Refers to the morphological similarity with the genus, Belonopsis.

Differs from Belonopsis by superficial apothecia, which sometimes arise from dark-colored hyphal mass, observed as dark spots in superficial view, flattened in section (scutum, pl. scuta), wholly brownish receptacle, and the absence of crystals in the medullary excipulum. Differs from Trichobelonium in lacking crystals in the medullary excipulum and anchoring hyphae connecting the basal apothecia and subiculum. Differs from Mollisia by longer ascospores with (0–)1–3 septa, the color contrast between white hymenium and dark receptacle, and its preference for graminicolous habitats such as the culms of Sasa spp. and Miscanthus sinensis Andersson.

Figure 3. 

Neobelonopsis acutata (TNS-F-86648, holotype) A fresh apothecia on the decaying culm of Miscanthus sinensis B dried apothecia C stromata with sparse subiculum D one month old colony on PDA E vertical section of the apothecium (in LA) F refractive vacuoles in fresh marginal cells (in water) G asci with ascospores (in CB/LA) H paraphyses (in CB/LA) I paraphyses with a long refractive vacuole (in water) J blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) K croziers at the base of ascus (in CB/LA) L ascospores (in CB/LA) M conidia (in water) N, O conidiophores (in water) P bulbile (in CB/LA) Q subicular hyphae (in CB/LA). Scale bars: 1 mm (A, B); 0.2 mm (C); 25 μm (E, Q); 10 μm (F–P).

Neobelonopsis multiguttata Itagaki & Hosoya.

Apothecia scattered to gregarious, superficial, sometimes developed from scuta developed from poorly developed subiculum, globose to pulvinate when immature, discoid to saucer-shape when mature, flat to concave, sometimes seated on thinly subiculum, sessile, with brown to blackish receptacle; disc entire to sinuate, without hairs, waxy, often white to pale gray when fresh (rarely reddish orange), turning yellowish when dried. Ectal excipulum textura globulosa to angularis, not gelatinized, without crystals or exudates, composed of 2–3 cell layers of brown thick-walled cells, brown, becoming darker toward the cortical cells; medullary excipulum textura intricata to prismatica, composed of loosely interwoven hyphae, thin-walled hyphae 2–3 µm diam, hyaline. Asci cylindrical clavate, 8-spored, with a thick-walled conical apex. Ascospores ellipsoid to fusiform(-subcylindrical), with obtuse-subacute(-acute) extremes, straight to slightly curved(-sigmoid), thin-walled, 0–3(–4)-septate, with or without guttules, hyaline. Paraphyses cylindrical to slightly clavate, straight to curved, branched to simple, thin-walled, hyaline, apical cell containing long refractive vacuoles when mounted fresh in water. Conidiogenesis phialidic (resembles that of Phialocephala or Cadophora) when present.

Named after the acute apices of ascospores.

Characterized by 3-septate ascospore with acute extremes and conidiophores densely aggregated in clusters. The present species resembles N. multiguttata. See Diagnosis in N. multiguttata for diagnostic characters.

TNS-F-86648, Yugashima, Izu City, Shizuoka Pref. Japan, 15 October 2021, on decaying culm of Miscanthus sinensis; ex-holotype culture NBRC 115570.

Apothecia arising from scuta. Scuta superficial, scattered to gregarious, flat discoid, blackish brown (C80M100Y80–100K60), 0.1–0.3 mm diam., textura epidermoidea, composed of closely packed thick-walled cells. Apothecia 0.1–0.2 mm high, seated on subiculum, with grayish brown (C0–30M30Y40K60) to black receptacle; disc 0.25–1.4 mm diam., white to pale gray (K10) when fresh, shrunk to 0.2–1 mm diam., turns pale yellow (Y10) when dried. Ectal excipulum 25–40 µm thick at base, 15–20 µm thick at the upper flank to margin; cortical cells hemispherical to pyriform, 14–16 × 9–11 µm at base, becoming smaller to 10–12 × 7–9 µm toward the upper flank to margin, containing refractive vacuoles in the protruding cells when mounted fresh in water. Medullary excipulum 25–50 µm thick. Asci (50–)65–82(–85) × 5–9 µm, arising from croziers, with MLZ + apical pore. Ascospores 15–22(–27.5) × 2.5–3.5 µm, long fusiform, with acute apices, (1–)3(–4)-septate, containing abundant guttules, often 2–3(–4) large guttules and several smaller ones. Paraphyses (65–)70–85(–93) × 2.5–3 µm, simple, 2–3-septate, apical cells containing long refractive vacuoles when mounted fresh in water. Subiculum thinly covering the surface of substrates in patches, sparse to moderately abundant around the scuta and apothecia, shiny brown; subicular hyphae straight to curved, usually 3–5 µm diam., with 0.5–1 µm thick-walls, septate every 15–25 µm, perpendicularly branched, covered by gelatinous substance, forming bulbils of 30–45 µm across in the middle or tip, composed of densely aggregated globular or moniliform thick-walled cells, dark brown. Colony of NBRC 115570 on PDA moderately undulate, superficial, cottony to hairy, brownish gray (C20–40M40Y40K60) from the surface, zonation only observed from the reverse, without soluble pigment and crystals; aerial mycelium densely fascicular, white. Conidiophores aggregated in inconspicuous clusters on aerial hyphae, (semi-)macronematous, constricted, arising vertically or laterally from hyphae, pale to dark brown, smooth, thick-walled, frequently branched; phialides ampulliform with determinate collarettes, up to 15 µm long, approximately 4 µm width at base, discrete to integrated, terminal or intercalary, pale brown, thick-walled, with cylindrical to wide funnel-shape collarettes of 5–8 × 2.5–3 µm; conidia aseptate, spherical to subspherical, 2–2.5 µm diam., hyaline, thin-walled.

