Rep. Tottori Mycol. Inst. 33: 7-13, 1995. A preliminary account of some unusual Onygenalean fungi from decayed wood T. C. Lumtey* and R. S. CuRRAH* Abstract Three Onygenalean fungi (Onygenales, Ascomycotina), Gymnascella littoralis, Myxotrichum ochraceum, and Uncinocarpus uncinatus, were is lated from well-decayed wood in Alberta, Canada. Gymnascella littoralis and U. uncinatus were isolated from Picea glauca using a specific medium (c} tremuloides on a non-specifi loheximide) and M. ochraceum was isolated from Populus , malt extract medi m, Descriptions for all three are given. These records are significant because the organisms are only rarely isolated and have not been described previously from decayed wood in nature. Key Words: ascomycetes, Onygenales, Gymnascella littoralis, Myxotrichum ochraceum, Uncinocarpus uncinatus, decayed wood, cycloheximide. Introduction The order Onygenales (sensu Currah, 1985) comprises four families that are distinctive among the ascomycetes because many of the species produce ascomata that have mesh-like peridia and an anamorph with thexolytically dehiscing arthro- and (or) aleurioconidia. The Arthrodermataceae and Onygenaceae form a well-defined clade within the order because of their ability to break down keratin. The Myxotrichaceae are similar to these families because conidia, when formed, are produced in chains of alternate arthroconidia or are repre~ sented by a distinctive Oidiodendron or Geomyces state. The family differs from the Arthrodermataceae and Onygenaceae because it is cellulolytic rather than keratinolytic. The Gymmoascaceae is a heterogeneous assemblage of taxa with mesh-like peridia and unknown or poorly defined substrate preferences. Rhexolytically dehiscing conidia are produced in some taxa. Ecological studies of the Onygenales have focused primarily on the recovery of representa- tives of the Arthrodermataceae and Onygenaceae © Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada7.0. Lumuey and R.S. Curran from soil, keratinous substrates, and keratin- enriched debris. The most common methods employed to isolate these fungi from non-clini- cal sources are baiting methods that use native keratin, such as hair and feathers, and selective media containing cycloheximide. Using these methods, reports of species of Aphanoascus, Arthroderma, Trichophyton, and Chrysosporium from soil and dung have become common (Currah, 1985). Most other taxa in the Onygenales are only rarely reported and little is known about their natural habitat or the frequen- cy of their occurrence in nature. Recently, during a study of the microfungi of fed logs in the boreal forest of Alberta, three species belonging to the Onygenales were isolat- ed. Gymnascella litoralis (Orr) Currah (Gymno- ascaceae) and Uncinocarpus uncinatus (Eidam) Currah (Onygenaceae) grew out from chunks of well-decayed wood that had been placed on plates of agar containing cycloheximide, and Myxotrichum ochraceum Berk. & Br. (Myxotrichaceae) appeared on a non-selective medium. Because these species have been reported so rarely in the literature, a preliminary account of their morphology is provided here along with some discussion of decayed wood as a habitat for Onygenalean fungi Materials and Methods Well-decayed wood of Picea glauca (Moench) Voss and Populus tremuloides Michx. was collected in July, 1995 from a for- st in Elk Island National Park, Alberta. Wood pieces (Iem') were surface sterilized by flam- ing briefly using a Bunsen burner and then bro- ken up across a plate of Mycobiotic Agar (Difco) or Malt Extract Agar (1.5% Difco agar, 1.5% Difco malt extract, w:v). Plates were incubated at room temperature. Ascomata developed after 14 to 18 weeks and were used {0 start pure cultures. Ascomata were mounted from primary isolations in polyvinyl alcohol