Three Species of Hypocrea with Stipitate Stromata and Trichoderma Anamorphs

Three species of Hypocrea with stipitate stromata and Trichoderma anamorphs

Gary J. Samuels The Hypocrea species discussed in the present work United States Department of Agriculture, Agriculture could be, or have been, included in Podostroma or Research Service, Systematic Botany and Mycology Podocrea (Sacc.) Lindau on account of their having Laboratory, Rm. 304, B-011A, BARC-W, Beltsville, Maryland 20705 stalked stromata. Saccardo (1883) included H. brevi pes in Hypocrea subg. Podocrea Sacc., and later (Sac D. J. Lodge cardo and Saccardo, 1905) included both H. brevipes Centerfor Forest Mycology Research, United States and H. poronioidea in Podocrea. Doi (1975, 1979) in Department of Agriculture, Forest Service, Forest Products Lab., PO Box B, Palmer, Puerto Rico 00721 cluded H. brevipes in Podostroma. Moreover, a syn onym of H. poronioidea is P. orbiculare. In Podostroma stromata comprise a stipe surmounted by a distinct, Abstract: Hypocrea brevipes, H. poronioidea and H. fertile cap. In Podocrea the stroma is cylindrical and capitata, a new species, were grown in pure culture not capitate. Atkinson (1905) synonymized the later from individual part-ascospores. Each produced a genus, Podocrea, with PodostromaJ and this synonymy Trichoderma anamorph, but none of the anamorphs has generally been followed. Rogerson (1970) ac were referable to any named Trichoderma aggregate cepted Podostroma. Doi (1975, 1979) provisionally ac species. Stromata with fertile ascospores formed in cepted Podostroma, doubting that it could be main cultures derived from single part-ascospores of H. po tained as a genus distinct from Hypocrea (see Doi, ronioidea. In addition, a hyphomycetous, Acremon 1975). ium-like synanamorph with hyaline conidia borne in Within Hypocrea there is nearly continuous varia slime formed in cultures of H. poronioidea. The dis tion in form that includes stromata that are indefi tribution of H. poronioidea is extended from the nitely effused, pulvinate but broadly attached, discoi Americas to Africa. These Hypocrea species and their dal and attached at a central point, or clavate to stip anamorphs are described. itate-capitate. Among the species that have Trichoder Key Words: Hypocreales, Podocrea,Podostroma, sys ma. anamorphs, there is no obvious morphological or tematics biological hiatus that could support generic separa tion on the basis of stromal form, and the clavate or stipitate condition per se does not imply a dose rela INTRODUCTION tionship among those species that posess it. Although stromata of H. poronioidea, H. capitata, and H. brev Three Hypocrea Fr. species having stipitate-capitate ipes are morphologically similar, stromata of H. brev stromata were collected recently in tropical habitats. ipes are anatomically distinct from the others. Hypo These species are H. brevipes (Mont.) Sacc., H. poron crea brevipes and Podostroma eperuae Rogerson & Sa ioidea Möller, and the new species H. capitata. Hy muels (Rogerson and Samuels, 1992), a species with pocrea poronioidea has been reported once since its a clavate stroma, share the feature of unusual elon original description (Möller 1901), as Podostroma or gated or brick-like cells at the stroma surface, but biculare Chardón (Chardón, 1921), but has not been otherwise have little in common. While our under redescribed in modern terms. Hypocrea brevipes was standing of stromal anatomy in Hypocrea is far from redescribed by Doi (1975, 1979). Trichoderma Pers. anamorphs developed in colonies derived from as complete, we suspect that these brick-like cells have cospores of each. In addition, a synanamorph having resulted from the extension of the stroma and do not hyaline conidia borne in colorless slime formed in indicate any relationship. cultures of H. poronioidea. Neither H. brevipes nor H. Anamorphs have suggested relationships among poronioidea have previously been linked to ana genera and species of the Hypocreales (Samuels and morphs. These Hypocrea species and their anamorphs Seifert, 1987). Few species of Podostroma have been are described, and the relationship of Hypocrea to Po grown in pure culture and linked to anarnorphs. At dostroma P. Karst. is discussed. kinson (1905) obtained stromata of P. alutaceum (Pers.) G.F. Atk. in agar culture tubes, but did not Accepted for publication October 31, 1995. note the presence of an anamorph. Tubaki (1958)

