae strains and other yeast species in the wine industry. Am. J. Enol. Vitic. 88, 1987b, 161-164.
van Uden, N. - Ethanol toxicity and ethanol tolerance in yeasts. Ann. Rep. on Ferment. Proc. 8, 1985, 11-58.
42
Viljoen, B.C.; Kock, J.L.F. and Britz, T.J. — The significance of long-chain fatty acid composition and other phenotypic characteristics in determining relationship among some Pichia and Candida species. J. Gen. Microb. 184, 1988, 1893-1899.
Viljoen, B.C.; Kock, J.L.F.; Muller, H.B. and Lategan, P.M. — Long-chain fatty acid compositions of some asporogenous yeasts and their respective ascosporogenous states. J. Gen. Microb., 188, 1987, 1019-1022.
Viljoen, B.C.; Kock, J.L.F. and Lategan, P.M. — Fatty acid composition as a guide to the classification of selected genera of yeasts belonging to the Endomycetales. J. Gen. Microb. 132, 1986, 2397-2400.
van der Walt, J. - The typological yeast species, and its delimitation. Jn “The Yeast”, vol. 1 (Rose, H. and Harrison, J., ed.). Academic Press, London, 1987, 95-121.
Yarrow, D. -— Torulaspora Lindner. In “The Yeasts, a taxonomic study”. (Kreger-van Rij, ed.), Elsevier Science Publishers, Amsterdam, 1984, 434-439.
MYCOTAXON
Vol. XXXVI, No. 1, pp. 43-45 October-December 1989
DESCRIPTION OF THE ANAMORPH OF VALSEUTYPELLA MULTICOLLIS IN CULTURE
JULIA CHECA
Department of Plant Biology (Botany). University of Alcala de Henares (Madrid), Spain.
AND ANGEL T. MART{NEZ
CIB, CSIC, Velazquez 144, 28006 Madrid, Spain.
INTRODUCTION
The genus Valseutypella Hodhnel is characterized by its receptacle-shaped stroma formed by pseudoparenchymatous and sclerotial cells.
This genus had remained monospecific until the description of V. multicollis Checa, Moreno & Barr (Checa et al., 1986), which differs from the type species V. tristicha (de Wot.) Hohnel, by its larger stromata containing numerous perithecia, as well as by the size of its ascospores and its habitat. Valseutypella tristicha is host specific on Rosa spp., whereas the Spanish species occurs on Quercus flex ssp. rotundifolia.
The anamorph of V. tristicha belongs to the genus Cytospora (Hubbes, 1960) and has not been described in detail. The anamorph of V. multicollis also belongs to Cytospora.
METHODS
With ascospores of V.multicollis, obtained from stromata grown on wood of Quercus ilex ssp. rotundifolia, we started cultures in PCA (potato carrot agar) with antibiotic. The fungus is conserved at the fungal culture collection of the CIB (IJFM) as IJFM A-522. After several months of incubation at room temperature, we observed the production of pycnidia, more quickly and abundantly on MEA (malt agar),
45
DESCRIPTION
Colonies on MEA growing quickly with scarce aerial myceliun. Producing a diffusible dark brown pigment and forming abundant pycnidia in concentric zones. Pycnidia black, variable in size (300- 700 pm),stromatic, globose to pyriform, with one or several ostioles, which disperse the conidia by means of drops of exudate. Phialides cylindrical (Figs. 1-2) sometimes narrowed at the base, 68-18 x 1,5-2 pm, in verticils of 3-5, on branched hyphae which grow filling up the greater part of the pycnidial cavity (Fig.3). Conidia hyaline, cylindrical-oval in front view (Figs.4-5), allantoid in side view (Fig.6) and becoming ellipsoidal (4-6 x 2,5 wm) and apiculated on one end. None of the cultures developed perithecia.
The comparison with the anamorph of V. tristicha (CBS 465.59) was not possible because this strain did not produce pycnidia in culture.
DISCUSSION
This pycnidial form must be placed in the genus Cytospora because of its stromatic pycnidia with dark walls, cylindrical and verticillated phialides not restricted to the base of the pycnidial cavity, and particularly because of the presence of allantoid conidia. However it includes ellipsoidal forms too, thus being different from the greater part of the species of Cytospora.
The definitive clarification of the systematic position of the anamorphs of Valseutypella continues the revision of the genus Cytospora, which includes anamorphs of Valsa (Spielman, 1985) and Leucostoma, as well as numerous anamorphic species whose variability in culture has to be determinated (Sutton, 1980).
We thank Dr. M.E. Barr of the University of Massachusetts, for reviewing the manuscrip and Mr. M. Heykoop for the English corrections.
LITERATURE CITED
CHECA, J.; G. MORENO & M.E.BARR (1986). Valseutypella multicollis sp. nov. Mycotaxon 25: 523-526. HUBBES,M. (1960). Untersuchungen tiber die Valsaceengattung Valseutype- lla v.H. Phytopathol.Z. 39: 389-400. SPIELMAN, L.J. (1985). A monograph of Valsa on hardwoods in North America. Canad.jJ.Bot. 63:1355-1378.SUTTON,B.C. (1980). The Coelomycetes. Commonwealth Agricultural Bureaux, Farnham Royal.
C9 OOM aS cw OL 10 0) 1, 18 0718) 0) OO 8 aye: (0. 0, 0: 16, 6 wy) EO fe) a0. 0:4 (8) eee *) © 0 te: 01-6 0), 0) 0) \e, 0) © Ole: Oe: 6) 0 mols, ne) 8 0 6! ve
Figs.1-6: Cytospora sp. 1-2.- Cylindrical phialides, verticils, on branched conidiophores. 3.- Branched conidiogenous hyphae in the pycnidial cavity. 4-5.- Oval to ellipsoidal conidia. 6.- Allantoid conidia. The bars indicate 10 pm.
MYCOTAXON
Vol. XXXVI, No. 1, pp. 47-61 October-December 1989
STUDIES ON J. B. CLELAND’S FUNGAL HERBARIUM - 2: CORTINARIUS SUBGENUS MYXACIUM (CORTINARIALES )
CHERYL A. GRGURINOVIC
51 Vardys Road, Kings Langley, N.S.W., Australia 2147.
Abstract
Collections of Cortinarius, subgenus Myxacium in the J.B. Cleland herbarium have been re-examined. Of the seven species described and the one recorded from South Australia, five are accepted, two are treated as synonyms, and one new_- species is described: Cortinarius bundarus.
Introduction
Cortinarius subgenus acium is distinguished from other subgenera by the gelatinizing universal veil. This subgenus contains small to large fleshy species with brown, yellow, blue or vinaceous-red pigmentation. Pigment is epimembranal inter- cellular or plasmatic. Hyphae of the cuticle are 7-20 pm wide, the spores are verrucose, subglobose to amygdaliform or lemon-shaped. Cheilocystidia are present or absent. (Singer, 1986:635).
J.B. Cleland described or recorded eight species within this subgenus. Fresh material could not’ be obtained to supplement the macroscopic descriptions provided by Cleland or to amplify distributional data. The macroscopic descriptions in this paper are adapted from Cleland’s handbook O19 o4—1995)) ' his “papers ((hI26 “Sl 933) cand) the collection notes accompanying each collection. Cleland recorded colours using the colour handbook Of Ridgway (1912). In this study, microscopic data were recorded from fragments of basidiome stained with ammoniacal Congo Red and then mounted in a 3% aqueous solution of potassium hydroxide.
48
KEY TO THE INCLUDED SPECIES OF SUBGENUS MYXACIUM
li) BaSra Gvomesrednes Wet cue ie: se eetclevere C. erythraeus 1]. 1*.. ‘Basidiomerdif ferent] y; coloured =i. 3. vacm eee rere aie 2. ‘BasidilomenveVLoOwisn Drown serch sss cae Sbete) oclamer were 3s 2%. Basidiomeiwith WL lac, Or, vioLlet, Cints i... om 4. 3. Spores approaching subfusiform, 12.2-18.0 x 7.0 = DOTA et wi eke steno ReRebetetNete «oie C. subarvinaceus 2.
3*. Spores ellipsoid; 6,8—-97.4x- 4.6 —S che pie ereeaeee ans sbetauenens isilen.s ie ewer cred sea eememel oneness) cu shone es C. sinapicolor 735
4. Pileus whitish to light buff; lamellae ochraceous buff; stipe with a trace of violet; odour of fenugreek WNENWE ic. toe cool izene
4x. Pileus violet-brown or violet becoming brown; lamellae violet-tinted; stipe pallid to pale VIiOLCC LOM VIG LOG. sirtecsichstoderenstels shone (clonulemen oleae De
5. Stipe with a distinct annulus, deep dull bluish violet below, pallid above; spores large, amygdal- iform, 11.4-15).8 .[-—17...6)\ x 6.4-876 0-9 34). patois ie became sine) hackers) Meiodis Aterb cumuases\ a Re kiotin torte a A Rivikc tas C.. archerimeor
5*. Stipe with a cortina, paler than above; spores smaller, short ellipsoid to elongate ellipsoid,
Up "tO uDL) GY yam LON Gee. the reteuene teste ee 0, «stele ot emeeente Gi: 6. Spores short ellipsoid to ellipsoid, 6.4-8.6 x
A, OO, SP ULL ouaie: oo skeh on Walton eke fen eshievts C.microarcheri=6o= 6*. Spores ellipsoid to elongate ellipsoid or
approaching cylindric, longer than 8.6 pm....7. 7. Spores elongate ellipsoid, sometimes approaching
cylindric, 8.6-2176 7x 4). 8-604) im ois oe ee ematee sip vehetein: mney ons tbiish seus eMemeTaueuchie esis tehioue inanas C. subarcheri 7. i*., Spores ellipsoid, .9..4-10.8 [-11.6])x 6.0-7 733)am mletadapete jercieite « ou<obemeetriacl sta vene tens ehclar elem C. bundarus 8,
1. Cortinarius erythraeus Berk. Fig. 1 A-B. Cortinarius erythraeus Berk. in Lond. J. Bot.4:48, 1845.
C.. ruber Clel. in, Trans.) R../Soc.4S. Australia od: 303-304, 1927:.
PILEUS up to 51 mm diam., convex, then irregularly wavy and convex, subgibbous, viscid, near Dragon’s Blood Red (XIII) passing into Rufous (XIV); flesh white, moderately thick over the stipe, thinning outwards. LAMELLAE slightly Sinuately adnexed, moderately close, a little ventricose, Tawny Olive (XXIX). STIPE up to 38 mm long, rather short and
49
stout, bulbous, to 19 mm diam. below and to 13 mm diam. above, viscid, solid or somewhat hollow, whitish and slightly striate above, concolorous with the pileus below the remains of the veil, with some yellowish mycelium at the base and whitish rooting mycelial strands below, flesh discoloured. CORTINA glutinous, yellowish red or red.
BASIDIOSPORES (55/6)+, 8.0 - 11.0 (#=9.8) x 5.9 - 8.3 (%=6.9) wm, L/B=1.44% ellipsoid, finely warty rough, slightly thick-walled, pale yellowish brown in 3% aqueous solution of potassium hydroxide. PASI DUAR C27/5)%) 02322-9032 . 0S- 851 220 xs36.9 )o x8 B46 = 13.0 (x=10.6) pm, with sterigmata up to 4.8 pm long, four-spored, clavate. CYSTIDIA not observed. UNIVERSAL VEIL of filamentous, smooth, gelatinized hyprae we (S872) ~ a2 04-1734 e¢x—5 . 0) eam idiam.) CLAMP CONNECTIONS present.
HABIT, HABITAT AND PHENOLOGY - gregarious, occasionally solitary or subcaespitose on the ground under eucalypts. Specimens collected in June and July.
MATERIAL - SOUTH AUSTRALIA: Kinchina, 7.vii.1923, AD 4291, Miss J. Buxton, watercolour no. 12 (syntype of C. ruber). Belair Recreation » Park, 21.V1i.1923, AD 4290: 5.vii.1924, AD 4289 (syntype Crem case Duy DeY ).*iyl 2. viel 952), AD? 4287.." 4 Morialtay BEV As 9 oo, peD.4 264+. 3. Vii 9387) AD 4280. Mto?Bold; OEMs 9394 ADS 4286".
The orange to blood-red glutinous universal veil indicates that this species belongs in Section Pyromyxae (Moser & Horak, 1975: 227, 229, 574; Singer, 1986:636).
il
Con ,6), fifty-five measurements from six collections (see Bas, C. p. 290 (1969)). 2
L/B., length-breadth ratio
50
2. Cortinarius subarvinaceus Clel. Fig. 2 A-B.
;Cortinarius subarvinaceus Clel. in Trans. R. Soc. S.! Australia’ 512304719277);
PILEUS 46 - 87 mm diam., convex sometimes repand, finally irregularly revolute, edge a little involute, sometimes substriate round the _ edge, very viscid, Mars Yellow (III), Ochraceous Tawny (XV), becoming much darker and shining in the centre; flesh slightly brownish, when old becoming semi-translucent, thick over the disc, /ithin outwards, cuticle thick and dark brown. LAMELLAE adnate or subsinuate, moderately close, slightly ventricose, pallid greyish cinnamon then Raw Sienna (III), Sayal Brown (XXIX) or Tawny Olive (XXIX) and darker. STIPE 37 - 75 mm long, stout, Powel 7) am hdiamen cylindric or base sometimes slightly bulbous, mealy, fibrillose, base viscid, whitish becoming brownish. SPORE PRINT near Tawny Olive (XXIX).
BASIDIOSPORES ((70/4),,; 12.2: = 18.00)¢x=14..6) x7))0n— 9.9% «(x=8.4) ») gm, (L/Bel. 7, approaching) | subfusitormm with the apices often drawn into obtuse mucrones, finely warty rough, slightly thick-walled, yellowish brown in 3% aqueous solution of
potassium hydroxide. BASIDIA (27/3), 34.4 - 50.0 CA Si9 wx) 2 ~ 16.0 (X=13).7) ym, with sterigmata up to Nard yum long, four-spored, clavate. CYSTIDIA absent. UNIVERSAL VEIL of repent, filamentous, loosely tangled hyphae, (40/1), 2.4 - 7,6 (x=4.3) pm diam.,:) gelatinized;,
hyaline, cuticle to 460 jm deep, subcuticular hyphae with pale reddish brown internal pigment, oleiferous hyphae scattered, cylindries;sy dark reddish brown. CLAMP CONNECTIONS not observed.
HABIT, HABITAT AND PHENOLOGY - gregarious, occasionally solitary or subcaespitose on the ground under eucalypts. Specimens collected from April to July.
MATERIAL - SOUTH AUSTRALIA: Mt Lofty, 25.iv.1921, AD 4323. Stirling West, 23ViLV 1927, “ADew4AS 2s (holotype). Encounter Bay, 24.v.1928, AD 4322. Mt Robinson, via Upper Willow Creek, Encounter Bay, 30'Jv .193977AD1:43 25). (Bchungay.:12 ov 41939) GAD V4a3 2G
51
Fig. 1. Cortinarius erythraeus.- A. Basidiospores. B. Basidia. Fig; IS on subarvinaceus.- A. Basidiospores. B. Basidia. Fig. 3. C. sinapicolor - A. Basidiospores. B. Basidia. Bars = 10 pm.
af
The large subfusiform spores and absence of clamp connections indicate that this species belongs in Section Defibulati, as defined in Singer (29362637):
3. Cortinarius ‘sinapiecolor Clel. =' Fig.3-A-B
Cortinarius sinapicolor Ciel. in. Trans: R® Soc: Ss AustraliarS7 21915, 29338.
C.. -ochraceus’ Clel. in Trans. R. Soc. S.; Australva bad 304, 1927) nen Peckr
PTILEUS, , up, to; 76¥ mmidiam., slightly \convex giro convex, sometimes umbonate, then repand, finally irregularly upturned, edge sometimes undulating, very glutinous, Mustard Yellow (XVI), Yellow Ochre (XV) or 4. Citrine.) Drab :(XL) .but>pwith ware Drowns tinge around the edge, with the centre darker or passing into Amber Brown (iP Et. flesh soapy-looking, thick over the disc’ ./raplady attenuated outwards. LAMELLAE adnate to’ sinuately adnexed, moderately close to close, to 8 mm deep, Buffy Brown (XL) when young, then near Sudan Brown (IIT) or Buckthorn: Brown (XV)) STIPERup to. 7 6am long, moderately slender to moderately stout, to 13 mm diam., flexuous, base a little bulbous with white mycelial threads, then cylindric, striate or fibrillose above, very glutinous, pallid becoming yellowish brown or tinted yellow, flesh turning slightly watery yellowish. CORTINA pale yellowish.
BASIDIOSPORES *)(50/1)), (6.8: +09 .46¢(x=8 .0 ox) 42 6a 5.7 (x=5.0) pam, L/B=1.6, ellipsoid, finely warty rough, slightly thick-walled, yellowish brown in 3% aqueous solution of potassium hydroxide. BASIDIA:, O25/1)) jue2. 4 |, 32.8 “CX=20% 500% 166 0 ee (X=8.6) jim; “with: <sterigmata Upto >.6) 71m) tong, four-spored rarely two-spored, clavate. CYSTIDIA not observed. UNIVERSAL VEIL of repent, filamentous hyphae, (31/1), 2.8 - 7.0 (x=4.6) jm diam., radially arranged, loose, gelatinized, with abundant epimembranal pigment. CLAMP CONNECTIONS present.