and photographed using an Olympus BH-2 microscope with a PM-1OAK photosystem, Descriptive Part Gymnascella littoralis (Orr) Currah_ Figs. 1-4. Ascomata tiny, semi-globose, yellow with some orange tints, 95-125 jim in diameter. Ascospores pale yellow, smooth, thick-walled, oblate with an equatorial rim, 3.6-4.2 um x 2.3-3.0 um. Asci evanescent and not observed Peridial hyphae pale to dark yellow, smooth, thick-walled, 1.6-2.6 pm in diameter, some- what branched, the terminus of some hyphae dichotomizing to form short, boat-hook-like appendages. Anamorph composed of smooth, thick-walled arthro- and aleurioconidia, arthro- conidia 4.2 um x 2.0-2.4 um, aleurioconidia 3.9-5.2 um x 2.6-3.0 um, Habitat: in well-decayed wood of P. glauca collected at Elk Island National Park, Alberta Substrate unknown, This species was originally described in the monotypic genus Plunkettomyces by Orr (Orr, 1977). von Arx transferred the species to Arachniotus (A. litoralis (Ort) von Arx] in 1977 and subsequently to Gymnoascus (von Arx, 1986). Currah placed the species in Gymmascella in 1985 (Currah, 1985) based on the mesh-like peridium, the smooth, oblate ascospores with an equatorial rim, and the inability of named iso- lates to break down keratin in a hair plate assay. ‘The taxon produces chains of short, squarrose arthroconidia. The genus Gymnascella is a het- erogeneous assemblage of unrelated species and in need of revision, It was placed in the Gymnoascaceae by Currah (1985) (Briksson and Hawksworth, 1993). The isolate described above appears to fit well in this species but is assigned to G. lit- toralis with some reservation. With the excep- tion of one report of isolation from soil in ItalyCOnygenalean fungi from decayed wood Fics, 1-4. Gynnascella linoratis. Fig. 1. Ascoma, Fig. 2. Partial ascoma showing peridial hyphae (open arrows) and ascoxpores (closed arrows). Fig. 3. Ascospore with equatorial arrow). Fig. 4. Aleurio- (closed arrow) and 4. Pies. 5-7. Uneinocarpus wncinatus, Fig (arrows). Fig. 6 Appendage apex and conglobate ascospores (arrows). Fig. 7 Rough walled sleurioconidia. Seale bar = 100 ym for Fig, Sand 5 jm for Figs. 6,7.T.C. Lumuey and R.S. Cunat (Orr, 1977), G. litoralis (= P. littoralis Orr) is hitherto known from marine materials including gastropod and crab shells, sea anemone and insect remains, coral and red algae. Considering this, itis unusual to find this organism in well- decayed wood at a site that is far inland. However, the association with invertebrates ‘may be more important ecologically than the locations in which previous collections were made. It is possible that the organism has a predilection for chitinous materials of inverte- brate origin and its occurrence in well-decayed wood is due to the presence of the remains of invertebrate animals. More isolations and testing for enzymatic capabilities would be helpful in clarifying the ecological niche of this organism Uncinocarpus uncinatus (Eidam) Currah Figs. 5-7 Ascomata semi-globose, yellow to pale brown, 250-1000 jm in diameter including appendages. Ascospores yellow, oblate, 3.64.2 jim x 2.4-2.8 um. Asci globose, 8-spored, 5.5-7.2 ym in diameter. Peridial hyphae yellow to orange, mostly smooth with some roughened areas, with some clavate termini. Appendages yellow to brownish-yellow, uncinate. unbranched and thickest at the distal end, 3.9-7.2 wm in diameter and 80-300 pm in length, septate only at the base. Anamorph of minutely roughened, yellowish aleurio- and arthroconidia, 3.5-4.9 jum x 4.6-7.8 pm. Habitat: in well-decayed wood of P. glauca collected at Elk Island National Park, Alberta Substrate unknown but generally considered to be keratin-rich material Previous reports of this taxon are almost exclusively from dung (Currah, 1985) so the iso- lation of this strain from decaying wood is unusual. The species bears a superficial resem- blance to Auxarthron californiense Orr & Kuehn which bears similar uncinate appendages but differs in having quite rough ascospores. 