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SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 303 and Doi (1967, 1973) have connected Podostroma tions of 12 h darkness/12 h cool white fluorescent species to Trichoderma or to Verticillium-like ana light. Colony colors are taken from Kornerup and morphs, respectively. Wanscher (1978). The Trichoderma anamorphs of the three Hypocrea Dry specimens were rehydrated briefly in 3% (aq.) species discussed in the present paper cannot be KOH and the mounts were then flooded with water. placed into any of the aggregate species defined by Measurements and photographs were taken of ma Rifai (1969), although they bear some similarity to T. terial mounted in water with the following excep hamatum- aggr., or to species included in Trichoderma tions. Perithecial sections were mounted in 100% lac sect. Pachybasium Bissett (Bissett, 1991b) or section tic acid. The optical brightener calcofluor (Sigma Trichoderma (Bissett, 1991a). The anamorph of H. Chemical Co., 0.05% w/v in sodium phosphate buff capitata is characterized by having ampulliform to la er at pH 8) was used for fluorescence microscopy. geniform phialides clustered on a rather broad main Anamorph measurements are taken from CMD. axis; additionally, sterile branches sometimes extend Four types of microscopy were used in this study. from conidiophores (FIG. 55). This anamorph could These are indicated in the legends to the illustrations be placed in sect. Pachybasium. The anamorph of H. as brightfield (BF), fluorescence (EL), differential in poronioidea is morphologically more diverse. In part terference contrast (DIC), and phase contrast (PC). it could be included in sect. Pachybasium because of The conventions KOH+ and KOH- are used. its clustered, ampulliform phialides (FIG. 58). How KOH+ indicates that perithecia and/or stromata be ever, it also produces more loosely aggregated whorls come some color of red in 3% KOH and yellow in of lageniform phialides (FIG. 59) which indicate sect. 100% lactic acid. KOH- indicates that no red color Trichoderma. The Trichoderma anamorph of H. brev changes were observed in 3% KOH or 100% lactic ipes, with its loose verticils of lageniform phialides is acid. better placed in T. sect. Trichoderma. Various Hypo Representative isolates have been deposited in the crea species have been linked to each of these sec American Type Culture Collection (ATCC) and the tions (Bissett, 1991a,b; Doi and Doi, 1979; Domsch Centraalbureau voor Schimmelcultures (CBS). et al., 1980), but the sections-whichare derived from Rifai’s (1969) aggregate species-are based en DESCRIPTIONS OF THE SPECIES tirely on form. Our knowledge of Hypocrea and Trich oderma is not sufficiently advanced to enable predic 1. Hypocrea brevipes (Mont.), Saccardo, Michelia 1: tion of an aggregate species (Rifai, 1969) or section (Bissett, 1991a) of Trichoderma, or type of Hypocrea from, respectively, an unknown Hypocrea or Tricho Cordyceps brevipes Montagne, Syll. gen. crypt. derma. 201. 1856. A remarkable feature of cultures of H. poronioidea Podocrea brevipes (Mont.) Sacc. & D. Sacc., Syll. Fung. 17: 799. 1905. was the formation of a hyphomycetous synanamorph Podostroma brevipes (Mont.) Seaver, Mycologia in association with incipient stromata in self-fertile 2: 66. 1910. cultures. Conidia of this synanamorph are colorless and held in conspicuous drops of liquid at the tips Anamorph. Trichoderma sp. FIGS. 10-15 of Acremonium-like conidiophores. Conidia are ca pable of germination so that while these may have a Stromata capitate-stipitate to pulvinate and then spermatial role, they are also capable of dissemina centrally attached with margins free; stroma some tion. This synanamorph is similar to the anamorphs times appearing to have formed through the lateral formed by species of Hypocrea that have effused stro fusion of adjacent stromata with more or less conspic mata such as H. lactea (Fr.: Fr.) Fr., H. pulvinata (Ri uous fissures at points of joining of adjacent stromata, fai and Webster, 1966) and other similar species (Doi caps of individual stromata circular in outline, 2-8 mm 1972). diam, hemispherical, margin slightly inrolled or not, in colors of dark to grayish brown (6E-F7-8), KOH-, surface glabrous, wrinkled and perithecial elevations MATERIALS AND METHODS not evident or slightly tuberculate from perithecial el Single ascospores were isolated with the aid of a mi evations, ostiolar openings visible as small viscid dots; cromanipulator on cornmeal dextrose agar medium undersurface of cap concolorous, slightly velvety or (CMD: Difco cornmeal agar + 2 g/l dextrose) at 20 not with velvet continuing on stipe or not. Stipe cen C. Colony characters were taken from CMD, malt ex tral, 1.5-5 mm long, 0.75-1 mm diam, adjacent stipes tract agar (ME), and potato dextrose agar (PDA, Dif often appearing to have fused, with fissures demarking co). Colonies were grown at 20-21 C under condi- the individual stipes, KOH-. Cells of stromal surface, 304 MYCOLOGIA SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 305