53
HABIT, HABITAT AND PHENOLOGY - gregarious on the ground under eucalypts. Specimens collected in June and July.
MATERIAL - SOUTH AUSTRALIA: Mt Lofty, DIVER LIZ, AD 4246 (holotype of Cy ochraceus). Belair Recreation Park, 20.vi.1931, AD 4654 (holotype of
EC.osinapicolor).:
Moser and Horak (1975:574) recorded Cortinarius ochraceus Clel. non Peck as a.) Variéetyretvar: austratiensis) » (of 7C. pardochraceys: Moser.’ However, examination of the holotype of C. ochraceus Clel. non Peck has shown it to have smaller spores’ than those quoted for Or paraochraceus var. australiensis Moser: 6.8 - 9.4 (x=8.6) x 4.9 - 5.6 (Xoeec)) Sum! last comparediitoe 1855="10757xu4N3 —1 1522 pigeons therlatter variety. Moser ;andtHorak didinot examine the holotype of C. ochraceus Clel. non Peck, but rather another collection with larger spores, belonging to a different species (Mt bortry, Lo vihl921 SAD 4243):
The yellowish to ochraceous pileus, brownish lamellae, cylindric, non radicating stipe, and gregarious habit indicate that this species belongs in Section Pyromyxae, as defined in Singer C19 36:7636)%
4. Cortinarius austroalbidus Fig. 4 A-B
Cortinarius austroalbidus Clel. & Harris in Rec. Senust Mus + 79/354, 1948:
C. albidus Clel. in Trans. R. Soc. S. Australia 57: POA eI Ss non VET.
PILEUS 37 - 62 mm diam., convex becoming plane, subumbonate, glutinous, smooth, edge subfibrillose when old, white with occasional tints of Light Buff (XV), when old near Light Buff (XV); flesh thin, attenuated outwards, with a very faint tint of violet. LAMELLAE sinuately adnexed, moderately close, ventricose, up to 9 mm deep, Light Ochraceous Buff (XV) to near Ochraceous Buff (XV). STIPE up to 62 mm long, moderately stout, to 13 mm dyvam-ei. tccy lindrac or slightly bulbous’ below,
54
sticky, fibrillose, white with a very faint tint of violet. VEIL brownish. ODOUR of fenugreek when dry.
BASIDIOSPORES (52/1), 9.3 - 11.6 [-12.8] (x=10.4) KSB) i ae Cx G 5) am, b/Bal6 7) ellipsoid) ie elongate ellipsoid, finely warty rough, slightly
thick-walled, pale yellowish brown in 3% aqueous solution of potassium hydroxide. BASIDIA (24/1), 28.2) 40.0) (X=S8 Sno xn 824) LT 07 Ceao. 40am, wie sterigmata up to sys pm long, four-spored,
clavate. CYSTIDIA absent. UNIVERSAL VEIL of filamentous, gelatinized hyphae, (31/1), 2.4 - 6.0 (x=3.8) pm diam., loosely repent, undulating,
pigment not seen. CLAMP CONNECTIONS not seen.
HABIT, HABITAT AND PHENOLOGY - gregarious on the ground under eucalypts. Specimen collected in June.
MATERIAL - SOUTH AUSTRALIA: Belair Recreation Park, 29.vi.1932, AD 4113 (holotype).
The small to medium-sized basidiome, whitish pileus, pale lamellae, and clavate to somewhat bulbous stipe indicate that this species belongs
in Section Malvacei, as defined in Singer ClS86 3638): 5 Cortinartusiarcheri) Berk. )- "Fig. SiiA7B
Cortinarius archeri Berk. in Hook. f. Fl. Tasm. 2: 247) C. L8ly 7, 1860.
PILEUS 63 - 89 mm diam., deeply convex then convex becoming plane, finally often with upturned edges and irregular, or with deep depressions’ and bosses, very viscid, deep violet becoming brown (Verona Brown (XXIX) to Bistre (XXIX) with a violet!) tint));))/tlesh chan except over disc. LAMELLAE slightly sinuate to adnate, moderately close, to 10mm deep, sometimes with reticulated ridges on the sides, Snuff Brown (XXIX) with a violet tint, especially on the edges. STIPE 63 - 89 mm long, stout, 18 - 25 mm diam. or more, at first bulbous below, usually slightly attenuated in the middle, sometimes flattened, striate above, hollow below, Deep Dull Bluish Violet (XXIV) below the glutinous subdistant ring, paler lilac above.
55
e e
A. A
austroalbidus.-
Cortinarius
4.
Basidiospores.
Fig.
microarcheri
archeri. 10 pm.
C C.
De 6. Bars
Fig Fug.
Basidia.
Basidia. B.
B. Basidia. B.
Basidiospores.
Basidiospores. A.
56
BASIDIOSPORES (61/5), 11.4 - 15.8 (-17.6) (x=13-4)
x) 640 CS BGO 4a (x=7 26) um, \/ B=) sepa ay gare iform, finely warty rough, thick-walled, pale yellowish brown in 3% aqueous solution of potassium hydroxide. BASIDIA (20/2), 32.0 - 43.2
(x=37.4) x 8.2 - 12.0 (x=9.8) pm, with sterigmata up to 7.6 pm long, four-spored occasionally two- spored, clavate. CYSTIDIA not observed. UNIVERSAL VEIL glutinous, to 590 pm deep, of filamentous, loosely repent to ascending hyphae, (20/2), 2.0 - 4.2 (x=3.5) pm diam. PILEAL CUTICLE a narrow band of repent, filamentous to cylindric hyphae. CLAMP CONNECTIONS present.
HABIT, HABITAT AND PHENOLOGY —- “solitary, gregarious or caespitose on the ground. Specimens collected in March, May and June.
MATERIAL - TASMANIA: Cheshunt, LOS LV... DESO wae holotype, W. Archer. NEW SOUTH WALES: Mosman, Sydney, 20.v.1917, AD 4103. SOUTH AUSTRALIA: Mt Lofty. 25.v.19207,.7 AD) 41057" "29 titel 24) (AD 40 So. Belair Recreation Park, LG we bo Sale, AD 4101. Woodside, 19.vi.1946, AD 4102. Adelaide hills, v.1954, AD 4100, N. Atkinson.
This species can be distinguished from Cortinarius
subarcheri Clel. and C. bundarus Grgurinovic by its large, amygdaliform spores. Cortinarius
archeri constitutes the type of Section Archeriani. 6. Cortinarius microarcherj Clel. - Fig. 6 A-B
Cortinarius microarcheri Clel. in Trans. R. Soc. S'.1 Australia ids 7 3191 933),
PILEUS 18 - 62 mm diam., convex to nearly plane, edge sometimes striate, glutinous, deep violet or violet-brown, drying from the centre to _ earth- brown or light brown (Amber Brown, XL), flesh violet-tinted or whitish. LAMELLAE slightly Sinuate to adnexed, moderately close, pallid violet to violet-brown, (Buffy Brown (XL), with violet tints), then earth-brown (Snuff Brown, XXIX). STIPE 31 - 50 mm long, rather slender, base a little thickened, fibrillose, slightly hollow or solid, pallid or pale violet.
57
BASIDIOSPORES (54/6), 6.4 - 8.6 (x=7.1) x 4.8 - 8.3. .(x=5.3). jam, Ley Baas, short ellipsoid to ellipsoid, warty rough, slightly thick-walled, yellowish brown in 3% aqueous solution of potassium, hydroxide. BASIDIA s(1/7/2)), 27552) — 4132 (x333.7) x 624 - 9.6 (x=8.4) pm, with sterigmata ups .toV6.4 suamvlong, four-spored, clavate... CYSTIDIA absent. UNIVERSAL VEIL to 25:0 pm deep, of filamentous hyphae, (11/1), 3.1 - 4.8 (x=4.2) jm diam., gelatinized, lower portion of loosely ascending hyphae, upper portion of loosely repent hyphae. PILEAL’ CUTICLE to .96 jim “deep, of), repent; Eplanentous. hyphae; 1G//1),.3.60-%555 ) ¢x=4.8).) jim diam., with plasmatic pigment. CLAMP CONNECTIONS present.
HABIT, HABITAT AND PHENOLOGY - solitary, ‘to gregarious on the ground. Specimens collected in June.
MATERIAL - SOUTH AUSTRALIA: Mt Lofty, TO SVR INT, AD 4231 (lectotype, here designated); 19.vi.1921, AD 4219 (syntype); 23.vi.1928, AD 4224 (syntype); Sav wos OAD eae 222 Csyntype ys Suvi elosi. SAD* 4223
(syntype). Eagle on the Hill, 5.vi.1932, AD 4230 (syntype). Kersbrook, 25.vi.1933, ADY4227 74 Dr Rogers.
The violet pileus, violet-brown lamellae, and ellipsoid spores indicate that this species
belongs in Section Archeriani, Stirps Iodes (Moser ReHoreky 1 197525782, Singer, 1986:638)..
iw COrtinariusesyubarchergsCledag.] Fig. \7
Cortinarius svubarcher? Cletl, )ineTrans > -<Ri«Soc, i.) S: Auseralraws2g:220,, (2 928)pap.
PILEUS 25 - 60 mm diam., at first deeply convex then expanding to convex, occasionally with a depression an the centre, viscid, becoming
shining, violet becoming brownish violet to violet (Brownish Vinaceous to Deep Brownish Vinaceous (XXXIX) with tints of Deep Purplish Vinaceous (XLIV) or Mars Brown (XV) and darker towards’ the edge). LAMELLAE adnate or sinuate, close, narrow, becoming slightly ventricose, violet becoming
58
aa ae
we fo te ° * reek Suns een? ° ° © 9: S60 Cr fece ary i e se s O tte ° . ee ae 2% s +o 8 oes erce ete coe” ° ul Ahad ee J) sae . ° oe ead . 2 . ° ee ° . Po Leet ie n*e? Sus oot ft aeel et NaS STR) . Pe Ne « O42 0° < seme hig On 2 *,0° 2° eos este ° AP oe fe # Latin oe! io ote oe ee waa ®,e * A 2 eee, fe, eo fe we AO TACO eo. +. 4 ee ee ° uO OAT o.? ore * 0, te 80 6 Pay weer e eo, &e ry Pig Vial glis ® ° ClO ef ceo e * 4 Oxi er igs oe ieiiw eote . ee hits * ee ee Ce ana age Coe e* oe CR one fae o°e O%y Oy hey Aig. ER ore ee ° . CW Ae URGE ene. MS * eos ee Ce ° . Maar ee ai hele ee ees auto eye ° °. eC eT ee Pe 7 ale DRE NEA o.oo On é ore oan ee o* ye 8
Fig. 7. Cortinarius subarcheri.- Basidiospores. Fig. 8. Cortinarius bundarus.- A. Basidiospores. B. Basidia. Bars = 10 pm
Vinaceous Fawn (XI); Sorghum Brown (XXXIX), Prout’s ‘Brown (XV))or Apricot Buff) (XIV). STIPE 35 - 51 mm long, stout, cylindric or base _ slightly bulbous’, / root!) (rather) conical, 25 5=)'25). mm) 7aiane, fibrillose, solid, sticky when moist, whitish to violet (Pale Lobelia Violet, XXXVII, Grayish Lavender to Deep Grayish Lavender, XLIII). FLESH white with a faint violet tint in stipe, or paler than Grayish Lavender. VEIL white.
BASIDIOSPORES) (94/4) 7184/00 216) ) (X=9705)) x4 on 6.1 (x=5.3) pm, L/B=1.8, elongate ellipsoid, some- times approaching cylindric, finely warty rough,
slightly thick-walled, yellowish brown in /3% aqueous solution of potassium hydroxide. BASIDIA (8/1951) 34.7037 6x Po] 88. 7 rpm wEth ster igmace up to 2.4 pm long, four-spored, clavate. CYSTIDIA not observed. UNIVERSAL VEIL to 354 pm deep, of loosely arranged, filamentous, gelatinized hyphae, (10/1), 2.4.-'4.3 (x=3.5) am diam. PILEAL CUTICLE
09
a narrow band of filamentous hyphae, with brownish pigment. CLAMP CONNECTIONS present.
HABIT, HABITAT AND PHENOLOGY - solitary, usually gregarious on the ground under Eucalyptus baxteri (Benth. ) Maiden & Blakely, etc. Specimens collected in May and June.
MATERIAL - SOUTH AUSTRALIA: Mt Lofty, 16.vi.1917, AD 4220; 16.vi.1917, AD 4225; 19.vi.1920, AD 4226. Kinchinaly; (7. viiit923'7) (AD) |) 46602 00) Mt) Burn) Porest Reserve, 30.v.1928, AD 4316 (lectotype, here designated). Morialta, ie OW, gr Ws ips Be 6 Yi AD 4311. Willunga Hill, VeibIS2y AD 4313. Echunga, 12.vi.1939, AD 4662.
The violet-brown pileus and lamellae, and the elongate ellipsoid spores indicate that this species belongs in Section Archeriani, Stirps Iodes (Moser & Horak, 1975:578; Singer, 1986:638).
8. Cortinarius bundarus Grgurinovic, sp.nov. - Fig. 8 A-B
Cortinarius "subarcheri Clels in Transi'R.Soc..()'Se mRustraivayo2 =: 220), 1928) p.p.
Pileus usque ad 89 mm diametro, irregulariter convexus, violaceus deinde versus apicem pallide brunneolus. lLamellae sinuatae, moderate confertae, ex violaceo caryophyllaceo-cinnamomeae. Stipes usque ad 38 mm longus, crassus, fibrillosus, ex violaceo pallidus. Sporae 9.4 ULI SCO ine lee Ly kaiOi § CKeLOR Ax 6 Oa eS exe ebi7 uM, el Mipnsoidegey, verrucosae. Basidia 32.0 - 44.4 (x=37.5) x 8.0 - 10.8 (x=9.4) pm, clavata, 4-sporigera. Holotypus: South Australia, Bundaleer State Forest, vi.1928, AD 4329.
PILEUS up to 89 mm diam., irregularly convex, violet, becoming pale brownish in centre. LAMELLAE Sinuate, slightly toothed, moderately close, Pinkish Cinnamon (XXIX) tinged with violet. STIPE Rey toy o38)\cmm) long, stout, tol. 38 *mPiidiam.; downy-fibrillose, violet-tinted. FLESH with violet Srite lac tints.
60
BASIDIOSPORES (50/1), 9.4 - 10.8 [-11.6] (x=10.1)
X46) ONY oars (x=6.7) pm, L/B=1.5, ellipsoid, coarsely warty rough, slightly thick-walled, yellowish brown in 3% aqueous solution #08
potassium hydroxide. BASIDIA (15/1), 32.0 - 44.4 (x=37.5) x 8.0 - 10.8 (X=9.4) pm, with sterigmata up to 4.0 pm long, four-spored, clavate. CYSTIDIA not observed. UNIVERSAL VEIL to 230 pm deep, of filamentous, gelatinized hyphae, (11/1), 1.9 - 4.3 (x=3.1) pm diam., loosely repent to somewhat ascending. CLAMP CONNECTIONS present.
HABIT, HABITAT AND PHENOLOGY - gregarious on the ground. Specimen collected in June.
MATERIAL - SOUTH AUSTRALIA: Bundaleer State Forest, vi.1928, AD 4329 (holotype).
This species is distinguished from Cortinarius Ssubarcheri by its ellipsoid and slightly larger spores. The violet pileus, violet tinged lamellae and ellipsoid spores indicate that this species belongs in Section Archeriani, Stirps Jodes, as defined in Singer (1986:638). The specific epithet is derived from the aboriginal word "bundara", meaning a "clump of trees".
ACKNOWLEDGEMENTS Ir, would. like... toe. thank Mr. J«) Simpsonyfor-whis
helpful advice and Dr R. Watling for reviewing the manuscript.
BIBLIOGRAPHY
Bas; .C..((1969)% Morphology and subdivision” of Amanita and a monograph of its section Lepidella. Persoonia 5:285 - 579.
Cleland, J.B. (1928). Australian fungi: notes and descriptions sw ii NOs: 17%. ei rans:.y tRewi SOCin for Australia 52:217 - 222.
ad haters (1933). Australian fungi: notes and descriptions ))i-iyNosawv9u) eTransem) Reo (SOCimaS
Australia 57:187 -194.
61
a (1934-1935). Toadstools and Mushrooms and Other berGcrerungivof South Australian vol. 1 & 2. Government Printer, Adelaide, South Australia.
Moser, M. & Horak, E. (1975). Cortinarius Fr. und nahe verwandte Gattungen in Sudamerika. Beih.
Nova Hedwigia H. 52.
Ridgway, R. (1912). Color Standards and Color
: Nomenclature. Published by tha author. Washington, | Bet Os
Singer, R. (1986). The Agaricales in Modern Taxonomy, 4th edn, Koeltz Scientific Books, Federal Republic of Germany.
lel
sii IX ti
MY COTAXON
Vol. XXXVI, No. 1, pp. 63-72 October-December 1989
STUDIES ON GALEROPSIS AND GASTROCYBE (BOLBITIACEAE, AGARICALES)
G. MORENO, M.HEYKOOP and C. ILLANA* Department of Plant Biology (Botany). University of Alcala de Henares, Madrid, Spain.