TNS-F-86671, Kawazu City, Kamo County, Shizuoka Pref., 16 October 2021, on decaying culm of M. sinensis, culture NBRC 115666.

Neobelonopsis acutata resembles Belonopsis graminea (P. Karst.) Sacc. & P. Syd., which has a whitish disc that turns yellowish when dried, asci, ascospores and paraphyses with overlapped biometry (Karsten 1871). However, N. acutata differs from B. graminea in its amyloid asci. Belonopsis graminea produces densely aggregated conidiophores (approximately 0.2 mm across, “spermogonium” sensu Karsten) and cylindrical to elongated fusiform conidia (8–10 × 1.5 µm, “spermatia” sensu Karsten) (Karsten 1871), while N. acutata has sparsely aggregated conidiophores and spherical conidia (Figs 3M, 14A).

Named after the two-color variability observed among the apothecia in a single population.

Characterized by apothecia that occur only on woody substrates, 2-celled ascospores, and monilioid hyphae surrounded by a gelatinous sheath that form on artificial media.

TNS-F-86605, Kagawa Town, Muroran City, Hokkaido, Japan, 3 August 2021, on decaying wood of Betula sp., ex-holotype culture NBRC 115569.

Apothecia superficial, without subiculum and scuta, 0.1–0.5 mm high, with blackish brown (C80M100Y80–100K60) to black receptacle; disc 0.8–1.5 mm diam., white to pale gray when fresh, shrunk to 0.5–1.2 mm diam., buff (M10Y30–40) or bluish gray (C30–40M20Y10–20K60) when dried. Ectal excipulum 40–50 µm thick at base, 25–40 µm thick at the upper flank to margin; cortical cells hemispherical to short clavate, 13–17 × 7.5–12 µm at base, becoming slender and smaller, moderately packed toward the margin. Medullary excipulum 10–25 µm thick, hyaline to pale brown. Asci (60–)67–80(–83) × 5–7.5 µm, arising from croziers, with MLZ + apical pore. Ascospores (10–)12–15(–17.5) × 2.5–3 µm, ellipsoid to fusiform with obtuse to subacute extremes, rarely constricted at the septum, (0–)1-septate, frequently containing two large guttules. Paraphyses (60–)62–77(–87.5) × 2.5–3(–4) µm, simple, rarely branched, 2–3-septate. Colony of NBRC 115569 on PDA convex, undulate, pulvinate, cottony to floccose, entirely pale gray (K10–40), darker from the reverse, without soluble pigment; crystals regular octahedron, 10–12.5 µm on a side, hyaline, forming on colony surface; aerial mycelium dense, white to pale gray.

TNS-F-86357, Mt. Yamizo, Daigo City, Kuji County, Ibaraki Pref., 24 May 2021, on decaying wood of Fraxinus sp., culture NBRC 115658; TNS-F-86606, Kagawa Town, Muroran City, Hokkaido, 3 August 2021, on decaying wood of Phellodendron amurense, culture NBRC 115663; TNS-F-86664, Yugashima, Izu City, Shizuoka Pref., 15 October 2021, on decaying wood of Zanthoxylum ailanthoides, culture NBRC 115665; TNS-F-86666, Mt. Amagi, Izu City, Shizuoka Pref., 15 October 2021, on decaying wood of Cornus controversa.

Figure 4. 

