J. Gen. Appl. Microbiol., 18, 433-454 (1972) COPROPHILOUS PYRENOMYCETES FROM JAPAN I KOUHEI FURUYA AND SHUN-ICHI UDAGAWA* Fermentation Research Laboratories, Sankyo Co., Ltd., Hiro-machi 1-chome, Shinagawa-ku, Tokyo 140 and *Department of Microbiology , National Institute of Hygienic Sciences, Kamiyoga 1-chome, Setagaya-ku, Tokyo 158 (Received July 13, 1972) For the purpose of these series of mycological survey, 220 dung samples of wild and domestic animals for determination of species of pyrenomycetous Ascomycetes were collected from various geographic regions of Japan, in- cluding Ryukyu and Bonin Islands. Fifteen species of Podospora (the Sor- dariaceae) from numerous collections are described and illustrated. Most of them were also obtained in living cultures. All species are new records in Japan. Generally speaking, animal dungs contain very rich nutritive components which may serve as growth factors for various types of microorganisms. The coprophilous fungi, one of such dung inhabitants, comprise a special group made up of members of several classes ranging through Myxo- mycetes to Basidiomycetes. In their pioneering studies, MASSEE and SAL- MON (1, 2) emphasized that 187 genera and 757 species from coprophilous occurrence had been listed in SACCARDO's Sylloge Fungorum in the early of the present century. These fungi have long attracted many mycologists for more than a hundred years, and a considerably large number of taxonomic papers have been published on them from various areas of the world. Investigations on this fascinating group of fungi were relatively few in Japan. The most extensive work is that of TUBAKI (3), who isolated 16 species belonging to the Hyphomycetes from dung sources in Japan. SONEDA (4, 5), in more recent reports, also listed 22 species of coprophilous yeasts, In two investigations resulted in the primary work of the Japanese Mucorales, some dung-inhabiting fungi were described by Indoh (6, 7). These contributions, however, provided only a little knowledge of their geographical distribution, especially on the occurrence of Japanese coprophilous Ascomycetes. Two recent studies on the discomycete flora, initiated by OTANI and KANZAWA (8, 9), have led to a suggestion that a majority of the coprophilous Ascomycetes found in Japan are common among other 433 434 FURUYA and UDAGAWA VOL. 18 countries, though they are little localized geographically. The present paper is the first attempt to give a survey of the Japanese members of detached group of Ascomycetes usually referred to as the coprophilous Pyrenomycetes. MATERIALS AND METHODS A total of 220 dung samples were collected from different localities in Japan from September 1966 to November 1971. These localities included Ryu- kyu Islands (now Okinawa Prefecture) and Bonin Islands (now Ogasawara Is- lands, Tokyo). The collections comprised of the droppings of following wild and domestic Mammalias : Brown bear (Ursus arctos), covey (Ochotona hyperborea), deer (Cervus nippon), fox (Vulpes vulpes), Japanese hare (Lepus brachyurus), marten (Martes melampus), monkey (Macaca fuscata), raccoon dog (Nyctereutes procyonoides), squirrel (Sciurus vulgaris and Tamias sibiricus), cattle, horse, goat, sheep, and others. The samples of dung collected in the field were held in moist chambers at room temperature, observing frequently for a period of one to four weeks or longer for development of marked fungi. Isolation was usually attempted on several media such as Leonian's yeast extract agar, oatmeal agar (with wheat germ powder), cellulose agar, potato-carrot agar and potato-dextrose agar. With some exceptions, the fungi mentioned in these series may be grown with great difficulty on the above laboratory media. In most cases, therefore, microscopical observations were carried out using living materials on the dung. Lactophenol preparations and dried specimens of all species have been placed in the Mycological Collection of the National Institute of Hygienic Sciences, Tokyo. RESULTS AND DESCRIPTIONS Among the total collection of 52 species of Pyrenomycetes examined, the order of occurrence for the chief genera was Podospora, Sporormiella, Conio- chaeta, Delitschia, Sordaria, and Chaetomium. The selected publications used for most of the identifications were GRIFFITHS (10), CAIN (11, 12), MOREAU (13), BOEDIJN (14), MIRZA and CAIN (15), ARMED and CAIN (16), and LUNDQVIST (17). Thus, it is now possible to report that 15 species of Podospora are known to occur in Japan. The remainder will be described in subsequent papers. Podospora appendiculata (Auersw.) Niessl, in Hedwigia, 22, 156 (1883) ; Mirza and Cain, in Can. J. Bot., 47, 2007 (1969) ; Lundqvist, in Symb. Bot. Upsal., 20, 129 (1972). (Fig. 1) Sordaria appendiculata Auersw., 1872. For additional synonymy see MIRZA and CAIN (15) and LUNDQVIST (17). Perithecia superficial, scattered or aggregated in small clusters, ovate to 1972 Coprophilous Pyrenomycetes I 435 Fig. 1. P, appendiculata (Auersw.) Niessl. a, Perithecia ; b, Ascus ; c, Ascus gores. somewhat conical, 700-1,000 x 380-800 p, dark brown to nearly black, sparingly hairy, neck