ABSTRACT
After we have. studied the holotype of Galeropsis desertorum Velen. & Dvorak, type specimen of the genus Galeropsis, the presence of a hymeniform epicutis is proved contrary to the traditional interpretation as a cutis. The same happens with G. bispora Vasil’kov and G. andina Singer. The genus Gastrocybe Watling is considered
as synonymous of Galeropsis Velen. We propose the following new combinations: Galeropsis lateritia (Watling) comb. nov., G. deceptiva (Baroni) comb. nov. and 4G. desertorum var. bispora (Vasil’kov) comb, et
Status nov. INTRODUCTION
The genus Galeropsis Velen. was described by VELENOVSKY (1930) and in his original description the following microscopic characters were specified: "Basidiis Subglobosis, quadristerigmatosis. Pulvere fusco. Sporis ovato-ellipticis, ochraceo-luteis, glabris. Cystidiis nullis.", without any reference to the structure of the epicutis.
Later on, SINGER (1963, 1986), WATLING (1968), SINGER & PONCE DE LEON (1982) and MORENO & a). (1987) considered the epicutis of Galeropsis as a cutis. On the other hand WATLING (1968) described the genus Gastrocybe with a hymeniform epicutis as the main differentiating character,
However, there are some references in the literature on the presence of a hymeniform epicutis in the genus Galeropsis, e.g. the comments given by Singer and compiled by HEIM (1950) in relation to G. Plantaginiformis (Lebedeva) Singer which we reproduce he-
* This paper was presented at the "X Congress of European Mycologists" celebrated in USSR (20-25 August 1989), and at the "VIII Simposios de Ciencias Criptogamicas" celebrated in Melilla (23-26 September 1989).
64
re: "Le péridium est composé de deux couches, l’enveloppe extréme consiste en cellules en palissade, claviformes, brunes ou Nhyalines (10-11,5 wpm de diam.), lautre plus épaisse, devient vers Jl intérieur une couche de jonction orientée transversalement, généralement plus dense vers lintérieur",
WASSER (1979) described the following: "Epicutis of the cap consists of brownish and colourless cells (17-22 D4 5-10 ym) and globate hyphae (10-42 ym diam.). Dermatocystidi absent".
The studies on the epicutis, very important in the differentiation of these genera, have not been mentioned neither in the studied species by HEIM (41950), nor in the following taxa described as new to science: Galeropsis allospora Singer, G. angusticeps (Peck) Singer, G. Jiberata (Kalchbr.) Heim, G. madagascariensis Singer and G. mitraeformis (Berk.) Heim.
However SINGER (1963), when he described Galeropsis andina Singer, pointed out: “epicutis of the peridium not @elatinized, of elongated hyphae which forms ae cutis", which makes us think on the possible existence of taxa without a hymeniform structure in their epicutis, conclusion which is wrong, as we will show further on /( the revision of G. andina has proven that this species has a hymeniform epicutis).
The presence of pileocystidia has not been described for Galeropsis desertorum, G. Plantaginiformis and G. bispora. However, they have been confirmed in Gastrocybe iberica (MORENO & a41., 1987).
Material examined: Galeropsis desertorum (holotypus) PR 448508; Galeropsis bispora (isotypus) PR 678864; Galeropsis Plantaginiformis (isotypus) PR 678867; Galeropsis desertorum var. bDispora (as G. desertorum), on roots of Poa bulbosa, Iran (Kabus), leg. T.F. Hewes, MA- Fungi 16932. Galeropsis andina Singer (isotypus) BAFC 31544.
Conclusions: We think that Galeropsis desertorum Velen. & Dvorak (Figs. 1-7), type specimen of the genus Galeropsis Velen., is very variable in the size of its carpophores and the measurements of its spores; this species presents lageniform pileocystidia (sometimes difficult to observe if the material has been badly dried or colapssed), and a hymeniform epicutis formed by a Single layer of cells instead of a cutis as traditionally Was thought. The latter makes that we believe the genus Galeropsis must be emended and, besides, that the genus Gastrocybe is synonymous of Galeropsis and therefore its described species have to be transferred to the = genus Galeropsis.
The bisporic species Galeropsis bispora Vasil’Kov (Figs. 46-24) and G. plantaginiformis (Lebedeva) Singer (Figs. 68-15) have also been revised, and it turns out that we consider G. bDispora as a _ bisporic variety of G.
Figs. 1-7. Galeropsis desertorum. Basidiocarps,
epicutis, pileocystidia and spores. PR 148508)
(Holotypus,
65
66
desertorum, being the main character to differentiate it the constancy of its bisporic basidia, without having observed any tetrasporic. On the other hand, the fact it has spores with a Digger size than 4G. desertorum is logical in a_ bisporic variety. The epicutis and the presence of pileocystidia are similar to those of G. desertorum, and they were not described up to date.
The species Gastrocybe iberica Moreno, Iillana & Heykoop (Figs. 25-29) is considered by us aS Synonymous of Galeropsis bispora (vide comparative table).
The typus of Galeropsis plantaginiformis presents tetrasporic basidia and very uncommonly some 0bisporic. The morphology of the spores (spore wall, germ pore) and the measurements are similar to those of G. desertorum, therefore both species are considered as synonymous by us as well as did PILAT (1948) and VASIL’KOV (1954), contrary to the opinion of SINGER (1936, 1954, 1956), HEIM (1950) and KOTLABA & POUZAR (1959).
After studying an tsotypus of Galeropsis andina we have observed that it presents a hymeniform epicutis (Figs. 30-35), formed by a single layer of cells, and not a cutis as pointed out by SINGER (1963). The morphology of the spores and basidia agree with the description given by this author.
GFALE ROP SITS Velenovsky, MykKologia, Praha 7:105 (1930. GASTROCYBE Watling, The Michigan Botanist 7:20 (1968). PSAMMOMYCES Lebedeva, Tr. Zasc. Rast. 5(1):116 (1932). CYTTAROPHYLLUM (Heim) Singer, Beih. bot. Cbl. 56/B:147 (1936).
Galeropsis tlateritia (Watling) Moreno, HeykKoop & Illana comb. nov.
= Gastrocybe Jateritia Watling, The Michigan Botanist 7:20 (1968).
Galeropsis deceptiva (Baroni) Moreno, HeykKoop & Illana comb. nov.
= Gastrocybe deceptiva Baroni, Mycologia 73:161-182 (1981).
Galeropsis desertorum Velen. & Dvorak var. bispora (Vasil’Kov) Moreno, HeyKoop & fIllana comb. et status nov. = Galeropsis bispora Vasil’Kov, Proc. Bot. Inst. Acad. Sci. USSR, ser. 2. 9:463 (1954).
= Gastrocybe iberica Moreno, Illana & Heykoop, Cryptogamie, Mycol. 68:323-324 (1987).
On the following table we compare the epicutis, basidia and basidiospores of Galeropsis desertorum Velen. & Dvorak, G. Pplantaginiformis (Lebedeva) Singer, G. bispora Vasil’Kov and Gastrocybe iberica Moreno, Illana & Heykoop.
110pm
Figs. 8-15. Galeropsis plantaginiformis. Basidiocarp, epicutis, pileocystidium, basidia and spores. (Isaty- pus, PR 678867).
| Meere 2b
*
Figs. 16-24. Galeropsis bispora. Basidiocarp, epicutis, pileocystidia, basidia and spores. (Isotypus, PR 6783864).
F e
¢
D me
g i Q
Cc h
. 20-29. Gastrocybe utis, pileocystidia, otypus, H.AH 9990).
i[berica. Basidiocarps,
asidium and spores.
69
70
Figs. 30-35. Galeropsis andina, Basidiocarps, epicutis and spores. (Isotypus, BAFC 31514).
Galeropsis desertorum
Epicutis Hymeniform with lageni- form pileocystidia
Basidia Tetrasporic
Spores 11-14 x 7-8 pm (KOTLABA & POUZAR, 1959)
Galeropsis bispora
Epicutis Hymeniform with lageni- form pileocystidia
Basidia bisporic
Spores 14-16 x 10-114 pm (KOTLABA & POUZAR, 1959) 12,8-15(17,5) x (10,5) pm sion)
7,5-9,8
Galeropsis
(personal revi-
Epicutis Hymeniform with lageni- form pileocystidia
Basidia Bisporic (SINGER, 1963) tetrasporic predominate (personal revision)
Spores 1051 2 OX hoy Omoro! a sin
(HEIM, 1950)
Gate Srey cso igs 0. UE (KOTLABA & POUZAR, 1959) 10/593 ek! VORTCSe “pm (personal revision)
Gastrocybe iberica
Epicutis Hymeniform with lageni- form pileocystidia
Basidia bisporic
Spores 45-20(23) xX MORENO & al., 1987)
ACKNOWLEDGMENTS
We wish to express our references, to Dr. Vv. Raitviir and Dr. R.
Singer, Dr. are also very grateful
Plantaginiformis
9-13(18) pm
71
gratitude, for supplying us with Antonin, Dr. R. Watling.
A. to
Dr. M. Svrtéek for the loan of the types of Velenovsky and
Vasil’Kkov which are Kept
for the loan of an isotype
Kept in BAFC,
in’ PR, and to Dr. J. E. Of Galeropsis andina which
REFERENCES
-Heinm, R. (sCyttarophyllum Gastérales. rev. Mycol. -Kotlaba, F.
steppe fungus,
(1950). Le Heim), trait
genre dunion entre GCParis) “15: 03-26. (1959), A new find of a rare desertorum
& Pouzar, Z. Galeropsis
Galeropsis Agaricales
Velen, et Dvor.,
Wright is
Velenovsky He
in
72
Czechoslovakia with notes on the genus Galeropsis Velen. CesKa MyKol. 13: 200-211.
-Moreno, G., Illana, Cc. & Heykoop, M. (1987). Gastrocybe iberica sp. nov. in Spain (Bolbitiaceae, Agaricales). Cryptogamie, Mycol. 8: 321-327.
-Pilat, <A. (1948). On the genus Galeropsis Velenovsky. Studia Bot. Cechoslovaca 9: 177-185.
-Singer, R. (1936). Galeropsis ein Gasteromycet! Bein. zum Bot. Centralbl. 56:147-149.
-Singer, R. (41954). The Agaricales (Mushrooms) in Modern Taxonomy. Lilloa 22:1-839.
-Singer, R. (1956). Type studies on Basidiomycetes. Gasteromycetes. Sydowl!a 8:427-431. -Singer, R. (1963), Notes on Secotiaceous fungi: Galeropsis and Brauniella. Proc. Fon. Ned. Akad. Wetenscn., Cc, 66:106-117.
-Singer, R. (1986). The Agaricales in modern taxonomy.
Koenigstein (RFA), Koeltz Scientific Books.
-Singer, R. & Ponce-de Leén, P. (1982). Galeropsidaceae west of the rocky mountains. Mycotaxon 14:82-90, -Vasil’kov, B.P. (14954). Some interesting and new = species of Gasteromycetes in the USSR. Proc. of Bot. Inst. Acad. Sci. of the USSR, Ser 2, 9: 447-464.
-Velenovsky, J. (41930). Galeropsis gen. nov. MyKologia (TO Sia Oo
-Wasser, S.P. (1979). Fungorum Rariorum Icones Coloratae 10:1-3e. J. Cramer. Vaduz.
-Watling, R. (1968). Observations on the Bolbitiaceae IV. A new genus of gastromycetoid fungi. The Michigan Botanist 7:19-24
MYCOTAXON
Vol. XXXVI, No. 1, pp. 73-90 October-December 1989
PHYTOPHTHORA ERYTHROSEPTICA H.H. Ho’ and 8.C. Jong’
‘Department of Biology, State University of New York, New Paltz, New York 12465
*Mycology & Botany Department, American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852
ABSTRACT
Phytophthora erythroseptica was established by Pethybridge in 1913 as the fungus causing pink rot of
potato tubers. It was characterized by formation of nonpapillate sporangia in water and the production in single cultures of abundant relatively large oogonia with amphigynous antheridia. Recently, several authors questioned the species concept of P. erythroseptica by including isolates that produced both amphigynous and paragynous antheridia. The present study was undertaken to compare 14 isolates of P. erythroseptica from various parts of the world in order to re-define the species based on morphological and cultural characteristics. It was concluded that isolates producing paragynous antheridia should not be assigned to P. erythroseptica.
INTRODUCTION
Phytophthora erythroseptica Pethyb. was named by Pethybridge (1913) as the incitant of pink rot of potato tubers. It was characterized by the formation in water OfisCONnLdLa., 44% more or less ovate or inversely pear- shaped, with a rather blunt or even somewhat flattened apex" and the production in single culture of abundant sex organs so that "the oogonial incept enters the antheridium at or near its bases, grows up through it and out at the top, expanding there to form the oogonium proper in which the oospore develops". This unique method of sexual reproduction in P. erythroseptica was confirmed by Murphy (1918) who described the antheridium as “amphigynous" in contrast to the "paragy-nous antheridium which grew up to the side of the oogonium".
Since the erection of P. erythroseptica, three varieties have been proposed: var. atropa on Atropa
74
belladonna (Alcock, 1926), var. pisi on pea (Bywater & Hickman, 1959) and var. drechsleri as a new combination for P. drechsleri Tucker (Sarejanni, 1936). They were all rejected by Waterhouse (1963) who considered var. atropa not significantly different from var. erythroseptica and var. drechsleri synonymous with P. drechsleri which she retained and var. pisi, invalidly published for failing to cite the type specimen. But she retained var. pisi in her key and considered P. himalayensis (Dastur, 1948) identical to P. erythroseptica. Regardless of their taxonomic status, members of these taxa had one character in common: the production of entirely amphigynous antheridia.
However, Cooper (1928) studied the type culture of P. erythroseptica and found no amphigynous antheridia. Instead, he found paragynous antheridia closely appressed at the base of the oogonia. Converse and Schwartz (1968) identified the causal agent of root rot of red raspberry as P. erythroseptica sensu lato even though paragynous antheridia were produced in addition to the abundant amphigynous antheridia. Savage et al. (1968) also found that some isolates of P. erythroseptica produced paragynous antheridia and amphigynous antheridia in varying proportions. They pointed out the similarity between P. erythroseptica and P. megasperma Drechs. in producing nonpapillate sporangia and both antheridial types in single cultures as well as pathogenicity to potato tubers. Recently, Pratt (1981) identified some fast growing isolates from arrowleaf clover tentatively as P. erythroseptica although the antheridia were predominantly paragynous in agar but mostly amphigynous
in broth cultures. He also questioned the distinction between P. erythroseptica and P. megasperma based on the antheridial type. Thus, Erwin (1983) listed P.
erythroseptica as one of the few species of Phytophthora where particular nomenclature problems exist requiring critical re-evaluation.
The present study was undertaken to compare colony morphology and culture characteristics of a wide variety of isolates of P.erythroseptica under uniform conditions in order to define the species more precisely.
MATERIALS AND METHODS
Isolates and media: Specific information on the isolates of P. erythroseptica used is given in Table l. All isolates were obtained from the American Type Culture Collection (ATCC), Rockville, Maryland. Unless otherwise stated, clarified V-8 juice agar medium (Ribeiro, 1978) supplemented with sitosterol (30 mg/l) (CV8) was used for culture.
Morphology: Colony characteristics on CV8 were compared after incubating the cultures in darkness at 20C
Table 1. Sources of Phytophthora erythroseptica isolates
ATCC Host Origin Source
10923(a) Unknown USA Jeffers
10924 Potato Unknown Jeffers
16698 Unknown USA Gallegly N65 16699(T*) Potato India Gallegly N117=CMI 17028 28766 Potato Scotland Pitt 38
36302 Potato USA Rowe 100
46725 Potato Tasmania Zentmyer P340 53014(T) Potato Ireland Ho H14.1 = CMI 34684 64047 Wild rice USA Ho H14.12 Gunnell 64127 Unknown Unknown CBS 233.30
64128 Tulipa Unknown CBS 380.61
64155 Unknown United Kingdom CMI 146453
64156 Potato United Kingdom CMI 181716
64861 Potato United Kingdom UCR P3513
(a) Received as P. drechsleri (T*) Type of P. himalayensis
(T) Type Culture
ATCC -- identified by American Type Culture Collection accession number
75
CMI -- Commonwealth Mycological Institute, Kew, Surrey, England UCR -- University of California, Riverside
for 7 days. The colony diameters were measured at right angles through the inoculum, and the width of primary hyphae measured using a light microscope. The minimal and maximal temperatures for growth were tested by growing the isolates at 5 C and 35 C. Sporangia formation was induced by incubating small mycelial agar discs on CV8 in freshly collected stream water that had been filtered through 0.45 um pore size membrane discs and incubating the produced discs under light at 20 C. Sex organs in single cultures were examined periodically by microscopy through the bottom of the petri dish. If the isolate failed to produce sex organs in single culture, it was paired with the appropriate mating types of P. nicotianae (ATCC 38606, A, and ATCC 38607, A,). In case of a successful mating, the ability of the isolate to produce sex organs by selfing was confirmed by pairing it with the compatible
76
strain across a polycarbonate membrane to prevent physical
contact between the cultures (Ko, 1978). They were examined for sex organs after 2-3 wk incubation in the dark Vat e200C. If these attempts failed to induce
formation of sex organs, the isolate was grown on tryptophan medium (Ribeiro, 1978) and sterilized oat grains (Van der Zwet & Forbes, 1961) and examined after 4-5 months.