FIGS. 10-13. Hypocrea brevipes Trichoderma anamorph on CMD. From van der Gucht & L. De Meester 92-14325b. Scale bars: 10-12 = 50 µm; 13 = 10 µm. in surface view, angular, 10-15 µm diam, walls 1 µm > 9 cm diam within 6 da; aerial mycelium lacking, thick, KOH-. Stromal surface region ca. 50 µm wide, diffusing pigment lacking; conidia forming in a sin cells pseudoparenchymatous, 6-10 µm diam, walls gle broad, continuous band around the margin, or 1.0-1.5 µm thick, pale yellow, colorless in lactic acid. in 2-3 concentric rings. Aggregates uniformly cot Tissue below the stromal surface of loosely inter tony with Fertile branches protruding, easily removed twined, 3 µm wide, thin-walled hyphae. Cells at sur from the agar surface; deep green (27E8) or grayish face of stipe brick-like and elongated parallel to the green (28D6) fading through gray to near white at long axis of the stipe o not elongated, 15 µm long, the margin. Colonies on PDA > 9 cm diam within 6 7.5 µm wide, walls 1.5-2.0 µm thick. Perithecia crowd da, no diffusing pigment formed; conidia formed ed below the stromal surface, elliptic, (n = 13) 245 profusely in dense concentric sings alternating with 328 µm high, 128-214 µm wide; cells at perithecial mycelial. production, grayish- to deep green (28D-E7 apex around ostiolar opening small and pseudopar 8) in the middle, progressively lighter green toward enchymatous, thin-walled, yellow in lactic acid; ostiolar the margin where conidia are nearly white. Colonies canal 62-104 µm long, periphysate. Tissue below per on ME > 9 cm within 6 da, no diffusing pigment ithecia of vertically oriented, brick-like cells to 50 µm formed; conidial production much as on PDA but long, 10 µm wide, walls ca. 1 µm thick, nodose ele conidia developing more slowly on ME and some ments lacking, colorless. Asci cylindrical, (n = 66) times with yellow pigment in conidia formed around (60.0-) 71.8-87.6 (-97.7) × (3.0-) 4.2-5.6(-6.6) µm, apex the center of the colony. Main axes of conidiophores thickened and with a ring; 8-spored, ascospores uni formed on CMD 3-4 µm wide, fertile branches 24 seriate and often with overlapping ends, completely 35 µm long, infrequently rebranched; phialides aris filling each ascus. Part-ascospores dimorphic, distal ing singly along the length of branches and in cru part subglobose to conic, (n = 71) (3.0-) 3.5-4.3 (-4.7) ciate whorls of ca. 3 at branch tips. Phialides lageni × (2.4) 3.1-3.9 (4.7) µm, proximal part wedge-shaped, form, (n = 30) (3.6-) 5.2-8.2 (-10.5) µm long, (2.4-) 2,7 (3.0-) 3.6-5.0 (-6.1) × (2.2-) 2.2-3.2 (-3.4) µm; hyaline, 3.5 (-4.3) µm wide in the middle, (1.5-) 1.9-2.5 (-2.9) finely spinulose. µm wide at the base. Hypha or branch immediately A few ascospores germinated within 48 h. On CMD subtending each phialide (n = 30) (2.0-) 2.4-3.4

FIGS. 1-9. Hypocrea brevipes. 1. Stroma. 2. Median longitudinal section through a mature perithecium. 3. Cells at stromal surface. 4. Detail of perithecial apex. 5. Detail of stromal suface between perithecia. 6. Internal tissue of stroma below perithecia. 7. Cells at surface of stipe. 8, 9. Asci; ascal apex visible in 9. Fig. 1 = BF 2-9 = DIC; Fig. 9 stained with cotton blue to reveal ascal apex. Figs. 10-13 = PC. All from van der Gucht & L. De Meester 92-14325b. Scale bars: 1, 8, 9 = IO µm; 2 = 100 µm; 3-7 = 50 µm. 306

FIGS. 14-16. Hypocrea brevipes. 14, 15. Trichoderma anamorph on CMD. 16. Ascus and ascospores. FIGS. 17, 18. H. capitata. 17. Ascus and ascospores. 18. Trichoderma anamorph from CMD. 14, 15 from van der Gucht & L. De Meester 92-14325b; 16 from the type; 17, 18 from Rossman 3298. Scale bars = 10 µm.

(-2.9) µm wide. Conidia oblong to ellipsoidal, (n = Known distribution. England (Doi, 1975), French 31) (2.9-) 3.1-3.9 (-4.7) × (2.0-) 2.2-2.8 ()-2.9) µm, lack- Guiana, Japan (Doi, 1975), New Guinea, Puerto Rico. ing a visible basal abscission scar, smooth, green. HOLOTYPE. FRENCH GUIANA. [locality and Chlamydospores not seen. date unknown], [on decorticated wood], Leprieur Habitat. Decaying wood. 1073 (PC! isotype BPI-Lloyd 715550!). SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 307