10 The genus Uncinocarpus was described by Sigler and Orr (in Sigler and Carmichael, 1976) to accommodate an ascomycete frequent in soil and keratin-rich substrata, It forms ascomata with uncinate appendages and minutely pune~ tate oblate ascospores. The anamorph is repre- sented by a Malbranchea state. Uncinocarpus uncinatus was originally described by Eidam (1880) as a species of Gymnoascus. Gymno- ascus has smooth ascospores, no conidial state and is not able to degrade keratin. Currah (1985) transferred the species to Uncinocarpus because of the rough or pitted ascospores, the arthroconidial anamorph and the ability to degrade keratin, The striking uncinate appendages unify U. uncinatus and U, reessii Sigler & Orr, the two species in the genus. The isolation plates that yielded this species were unusual in that the distinctive colonies developed from many of the wood fragments The abundance of U. uncinatus in this wood sample is difficult to explain based on its place- ment in the Onygenaceae (Currah, 1985), More detailed studies concerning the substrate prefeer- ences of this fungus are necessary to clarify its role in the late stages of wood decay. Myxotrichum ochraceum Berk. & Br. Figs. 8-11 Ascomata dark brown, more or less globose, 400-1000 pm in diameter, including appendages. Ascospores hyaline to pale yellow, fusiform, 1.3-2.0 pm x 4,0-5.0 im. Asci 8- spored, hyaline, stipitate, 5.2-7.2 ym in diame ter and up to 13 um in length including stipe. Peridial hyphae dark brown, thick-walled and branching at acute angles, similar in overall morphology to appendages. Appendages straight and blunt, dark brown, 2.2-2.6 pm in diameter at the base, narrowing at the tip to 1.6-2.4 jum and up to 250 tim in length, branch- ing in the proximal region and often lighter in colour at the apex. Anamorph not observed.‘Onygenalean fun from decayed wood Figs, 8-11, Myxotrichum ochraceum. Fig. 8. Ascoma with appendages (arrows). Fig. 9, Stipitate ascus. Fig, 10. Thick- walled, fusoid ascospores. Fig. 11. Branches on the proximal portion of appendages. Seale bar = 100 um for Fig. 8 and 5 tum for Figs. 9-11 Habitat: decayed log of P. tremuloides. Substrate: cellulose. Currah (1985) placed Myxotrichum, Byssoascus and Pseudogymnoascus in the Myxotrichaceae. Gymnostellatospora japonica Udagawa, Uchiyama & Kamiya was added to the family in 1993 (Udagawa et al., 1993). All of the taxa produce fusoid or boat-shaped ascospores. Myxotrichum ochraceum is only rarely reported in the literature. Specimens have been observed from bark, rotten wood, cardboard. and straw and from a grass basket. Recently, Uchiyama et al. (1995) described a new variety (M. ochraceum var. frigidum) from Japanese soil that differs from the specific form in hav- ing shorter appendages. Decayed wood is not a surprising place to find M. ochraceum, given the well-known predilection of this group for cellulosic sub- " strates. It might be expected that Myxo- trichaceae will be found to be more abundant in the boreal forest because of the taxon’s tenden- cy to be represented in cold habitats and the role of Oidiodendron species as ericoid mycor- thizal fungi. Ericoid mycorrhizal plants are often among the first vascular plants to become established on decaying fallen logs in the bore- al forest. Myxotrichum species and anamorphs may be co-opted in this situation as mycor~ thizal fungi by these early plant colonists. The role this species plays in the decay sequence is unknown. Conclusion In general, Onygenalean fungi are not known to be associated with wood although a few collections of some taxa exist from this