RESULTS
Colony morphology
Isolates of P. erythroseptica grew well on CV8 (4- 8 mmn/day) at. 20°C)’ forming uniform; (‘non-flutfy? to moderately fluffy colonies with a diffuse and slightly irregular outline and no special growth pattern. ATCC 64047 colonies were distinctly fluffy with tall, erect aerial hyphae almost reaching the cover of the petri plate. ATCC 16699 colony radial growth was exceptionally slow (2mm/day), whereas ATCC 58104 grew faster than the others (11 mm/day). No isolates grew at 35 C, while some grew at 5 C (Table 2). The leading hyphae were uniform and fine, measuring 5-6 um wide. In most isolates, small spherical to angular swellings in clusters or in chains were produced when mycelial agar discs were transferred to water although ATCC 46725 produced them also in old cultures.
Sporangia
All isolates produced sporangia readily in water within 24-48 hr, except ATCC 16698 and ATCC 64861 which formed them sparsely only in non-sterile stream water. The sporangia were non-deciduous, non-papillate and borne terminally on short sympodial branches of a slender sporangiophore. They were mostly 40-55 x 25-30 um (Table 2), obpyriform to ovoid with rounded base although elongated ellipsoidal sporangia with tapering bases were also found, especially in ATCC 36302, ATCC 46725 and ATCC 58104. ATCC 64047 produced distinctly larger sporangia (68+11 x 35+7 wpm). The apex of the sporangium in all isolates flattened easily when prepared for microscopy, and the empty sporangium partially collapsed after oospore release. The base of the sporangium was often plugged, and the ability of sporangia to proliferate internally varied with the isolate. Internal proliferation of sporangia was rare in ATCC 16699, ATCC 28766, ATCC 36302, ATCC 46725 and ATCC 58104 and occurred mostly in nonsterile stream water only, but it was commonly found in ATCC 16698 and especially ATCC 64047. Repeated emergence of zoospores was observed only in ATCC 46725.
Sex organs
Most isolates of P. erythroseptica produced abundant
77
e.Buesods jo AyLoned 03 anp sjuaueuNseaw Og UeYY ssa] (9)
SjuawauNseaw QG UO paseq ‘JOJJa puepueIs + UeaW (q)
uLeyd UL JO pasaysn}9 ‘yeynBue ‘})ewWs (e)
ee
as + + 2°0+9"4 £+92 2+6¢ -/+ -/+ -/+ L°O+S"L e+Le +19 -/+ -/+ -/+ L°O+9°L £+92 S+L9 ad -/+ a 2°0+2°L 9+0¢ 9+1LS => -/+ = 2°0+9°L £+0¢ 9+89 + > -/+ S°0+0°2 2+S¢ LL+89 -/+ + + 7°0+8°L £+9?2 8+c7 -/+ + + 9°0+0°2 S+Z2 2+69 -/+ -/+ -/+ L°0+0°2 £+82 2+9S -/+ -/+ Se £°0+8°1 S+L¢ 9+9S hes <= = £°O+2°L €+¢2 (9)G+8E -/+ = L°O+2°L £+82 S+L9 -/+ + -/+ £°0+8°1 9+92 8+l4 -/+ -/+ -/+ 2°0+9" 1 9+LE (9)94+84 uolzesaytjoud xade UO13914}SUu0) (un) (ur) yeusa3ul 3e)4 ue [paw 8/1 9) 43peasg(7)y36ua7 wn t6uesods
+ -- - IN 19879
- -- + 4N 9SL99
- -- + IN SSLY9
- -- -/+ dW 82L49
- -- + IN L279
-/+ -- -- | 29099
+ -- + 4S 7018S
+ -- + 4S G2299
+ -- -- dS c0L9¢
-/+ -- -- 4S 99l82
+ -- -- dW 66991
+ -- -- dW 86991
+ -- -- dW 92601
+ -- -/+ Fr] £2601
(@)s6ut) 24S 3S6€ 396 Auojo9 J91V yeuddy 3@ YyIMOI
eo13dasouyyAsa esoyyydoyAyg yo so1ysisajzoeseyo ye1BuesJods pue y3M0JN *2 a)qQe1
78
sex organs readily either in water immediately following sporangial production or in single cultures on agar plates an c2=3 wke The spherical oogonia were rather large, mostly 35-40 um diam (Table 3), narrowing abruptly to a tubular stalk and were non-pigmented, although they eventually became faintly yellow or straw-colored with age. The oospores were aplerotic to markedly aplerotic with oospore wall 2-4 um thick. The antheridium was unicellular, amphigynous and cylindrical. Occasionally, structures resembling paragynous antheridia, as described by Ho et al. (1983), were found in addition to the amphigynous antheridium. The dimensions of the sex organs are summarized in Table 3.
Table 3. Dimensions of sex organs of Phytophthora erythroseptica
Antheridium
Oogoni um Oospore Free oogonial ATCC diameter (Um) diameter (um) space (a) Type Length(um) Width()m) 10923 © 34#3 (b) 28+3 32% A 1442 1342 10924 37+4 3143 30% A 11+2(b) 12+) 16698 40+3 3145 40% A 1242 1641 16699 ik No sex organs 28766 38+2 32+2 29% A 12+2 12+2 36302 38+3 32+3 29% A 12+2 13+1 46725 3843 3143 33% A 1443 1342 58104 No sex organs produced in single culture 64047 41+1 31+4 43% A 17+3 1442 64127-3743 3143 30% A t20o! |) eee 64128 3742 30+2 34% A 13+2 12+1 64155 3444 273 37% A 1142 1341 64156 3742 31+2 30% A 12+2 12+4 64861 No sex organs produced in single cell
(a) Free oogonial space = Oogonium diam - oospore diam X 100 Oogonium diam
(b) Mean + standard error, based on 50 measurements A = Amphigynous
ATCC 46725 produced sex organs only in old cultures or on oat grains, whereas ATCC 16699 never produced sex organs. Sex organs were produced by ATCC 10924 only once in water but were consistently induced by pairing it with Al mating types of P. nicotianae and other heterothallic species of Phytophthora (Table 4). Similarly, ATCC 58104 never produced sex organs in single cultures but produced them readily when crossed with an A2 mating type of P. nicotianae and other heterothallic species. None of the other isolates of DP. erythroseptica demonstrated heterothallic behavior. However, the oogonia of ATCC 10924 and ATCC 58104 formed heterothallically by selfing on membranes were smaller, mostly 29-33 um diam and pigmented (brown), different from those normally formed in single cultures.
He)
Table 4. Production of sex organs in crosses between ATCC 10924 and ATCC 58104 Phytophthora erythroseptica and other species of Phytophthora
P. erythroseptica
Name ATCC —- Mating Type ATCC 10924 ATCC 58104 P. cinnamomi 32992 A2 --- +++ 32993 Al +++ ook P. cryptogea 46721 Al +++ Bee 52403 A2 ae ee P. nicotianae 38606 A2 --- +++ 38607 Al +++ AU oKe P. palmivora 26200 A2 --- +++ 26201 Al +++ Aes P. erythroseptica 10924 --- +++ 58104 +++ ae
Chlamydospores
No chlamydospores was produced by any isolate of P. erythroseptica under the stated cultural conditions.
DISCUSSION
Phytophthora erythroseptica is well known as the causal agent of pink rot of potato tubers throughout the world (Hickman, 1958; Stamps, 1978) although it has minor hosts in belladonna (Alcock, 1926), tulip (Buddin, 1938), calla (Tompkins & Tucker, 1947,1950), sugarcane (Steib & Chitton 21°50; 4Sangh, 1955) onron, (Hickman, 1953) 7) pea (Bywater & Hickman, 1959), tomato (Walker & McLeod, 1970), cineraria (Lucas, 1977), and calceolaria (anonymous, 1980). The disease in potato is characterized by formation of a deep salmon-pink coloration when affected tissue is exposed to the air due to a tyrosinase reaction (White, 1946) and was initially considered as a diagnostic symptom. It was later found that similar or identical symptoms could be induced by other species of Phytophthora by artificial inoculation, and in nature P. megasperma, P. cryptogea and P. drechsleri were also isolated from diseased potato (Tucker, 1931; Carnes & Muskett, 1933; Goss, 1949; Rowe & Schmitthenner, 1977). Thus, the precise identification of P. erythroseptica is critical to identification of the pathogen that causes pink rot of potato in the field. However, there have been controversies over the species concept of the pathogen. For the sake of discussion, the published data on P. erythroseptica are summarized in Table 5.
80
at ral v Al} 2b v aL St Vv v v v 6L-2t 6l-2b OV Vv Vv Vv v v v v Vv Vv Vv v Vv Vv Vv Vv v v v v (wr) (u r) yapim y3z6ue7 = adAL wn ipl sayzuy
£-2
7-2
£
paonposd uebso Xas ON
de 9e 9e
2£-62 0£-62 Le
62
62-S2 Le £e ef Lg Lg be ek ef £e 2g ef Le 2 o£ be
2-62 £e 9
11M (wrpwe tp
auodsoo
sl 9 vil va 8° L £¢ Zt 9 OL
9E-FE 9
2£-0£ 9°
9
8s
(wwe tp 8/1 wn 1u0600
92 89 g 27 ie 2s of 2s Ze 19 02 rh
9£-22 0S-62 02 ce
L2 ce
of 69
of 8”
92 4
82 SY
02 2g
(wr) y3peesg (wr) y36u87
wn i 6uesods
s6ut}]]emMs yeydAH
st S st S sf S Se S 82 g auesuebns 03e830d (q)03820d (q)03830d
2))e9 33144 G2. eS 81189 yUld
"¢ 8°47 03840d dyn (eyedoszy 03840d o£ Sl 03e810g edosiy edosiy 03830d JE) "xen ULW 3SOH aunyesedwe | y3Mos5
79 99 7
“324
A} 189160] 0u0sYyo 34nzesaz1) Ul papsozeJ B2tydasosyzAua esoyzydozAyd 4O S2t3StyazIeVeYD “GS 91981
81
tsid "yea ed1ydasosy Asa “4d (2)
Sl Sl v ce pe3ebu0)}3 = v 7-Gal acs: el OL v 6£ 2t-St Vv yeotupuryAd VW S£-02 <b vA v 9L-£b v Sl v 8L-OL O2-LL v 0£-62 9 dt v v3 eb £l v 92 cb Sl v bg £b £l v 82 £b Sl aN, be £b Sl v o£ £b SL v Le 2 vA v bg £l el v le 91 Sl v 82 £b vA! v £b vai v v 62 gL 6L v 92 call VAs oe o£ (wi rt) (w 1) y3pim =: y3Buay— aA 11eM(urpwe tp wniptsayjuy asodsog
LY 8°L Ze 91 ev 91 07-02 0S-82 S£-0£ Z°b c¢ OL 9° L-9°L ray] 9° 9¢ a 5 S¢ ot S¢ SL SE Lb SE 971 5 9 91 SE SL 2% aL 91 SL s¢ aL 89-249 SL G¢-0£ 2b L°2 dg S2 o£ ol (wy{we ip 8/1 wn tuo609
(edtydasosy3Aua *d)
= StsuaAe)]ewl
ug "9 yeoi3diy)3
of 99 is ”
8£-8L $9-82 92 £4 22 9
0S-S 02 ze 22 £7 82 29 92 £7 92 Ze 92 99 22 £7 22 £0 22 27 92 27 Ls LS o¢ 89 62 LY is ef 92 £7 62 £9 Se 98 33 £S
(url) y3peaug (wr) y36ua7
wnn i Buesods
S£-0£ 9%
+
S£-ek
o£
s6u1]]9mMS
yeudduy
OL-S 6
4
me) 09.
“xeW aunzesedwa |
“ULW
yamou9
201d PTIA 04e40d 03830q 038304 oewWo!
eLyesaulg 038}0g
03830q
03820q
03830q
03840d
(2)e8aq (q)03830q
3SOH
Yd (qQ) edosje “yea eatydasosyyAua “gq (e)
8s
“32a
(panuijuo2) *¢ 3]qe1
82
Our study confirmed the findings of previous workers concerning the cultural and hyphal characteristics of P. erythroseptica. Isolates of P. erythroseptica grew well on common agar media, producing non-fluffy CO Telurfy colonies with uniform and fine hyphae. ATCC 16699 which grew exceptionally slow, showed signs of degeneration probably as a result of long-term storage. In literature, P. erythroseptica var. piSi was unusual in having irregular, tortuous hyphae, whereas the luxuriant, tall aerial mycelium of ATCC 64047 in the present study distinguished itself from the other isolates. Small angular to spherical hyphal swellings, in clusters or in chains, as reported earlier by some researchers, were also produced by some isolates in the present study, usually in water culture. Chlamydospores were never observed. Previous authors also failed to record chlamydospores in P. erythroseptica although Ershad (1971) and Krober (1985) reported chlamydospores, whereas hyaline spherical structures were interpreted as "poorly differentiated chlamydospores" by Tompkins & Tucker (1947) or "chlamydospore-like bodies" by Novotelnova (1974). As noted by previous workers, no isolates Offa: (BP. erythrospetica grew at 35 C. The sole report of good growth at 35 C by P. erythroseptica (Van der Zwet & Forbes, 1961) needs to be confirmed. Tucker (1931) considered the rather narrow temperature range (15-30 C) a diagnostic character of P. erythroseptica, but this could neither be confirmed in the present study nor by most authors in the literature.
In the species description of P. erythroseptica by Waterhouse (1963) and Stamps (1978), the sporangia were described as variable in shape, ellipsoid or obpyriform (43 - 26 um; L/B=1.65), often constricted near the middle, sometimes tapering to the sporangiophore. [In our study, the overall mean length(L) and breadth(B) of the sporangia for all isolates were 48 + 7 x 28 + 3 wm with L/B=1.7 + 0.2, agreeing well with those reported in literature (46 ree 4pm = lage linia: vey U3 Bese PY ahs Be Tui sua Oey ee ATCC 64047 produced distinctly larger sporangia that were similar to those reported for P. erythroseptica var. pisi although not as narrow. Median constrictions of sporangia were not mentioned in the original description (Pethybridge, 1913) and have been reported to occur only occasionally by Bywater & Hickman (1959) and Vargus & Nielson (1972). They were observed in some isolates in the present study, and were especially common in ATCC 46725 and ATCC 58104 which produced mostly elongated sporangia with tapering bases. The elongated sporangia of the type culture (ATCC 58104) differed from the obpyriform sporangia diagramed by Pethybridge (1913) for P. erythroseptica. It is not clear if the type culture has undergone changes accounting for the difference which could conceivably be due to different culture conditions. Similarly, ATCC 16699, the sporangia of the type culture of P. himalayensis (=P. erythroseptica) were oval to ellipsoidal instead of
83
"slipper-shaped" as described by Dastur initially (1948). The broad apex of sporangium of P. himalayensis could be due to its flattening upon mounting.
As reported previously, the sporangiophore of P. erythroseptica showed distinct sympodial branching, but we found that in most cases, the branching was close rather than "lax" (Waterhouse, 1963), and our observation seemed to agree with those of other authors based on their diagrams and/or photographs. Waterhouse & Blackwell (1954), Hickman (1940) and Waterhouse (1963) stated that internal proliferation of sporangia was rare in P. erythroseptica, whereas Gerrettson-Cornell (1981) failed to observe it in an Australian isolate from potato. In our study, the ability of the sporangium to proliferate internally varied with the isolate but was usually not as common as in other related species of Phytophthora with non-papillate sporangia. Many sporangia were plugged at the base after zoospore release. In fact, the sporangia of most isolates of P. erythroseptica, did not release zoospores easily unless the sporangia were chilled (Vujae1c eS). Colhoun;,!))1966) ; Pethybridge (1931, 1914) experienced difficulty in inducing zoospore release in P. erythroseptica, whereas Dastur (1948) reported that the sporangia of P. himalayensis (=P. erythroseptica) germinated only "conidially". Neither of them noted internal proliferation of sporangia.
We have also confirmed previous observations that P. erythroseptica produces readily in single culture abundant rather large, smooth spherical oogonia narrowing abruptly to a tubular stalk. Apparently, some isolates of P. erythroseptica have lost their self-fertility due to prolonged storage and/or repeated subcultures. It is interesting to note that ATCC 10924 and ATCC 58104 behaved heterothallically instead, mating successfully with A, and A, mating types, respectively, of various Phytophthora species. This may be considered a case of "secondary heterothallism" derived from a homothallic precursor, lending support to the hypothesis that heterothallism probably evolved from homothallism (Brasier, 1983; Ho, 1986). Heterothallic isolates of normally homothallic P. megasperma have also been reported (Barr, 1980). However, the oogonia formed by ATCC 10924 and ATCC 58104 as a result of selfing on membranes when crossed with the appropriate mating strain of Phytophthora were smaller and brown in color, different from the larger, usually non- pigmented to pale yellow oogonia formed in single cultures of P. erythroseptica. It is difficult to assess the differences in oogonia characteristics without knowing the changes that have occurred during storage; the heterothallic behavior should not be considered typical for the species. ATCC 16698 never produced any sex organs alone or in pair cultures. The loss of sexuality by isolates of P. erythroseptica in artificial culture was noted by Bywater (1956), Carnes & Muskett (1933) and
84
Ribeiro et al. (1975). Waterhouse (1963) and Stamps (1978) described the oospores of P. erythroseptica as "nearly filling oogonium", but we found that the oospores in most isolates largely aplerotic. The overall mean values of oogonial diameters and oospore diameters for all isolates in our study were 37 + 2 and 34 + 4 un, respectively, agreeing well with those in the literature (39 + 3 and 34 + 5 um, respectively).