Additional specimens examined. NEW GUINEA, gion of the underside of the cap and of the stipe MADANG PROV.: Ramu Region, road to Bundi, circular to angular, ca. 7 µm diam, thin-walled, pro 05°44'N, 145°19'E, on decayed wood, 27 May 1992, ducing hyphae to 100 µm long, ca. 5 µm wide, sep K. van der Gucht & L. De Meester 92-14325b (Sa tate, unbranched through most of the length and muels culture 92-76) (BPI, GENT). PUERTO RICO. with many free ends, reddish brown, not changing El Yunque, [host unknown], 4 Dec. 1912, F J. Seaver color in lactic acid. Perithecia crowded below the (BPI 630179). stromal surface, oblong, (n = 50) (200-) 237-281( Notes. Hypocrea brevipes was cultured from the re 300) µm high, (76-) 89-131 (-162) µm wide; cells at cent collection from New Guinea. The above descrip perithecia1 apex around ostiolar opening inter tion is drawn from the three collections cited above. twined, short, and conspicuously thick-walled, form The stromata of the New Guinea and Puerto Rican ing “shoulders” on the perithecium as seen in lon collections appear to have resulted from the fusion gitudinal section, yellow in lactic acid; ostiolar canal of two or more adjacent stromata. The type collection 22-79 µm long, periphysate. Tissue of stromal inte consists of only a single stroma that has a longitudi rior below perithecia densely disposed 5-6 µm wide nally furrowed stipe, but that is not obviously com hyphae with walls conspicuously thickened, ca. 1.5 pound. The undersurface of the cap, and the stipe µm; swollen or nodose elements lacking. Asci cylin of the type collection are slightly velutinous whereas drical, (n = 172) (41-) 51-69 (-85) × (2.5-) 3.0-4.6 ( the stromata of the other two collections are either 6.4) µm, apex thickened and with a ring; 8-spored: glabrous or, at most, slightly velutinous. ascospores at first uniseriate but at maturity becom Doi (1975) accepted a wide range in size and mor ing transversely oriented or overlapping, completely phology for H. brevipes, reporting the species from filling each ascus. Part-ascospores dimorphic, distal temperate and tropical latitudes, part globose to subglobose, (n = 185) (1.9-) 2.6-2.9 (-3.2) × (1.5-) 1.9-2.5 (-2.8) µm; proximal part ob 2. Hypocrea poronioidea A. Möller, Phycom. und As long, (2.0-) 2.5-5.3 (-4.5) × (1.4-) 1.7-2.1 (-2.5) µm; ma turing slowly in the ascus, hyaline, ultimately becom com. p. 295. 1901. FIGS. 19-40 ing finely spinulose. Podocrea poronioidea (A. Möller) Sacc. & D. Ascospores germinating in low percentage within Sacc., Syll. Fung. 17: 799. 1905. 36 h. Colonies on CMD and ME > 9 cm diam within = Podostroma orbiculare Chardón, Mycolegia 13: 6 da; aerial mycelium and pigment lacking, conidia 286. 1921. forming only after 10 da on CMD, in 1-3 mm diam pulvinate tufts at the colony margin. Colonies on Anamorph: Trichoderma sp. FIGS. 28-40 PDA > 9 cm diam within 6 da; aerial mycelium scant, Stromata capitate-stipitate to pulvinate and then pigment lacking; conidial production beginning centrally attached with margins free; cap circular in within 6 da. Conidia on ME and PDA forming in outline, (2.0-) 3.0-6.5 (-10.0) mm diam, slightly con more or less uniformly distributed pulvinate tufts that vex and margin slightly inrolled, in colors of brown are cottony and that have protruding branches that (6E8, 7E7), yellowish brown (5D8, 5E8), light brown are fertile to the tip as well as with long protruding (6D8), or brownish gray (6F8), KOH-, surface gla branches that are fertile only at the tip; on CMD, ME brous, plane and perithecial elevations not evident, and PDA deep green (28D8 to 30D8) or pea green ostiolar openings visible as dark brown or black dots (29D5); conidial production on CMD is considerably against the lighter brown background; undersurface less than on ME or PDA. Main axes and lateral of cap often lighter or darker in color, apricot yellow branches 1.8-3.2 µm wide; phialides arising singly to Indian yellow (5B6-7), linoleum brown (5E7), or from the main axis or lateral branches, or arranged brown (6E8), KOH-, velvety with velvet continuing on in ‘cruciate’ whorls of 3-5 at the tips of short branch stipe. Stipe central, to 7 mm long, 1.0-2.5 mm diam, es, or more densely clustered, tending to be botryose. or lacking, concolorous with underside of cap, vel Phialides lageniform and widest in the middle, (n = vety. Cells of stromal surface, in surface view, lacking 40) (3.6-) 4.2-7.4 (-10.0) µm long, (1.9-) 2.1-2.7 (-3.1) a definite outline or angular, walls ca. 1.5 µm thick. µm wide in the middle, (1.0-) 1.2-l.8 (-2.3) µm wide In section, stromal surface clearly differentiated from at the base. Hypha or branch immediately subtend underlying tissue, crustose, 15-25 µm wide, cells an- ing each phialide (n = 30) (1.8-) 2.2-3.0 (-3.2) µm gular and ca. 7 µm diam, or almost brick-like and 4- wide. Conidia ellipsoidal, (n=48) (1.9-) 2.3-2.9 (-3.0) 6 × 3 µm, walls 1-2.5µm thick, yellow in lactic acid. × (1.5-) 1.6-2.2 (-2.4) µm, lacking a visible basal ab- Tissue immediately below the stromal surface of scission scar, smooth, green. Acremonium-like conidi loosely intertwined, 2-3 µm wide, branching and sep- ophores arising from cottony, incipient stromata; tate, smooth-walled hyphae. Cells of the velvety re- conidiogenous cells 5-30 µm long, straight, smooth, 308 MYCOLOGIA