material (e.g., Pseudogymnoascus roseusT.C. Lumuey and R.S. Cunnat Raillo, Amauroascus aureus (Eidam) von Arx, Auxarthron reticulatum (Zukal) Orr & Plunkett). The specific niche that would be occupied by Onygenalean fungi in rotting wood is unclear. Members of the Myxo- trichaceae have cellulases that allow them to be involved in the active degradation of wood components, and some of these taxa have been shown to form in vitro. mycorthizas with erica- ceous plants (Dalpé, 1991). Taxa within the Gymnoascaceae show no consistent substrate preference, although some may be cellulolytic. The presence of U. uncinatus in rotting wood is anomalous in that Onygenaceae are normally found associated with keratin-rich substrata. Many species from the Onygenaceae have been isolated from soil and it is probable that rotten ‘wood provides a similar habitat. However, U. uncinatus is not known from soil and the re: son for its presence in a rotting log is a matter for speculation. The presence of G. littoralis is also puzzling although, as mentioned above, it is possible that it is a saprophyte on chitinous remains of invertebrates. Studies of fungus communities in decaying wood, with a few exceptions (Hallaksela, 1993; Hutchison and Hiratsuka, 1994; Rayner, 1977), deal mainly with macrofungi (e.g., Niemeki et al., 1995). This paper outlines some pretiminary results from an ongoing survey of Onygenalean and other microfungi from decaying logs. More extensive surveys are underway in the Boreal Forests of Alberta to provide a checklist of these organisms in natural rotting logs and to deter- mine their role in the forest ecosystem. References Currah, R.S. 1985. Taxonomy of the Onygenales: Arthrodermataceae, Gymno- ascaceae, Myxotrichaceae, and Onygenaceae. Mycotaxon 24: 1-216. Dalpé, Y. 1991. Statut endomycorhizien du genre Oidiodendron. Can. J. Bot. 69: 1712-1714, 12 Eidam, E. Gymnoascaceen Pflanzen 3: 267-305, Eriksson, O.E. and Hawksworth, D.L. 1993. Outline of the Ascomycetes-1993. Syst Ascomycetum 12: 51-257. Hallaksela, A.M. 1993. Early interactions of Heterobasidion annosum and Stereum san- guinolentum with non-decay fungi and bac teria following inoculation into stems of Picea abies. Eur. J. For. Pathol. 23: 416-430, Hutchison, LJ. and Hiratsuka, Y. 1994. Some wood-inhabiting yeasts of trembling aspen (Populus tremuloides) from Alberta and northern British Columbia, Mycologia 86: 386-391 Niemeki, T., Renvall, P. and Penttla, R. 1995. Interactions of fungi at late stages of wood decomposition. Ann. Bot. Fennici 32 141-152. Orr, G.F. 1977. New Gymnoascaceae Mycotaxon 6: 33-42. Rayner, A.D.M. 1977. Fungal colonization of hardwood stumps from natural sources. I. Non-basidiomycetes. Trans. Br. Mycol. Soc. 69: 291-302, Sigler, L. and Carmichael, I.W. 1976. Taxonomy of Malbranchea and some other hyphomycetes with arthroconidia Myco- taxon 4: 349-488, Uchiyama, S., Kamiya, S., and Udagawa, S. 1995. A new variety of Myxorrichum ochraceum from Japanese soil, Mycoscience 36: 289-292 Udagawa, S., Uchiyama, S., and Kamiya, S. 1993. Gymnostellatospora, a new genus of the Myxotrichaceae, Mycotaxon 48 157-164. von Arx, J.A. 1977. Notes on the Gymno- ascaceae. Persoonia 9: 393-400. von Arx, J.A. 1986. The ascomycete genus Gymnoascus. Persoonia 13: 173-183. 1880. Beitrag zur Kenntnis der Cohn’s Beitr. Biol‘Onygenalean fungi from decayed wood a E BAD 5 MLE NRK SAO 3 HEIST T.C. Luwey - R.S. CuRRAH AP IOT MN FHILBUT, HOD te DML EAM DS EO 9 MMARATED (Onygenales) (27H 3 ff (Gymnascella littoralis, Myxotrichum ochraceum 43 £ U Uncinocarpus uncinatus) @ 508, EHEROMI OWT RL te FOI, G. lintoralis BE OU, uncinatus (Eb YE BOM Picea glauca PSY TONEY ERROR EMOT. EH M, ochraceum (9:2 $¥ RO fi Populus tremuloides & BIL ASH % ACIP L LAL 3 BIRR MEA HDT BM. Cab. ET. BIR ARAE FOIE L ACHE b SMES He Hada 13