We are also in agreement with those authors who believed that P. erythroseptica produces solely amphigynous antheridia (Hickman, 1958; Tucker, 1931; Waterhouse & Blackwell, 1954; Waterhouse, 1963; Stamps, 1978; Gerrettson-Cornell, 1985). The brief report by Cooper (1928) that P. erythroseptica produced only paragynous instead of amphigynous antheridia should be questioned. It is also our opinion that Phytophthora species identified as P. erythroseptica in the past producing both amphigynous and paragynous antheridia should be reclassified because the identification was either debatable or provisional. Thus, the causal agent of root rot of red raspberry was identified as P. erythroseptica by Converse & Schwartz (1968) is sRs megasperma by Duncan et al. (1987) and P. fragariae by Wilcox (1989). Pratt (1981) identified some fast growing isolates from arrowleaf clover tentatively as P. erythroseptica, but a thorough search of the literature and a comparative cultural study under similar conditions showed that these isolates are within the range of intraspecific variability for P. megasperma and could be identified as such (Ho, unpublished). Morgan & Johnson (1965) identified the pathogen from vetch as P. erythroseptica because it produced only amphigynous antheridia. However, paragynous antheridia were later found in their isolate (Savage et al., 1968), and it should be re-classified as P. megasperma which it resembled closely based on the published photographs. There is no doubt that the antheridial type, along with the sporangial papillation, is the most important taxonomic criterion in Phytophthora (Tucker, 1931; Waterhouse, 1983; Gerrettson-Cornell 1985). It would be unwise to cloud the species concept of P. erythroseptica by including controversial isolates that produce both amphigynous and paragynous antheridia (Savage et al., 1968; Pratt, 1981). Gerrettson-Cornell (1985) considered the occasional presence of two or three antheridia (amphigynous) to a single oogonium a _ characteristic feature of P. erythroseptica but we could not confirm this observation in present study.
Since Pethybridge erected P. erythroseptica as a new species (1913) Leonian & Geer (1929) suggested combining P. erythroseptica and mexicana Hotson & Hartge, but this was promptly rejectea by Tucker (1931). Bywater & Hickman (1959) proposed that P. cryptogea, P. drechsleri, P. erythroseptica, P. himalayensis and P. richardiae Buis
85
should be grouped together as one species: P. erythroseptica. Although we agree with Bywater (1956) that these species have much in common especially in growth-temperature relations and sporangial characteristics, P. richardiae seems distinct enough to be retained as a separate species (Ho, unpublished), whereas P. cryptogea (P. drechsleri) can be differentiated based on its heterothallism and usually smaller, brown
oogonia with plerotic oospores. The merging of P. Gdrechsleri with P. cryptogea, which has priority, is favored by many workers (Ho & Jong, 1986). Sarejanni
(1936) treated P. drechsleri as P. erythroseptica var. drechsleri, but it was not well received. P. himalayensis should be treated as synonymous with P. erythroseptica as suggested by Waterhouse (1963). Thus, P. erythroseptica is characterized by non-papillate sporangia, homothallism and entirely emphigynous antheridia. Based on this broad species concept, the pea isolate was treated as a variety of P. erythroseptica: P. erythroseptica var. pisi (Bywater & Hickman 1959) even though initially it was considered different enough to warrant a new species epithet: P.
TSic (Bywater, 1956)7. The acceptance of P. erythroseptica var. pisi as a validly published taxon is debatable. Article 37 of the International Code of
Botanical Nomenclature (Voss, 1983) stipulates that "publication on or after 1 Jan. 1958 of the name of a new taxon of the rank of family or below is valid only when the nomenclatural type is indicated". Waterhouse (1963) considered P. erythroseptica var. pisi, which was published in 1959 without specifically citing a type specimen according to recommendations 37A and 37B, to be invalid. On ‘the other: (hand, “i2t. is) trueithat the nomenclatural type was clearly indicated in Bywater & Hickman's paper. We believe that this well-defined and well-described taxon should be retained. The isolate from wild rice is of special interest. Although it was identified as P. erythroseptica sensu lato, (Grunnel & Webster, 1988), it is different from the other isolates in its distinctly fluffy colony, larger sporangia, poor growth on malt extract agar medium, inability to hydrolyze starch, sensitivity to HMI and lack of pigment production on casein-hydrolysate medium (Ho, unpublished) and may be considered as a new variety. ATCC 10924, received originally as P. drechsleri (Ho & Jong, 1986), is now re- classified as P. erythroseptica based on its homothallism. The isolate of P. erythroseptica from sugarcane was identified primarily based on the production of oogonia with amphigynous antheridia in single cultures (Steib & Chilton,-129507)) Van “der cZwet. -& "Forbes, \\196L)%" but: a detailed description of the fungus was not published. Based on the data and photographs in Singh's Ph.D. Thesis (L955)), ~the unusually large oogonia. 3(46.75 (494019 om diam), the rather large obypyriform sporangia (60.94 + 6.25 X 37.38 + 3.66 wm; L/B=1.6) and the spherical hyphal swellings spaced evenly in chains suggest greater Similarity with the large-spore type of P. megasperma
rather than P. erythroseptica sensu stricto. Perhaps, the antheridial configuration of the sugarcane isolate should be re-examined to determine if it produces any paragynous antheridia. Erwin (1965) initially identified the alfalfa isolate as P. cryptogea on account of its amphigynous antheridia but later renamed it as P. megasperma when paragynous antheridia were found.
ACKNOWLEDGEMENTS
This work was supported in part by a grant-in-aid from the Whitehall Foundation to H.H. Ho and NSF Grant BSR 8413523 to S.C. Jong. The authors thank Dr. W. Chris Dermody, Dr. Larry McDaniel and Mr. Elmer E. Davis for reviewing the manuscript.
REFERENCES
1. Alcock, N.L. 1926. A preliminary note ona Phytophthora on Altropa belladonna. Pharmaceut. J. D638) 23:2".
2. Anonymous. 1980. New records of disease and of fungi. Report, Botany Department, Victoria University of Wellington, No. 10. 20 pp In: Rev. Pl. Pathots) S93 easi73
3. Barr, D.J.S. 1980. Heterothallic-like reaction in the large-oospore form of Phytophthora megasperma. Cans ):d. Pl.’ Pathol.)'23) 116-118.
4. Brasier, C.M. 1983. Problems and prospects in Phytophthora research. Pages 351-364 in: Phytophthora: Its Biology, Taxonomy, Ecology, and Pathology. D.C. Erwin, S. Bartnicki-Garcia and P.H. Tsao, eds. Am. Phytopathol. Soc., St. Paul, MN. 392 Pp-
5. Buddin, W. 1938. Root rot, shoot rot and shanking of tulip caused by Phytophthora cryptogea Pethybr. & Laff. and P. erythroseptica Pethybr. Ann. Appl. Biol. 292 705-729..
6. Bywater, J. 1956. Studies on organisms causing root and stem diseases of peas. Doctoral Dissertation, Univ. Birmingham, Birmingham, England. 169 pp.
7. Bywater, J. & C.J. Hickman. 1959. A new variety of Phytophthora erythroseptica, which causes a soft rot of pea’ roots." Trans.) (Br. (Mycol. Soc 42 e /S13-924:.
8. Cairnes, H. & A.E. Muskett. 1933. Pink rot of the potato. Ann. Appl. Biol. 20: 381-403.
9.) Christ) (Bodie 5 eis Kam SD. Ne (OLSONL 1987.0 pPankeTot of potato caused by Phytophthora erythroseptica in Pennsylvania (Abstr.) Plant Dis. 72: 747.
10.Converse, R.H. & C.D. Schwartz. 1968. A root rot of red raspberry caused by Phytophthora erythroseptica. Phytopathology 58: 56-59.
11.Cooper, D. 1928. Paragynous antheridia of Phytophthora spp. (Abstr.). Phytopathology 18:149-150.
87
12.Dastur, J.F. 1948. Phytophthora spp. of potatoes (Solanum tuberosum L.) in the Simla hills. Indian Phytopathol. 1: 19-26.
13.Duncan, J.M., D.M. Kennedy, & E. Seemuller. 1987. Identities and pathogenicities of Phytophtora spp. causing root rot of red raspberry. Plant Pathology 36; 276-289.
14.Ershad, D. 1971. Beitrag zur Kenntnis der Phytophthora arten in Iran und ihrer phytopathologischen Bedeutung. Mitt. bio. BundAnst. Ld. u. Forstw. 140. 84 pp.
15.Erwin, D.C. 1965. Reclassification of the causal agent of root rot of alfalfa from Phytophthora cryptogea to P. megasperma. Phytopathology 55: 1139- 1143.
16.Erwin, D.C., G.A. Zentmyer, J. Galindo, & J.S. Niederhauser. 1963. Variation in the genus Phytophthora. Annu. Rev. Phytopathol. 1: 375-396.
17.Gerrettson-Cornell, L. 1981. Note on Phytophthora erythroseptica Pethybridge. (Abstr.) Phytophthora Newsletter 9: 26.
18.Gerrettson-Cornell, L. 1985. A working key to the species of Phytophthora de Bary. Acta Bot. Hung. 31: 89-97.
19.Goss, R.W. 1949. Pink rot of potatoes caused by Phytophthora erythroseptica Pethyb. Univ. Nebraska Agr. Exp.'Sta.| ‘Res. Bull. (1603 1-27.
20.Grunnell, P.S. & R.K. Webster. 1988. Crown and root rot of cultivated wild rice in California sensu lato. Plant Dis. 72: 909-910.
21.Hickman, C.J. 1940. The red core root disease of the strawberry caused by Phytophthora fragariae sp. n. J. Hort. Sci. 18: 89-118.
22.Hickman, C.J. 1958. Phytophthora - plant destroyer. Trans. Br. Mycol. Soc. 413) 1-13.
23.Ho, H.H. 1986. Notes on the heterothallic behavior of Phytophthora megasperma from alfalfa. Mycologia 78: 306-309.
24.Ho, H.H., W.Y. Zhuang, Z.R. Liang & Y.N. Yu. 1983. Phytophthora cinnamomi on black locust (Robinia pseudoacacia) in Jiangsu Province of China. Mycologia 75: 881-886.
259.HO, "HH. & S.C. Jong. 1986. A comparison between Phytophthora cryptogea and P. drechsleri. Mycotaxon ZL V2e9e329'.
26.Ilieva, E. 1979. Pathogens of Phytophthora rot of glasshouse tomato. Hort. Vitic. Sc. 16: 67-74.
27.Jones, W. 1944. Pink rot disease of potatoes in British Columbia. Sci. Agr. 25: 597-600.
28.Jones, W. 1954. Pink rot of potato tubers on Vancouver Island. Can. J. Agr. Sc. 34: 504-506.
29.Ko, W.H. 1978. Heterothallic Phytophthora: Evidence for hormonal regulation of sexual reproduction. J. Gen. Microbiol. 107: 15-18.
30.Kouyeas, H. & A. Chitzanidis. 1968. Notes on Greek species of Phytophthora. Ann. Inst. Phytopath. Benaki,
N.S.16% 1127 5=192..
31.Krober, H. 1985. Experiences with Phytophthora de Bary and Pythium Pringsheim. Bundesanst fur Land-und Forstw. (Berlin-Dahlem) 225: 1-175.
32.Leonian, L.H. 1934. Identification of Phytophthora species. W. Virginia Univ. Agr. Exp. Sta. Res. Bull. 2625 42-363
33.Leonian, L.J. & H.L. Geer. 1929. Comparative value of the size of Phytophthora sporangia obtained under standardized conditions. J. Agr. Res. 39: 293-311.
34.Lucas, R. 1977. Phytophthora erythroseptica Pethy. wilt of cineraria (Senecio cruentus D.C.) cultivars. N.Z.J. Agr. Res. 20: 253-254.
S5eLu0, (“Lb bHsie “HO; aa S.C. Jong. 1988. Study on the physiological characteristics of the genus Phytophthora. Mycotaxon 32: 199-217.
36.Morgan, F.L. & H.W. Johnson. 1965. Phytophthora root and crown rot of vetch. Pl. Dis. Rep. 49: 84-88.
37.Mundkur, B.D. 1949. Morphology and cytology of development of the sex organs of Phytophthora himalayensis Dastur. Bot. Gaz. 110: 475-486.
38.Murphy, P.A. 1918. The morphology and cytology of the sex organs of Phytophthora erythroseptica Pethyb. Anns, Bot. 5322 2—153%
39.Newhook, F.J., G.M. Waterhouse, & D.J. Stamps. 1978. Tabular Key to the Species of Phytophthora de Bary. Mycol. Pap. 143, Commonw. Mycol. Inst. Key, Surrey, England. 20 pp.
40.Novotel'nova, N.S. 1974. The Genus Phytophthora Sov. Acad. Sci. Bot. Inst. Lening. U.S.S.R. 206 pp.
41.Pethybridge, G.H. 1913). On the rotting of potato tubers by a new species of Phytophthora having a method of sexual reproduction hitherto undescribed. Sc. Proc. R. Dubl. Soc. 13: 529-564.
42.Pethybridge, G.H. 1914. Further observations on Phytophthora erythroseptica Pethybr., and on the disease produced by it in the potato plant. Sc. Proc.’ R.iDubl.) ‘Sec. 179-198.
43.Pratt, R.G. 1981. Morphology, pathogenicity, and host range of Phytophthora megasperma, P. erythroseptica, and P. parasitica from arrowleaf clover. Phytopathology 71: 276-282.
44.Ribeiro, O.K. 1978. A Source Book of the Genus Phyph- thora. J. Cramer, Vaduz, Germany. 417 pp.
45.Ribeiro, O.K., Erwin, D.C., & G.A. Zentmyer. 1975. An improved synthetic medium for oospore production and germination of several Phytophthora species. Mycologia 67: 1012-1019.
46.Rosenbaum, J. 1917. Studies on the_- genus Phytophthora. J. Agric. Res. 8: 233-276.
47.Rowe, R.C. & A.F. Schmitthenner. 1977. Potato pink rot in Ohio caused by Phytophthora erythroseptica and P. cryptogea. Pl. Dis. Rep. 61: 807-810.
48.Sarejanni, J.A. 1939. La pourriture du collet des Solanees cultivees et la classification du genre Phytophthora. Ann.Inst.Phytopath. Benaki 2: 35-52.
89
B9,SaVvage, (bed. ) C.Ws. Clayton, (Ji. (hunter, JA. Brenneman, C. Laviola, & M.E. Gallegly. 1968. Homothallism, heterothallism, and interspecific hybridization in the genus Phytophthora. Phytopathology 58: 1004-1021.
50.Schwinn, F.J. 1959. Untersuchungen zur systematik der gattung Phytophthora de Bary. Arch. Mikrobiol. Bos 2es~-20e.
51.Singh, ‘K.1) 1955... ‘Phytophthora rot of) sugarcane. Ph.D. Dissertation, Louisiana State Univ., Baton Rouge, Louisiana. 147 pp.
52.Stamps, D.J. 1978. Phytophthora erythroseptica. C.M.I. Descriptions of pathogenic fungi and bacteria NO@wo 93." 72>) pp.
53 -sceib, uB.d. & S.d.P. Chilton: 1950. The Phytophthora rot of sugarcane seed pieces in Louisiana. Proc.Intern.Soc.Sugar Cane Technologists, 7th Congr.: 524-528.
54.Tompkins, C.M. & C.M. Tucker. 1947. Leaf blight of pink calla caused by Phytophthora erythroseptica. Phytopathology 37: 382-389.
55.Tompkins, C.M. & C.M. Tucker. 1950. Rhizome rot of white calla caused by Phytophthora erythroseptica. Agr. Res. 40: 712-714.
56.Tucker, C.M. 1931. Taxonomy of the genus Phytophthora de Bary. Univ. Mo. Agr. Exp. Sta. Res. BUDD. Ds . 81-208
57.van der Zwet, T. & I.L. Forbes. 1961. Phytophthora megasperma, the principal cause of seed piece rot of sugarcane in Louisiana. Phytopathology 51: 634-640.
58.Vargas, L.A. & L.W. Nielsen. 1972. Phytophthora erythroseptica in Peru: its identification and pathogenesis. Am. Potato J. 69: 309-320.
59.Voss, E.G. (Ed.) 1983. International Code of Botanical Nomenclature. Reg. Veg. 111: 1-427.
60.Vujicic, R. & J. Colhoun. 1966. Asexual reproduction in Phytophthora erythroseptica. Trans.Br.Mycol.Soc. 49: 245-254.
61.Walker, J. & R.W. McLeod. 1970. New records of plant diseases in New South Wales, Agr. Gaz. New South Wales 81: 452-458.
62.Waterhouse, G.M. 1963. Key to the species of Phytophthora de Bary. Mycol. Pap.92. Commonw.Mycol. Inst., Kew, Surrey, England, 22 p.
63.Waterhouse, G.M. & E.M. Blackwell. 1954. Key to the species of Phytophthora recorded in the British Isles. Mycol. Pap. 57. Commonw. Mcol. Inst., Kew, Surrey, England. 9 pp.
64.Westerdijk, J. & A.V. Luijk. 1920. Phytophthora erythroseptica Peth. als parasit von Atropa belladonna. Meded. Phytopath. Lab."Willie Commelin Scholten" 4: 31-32.
65.White, N.H. 1946. Host parasite relations in pink rot of potato.. J. Austr. Inst. Agr. Sc. 11: 195-197.
66.Wilcox, W.F. 1989. Identity, virulence, and isolation frequency of seven Phytophthora spp.
