JAKLITSCH ET AL. 2. <strong>Hypocrea</strong> <strong>viride</strong>scens Jaklitsch & Samuels, sp. nov. MycoBank MB494775. Figs 4–6, 7 g–l, 8 f–h. Etymology: <strong>viride</strong>scens, turning green, in reference to the anamorph. <strong>Hypocrea</strong>e <strong>rufa</strong>e similis sed anamorphosis differt. Ascosporae: pars distalis (3.3–)4.1–5.2(–7.4) × (3.2–)3.8–4.6(–5.4) μm; pars proxima (3.4–)4.4–5.8(–7.9) × (2.7–)3.3–4.0(–5.3) μm. Anamorphosis <strong>Trichoderma</strong> <strong>viride</strong>scens (A.S. Horne & H.S. Williamson) Jaklitsch & Samuels. Conidia subglobosa vel ellipsoidea, levia, (2.8–)3.5–4.5(– 8.5) × (2.3–)3.0–3.7(–4.7) μm, L/W (0.9–)1.1–1.4(–2.0). Anamorph: <strong>Trichoderma</strong> <strong>viride</strong>scens (A.S. Horne & H.S. Williamson) Jaklitsch & Samuels, comb. nov. MycoBank MB501049. ≡ Eidamia <strong>viride</strong>scens A.S. Horne & H.S. Williamson, Ann. Bot. 37: 396. 1923. Stromata when fresh (Fig. 4a–g) ca. 0.5–3.5 mm diam, ca. 0.5–1.5 mm thick, single, loosely gregarious or densely aggregated in lines, young downy, covered with yellowish to rust-coloured hairs, first with white inconspicuous margin; later glabrous, pulvinate, centrally attached, margins free, outline circular, angular to irregular, ostioles invisible to inconspicuous or light to dark dots, surface smooth to finely granular by perithecial contours, young light brown 5B5–6 to 6CD5–6, yellow-brown to nearly orange-brown 6CD7- 8, intensely orange-rust-reddish brown 8(B)C7–8; mature mostly dark reddish brown, 7DE7–8, 8EF6–8 (to 9F7–8, 10CD7–8, 10F7–8), margin usually dark; rarely rosy-brownish with dark spots to reddish brown 8CD5-6, more rarely 6A6–7 to 7A5–6. Stromata when dry (Fig. 4h, i) (0.2–)0.6–1.6(–3.6) mm (n = 277) diam, 0.2–0.5(–0.8) mm (n = 33) thick, unchanged or slightly darkened in 3 % KOH; young thin, semi-effuse to effuse, hairy, white to yellowish brown to rust-brown, 4A4, 5BD4–7 to 6–7CD5–8, with white to rust margin; later effluent, discrete <strong>and</strong> pulvinate with circular, angular to irregular outline; surface uneven, tubercular to wrinkled; ostioles invisible or appearing as light dots with darker marginal rings, not or slightly projecting, convex; colour in the great majority of stromata deep <strong>and</strong> dark reddish brown 7–9EF5–8. Stroma anatomy: Cells of stroma surface in face view (Fig. 4 l) glassy, isodiametric to slightly elongated, (2–) 3.5–8.5(–19) × (1.5–)2.5–6(–10) µm (n = 313, 210), walls thickened <strong>and</strong> pigment not uniformly deposited. Hairs arising from cells of the stroma surface, usually abundant when young, scant on mature stromata, 1–4celled, thick-walled, (5–)10–24(–47) μm (n = 240) long, (2–)3.2–5.0(–6.5) µm (n = 83) wide, base to 7 µm wide, apically rounded, pale brownish, smooth to slightly verruculose. Ostiolar area of dry stromata (24–)34–64 (–79) µm (n = 33) diam. Cortical region of stroma (Fig. 4k) (15–)18–36(–60) µm (n = 63) thick, around the entire stroma except the point of attachment, reddish brown to yellow-brown, cells in section of textura angularis, (2.0–)3.5–7.5(–17) × (1.7–)2.8–5.0(–8.5) μm (n = 279, 150), with walls thickened in the outer part, thinner towards the interior. Cells immediately below the cortex comprising a mixture of intertwined, (3–)4–6(–6.5) µm wide hyphae (n = 15), vertical <strong>and</strong> parallel between perithecia, <strong>and</strong> hyaline, subglobose to angular, mainly 156 isodiametric cells, (3–)5–9.5(–13) µm (n = 30) diam, flanking the ostioles. Perithecia flask-shaped, often somewhat narrowed towards the base, sometimes ellipsoidal to globose, (136–)220–287(–337) μm high, (72–)150–224(–280) μm diam (n = 149); peridium at the base (17–)19–26(–30) µm, laterally (12–)13–20 (–22) µm (n = 15) thick, hyaline. Ostiolar canal (53–) 70–98(–130) μm (n = 138) long, (30–)33–49(–57) µm (n = 15) wide at the opening, the