FIGS. 19-27. Hypocrea poronioidea. 19. Habit of two stromata. 20. Cells at stromal surface in the area of the ostiolar opening (arrow). 21. Median longitudinal section through a mature perithecium. 22. Detail of stromal surface between perithecia. 23. Internal tissue of stroma below perithecia. 24. Detail of perithecial apex. 25. Surface of stipe. 26, 27. Asci. Fig. 19 = BF; 20-22, 24-27 = DIC 23 = FL. Figs. 19, 26 from Rick 220 (S-Rehm), 20, 23-25 from “authentic” (FH-GEN), 21, 22, 27 from GJS 92-29. Scale bars: 19 = 10 mm; 20-25 = 50 µm, 26, 26 = 10 µm. SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 309

FIGS. 28-35. Hypocrea poronioidea, synanamorphs. 28-32.Trichoderma synanamorph. 33-35. Acremonium-like synanamorph from incipient stromata. Figs. 28, 30 = FL; 29, 33, 34 = PC; 31, 32, 35 = DIC. Figs, 28-30from SNA, 31, 32 from PDA, 33 35 from CMD. Scale bars: 28-30, 33, 34 = 10 µm; 31, 32, 35 = 50 µm

tip 1 µm wide, lacking periclinal thickening, not 1921, Rick s. n. (BPI-Lloyd 6106); São Leopoldo, in flared, base 2-3 µm wide. Conidia oblong, 7-13 × 3-4 ligno frondoso, 1907, Rick Fungi Austro-Americani µm, unicellular, lacking a basal abscission scar, hya 220 (HBG-Magnus, HBG-Hamburgense, S-Rehm); line. Chlamydospores not observed. São Leopoldo, ad ligno, Rick 109b (S-Bresadola). Habitat. On bark. DOMINICA. Springfield Estate, on wood, 21 Jun. Known distribution. Brazil, Dominica. Puerto Rico, 1970, A. Y. Rossman 246 (BPI 630172, CUP-DO 42). Uganda. PUERTO RICO. Mayaguez, on logs, 11 Dec. 7915, B. TYPE. BRAZIL.. Sta. Catharina pr. Blumenau, in lig Fink 239 (BPI 630180, HOLOTYPE of P. orbiculare); no putrido, leg. A. Möller (FH-GEN; specimen an on log, 12 Dec. 1915, B. Fink 922 (BPI 630181, as P. notated as “authentic”). orbiculare); Luquillo Mts., Caribbean National Forest, Additional specimens examined. BRAZIL. Estrella, El Verde Research Area, on log, 11 Jul. 1992, D. J. 310 MYCOLOGIA

FIGS. 36-40. Hypocrea poronioidea. 36, 37. Trichoderma synanamorph from PDA. 38. Acremonium-like synanamorph from incipient stroma on CMD. 39. Median longitudinal section through a whole stroma (diagramatic). 40 Ascus and ascospores. Figs. 36, 38 from GJS 93-29, 37 from GJS 92-29, 39 from Lloyd 6106; 40 from “authentic” (FH-GEN). Scale bars = 10 µm except 39 = 2 mm.