90
causing root rot of raspberry in New York. Phytopathology 79: 93-101.
MY COTAXON
Vol. XXXVI, No. 1, pp. 91-94 October-December 1989
DEUTEROMYCOTINA FROM ANTARCTICA
NEW SPECIES OF HYPHOMYCETES FROM DANCO COAST, ANTARCTIC PENINSULA
Marta N. Cabello* Instituto Spegazzini, 53 N2 477,1900 La Plata,Argentina. SUMMARY
Three new species of Hyphomycetes have been found recently in the Antarctic Continent, growing on rhizosphere soil of Colobanthus guitensis (Kunth) Bartl. (Caryophylla- ceae) and Deschampsia antarctica Desv. (Gramineae).
The new species here proposed are: Acrodontium antarc- ticum nov. sp., Ghalara antarctica nov. sp. and Phialophora dancoii nov. sp.
These new species were found during the "Campafia Antartica Argentina de verano 1989" in Danco Coast, Base Primavera (64210'S, 60957'W). The same locality is indica- ted in a map by Gamundf and Spinedi (1987).
The fungal isolation was carried out using the soil washing technique (Parkinson and Williams, 1961).
The following descriptions concerns observations in pure culture.
ACRODONTIUM ANTARCTICUM Cabello nov. sp. Fig. 1.
Coloniae in vitro post 20 dies 15 mm diametro, floccosae vel funiculosae, primum grisae ad olivaceae, mox nigrescentes. Reversum nigrum. Hyphae hyalinae ad pallide olivaceae, glabrae. Cellulae coni- diogenae polyblasticae, integratae vel discretae, sympodiales, pallide brunneae, ad apicem pallidiores, plerumque 9-12 x 2.4-3 um et rachide tenuiter denticulata, recta vel flexuosa, 0.9=-1 um crassa, ad 2.5-6.5 longa. Conidia acropleurogena, simplicia, hyalina, laevia, ovoidea basi apiculata, 2.5-3.5 x 1.5=2 um.
HOLOTYPUS: Antarctica, Peninsula Antarctica, Terra de Danco, Base Primavera, leg. M.N.Cabello, 31-I-1989. Ex solo. LPS 44594.
Colonies in vitro growing rather slowly, attaining a diam. of 15 mm in 20 days, appearing floccose or somewhat funiculose, at first grey to olivaceous, soon becoming blackish. Reverse black. Hyphae hyaline or pale olive,
*Researcher of the Comisidn de Investigaciones Cientificas de la Provincia de Buenos Aires (CIC), Argentina.
92
smooth. CGonidiogenous cells polyblastic, integrated or discrete, sympodial, pale brown, paler towards the apex, 9-12 um hight, 2.4-3 um wide at the base,rachis denticulate often not distinctly differentiated from the- basal part, straight or flexuous up to 2.5-6.5 um long and 0.9-1 um wide. CGonidia acropleurogenous, hyaline, simple, smooth, ovoid, with an apiculate base, 2.5-3.5 x 1.5-2 um.
Our species differs from A. crateriforme (van Beyma) de Hoog (de Hoog, 1972) by the smaller size of the conidio- genous cells and the denticulate rachis which in A. crate- riforme can be up to 45 um while in our species is up to 6.5 um. It also differs by the lack of sharp denticles in our species.
Holotypus: Antartic Peninsula, Danco Coast, Base Primavera, leg. M.N.Cabello, 31-I-1989, rhizosfere soil of Colobanthus quitensis. LPS 44594.
CHALARA ANTARCTICA Cabello, nov. sp. Fig. 2-4.
Coloniae in vitro post 30 dies 50 mm diametro, floccosae vel tomentosae, brunneae. Reversum brunneum. Phialophora erecta, simpli- cia, 1-4 cellularia, brunnea, cum phialidibus 60 um longa. Phialides elongatae, haut inflatae ad basim, 20-25 x 3 um; cylindraceo vel conico, interne 2-3 coniidis munitae, 7-10 um longa. Conidia ellipsoi- dea, hyalinea, 3-4 x 2-2.1 um. Chlamydosporae absunt.
HOLOTYPUS: Antarctica, Peninsula Antarctica, Terra de Danco, Base Primavera, leg M.N.Cabello, 2-II-1989. Ex solo. LPS 44595.
Colonies in vitro attaining a diameter of 50 mm in 30 days, appearing floccose or tufted, brown. Reverse dark brown. Phialophores erect, simple, bearing phialides. Total lenght of phialophores with phialides 60 um with 1-4 septa. Phialides with a medium brown, slightly or not inflated venter, 20-25 x 3 um, and a cylindrical or obconical, light brown collar, cointaining 2-3 conidia, 7-10 um long. Conidia ellipsoidal, hyaline 3-4 x 2-2.1 um. Aleuroconidia or chlamydospores absent.
The more related species are C. microspora (Corda) Hughes and G. neocaledoniae Dadant ex Kiffer and Delon. Our species differs from C. microspora (Nag Raj and Ken- drick, 1975) by the size of the conidia (C. microspora 3-8.5 x 1-1.5 um, CG. antarctica 3-4 x 2-2.1 au). It) aleo differs from C. neocaledoniae (Kiffer and Delon, 1983) by the size of the phialide that is larger in this species. Holotypus: Antarctic Peninsula, Danco Coast, Base Primave- ra, leg. M.N.Cabello, 2-II-1989, rhizosphere soil of Colo- banthus quitensis and Deschampsia antarctica. LPS 44595.
ACRODONTIUM ANTARCTICUM. 1.Conidial structures and conidia.
CHALARA ANTARCTICA. 2. Conidiophore, phialide and conidia. 3. Phiali- des and conidia. 4. Conidia.
PHIALOPHORA DANCOII. 5. Phialides and conidia. 6. Conidia.
93
94
PHIALOPHORA DANCOII Cabello nov. sp. Fig. 5-6.
Coloniae modice lente crescunt, olivaceo-grisae, pulverulentae, tomentosae. Reversum olivaceo-nigrum. Hyphae vegetativae modice pig- mentatae, 1.2-2 um latae. Phialides simplices vel conidiophora compo- sita ramosa; phialides brunneae, 11-12 x 2-3 um, collare 1-2 um altum divergens. Conidia agregata, ellipsoidea, hyalina, laevia, 2.5-5 x 1.5-2 um.
HOLOTYPUS: Antarctica, Peninsula Antarctica, Terra de Danco, Base Primavera, leg M.N.Cabello, 30-I-1989. Ex solo. LPS 44596.
Colonies reaching 36 mm diam. in 24 days at 202C; olivaceous grey, powdery and tufted. Reverse olivaceous black. Vegetative hyphae slightly pigmented, 1.2-2 um wide. Gonidiophores consisting of simple phialides, someti- mes septate and branched. Phialides brown, 11-12 x 2-3 um, collarette 1-2 um, slightly divergent. Sympodial proli- feration absent. Conidia in heads, ellipsoidal, hyaline, smooth-walled, 2.5-5 x 1.5-2 un.
The species most closely related with P. dancoii is P. phaeophora Gams (Gams and Holubové-Jechova, 1976), but differs by the shape of the conidia which are dacryoid with a truncate base in Gams species. Holotypus: Antarctic Peninsula, Danco Coast, Base Primave- ra, leg M.N.Cabello, 30-I-1989. Isolated from rhizosphere
soil of Colobanthus quitensis. LPS 44596. AKNOWLEDGEMENTS
I thank Drs. J. E. Wright and I. J. Gamundi for their criticisms on reading the manuscript.
I acknowledge the Direccién Nacional del Ant&rtico, Instituto Antdrtico Argentino, for supporting the expedi- tion.
REFERENCES
GAMUNDI, I.J. & H.eA.SPINEDI, 1987. Sclerotinia antarctica nov. sp. the teleomorph of the first fungus described from Antarcti- cae Mycotaxon 29: 81-89.
GAMS, We & V. HOLUBOVA-JECHOVA, 1976. Chloridium and some other dema- tiaceous Hyphomycetes growing on decaying wood. Studies in Mycology 13: 1-99.
HOOG de, G S. 1972. The genera Beauveria, Isaria, Tritirachium and Acrodontium gen. nov. Studies in Mycology 1: 1-41.
KIFFER, E. & R. DELON, 1983. Chalara elegans (Thielaviopsis basicola) and allied species. II. Validation of two taxa. Mycotaxon 18(1): 165-1746
NAG RAJ, T.R. & Be KENDRICK. 1975. A monograph of Chalara and allied genera. W. Laurier Univ. Pr. Waterloo.
PARKINSON D. & S.T. WILLIAMS. 1961. A method for isolating fungi from soil microhabitats. Plant Soil 13: 347-355.
MYCOTAXON
Vol. XXXVI, No. 1, pp. 95-145 October-December 1989
STUDIES ON PHOLIOTA IN CULTURE
STIG JACOBSSON
University of Goteborg Department of Systematic Botany Carl Skottsbergs gata 22 413 19 Goteborg, Sweden
ABSTRACT
Cultural characters are reported for 22 species of Pholiota and allied genera. One or several polarity tests have been made for most species in order to determine their mating systems. One species (P. jahnit) is bipolar, the others tetrapolar but some are amphithallic. The culture characters reported here add new information of value for intrageneric delimitations. Intercompatibility tests are performed where the species limits are unclear. One new combination is proposed: Pholiota lignicola(Peck)S. Jacobss.
INTRODUCTION
In modern fungal taxonomy studies of mycelia in culture play an im- portant role. Culture characters have been described for a great number of species, especially those belonging to Polyporaceae and Corticiaceae s.1. Interfertility tests are increasingly used to delimit species and resolve dis- cussions of synonymy at the species level. Sometimes the use of culture characters and interfertility tests is the only way to solve such problems. In Agaricales s. 1. only fragmentary culture data are hitherto available but it is evident that they are valuable for a better understanding of the taxo- nomy. With earlier known methods only the saprophytic species have been cultured, thus many of the large, mycorrhizal genera are unknown in cul- ture.
Pholiota (Fr.)Kummer is a genus of saprophytic species. Most species in the genus are active wood destroyers and, sometimes, they are parasitic.
96
Some live on charcoal, soil or humus but no species forms ectotrophic mycorrhiza. Since lignicolous species often are relatively easy to culture members of Pholiota were among the first agarics to be studied in that way.
Vandendries (1933 and 1934) and Vandendries & Brodie (1933) studied some species of Pholiota in culture and performed polarity tests. Kiihner (1946) made morphological and caryological studies on P. gummosa. Denyer (1960) studied Pholiota alnicola in culture. The mating system in P. adiposa was studied by Arita & Mimura (1969) and later Arita (1979) published a comprehensive thesis on the cytology of P. adiposa and P. nameko. Farr, Miller & Farr (1977) carried out biosystematic studies in the adiposa group by using interfertility tests. Hiibsch (1978) investigated the’ different types of conidia (accessory spores) in some species.
However, the published papers based on cultural studies of Pholiota are still rather few and only certain species are involved. One is P. adi- posa, but as this name is interpreted in different ways (cfr Jacobsson 1987) it is not always clear which species had been studied.
The aim of this study is to get additional viewpoints for a better un- derstanding of the systematic relationships within Pholiota by the aid of culture characters and interfertility tests. Twentytwo species belonging to Pholiota or allied genera have been cultured and studied in different ways.
MATERIAL AND METHODS
Collecting has taken place in various parts of Sweden and Denmark during 1979 - 1988. Basidiospores from collected basidiocarps were, as soon as possible, dispersed on plastic petri dishes with common malt agar for germination. From the germinated spores of each collection a number of monosporous mycelia were isolated. If a sufficient number of single spore (ss-)isolates were obtained, the breeding ability (the ability to form clamps in matings) and the mating-type was investigated by pairing all isolates with each other (polarity tests). The presence or absence of clamps, occurrence of accessory spores and other characters were examined when the mycelia had been in contact with each other for at least three or four weeks.
For some species no Swedish collections were available in culture. Mycelia of Pholiota albocrenulata and P. lucifera were obtained from CBS Baarn, the Netherlands. Compatibility tests between European and Ameri- can specimens have been performed between Swedish material and some isolates of P. alnicola, P. limonella and P. squarrosoides, which were ob- tained from DAOM, Canada.
After the polarity tests, compatibility tests between different strains of the same species were performed. Compatibility tests between different
97
forms were also made as a method to delimit closely related species or to confirm infraspecific variation. Intercompatibility is accepted as a strong support for conspecificity. Negative results consequently indicate different species but it is well known that there exist other explanations for negative pairings between specimens within a biological species, e.g. a reduction in mating capacity due to degenerative mutations or senescence (cfr Boidin, 1986).
Sometimes only polyspore (ps-) cultures with clamped hyphae were available for compatibility tests. They were then used in di-mon matings. In this type of crossing, a piece of a ps-mycelium is inoculated at the edge of a malt agar dish, while pieces of ss-cultures are inoculated in a half- circle around the ps-culture. After being in contact for four weeksor more the mycelium on the outside of the half-circle is checked for the occur- rence of clamps (Buller phenomenon). Occurrence of clamps there is a strong support for conspecificy.
When partial compatibility or unexpected results appeared, the matings were repeated. There is a considerable variation between different species in many characters of the cultured mycelia, e.g. growth rate, appearance of the mycelial mat, width of the hyphae etc and accessory spores (conidia) of various kinds. In Polyporaceae and Corticiaceae a coded sys- tem developed by Nobles has been used for a long time to describe the morphology of polyspore cultures. This system is used herein to describe the Pholiotas. The codes are from Nobles (1965) with emendations by Boidin & Lanquetin (1983).
The production of the extra-cellular enzymes laccase and tyrosinase is studied by using several phenolic compounds in drop-tests. This method, the nature, effectiveness and reliability of different reagents is described in detail by Marr (1979). The procedure in this study follows Marr’s direc- tions. Small slices of tissue, about 10 mm diam and 5 mm thick, were re- moved from the mycelial colonies. The slices were placed in cavities of distilled- water rinsed, porcelain, depression plates. Systematically, the six reagents used (Syringaldazine, 1-Naphtol, Guaiacol, Gum Guaiac, P-cresol, L-tyrosine) were added to the tissues, each tissue submerged in several drops of one reagent. Syringaldazine and 1-Naphtol reagents are laccase- specific, p-Cresol and L-tyrosine are tyrosinase-specific.
Also nuclear staining in spores and mycelia has been made with giemsa according to Boidin (1958) in order to study the nuclear behavior in different stages. The terminology in this respect follows Boidin & Lan- quetin.
The cultures are stored in the culture-collection at the Department of Systematic Botany, Gothenburg University. The GB-numbers refer to the culture-collection. The original specimens can be identified by the same number and are kept in the herbarium at Gothenburg (GB).
98
RESULTS
The basidiospores of many Pholiota species germinate easily in culture. However, the ability to germinate on common malt agar without any spe- cial arrangements varied within the genus. Spores from three species, P. flammans, P. nematolomoides and P. astragalina, never germinated in spite of several attempts. These species also have in common to grow on decayed coniferous wood. Also spores of P. tuberculosa were difficult to germinate, but eventually an attempt succeeded. In P. highlandensis only a few spores germinated. In most other species numerous spores germinated, but the vitality decreased rapidly after some days in storage. Spores stored more than a week generally did not germinate. In P. alnicola and P. pini- cola the spore-prints had to be stored about two weeks in a freezer (ca - 5 C) before germinating occurred. Some attempts to culture these species without this treatment did not succeed.
Primary mycelia of most species of Pholiota grow rather rapidly and usually the two mycelia paired on a dish grew together within two or three weeks. Then they became completely interwoven or formed a more or less distinct barrier. A distinct barrier was usually combined with absence of clamps but exceptions to this rule occurred. Incomplete clamps were some- times noted, generally in dishes where also genuine clamps were present. False clamps were rarely seen. In certain dishes, clamps were noted only on one side of the confrontation line (unilateral dikaryotization).
The positive pairings were compiled in pairing-tables to determine the mating-systems. However, the interpretation of the pairing tables were often more complicated than expected. Certain tables indicated bipolarity, others tetrapolarity, sometimes for the same species. Frequently irregulari- ties appearad, failures to form clamps where expected but also clamps in combinations which at first sight seemed to be impossible.
CYTOLOGY:
The basidia in all species of Pholiota are normally four-spored. Most basidiospores in all species investigated contained two nuclei, which is the normal condition in Strophariaceae and other chromosporic Agarics (Kiihner 1980). Certain spores seemed to contain one or more than two nuclei. Two nuclei in each spore is a result of three successive divisions of the fusion nucleus, the meiosis and two mitotic divisions, which yields eight nuclei. The third division occurs after the sterigmata are formed but the location in which it takes place is variable. Sometimes it occurs in the basidia, during the passage of the nuclei through the sterigmata or in the spores. Arita (1979), who studied the nuclear behaviour in P. "adiposa” (=P. jahnii?) and P. nameko, found that the third division took place in the spores in P. adiposa but this varied and frequently took place in the basidia in P. nameko.
99
The results of the polarity tests in this investigation show that it is common for the nuclei in certain basidiospores to contain different mating factors. In most pairing tables there occur positive pairings that can be ex- plained only if the spores contain nuclei with two noncomplementary fac- tors. Such heterocaryotic spores have been recognized in Psathyrella can- dolleana by Galland (1971). She showed that certain monosporic mycelia contained two types of nuclei incompatible because they possess the same allele for one of the mating type factors, A or B. Thus nuclei A2B2 and A2B1 may be found in the same primary mycelium. Undoubtedly the same conditions are common in most species of Pholiota. A tendency to "illegitimate copulations" between haplonts of the same strain occur also in Coprinus (Lange 1952). Future studies will reveal if this phenomenon is common also in other chromosporic genera.