small, papilliform to broadly conical, usually 50-150>< 120-160 p, roughened with black papillae. Hairs short, straight or slightly flexuous, septate, 30-150 p long, 4-5 p wide and brown at the base, with rounded or sometimes pointed, hyaline tip. Peridium fairly thick, coriaceous, dark brown and opaque, consisting of irregular cells, measuring 11.5-22.5 in diam. Asci 8-spored, clavate, 180-300 x 25-45 p, contracted and narrowly rounded above, with small thickened ring at the apex, tapering below into a slender stipe, measuring 70-100 p long ; paraphyses hyaline, thread-like, evanescent. Ascospores biseriate in central portion, hyaline and clavate when young, becoming dark brown and ellipsoid at maturity, 25-30 x 12.5-15 p, rounded above, somewhat truncate at the base, with apical germinal pore ; primary appendage cylindrical to conical, 12.5-19x4.5-6.5 p, hyaline, fugacious ; lower secondary appendage lash-like, hyaline, gelatinous, 25-60 p in length, attached 436 FURUYA and UDAGAWA VOL. 18 to the end of primary appendage ; upper one lash-like, 60-100 p in length, hollow at the base, symmetrically attached at the apex of the ascospore. Cultures on oatmeal agar growing slowly, with a thin mycelial felt, more or less funiculose with the development of a few aerial hyphae, perithecia tardily produced, grayish brown to dark brown ; reverse dark brownish gray. Habitat : On hare dung, Fujiyoshida-shi, Yamanashi-ken, April 26, 1967, NHL 22385, Kazikazawa-machi, Minamikoma-gun, Yamanashi-ken, Nov. 4, 1966, Kamikawa-machi, Kamikawa-gun, Hokkaido, July 28, 1967, Sanada-machi, ChIsagata-gun, Nagano-ken, Aug. 20, 1967, Ebino-machi, Nishimorokata-gun, Miyazaki-ken, April 19, 1971, NHL 22542; and on deer dung, Nara-shi, Nara ken, Oct. 16, 1966, NHL 22543. P. appendiculata is characterized by large ovate perithecia with pseudo bombardioid peridium that is somewhat suggestive of members of the genus Cercophora. Podospora australis (Speg.) Niessl, in Hedwigia, 22, 156 (1883) ; Mirza and Cain, in Can. J. Bot., 47, 2009 (1969) ; Lundqvist, in Symb. Bot. Upsal., 20, 203 (1972). (Fig. 2) Hypocopra australis Speg., 1880. For additional synonymy see MIRZA and CAIN (15) and LUNDQVIST (17). Perithecia scattered, superficial or partially immersed, dark brown to nearly black, pyriform, 900-1,600 x 600-700 p, densely covered on the exposed part with long, septate, branched, olivaceous brown hairs measuring 2.5-3 p in diam. ; neck black, papilliform to cylindrical, 280-700x180-280 p, covered with short, straight, septate, hyaline-tipped hairs measuring up to 55 p long and 4 p wide at base, or almost bare. Peridium dark olivaceous brown,, membranaceous to rather coriaceous. Asci 4-spored, cylindrical, 300-380 x 40 -45 p , tapering below into a slender stipe measuring 80-150 p in length ; paraphyses filiform to ventricose. Ascospores uniseriate, dark brown and opaque, ellipsoid, broadly rounded at the both ends, (52-)56-60(-70) x (28-) 32-34(-40) p ; exospore 1.5-2 p thick ; primary appendage reduced to a small basal apiculum, hyaline, 3.5-4 p wide at the base ; lower secondary appendage gelatinous, long, attached to the basal end of spore, completely covering the apiculum, appearing hollow and taeniate ; upper secondary appendage similar, 50-80 (-100) x 8-15 p, attached eccentrically ; germinal pore apical. Habitat : On goat dung, Sanada-machi, Chisagata-gun, Nagano-ken, Oct. 3, 1969, NHL 22548. This species is quite distinctive and can be identified by its large asco- spores with hyaline, small, basal apiculum and two taeniate gelatinous ap- pendages. Podospora communis (Speg.) Niessl, in Hedwigia, 22, 156 (1883) ; Mirza and Cain, in Can. J. Bot., 47, 2012 (1969) ; Lundqvist, in Symb. Bot. Upsal., 1972 Coprophilous Pyrenomycetes I 437- Fig. 2. P. australis (Speg.) Niessl. a, Perithecium ; b, Ascus ; c, Ascospores. 20, 171 (1972). (Fig. 3) Hypocopra communis Speg., 1880. For additional synonymy see MIRzA and CAIN (15) and LUNDQVIST (17). Perithecia scattered, semi-immersed to nearly superficial, olivaceous brown to dark brown, pyriform, 650-820x325-400 u, covered on the basal part with pale olive, long, flexuous, septate hairs measuring 1.5-2 t wide near the base, sometimes nearly glabrous ; neck cylindrical, 170-400 x 100-135 ~c, roughened with small, black papillae. Perithecial peridium thin, mem-- branaceous, semitransparent, consisting of yellowish olive, angular cells. Asci 8-spored, clavate, 180-220x36-40 p, narrowly rounded above, with no discharged structure visible in apex, gradually contracted below into a short stipe up to 40 in length ; paraphyses ventricose, indistinct. Ascospores biseriate, dark olivaceous brown and opaque, ellipsoid, 28-36(-40) x 18-22 t ; primary appendage hyaline, long, cylindrical, 25-32 x 4-6 ~c, tipped at the lower end with 3 or 4, short, divergent, gelatinous secondary appendages ; apex of the spore with 4 similar secondary appendages ; germinal pore apical, small, about 1.5 in diam. Cultures on oatmeal agar growing rapidly, thin, loosely floccose, dark . greenish gray, conidial structures limited in number ; perithecia lacking; reverse greenish black.