Lodge PR-913 (BPI 802499), El Verde Research Area, 1992, D. J. Lodge s.n. (Samuels culture 92-29, BPI); trail to the Radiation Center, on log, 23 Mar. 1993, on log, 22 Nov. 1992, S. M. Huhndorf 49 & D.J- D. J. Lodge PR-1026 (BPI); El Verde Research Area, Lodge PR-1081 (BPI, NY); El Verde Research Area, along La Prièta Trail, on fallen tree with bark, 1 Mar. trail to Río Sonadora, on right side just after leaving SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 311 the El Verde Field Station, elev. 350 m, on log, 15 self fertile colonies. Perithecia formed on CMD and Dec. 1992, D. J. Lodge PR-961 (Samuels culture 93 PDA in unsealed petri dishes incubated at room tem 29, ATCC 90748, CBS 37494; BPI). UGANDA. Kibale perature (ca. 21 C); they did not form in petri dishes Natl. Park, Ngogo Station, on decorticated wood, 2 incubated in an incubator at 20-21 C with alternating Aug. 1995, K.T. Hodge U0267 (BPI, Samuels culture (21 h/12 h) darkness and cool white fluorescent 95-133). light. Study of the genetics of sexuality in this species Illustrations. Möller (1901, Taf. II, Fig. 37). Char is continuing. dón (1921, Pl. 13, Fig, 11). Lloyd (1923, Pl. 233, Fig. 2386). 3. Hypocrea capitata Samuels, sp. nov. FIGS. 41-59 Notes. Three recent collections of Hypoma poron ioidea, two from Puerto Rico and one from Uganda, Stromata brunnea, breviter stipitata, capitata. Caputa were brought into culture. The original isolation of convex vel hemisphaerica, tuberculata, 1.5-3 mm diam. Per Samuels 92-29 was made from tissue of the stipe. As ithecia (190-) 197-219 (-241) µm alta, (127-) 132-162 (-182) cospores were subsequently isolated from stromata µm lata. Asci cylindrici, (43-) 46-55 (-59) × (2.7-) 3.3-4.3 (-4.9) that formed in culture. µm, ad apicem incrassati. Ascosporae bicellulares, ad sep tum disarticulatae; parte distali globosa vel subglobosa, (2.0 This species was originally described by Möller ) 2.1-2.7 (-2.9) × (1.8-) 1.9-2.6 (-2.9) µm parte proximi oblon (1901) from southern Brazil, and was redescribed by ga vel cuneiformi, (2.3-) 2.4-3.0 (-3.2) × (1.7-)1.8-2.2(-2.3) Chardón (1921) as Podostroma orbiculare from Puerto µm, hyalinae, minute verrucosae. Rico. The species has apparently neither been re Anamorphe Trichoderma sp. ported nor described under other names from out Holotypus in cortice, Guyana, leg. A. Rossman 3298 side of these areas (Rick, 1906; Theissen, 1911). (BPI). Hypocrea poronioidea is characteristic in that stro mata comprise a nearly circular, brownish cap that is Anamorph: Trichoderma sp. FIGS. 53-59 situated on a short stipe. This aspect is suggestive of stromata of the xylariaceous genus Poronia, hence Stromata capitate-stipitate to pulvinate and then the species epithet. Less frequently the stipe may be centrally attached with margins free; cap circular in reduced and thus the cap is sessile; however, in these outline, 1.5-3 mm diam, convex to nearly hemispher cases the stroma is attached at a central point and ical, margin slightly inrolled, in colors of brown (6E the margins are free. F8), KOH-, surface glabrous, slightly tuberculate Hypocrea poronioidea was included in Podocrea from perithecial elevations, ostiolar openings visible (Sacc.) Lindau, which is Podostroma (following Boe as darker brown dots against the lighter brown back dijn, 1938). While H. poronioidea, because of its flat ground; undersurface of cap slightly lighter in color, tened cap, does not fit in Podostroma in the sense of KOH-, velvety with velvet continuing on stipe. Stipe Boedijn, it is morphologically similar to P. brevipes. 0.5-1.0 mm long, 0.5-10 mm diam, central, concol Stromata of P. brevipes are larger in stature, the cap is orous with underside of cap, velvety. Cells of stromal tuberculate from the perithecial elevations, and the surface, in surface view, lacking a definite outline or ascospores are larger than those of H. poronioidea. On angular, walls to 3 µm thick. In section, stromal sur the basis of its stromal anatomy, H. poronioidea is re face clearly differentiated from underlying tissue, ferable to Hypocrea sect. Hypocrea subsect. Hypocrea crustose, 15-25 µm wide, cells angular and ca. 10 × (Doi, 1972). Within that subsection, however, the per 5-6 µm, walls 1.5 µm thick, yellow in lactic acid. Tis ithecial apex of H. poronioidea that is formed of dense sue immediately below the stromal surface of loosely ly interwoven, thick walled hyphae that are yellow in intertwined, 2-3 µm wide, branching and septate, lactic acid is distinctive, as is the internal tissue of the smooth-walled hyphae. Cells of the velvety region of stroma that is formed of thick walled hyaline hyphae. the underside of the cap and of the stipe circular to Single part-ascospores were isolated from asci of H. angular, ca. 7 µm diam, thin-walled, producing in poronioidea of the Puerto Rican and Ugandan collec conspicuous hyphal hairs < 25 µm long, septate, un tions. The collections from both countries were het branched. Perithecia crowded below the stromal sur erothallic. An analysis of mating-type segregation was face, oblong to elliptic in section, (n = 18) (190-) 197 undertaken with the Puerto Rican collections. Cul 219 (-241) µm high, (127-) 1.32-162 (-182) µm wide; tures derived from eight part-ascospores in each of cells at perithecial apex around ostiolar opening in several asci were self fertile, producing mature peri tertwined, short and conspicuously thick-walled, thecia in turbinate stromata at the periphery of the forming “shoulders” on the perithecium as seen in colony. Colonies derived from the other eight part longitudinal section, yellow in lactic acid; ostiolar ca ascospores produced conidia only. They did not form nal 43-60 µm long, periphysate. Tissue of stromal perithecia when mated among themselves or with the interior below perithecia densely disposed 3 µm wide 312 MYCOLOGlA SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 313