Kihner (1977) considers that the discrepancies in pairing tables of tetrasporic species at least partly depends on spores borne on occasional basidia with only two or three sterigmata. Such spores may contain one or two additional nuclei which are genetically different. However, the num- ber of spores with more than one mating factor appears to be higher than the corresponding number of basidia with less than four basidiospores. Perhaps spores with more than one mating factor germinate more easily. The occurrence of amphithallism may cause difficulty in interpreting some pairing tables. Mounce (1929) and Vandendries (1933) reported a bipolar mating type in Pholiota adiposa (=P. jahnii?) and P. aurivella respectively. However, Farr et al. (1977) found that although P. aurivella and P. limonella first appeared bipolar, a careful reexamination showed them to be tetrapolar. As already indicated by Ginns (1974) several cases of re- ported bipolarity were the results of misinterpretation. Rewriting of the original bipolar tables showed tetrapolarity. The reason for the misinter- pretations was that the authors did not attempt to explain the irregularities that showed up in the pairing tables, e.g. the formation of clamps in pairings with presumably identical factors or failures in clamp-formation between assumed compatible pairings. Most of the pairing-tables in this investigation which indicate bipolarity are artificial, but Pholiota jahnii and possibly Pholiota mutabilis really seem to be bipolar or have bipolar forms.
Isolates from basidiospores of Pholiota usually germinate to non- clamped mycelia. The exceptions (approx. 10 - 20 %) may partly depend on dicaryotization between two germlings before they were isolated but probably spores with two compatible nuclei exist. However, such spores are produced only in a low number by chance and no species of Pholiota seems to be "secondarily homothallic" (Whitehouse 1949, Raper 1966). This term is applied to a species that regularily produce a high percentage of basidiospores with two compatible nuclei.
In some cases, e.g. in certain isolates of P. gummosa and P. limonella,
100
clamps suddenly and unexpected have appeared in ss-mycelia months or years after germination. A spontaneous change of a mating type factor must have taken place. Such changes occur in Psathyrella candolleana (Galland 1971). A spontanoeous mutation of a mating factor may excep- tionally be the explanation of an unexpected positive mating in a pairing table.
MYCELIA AND HYPHAE:
The appearance of the mycelia varied considerably between different species or groups of species. The surface of the mycelial mat is smooth in many species but distinctly granular in others. In P. adiposa and P. limonella distinct strands run over the surface. A cottony aerial mycelium is present in most species but there is variation between different mycelia of the same species. The advancing zones are generally even and appressed, with a few exceptions. These characters are described in the species descriptions and photos were taken of the cultures after six weeks growth (Fig. 1 - 3). Also the growth rate appeared to be very different.
Most hyphae in a mycelial colony are generally rather narrow (1 - 5 jum). The widths vary between the species but also between different iso- lates of the same species. On average the primary are narrower than the secondary mycelia. In all species certain hyphae become consideraby wider and frequently also more irregular in shape. In many species certain hyphae contain a row of short, swollen cells, which give them a monili- form appearance. Such cells appear both apically and intercalary. Also the frequency of oil-drops and other characters is variable (cfr fig. 8, P. tuberculosa).
The appearance of mycelia and hyphae seems to be very typical and constant within each species and therefore of a taxonomic value. Closely related species, e.g. P. adiposa and P. limonella, P. spumosa and P. mixta, P. alnicola and P. pinicola, are very similar in these characters. ACCESSORY SPORES:
Most mycelia of Pholiotas, both ss- and ps-mycelia, form accessory spores in culture. Two types of accessory spores occur in the genus: arthrospores (oidia) and chlamydospores (aleuriospores). The arthrospores are generally formed on short, narrow branches (conidiophores) from cer- tain hyphae on the surface of a mycelial colony. They arise as two or more cells in the apical parts of a branch and start as protoplasmic contraction followed by dissolution of the walls of the emptied parts of the hyphae. The process frequently takes place repeatedly on the same conidiophore. Sometimes the arthrospore-forming branches are coiled, e.g. in certain mycelia of P. jahnii. The spore-forming branches appear simple (for instance in P. spumosa) or in different-looking ramifications. Possibly also protoplasmic contraction in ordinary hyphae may lead to the formation of arthrospores.
Chlamydospores are formed in several species. They are very frequent,
101
Fig. 1. Cultures after six weeks growth. a) Pholiota squarrosa, GB 1303, b) P. heteroclita, GB 1457, c) P. adiposa, GB 1071, d) P. limonella, GB 1456, e) P. jahnii, GB 1061, f) P. squarrosoides, GB 1458.
Fig. 2. Cultures after six weeks growth. a) Pholiota spumosa, GB 884, b) P. mixta, GB 948, c) P. lenta, GB 1060, d) P. lubrica, GB 1293, e) P. highlandensis, GB 1328, f) P. scamba, GB 1221.
Fig. 3. Cultures after six weeks growth. a) Pholiota gummosa, GB 1295, b) P. graminis, GB 1273, c) P. tuberculosa, GB 1692, d) P. mutabilis, GB 1304, e) P. alnicola, GB 1243, f) P. pinicola, GB 1359.
104
large and conspicuous (15 - 30 x 10 - 20 wm) in certain species, e.g. P. alnicola, P. pinicola, P. gummosa, P. graminis and P. squarrosa, but less numerous and rather inconspicuous in others. They are formed both later- ally and apically, frequently as the end-cell in a row of moniliform swellings. Chlamydospores occur anywhere in the mycelia but preferably in the submerged parts. Typical chlamydospores are not seen at all in some species, but probably any kind of swellings may loosen and then serve as a chlamydospore. Actually there is no fundamental difference between the two kinds of spores more than the size and shape. Of course chlamydospores are never sO numerous as arthrospores, as they are produced for survival and not for dispersal as arthrospores.
The type and shape of accessory spores is very constant within each species and they are a systematically valuable character. However, they are not always formed by all isolates of the same stock. Genetical differences are probably responsible for presence or absence of accessory spores but also environmental conditions may play a role. Many species form only one type of accessory spore but both arthrospores and typical chlamydospores are seen in the same mycelial colonies of Pholiota squarrosa and P. gum- mosa. Arthrospores are common e.g. in the adiposa and lubrica groups and in P. squarrosa and P. gummosa. Conspicuous, moniliform swellings are especially common in Pholiota tuberculosa and P. albocrenulata.
Hiibsch (1978) described the accessory spores of certain species of Pholiota and also illustrated some petri plates to illustrate their formation process. His results correspond well with those of this investigation. DROP-TESTS:
The results of the drop-tests are presented in table I. The reactions were recorded 5 min., 30 min. and 2 hours after application of the reagents. In all cases where a positive reaction was recorded, it was visible within 30 min.
A positive response is an effect of oxidation and implies occurrence of enzymes. According to the investigations of Marr (1979) and others syringaldazine and 1-naphtol are laccase-specific. L-tyrosine and p-cresol reagents are tyrosinase-specific. Guaiac and guaiacol are nonspecific for laccase and tyrosinase but more effectively oxidized by laccase than tyrosi- nase.
In this study eleven species contained both laccase and tyrosinase: Jahnii, adiposa, limonella, squarrosoides, graminis, gummosa, scamba, spumosa, mixta and mutabilis. Only tyrosinase is indicated in squarrosa, heteroclita and lenta. The reactions indicate only laccase in junonius, high- landensis, lignicola and tuberculosa. Neither laccase nor tyrosinase reaction is noted in the following species: albocrenulata, alnicola and pinicola.
105
Table I Result of drop-tests
+ = positive reaction, (+) = weak positive reaction, - = no reaction noted. Sy N Gl G p-C Ty - adiposa (+) . om eS MAES. a albocrenulata - ~ - = Ae = alnicola - - = = os bs graminis + + + + + rs gummosa + - (+) - - = heteroclita ~ - - cs 2 (+) highlandensis ~ + ~ (+) - . jahnii + + + + sh _ Junonius - (+) - (+) - e lenta - - = 2 = (+) lignicola + (+) (4+) + = e limonella - (+) - (+) + 2 lubrica - - cs yee 5 mixta + + + + + + mutabilis + . + + + + pinicola ~ - = = ‘i = scamba + + + + fe ag spumosa + - 4 + + + Squarrosa - - (+) - (+) (+) squarrosoides - = (+) ~ - (+) tuberculosa - + - (+) “= i
Abbrevations: Sy = syringaldazine, N = 1-Naphtol, Gl = guaiacol, G = guaiac, p-C = p-cresol, Ty = L-tyrosine.
The results of the spot tests in some species varied from the results in other studies. K44rik (1970) summarized results of spot tests and other culture characters for a large number of species, among them eight Pholiota species. She reported a strong laccase reaction in P. heteroclita and P. squarrosa, whereas no laccase was indicated in this study. She re- ported a weak tyrosinase reaction in P. gummosa and a strong laccase re- action in P. adiposa, which do not correspond with the result of this study. Marr (1979) studied P. squarrosoides and P. lenta. They were placed in the group of species with tyrosinase only, which corresponds with the results of this study.
Marr concluded that syringaldazine is the best reagent for laccase and L-tyrosinase or p-cresol the better reagents for detecting tyrosinase. It is therefore worth mentioning that laccase in P. tuberculosa and P. high-
106
landensis was indicated by a positive reaction with l-naphtol but not syringaldazine.
The deviating results of different studies may depend on the fact that the localization of laccase and tyrosinase varies either among species or during ontogeny. These variations are associated with other processes, e.g. pigmentation or fruiting. Marr and K 44rik performed their tests on tissues from basidiocarps, which may give other results than tests on mycelial colonies in culture. Also environmental factors, such as nutrition, tempera- ture, pH, etc. influence the production of the enzymes. Results of drop- tests therefore must be interpreted carefully and preferably compared with other methods before conclusions can be reached.
CULTURE CHARACTERS FOR THE SPECIES Pholiota squarrosa (Weigel:Fr.)K ummer
MATERIAL:
GB 165/ Fagus/Sweden, Halland, Tjoléholm (SJ 80261).
GB 1249/ Picea/Norway, Oppland, Ormtjernkampen (NH 8481).
GB 1299/ Quercus/Sweden, V4stergétland, V.Tunhem (SJ 84157).
GB 1303/ Quercus/Sweden, VAstergotland, Géteborg (SJ 84165).
GB 1535/ Abies/Romania, Neamt (NH 9201).
POLARITY: Only two polarity tests have been performed, both, however, with a rather small number of available isolates (Tab. 2). In 1249 only two positive pairings were observed. They indicate two different compatibility groups. The remaining isolates may belong to a third and same compatibi- lity group but the negative pairings may be due to reduced fertility. In 1535 three compatibility groups (5 = AIB1; 1, 2, 6 = A2B2; 3, 4, 7 = A1B2) is the best interpretation of the result. Tetrapolarity therefore is the only reasonable mating system in P. squarrosa.
GB 1249, GB 1299, GB 1303, GB 1535 were compatible with each other. CYTOLOGY: Astatocoenocytic. The cells of the ss-mycelia contain a variable number of nuclei, at least 2 - 25 have been noted. Multinucleate hyphal cells are distinctly wider than others and occur especially in the terminal part of the hyphae. Multinucleate cells are also noted in a secondary mycelium (GB 1303) though most of them are dicaryotic. CULTURE CHARACTERS (GB 1303): Growth moderately rapid, the dishes completely covered by mycelia in 3 - 4 weeks. Advancing zone even, appressed. Mycelial mat soon floccose, at first whitish, later yellowish with brown floccules (Fig. 1 A).
Reverse brownish. Hyphae usually 2 - 4 um wide, regularly branched, with a clamp at all septa. Young, apical cells are frequently somewhat wider (3 - 6 pm) and more irregular in outline. Numerous arthrospores (6
107
- 10 x 3 - 4 pm) are formed by short branches from certain hyphae in the floccules on the surface. In the submerged part of the mycelium also large
Tab. 2. Polarity tests in Pholiota squarrosa.
N (sa) N MINAS 4 UN NS 3 3 2 2 2 3 Soe oat RN eee a tS SU ay ale ale ie ata al aus Bead ea as a) —_ -_ ae oi © ie) ie) cb) iS) Oo $6 & & 8 BUSES / 1) a ya ta wen any ole SBMOES pI tee GB 1535/2 2G\) Uae Ge CB 1249/2 Fie . GB 1535/3 ROAD ALGH ER g CB 1249/3 aide lia GB 1535/4 Ay OE SOS GB 1249/4 aM GB 1535/5 re CB 1249/5 i GB 1535/6 ’
chlamydospores (10 - 16 x 8 - 12 wm) are rather common. Fig.4.
CODE: 2b, 3c, 26, 34, 35, 37, 39, 43-44, 49, 54, 60, 63. Oxidase reactions: weak reaction with guaiacol, p-cresol and L-Tyrosine, all other tests nega- tive.
Fig. 4. Hyphae, arthrospores and chlamydospores in Pholiota squarrosa (GB 1303).
The cultural characters of P. squarrosa have much in common with P. gummosa (e.g. the appearance of the mycelia and the accessory spores) and
108
indicate a relationship. According to K 4arik (1965), P. squarrosa belongs to the group of species which produces both laccase and and tyrosinase. In this investigation only tyrosinase is verified. Possibly the weak reaction with guaiacol may be an indication of laccase. Kaarik also reports a rather rapid growth rate for this species.
Pholiota limonella (Peck)Sacc.
MATERIAL:
GB 166/ Alnus trunk/Sweden, Géteborg, Naturparken (SJ 80286). GB 1438/ Betula/Sweden, Vastergétland, Langared (SJ 85056).
GB 1456/ cut trunk of Quercus robur/Sweden, VAstergotland,
Rada (SJ 85092).
GB 1508/ Abies alba/Romania, Neamt (NH 9200).
GB 1513/ Deciduous wood/Romania, Iasi (NH 9084).
GB 1597/ Substrate unknown/USA, VT 397 (Blacksburg, Virginia). GB 1717/ Populus/Sweden, Medelpad, Borgsjé.
GB 1736/ Alnus glutinosa/Sweden, Uppland, Billudden.
The compatibility between these specimens and with the closely re- lated Pholiota adiposa was dealt with in another paper (Jacobsson 1987). POLARITY: Tetrapolarity is indicated but there are several irregularities, such as failures of clamp formation in supposed compatible pairings and occurrence of clamps in unexpected combinations. In several cases the only explanation for compatible pairings is that certain spores must contain two or more incompatible factors. It is not always easy to interpret a pairing table with total certainty due to irregularities which make more than one solution possible. The interpretation with the lowest number of mating failures and without positive matings between isolates assigned the same factors is considered to be the most probable one, at least theoretically. The pairing tables are showed in tab. 3.
In GB 166 the most reasonable interpretation is: 1 = AIB1; 5, 6 = A2B2; 3, 4, 7 = AIB2; 2 = AIBI+A2B1; 8 = A2B2+A2B1. There are two failures with this explanation (2 x 4, 2 x 7).
GB 1438: 4, 8 = AIB1; 5 = A2B2; 3, 7 = A1B2; 1, 2 = A2B2+A2BI1. No 6 is excluded, as it did not result in any positive mating. This isolate is supposed to have a restricted fertility for some reason. With this interpretation there are two failures (1 x 7, 2 x 3), in other interpretations there are more failures.
GB 1456 is tetrapolar without failures: 1 = AIB1; 2, 4 = A2B2; 3, 5, 7 = A1B2; 6 = A2BIl.
GB 1513. Amphithallic tetrapolarity is the only possible interpretation. If 1, 2, 3, 7 = A2B1+AI1B1; 5 = AIBI; 8 = A2B2; 6 = A1B2; 4 = A2B1, there are no failures. Other interpretations are possible, but in those cases there are at least some failures.
109
GB 1717: This pairing table is difficult to interpret, but one interpre- tation indicates amphithallic tetrapolarity without failures: 1, 2 = AIB1; 3,
8 = A2B2; 5 = A1B2; 7 = A2B1; 10 = A1B1+A2B1; 9 = A2B2+A1B2.
Some polarity tests are excluded, because few isolates were available.
‘Tab. 3. Polarity tests in Pholiota limonella.
SRA Fomas Norma iS 866 686 8 8 8 GB 166/1 a = GB 166/2 F- =- + GB 166/3 - - - = + GB 166/4 SP) Gap ESE an GB 166/5 a=! aa GB 166/6 - - GB 166/7 ie PE et ae Urey a ge 888s:ss8 8s GB 1508/1 - Se ee San ae GB 1508/2 = + GB 1508/3 me 4 Seo me Ap GB 1508/4 es ae 2 ee GB 1508/5 a ad GB 1508/6 Ns, GB 1508/7 = ° dees eer ee $$ 86 8 8 8 8 GBatti7/iae = Bee eos ea gr GB 1717/2 a HER SAC aa GB 1717/3 teen ee are GB 1717/5 i bem te GB 1717/7 ey de A GB 1717/8 bate GB 1717/9 ae
GB GB GB GB GB GB GB
GB 1513/1
GB 1438/2
1438/1 1438/2 1438/3 1438/4 1438/5 1438/6 1438/7
GB 1513/2
GB 1513/2 GB 1513/3 GB 1513/7 GB 1513/6 GB 1513/8 GB 1513/4
(+) means that only a few clamps were found,
restricted to the confrontation line.