opening formed by a palisade of hyaline, apically elongate narrowly clavate cells, even with the stroma surface or projecting up to 15 µm (n = 15). Subperithecial tissue homogeneous, dense textura angularis to t. epidermoidea, cells (sub-)globose to elongate to subangular or curved, (3.5–) 4.5–15(–39) µm (n = 337) long, (2–)4.5–10 (–17) µm (n = 240) wide, hyaline to pale brownish, with ca. 0.5 µm thick walls, tissue not layered but cells gradually smaller <strong>and</strong> interspersed with few narrow, ca. 3–4 µm wide hyphae towards the base of the stroma; in the central part of the stroma (point of attachment) stratified by a palisade (Fig. 4n) of oblong refractive glassy cells, ca. 14–31 × 4–9 µm, above the smallercelled pseudoparenchyma, sometimes irregular brownish pigment in patches incorporated through the whole tissue; basally delimited by the reddish brown cortex. Asci (Fig. 4o) cylindrical, (56–)82–101(–118) × (3–)5–7(–9) μm (n = 314), including a stipe of (4–)6– 22(–33) μm (n = 31); apical pore minute. Ascospores hyaline, verruculose to verrucose with verrucae ca. 0.5 µm long <strong>and</strong> diam, dimorphic; distal part subglobose, oval to wedge-shaped, (3.3–)4.1–5.2(–7.4) × (3.2–) 3.8–4.6(–5.4) μm (n = 411); proximal part oblong to wedge-shaped, lower end broadly rounded, (3.4–)4.4 –5.8(–7.9) × (2.7–)3.3–4.0(–5.3) μm (n = 411). Colony characteristics: Optimal temperature for growth on PDA <strong>and</strong> SNA 25 °C; after 72 h in darkness colony radius on PDA 32–45 mm, on SNA 24–37 mm; typically slower at 30 °C than at 20 °C; not growing at 35 °C. Colonies grown on PDA for 1 wk at 25 °C with alternating light <strong>and</strong> darkness (Fig. 7g–j) developing conidia in 3 or 4 distinct concentric rings, but often remaining sterile or producing very few conidia. Conidial masses green (28D–E5–6). Diffusing pigment not noted or reverse only slightly yellowish (3A3–4 to 3B4). Coconut odour typically present. Autolytic activity more pronounced at 15 °C, coilings more frequent at 30 °C. Colonies grown on CMD (Fig. 7k; 25 °C) filling the Petri plate within 5–6 d, thin, hyaline, regularly circular, hyphae loosely arranged radially, no zonation. Autolytic activity inconspicuous, coilings abundant in some isolates. Aerial hyphae often scarce during fast growth, becoming abundant, particularly towards the margin, broad zone at the margin becoming downy. A diffuse greenish yellow pigment (1B2–6, 2A3–3B4 to 29A2–3) often diffusing through the whole culture. A strong coconut-like odour usually formed. At 15 <strong>and</strong> 30 °C slower development with less conidiation <strong>and</strong> less strong coconut-like odour formed, coilings more frequent at 30 °C. Conidia forming within 2–10 d at 25 °C under alternating light, typically effuse, conidial pustules uniformly dispersed throughout the colony, developing in a broad marginal
a d e f g h i b k l m n c HYPOCREA RUFA / TRICHODERMA VIRIDE Fig. 4. <strong>Hypocrea</strong>/<strong>Trichoderma</strong> <strong>viride</strong>scens teleomorph. a–g. Fresh stromata (a, d–e: immature, b–c, f–g: mature). h–i. Dry mature stromata. j. Surface of stroma reconstituted with water, showing ostioles <strong>and</strong> unevenly distributed pigment. k. Surface of stroma <strong>and</strong> ostiolar opening in section. l. Surface of stroma in face view, including unicellular hair. m. Subperithecial tissue in section. n. Palisade of cells above point of attachment of stroma in section. o. Asci with ascospores in cotton blue/lactic acid. Sources: a, l, o: WU 24025. b–c: WU 24027. d, f: holotype WU 24029. e: WU 24024. g, j, m–n: WU 24019. h: WU 24018. i: WU 24028. k: G.J.S. 98-182. Scale bars: a = 2.7 mm, b–c, e = 2 mm, d, g = 1 mm, f = 2.5 mm, h = 0.4 mm, i = 0.5 mm, j = 0.2 mm, k = 30 µm, l = 10 µm, m, o = 15 µm, n = 35 µm. j o 157
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