FIGS. 53-59. Hypocrea capitata, Trichoderma anamorph from CMD. Figs. 53, 54, 56, 57, 59 = PC; 55 = DIC; 58 = FL. All from Rossman 3298. Scale bars: 53, 54 = 50 µm, 55-59 = 10 µm.

FIGS. 41-52. Hypocrea capitata. 41, 42. Habit of stromata. 43. Median longitudinal section through a mature stroma. 44. Median longitudinal section through a mature perithecium. Detail of perithecial apex. 46. Stromal surface showing outlines of cells. 47. Detail of stromal surface. 48. Cells of stipe. 49. Internal tissue of stroma below perithecia. 50-52. Asci and ascospores. Figs. 41-43 = BF; 44-50, 52 = DIC; 51 = PC. All from Rossman 3298. Scale bars: 41 = 2 mm, 42 = 2 mm, 43 = 500 µm, 44-49 = 50 µm, 50-52 = 10 µm. 314 MYCOLOGIA branched and septate hyphae with walls 1 µm thick; cial elevations, whereas the stromal surface of H. pa swollen or nodose elements lacking. Asci cylindrical, ronioidea is plane. Ascospores and conidia of H. cap (n = 30) (43-) 46-55 (-59) × (2.7-) 3.3-4.3 (-4.9) µm, itata and H. poronioidea are smaller than those of H. completely filled with ascospores; apex thickened brevipes. and with an obscure ring; 8-spored, ascospores uni seriate or partly biseriate. Part-ascospores dimorphic, distal part globose to subglobose, (n = 30) (2.0-) 2.1 ACKNOWLEDGMENTS 2.7 (-2.9) × (1.8-) 1.9-2.6 (-2.9) µm; proximal part ob James Plaskowitz processed film and prepared the photo long to narrowly wedge-shaped, (22%) 2.4-3.0(-3.2) × graphic prints. Anita Phillips provided technical assistance. (1.7-) 1.8-2.2 (-2.3) µm; hyaline, ultimately becoming Katleen van der Gucht, Kathie Hodge and Amy Rossman finely spinulose. provided us with fresh collections of H. capitata and H. Ascospores germinating in low percentage within brevipes. Walter Gams corrected the Latin description. Ger 24 h on CMD at 20 C. Colonies on CMD > 9 cm ald Bills and Amy Rossman provided helpful critical com diam within 6 da, but growing slowly; aerial mycelium ment on the manuscript. We appreciate the loan of speci scant to lacking; a pale yellow pigment slowly devel mens from the following herbaria: B, FH, GENT, HBG, NY, oping after 6 da and spreading through the agar; PC, S. Conidia on CMD beginning to form in dense ‘balls’ or aggregates in the aerial myclium. Within 2 weeks LITERATURE CITED conidia 29D-E8 (deep green). Colonies on PDA > 9 cm diam within 6 da; aerial mycelium scant, cottony, Atkinson, G. F. 1905. Life history of Hypocrea alutacea. Bot. with a spreading yellow pigment (3B-C5-6: absinthe Gaz. 40: 401-417. yellow, mustard yellow, grayish yellow) in colony re Bissett, J. 1991a. A revision of the genus Trichoderma. II. verse; conidial production beginning within 6 da; co Infrageneric classification. Canad. J. Bot. 69: 2357 nidia slowly turning green, after 14 da conidia 30E8 2372. (parrot green) in the center to 30D6-8 (foliage . 1991b. A revision of the genus Trichoderma. III. Section Pachybasium. Canad J. Bot. 69: 2373-2427. green) at the margin. Colonies on ME > 9 cm within Boedijn, K. B. 1938. A new species of the genus Podostroma 6 da, much as on PDA but sterile; conidia developing from Africa. Sydowia 36: 314-317. slowly, eventually greenish yellow, after 14 da conidia Chardón, C. E. 1921. A contribution to our knowledge of 3C-D8 (olive yellow) to nearly white. Main axes of the pyrenomycetes of Porto Rico. Mycologia 13: 279 conidiophores formed on CMD 5-10 µm wide, fertile 300 + pls. 13, 14. branches 2.5-4.5 µm wide, to 15 µm long, tending Doi, N., and Y: Doi. 1979. Notes on Trichoderma and its to radiate in groups of 3-4 from a central point, each allies 1. A list of teleomorphic species with Trichoderma rebranching and bearing a penicillus of 3-4 phiali or its allied anamorphs hitherto known. Bull. Nat. Sci. des. Phialides lageniform to ampulliform, (n = 30) Mus. Ser. B (Bot.) 5: 117-123. (4.2) 5.1-6.9 (-7.9) µm long, (2.1-) 2.6-3.6 (-4.2) µm Doi, Y. 1967. A revision of the Hypocreales with cultural wide in the middle, (1.2-) 1.9-2.3 (-2.9) µm wide at the observations. III. Three species of the genus Podostro ma with Trichoderma or Trichoderma-like conidial states. base. Hypha or branch immediately subtending each Trans. Mycol. Soc. Japan 8: 54-57. phialide (n = 30) (2.6-) 2.9-3.9(-4.4) µm. Conidia ob . 1972. Revision of the Hypocreales with cultural ob long to ellipsoidal, (n = 35) (2.5-)2.6-3.2 (-3.7) × servations IV. The genus Hypocrea and its allies in Ja (1.5) 1.7-2.3 (-2.5) µm, lacking a visible basal abscis pan (2). Enumeration of the species. Bull. Nat. Sci. sion scar, smooth, green. Chlamydospores not seen. Mus. 15: 649-751. Habitat. Decorticated wood. . 1973. Revision of the Hypocreales with cultural ob Known distribution. French Guiana, known only servations. V. Podostroma giganteum Imai, P. cornu-da from the type. mae (Pat.) Boedijn and Hypocrea pseudogelatinosa sp. HOLOTYPE. FRENCH GUIANA. Route de Beli nov. Rep. Tottori Mycol. Inst. 10: 421-427. zon, 15 km from road N2, track to Montagne Tortue, . 1975. Revision of the Hypocreales with cultural ob 52°21'N, 04°25'W, on decorticated wood, 19 Feb. servations VIII. Hypocrea peltata (Jungh.) Berk. and its 1988, A. Y. Rossman 3298 (Samuels culture 88-11, allies. Bull. Nat. Sci. Mus. Ser. B (Bot.) I: 121-134. . 1979. Revision of the Hypocreales with cultural ob ATCC 96281) (BPI). servations XII. Additional note on Hypocrea peltata Notes. Hypocrea capitata, H. poronioidea and H. (Jungh.) Berk and its allied species. Bull. Nat. Sci. Mus. brevipes differ most conspicuously in their ana Ser. B (Bot.) 5: 37-49. morphs. The stroma of the only known collection of Domsch, K. H., W. Gams, and T.-H. Anderson 1980. Com H. capitata is considerably smaller than are the stro pendium of soil fungi. Vol. 1. Academic Press, London mata of the other two species. The stromal surface and New York. of H. brevipes is slightly tuberculate from the perithe Kornerup, A., and J. H. Wanscher. 1978. Methuen handbook SAMUELS AND LODGE: HYPOCREA WITH STIPITATE STROMATA 315