GB 1456/1 GB 1456/2 GB 1456/3 GB 1456/4 GB 1456/5 GB 1456/6
GB 1438/3
+
GB 1456/2
+
GB 1513/3
GB 1438/4
+
+
GB 1513/7
GB 1456/3
GB 1438/5
GB 1513/5
+ + + +
GB 1456/4
+
|
GB 1438/6
GB 1513/8
+ + + +
GB 1456/5
GB 1438/7
GB 1513/4
GB 1456/6
GB 1438/8
+
GB 1513/5
GB 1456/7
110
CYTOLOGY: Astatocoenocytic. The hyphal cells of a homocaryotic mycelial colony contain a various number of nuclei, 2 - 22 are counted. The highest number of nuclei is found in the youngest, terminal cell of a hypha. Most cells in the secondary mycelia are dicaryotic but multinucleate cells containing as many as 25 nuclei are also seen.
CULTURE CHARACTERS: Growth slow, dishes covered in 5 or 6 weeks. Advancing zone even, appressed. Aerial mycelium cottony-floccose, at first whitish, becoming yellowish brown in old parts. Conspicuous, branched strands run over the surface (Fig. 1 D). Reverse brownish. Hyphae | - 4 wm wide, regularly branched, with a clamp at most septa, hyaline or yellowish. Walls in old hyphae brown and incrusted. Numerous arthrospores (5 - 11 x 1,5 - 3 um), formed by short branches from hyphae on the surface. Chlamydospores (8 - 30 x 5 - 10 wm) occur in submerged mycelia, but not numerous. Old hyphal cells in the submerged mycelia fre- quently becoming filled with reddish brown necropigments and finally broken in pieces (Fig. 5).
CODE: 2a, 2b, 3c, 16, 26, 34, 35, 37, 39, 45-46, 49, 54, 60, 63.
OXIDASE REACTIONS: Positive reaction with guaiac (very weak), p- Cresol and 1-Naphtol (weak), all other tests negative.
Pholiota limonella is closely related to Pholiota adiposa (=aurivella auct.). It is not possible to separate the two species in the basidiocarps. A slight difference in the spore size is the only reliable morphological character. Since this species earlier has not been distinguished by any European author, it is quite possible that published culture data (Vandendries 1933, K4arik 1970, Hiibsch 1978 etc) of Pholiota aurivella (at least partly also adiposa and squarroso-adiposa) actually represent Pholiota limonella. However, so far no difference in the cultural characters have been found.
Pholiota adiposa (Batsch:Fr.)Kummer
MATERIAL:
GB 167/Fagus/Sweden, Halland, Grytsjén (SJ 80307).
GB 1072/Fagus/Sweden, Skane, Silvakra (SJ 83119).
GB 1291/Salix /Sweden, Skane, Kristianstad (SJ 84131).
POLARITY: GB 167 and GB 1291 were tested (Tab. 4). In GB 167 only
one positive pairing was obtained. In GB 1291 the only possible interpreta-
tion is amphithallic tetrapolarity: 8, 10 = A1B1; 1, 2, 5, = A2B2; 4, 6 =
A1B2; 7 = A2B1; 9 = A2B2+A1B2. This explanation gives no failures. The three mentioned specimens are intercompatible with each other.
Pholiota adiposa is very closely related to Pholiota limonella. The taxo-
nomy and the differences between them are published in an earlier paper
111
(Jacobsson 1987).
CYTOLOGY: Astatocoenocytic. Characters identical with those of P. limonella.
CULTURE CHARACTERS (GB 1072): Growth rate and general appear- ance identical with the closely related P. limonella. Advancing zone even, appressed. Later a cottony-floccose aerial mycelium is formed. Conspicous, branched strands run over the surface, at first whitish, soon yellowish to brownish (Fig. 1 C). Reverse brownish. Hyphae with a clamp at most septa, regularly branched, | - 4 wm wide. Numerous arthrospores (5 - 11 x 1,5 - 3 um), formed in the same way and of the same appearance as in P. limonella
CODE: (2ab), 3c, 16, 26, 34, 35, 37, 39, 45-46, 49, 54, 60, 63.
OXIDASE TESTS: All tests negative in GB 1072. In GB 167 a positive but weak reaction with syringaldazine and p-cresol.
Tab. 4. Polarity tests in Pholiota adiposa.
RAH ONHEN ae MMR Lakh Teg Rae tienen Co ann -- ne -- en -- et -- -- === Cee ste, LAs SE iets SETS CEN 9 ILENE WN NEN Nah SN Ss Wis eS oo 2 9 © GB 1291/1 os << UA AUt eae i ah A as (o> See a = GB 1291/2 aU FO Hi Se CON CON Con eS GB 1291/5 val SE ore Tyee pA Ese iene S GB 167/2 a ae GB 1291/6 Sav ce tN rea ah Tue GB 167/3 Teka ey GB 1291/7 VAN Th Rees BS il 714 ee ie GB 1291/9 BAO eg GB 167/5 = GB 1291/8 mes
This species is better known as Pholiota aurivella (Batsch:Fr.)Kummer. However, as this name has been recently questioned (Kuyper & Tjallingii- Beukers 1986) and Pholiota adiposa in its original sense undoubtedly is correct (cfr Jacobsson 1987) a name change to P. adiposa is unavoidable.
The confusing taxonomy in this group implies that published data on Pholiota aurivella should actually be referred to Pholiota adiposa or the almost indistinguishable Pholiota limonella. On the other hand, published data on Pholiota adiposa, at least in recent time (Hiibsch 1978, Arita 1979) should be transferred to Pholiota jahnit.
Pholiota "aurivella" is one of the most studied species of the genus. Martens & Vandendries (1932) and Vandendries (1933) reports bipolarity for this species, which seems unlikely when compared with later investiga- tions (Farr, Miller and Farr 1977, and those in this paper, cfr Pholiota jahnii). Hibsch (1968) noticed the occurrence of arthrospores and K aarik (1970) a strong reaction both for laccase and tyrosinase together with some
112
other characters. Only weak reactions for laccase and tyrosinase reaction were noted in this investigation. The difference may depend on the fact that Kaarik used basidiocarp tissue for her tests.
Sy eee Vip eg \ PIAS \7 } i Va
Fig. 5. Hyphae, arthrospores and chlamydospores in Pholiota adiposa (GB 1291).
Pholiota jahnii Tjall. & Bas
MATERIAL:
GB 83/Fagus/Sweden, Skane, Békebergsslatt (SJ 79231).
GB 164/Fagus/Danmark, Sjalland, Vordingborg (SJ 80210).
GB 1061/Fagus/Sweden, Skane, Silvakra (SJ 83118).
GB 1305/Fagus/Sweden, Blekinge, Sdlvesborg (L. Orstadius).
GB 1342/probably Picea/Sweden, Séddermanland, Overjarna (K.Jaederfeldt).
GB 2021/buried wood, probably Fagus/Sweden, Skane, Silvakra (SJ88061). POLARITY. Polarity tests have been performed with the specimens GB 83, GB 1061, GB 1305, GB 1342 and GB 2021 (Tab. 5).
In GB 1342 only two positive pairings were yielded, which indicates a restricted fertility for some reason. The result of the other four polarity tests clearly indicates bipolarity. However, there is one failure in GB 1061 (4 x 9).
All specimens are completely intercompatible with each other (Jacobsson 1987).
HIS
Tab. 5. Polarity tests in Pholiota jahnii.
Role Ca cee) WT i, a OS with oh A ey eva alata ioe 85323 Ba a te oo ye es es 4.9 2 3 3's GBBS/1 = HY HR + ae he GB IBOSh ie) has) Ven! occer od ae ay GB 83/2 RLY Me aes ge der ame GB 1305/2 Ni ie EMER, ks GB 83/6 es, tae aid mee sme GB 1305/3 ey ie Sh Nak GB 83/7 + + + + + GB 1305/5 a es GB 83/3 Se ft eit Be GB 1305/7 fey GB 83/4 con Es eats GB 1305/4 al GB 83/5 oe le S GB 83/8 . de a PE N N N N N N N N N rm 2@yese Se So) Gus. G 8: 8 238 83 8 GB 1362) eet ae tel ae ees eet Pane he UE GB 1342/2 ee ME me Z Cane Une fc. e GB 1342/3 Hie erie! Se GRRIO6I {Gf S/S ea CB 1342/4 Logis ee, \ Bed eee re GB 1061/7 - + + + GB 1342/5 ee ees ee GB 1061/8 + + + GB 1342/6 Cory Recut at), ees GB 1061/5 oan = GB 1342/7 ee CB 1061/6 CB 1342/8 ayrie GB 1342/9 = N Ga) —- Ya) Ne) ~ 2) i>) = BS eee ny ied Siege snag |S 888s: 8&6 8 & & BBR02 lee | fase yoga! Pac) ed eas tg GB 2021/2 Bey Mek ol, aeRO Sa sgn GB 2021/3 See men’ Senhes UR Rare GB 2021/4 eG lal bath el n Mist GB 2021/5 Be) | CES EP ee eg GB 2021/6 Aiea ie bed GB 2021/7 ae eee GB 2021/8 =: GB 2021/9 +
CYTOLOGY (GB 1061): Heterocytic. Terminal cells of ss-mycelia are multinucleate and wider than most intercalary cells, which generally con- tain two nuclei. In the secondary mycelium only clamped, binucleate cells are found.
CULTURE CHARACTERS (GB 1061): Growth slow, dishes covered in six weeks. Advancing zone even, appressed. Aerial mycelium cottony, whitish, but becoming slightly yellowish brown in patches in old parts (Fig. | E). Reverse somewhat brownish. Hyphae generally 1 - 3 um wide, regularly
114
branched, with a clamp at all septa. Numerous arthrospores (3 -6x1-3 um) are formed at the surface of the mycelial colony (Fig. 6). Chlamydospores (8 - 45 x 6 - 10 um) rather rare, only seen in certain mycelia. Many old hyphae are filled with dark reddish-brown necropig- ments and easily broken to pieces.
CODE: 2ab, 3c, 26, 34, 35, 37, 39, 46, 49, 54, 59, 63.
OXIDASE REACTIONS: Positive reactions noted for all tests except p- cresol.
Fig. 6. Hyphae, arthrospores and chlamydospore forming in Pholiota jahnii (GB 1061).
Pholiota jahnii is a recently created new name for the species earlier known as Pholiota muelleri (Fr.)Orton. It also includes Pholiota adiposa auct. (ss Lange etc, cfr Jacobsson 1987). Therefore most published data on Pholiota adiposa (e.g. Arita & Mimura 1969, Arita 1979) probably refer to Pholiota jahnii.
The most remarkable character of Pholiota jahnii is the bipolarity. Three polarity tests clearly indicate bipolarity, which result corresponds with earlier reports. Arita and Mimura (1969) reported Pholiota "adiposa" to be bipolar. Vandendries (1933) reports a perfect bipolarity in a test with 16 isolates of Pholiota “aurivella". The latter species is definitely not bi-
115
polar and it is more probable that he had collected P. jahnii, a species not known at that time.
The strong reaction for both laccase and tyrosinase corresponds well with the report of K4arik (1970).
Pholiota squarrosoides (Peck)Sacc.
MATERIAL:
GB 1458/on dead Populus trunk/Sweden, Uppland, Bondkyrka, (S.Ryman 8065).
GB 1680/Canada, Br.Col., Stamp Falls, polysporous & tissue cultures isolated from fruit body (2 cultures, but without clamps, DAOM 22113). POLARITY: GB 1458 is tested regarding the mating type (Tab. 6). Amphithallic tetrapolarity is the only possible interpretation. If 6, 7 = AIB1; 1, 5 = A1B2; 2 = A2B1; 4 = A2B2+A1B2; 3 = A2B2+A2BI1, there is only one failure (1 x 2).
Tab. 6. Polarity test in Pholiota squarrosoides.
N isa) ~— wy Ke) ~ = — - >) ~ ice} @ ie] ee) ce) (ee) wy wv va) Ww va) WwW Ba tie ao so xB ea) =s) oO co 1) Oo o>) ee) i) Oo
GB PA5G/ i Sire re ae
GB 1458/2 RE AA NG AM ae
GB 1458/3 fed (le bay ot
GB 1458/4 ae
GB 1458/5 eas
GB 1458/6 at
The Swedish and Canadian specimens turned out to be incompatible. However, other explanations than species differentiation are possible. The absence of clamps in DAOM 22113, in spite of the fact that it originates from a basidiocarp, indicates that a degenerative mutation has taken place and a restricted mating ability therefore is probable. These cultures were made in 1949.
CYTOLOGY (GB 1458): Probably heterocytic. All hyphae in the secondary mycelium are clamped and binucleate.
CULTURE CHARACTERS (GB 1458): Growth very slow, reaching S50 - 52 mm and not covering the dishes in six weeks. Advancing zone even, appressed. Aerial mycelium almost absent, but the surface of the mat with numerous minute granules, whitish -yellowish (Fig. 1 F). Reverse weakly brownish. Hyphae 1 - 5 ym wide, regularly branched, with a clamp at most septa. Arthrospores (3 - 8 x 1 - 3 wm, frequently curved) are formed by irregular, short branches in the granules on the surface (Fig. 7). A few
116
rather small and inconspicuous chlamydospores (9 - 15 x 6 - 8 um) are seen.
Fig. 7. Hyphae, arthrospores and chlamydospores in Pholiota squarrosoides (GB 1458).
CODE: (2ab), 3c, 26, 34, 35, 37, 39, 47, 49, 54, 60, 63. OXIDASE REACTIONS: All tests negative.
No data on culture characters for Pholiota squarrosoides are published previously. The species apparently belongs to the adiposa group, based on its characters, e.g. the occurrence of arthrospores, which are similar to those of the other species in the group. However, it differs from the other species in the negative reactions for laccase and tyrosinase.
Pholiota tuberculosa (Schff.:Fr.)Kummer
MATERIAL:
GB 1692/Tilia/Sweden, Vastergétland, Trollhattan (SJ 86019). POLARITY: 7 isolates of the specimen GB 1692 were mated in all possible combinations but no positive pairings appeared. Thus the mating type of this species is still unknown.
THA
POON DOS =—
one Tele /.Yargreyars
> o_O. 2S Fe
ae SEE Q
Fig. 8. Hyphae in Pholiota tuberculosa (GB 1692). A) in ps-mycelium, B) in ss-mycelium.
CYTOLOGY: Not studied.
CULTURE CHARACTERS: Growth rapid, the dishes completely covered in two weeks. Advancing zone somewhat fringed. Aerial mycelium cottony, slightly yellowish. Reverse brownish. Normal hyphae 3 - 5 um wide, regularly branched and with a clamp at all septa, densely filled with oil-drops. Certain hyphae have a moniliform appearance with short and very broad (10 - 30 um) cells. The ss-mycelia to a great extent consist of very wide ( - 20 um) hyphae, with very big, sometimes strongly yellow, oildrops (Fig. 8). Sometimes bladder-like cells seem to serve as chlamydospores.
118
CODE: 2a, 3c, 26e, 34, 37, 39, 42, 49, 54, 58. OXIDASE REACTIONS: Positive but weak reactions with guaiac and 1- Naphtol, otherwise negative.
Pholiota tuberculosa has not been studied in culture before. Some of the cultural characters such as the rapid growth rate and the very wide hyphae densely filled with oildrops differ considerably from other Pholiota species and indicate that this species takes a rather isolated systematic position in the genus.
The species seems to be difficult to culture, as spores from one speci- men only germinated. Some other attempts have been unsuccessful. It is desirable with more cultures to verify the results.
Pholiota lucifer (Lasch)Quél.
MATERIAL:
GB 1852/Abies/Austria/CBS 595.82 (1 ps-culture).
The culture characters of this strain appeared to be almost identical with those of P. adiposa - limonella and suggest that P. lucifer belongs to this complex. However, this seems unlikely, as the morphological characters are very different and more similar to those of P. tuberculosa. Compatibility tests by using Buller’s phenomenon have been performed with P. limonella (GB 1456) and P. adiposa (GB 1072). They were found to be negative. In spite ot that it is possible that the culture descends from a misidentified basidiocarp. No herbarium material is preserved to check the determina- tion. It is desirable to get the culture characters verified by another strain before conclusions.
Pholiota heteroclita (Fr.:Fr.)Quél.
MATERIAL:
GB 1457/Betula/Sweden, Halland, Lindome (SJ 85093).
POLARITY: 8 isolates of GB 1457 were mated in all possible combina- tions. However, the mycelia ceased to grow after a while and only in two combinations were clamps formed and then there were only a few. CYTOLOGY: Not studied.
CULTURE CHARACTERS: Growth rate very slow for the genus, in six weeks the mycelia had reached only 30 - 37 mm. Advancing zone some- what uneven and fringed. Aerial mycelium downy, slightly plumose, whitish in young parts, gradually becoming brownish. Reverse somewhat brownish. Hyphae 1 - 3 um wide, not very differentied, regularly branched, with a clamp at most septa. Numerous arthrospores formed in
119
the aerial mycelium by short branches in terminal or lateral, brush-like clusters (Fig. 9), rather different-looking compared with those in other Pholiotas. Occasionally small intercalary or terminal swellings appear. CODE: 2b, 3c, 7, 35, 37, 39, 47, 49, 54, 58.