of colour. Third revised edition. Eyre Methuen Ltd., , and G. J. Samuels. 1992. New species of Hypocre London, UK. ales (Fungi, Ascomycetes). Brittonia 44: 256-263. Lloyd, C. G. 1923. Mycological Notes 68. Mycological Writ Saccardo, P. A. 1883. Sylloge Fungorum 2: 1-815 + i-lxix + ings 7: 1181. 1-77. Möller, A. 1901. Phycomyceten und Ascomyceten Untersu , and D. Saccardo. 1905. Sylloge Fungorum 17: i-cvii, chungen aus Brasilien. Botanische Mittheilungen aus 1-991. den Tropen. Heft 9. Jena. Samuels, G. J., and K. A. Seifert. 1987. Taxonomic impli cations of variation among hypocrealean anamorphs. Rick, J. 1906. Pilze aus RiO Grande do Sul (Brasilien). Bro Pp. 29-56. In: Pleomorphic Fungi: The diversity and its teria 5: 5-53 + 6 plates. taxonomic implications. Ed., J. Sugiyama. Kodansha, To Rifai, M. A. 1969. A revision of the genus Trichoderma. My kyo, Japan. col. Pap. 116: 1-56, Theissen, F. 1911. Die Hypocreaceen von Rio Grande do , and J. Webster. 1966. Culture studies on Hypocrea Sul, Sudbrasilien. Ann. Mycol. 9: 40-74 + 3 plates. and Trichoderma III. H. lactea (= H. citrina) and H. Tubaki, K. 1958. Studies on the Japanese Hyphomycetes. pulvinata . Trans. Brit. Mycol. Soc. 49: 297-310. V. Leaf & stem group with a discussion of the classifi Rogerson, C. T. 1970. The hypocrealean fungi (Ascomy cation of the Hyphomycetes and their perfect stages. J. cetes, Hypocreales) . Mycologia 62: 865-910. Hattori Bot. Lab. 20: 142-244.