Neobelonopsis bicolor (TNS-F-86605, holotype) A fresh apothecia on decaying unidentified wood (TNS-F-86666) B dried apothecia with yellowish disc and blackish flask (TNS-F-86666) C dried apothecia with greyish discs D vertical section of the apothecium (TNS-F-86666, in LA) E hyaline medullary excipulum (TNS-F-86666, in LA) F brown medullary excipulum G ascospores (in CB/LA) H blue-stained apical pore of asci (in Melzer’s solution after 3% KOH pretreatment) I croziers at the base of asci (in CB/LA) J asci with ascospores (in CB/LA) K paraphyses (in CB/LA) L one month old colony on PDA M dark, gelatinous hyphal mass on CMA N monilioid hyphae surrounded by a gelatinous sheath (in diluted black ink) O octahedron crystals with monilioid hyphae on CMA (in water) P monilioid hyphae containing abundant guttles (in water). Scale bars: 1 mm (A, M); 0.5 mm (B, C); 50 μm (D–F); 25 μm (N, O); 10 μm (G–K, P).

Neobelonopsis bicolor shares biometry and morphology of ascospore with Belonopsis juncicola Graddon but differs in having larger asci (vs. 40 × 5 µm) and lignicolous habitat (vs. Juncus) (Graddon 1990).

Both TNS-F-86605 (holotype) and 86606, which were collected from the same location in Hokkaido on the same day in October, have bluish gray hymenium (Fig. 4C) and pigmented medullary excipulum (Fig. 4F). Other specimens collected from spring to summer (May to August) in Honshu (TNS-F-86357 and 86664) have whitish to yellowish hymenium (Fig. 4B) and hyaline medullary excipulum (Fig. 4E). In the phylogenetic tree (Figs 1, 2), specimens with the two-color variability of hymenium formed a well-supported identical clade. Further sampling and morphological comparisons are needed to clarify whether these morphological differences depend on geographic or seasonal variability.

Neobelonopsis bicolor produces dark gelatinous hyphal structures on the colony surface of CMA and 2% MEA (Fig. 4M). The hyphal structure is composed of monilioid cells hyaline to pale brown, 5–10 µm diam., containing abundant guttles and a thick-walls. The monilioid cells are arranged linearly or sympodially and branch vertically or laterally (Figs 4P, 14B). The monilioid cells are covered with a thick gelatinous sheath (Fig. 4N). No asexual stage observed in colonies on any medium.

Cinnabarina in Latin, referring to the remarkable color of disc.

Differs from all other Neobelonopsis species by reddish orange disc.

TNS-F-86682, Yuzawa Town, Minami-uonuma County, Niigata Pref., Japan, 31 October 2021, on decaying culms of Miscanthus sinensis, ex-holotype culture NBRC 115571.

Apothecia developed from scuta. Scuta superficial, scattered to gregarious, flat discoid, pale reddish brown (C30–60M80Y80–100K10) to dark brown (C40–60M80Y100K60), 125–375 µm diam., textura epidermoidea. Apothecia flat to cushion-shape, 0.2–0.5 mm high, with blackish brown (C100M100Y80–100K60) to greenish dark brown (C80M80Y80–100K60) receptacle, releasing magenta pigment (C40–20M100Y10–30K60) in 3% KOH; disc 0.6–2 mm diam., light orange (C0–30M80Y100K0) to reddish orange (C0–20M100Y100) when fresh, shrunk to 0.3–1.5 mm diam. Ectal excipulum 25–40 µm thick at base, 15–25 µm thick at the upper flask to margin; cortical cells clavate to pyriform, 14–18(–20) × 8.5–10 µm at base, becoming smaller toward the margin, 10–12 × 5–7 µm, containing yellow to orange cytoplasm which turns magenta in 3% KOH, containing guttules that disappeared in 3% KOH. Medullary excipulum 25–50 µm thick. Asci (56–)62–75(–83) × 6–7.5 µm, arising from croziers, with MLZ + apical pore. Ascospores 15–20(–22.5) × 3.5–4.5 µm, ellipsoid to subcylindrical, with rounded to subacute extremes, aseptate, hyaline or yellow when mounted fresh in water, containing 2(–4) large guttules. Paraphyses (50–)60–75(–80) × 2.5–3.5 µm, wider toward the apex up to 5 µm, simple, septum distance closer towards the base, containing long yellowish refractive vacuoles when mounted fresh in water, changed magenta in 3% KOH and showing color gradation (darker toward the tip). Subiculum thinly developed the surface of substrate, sparse to moderately abundant around the scuta and apothecia, shiny brown; subicular hyphae straight to curved, sometimes forming fascicules with 2–3 hyphae, 2.5–4 µm diam. with 0.5–1 µm thick-walls, branched at right angle, walls covered by a thick gelatinous substance. Colony of NBRC 115571 on PDA entire to slightly undulate, flat to slightly winkled, floccose to felted, brownish gray (C0–20M30–40Y40K30) from the surface, turning white at the edge, same color from the reverse, without soluble pigment; crystals ovoid to dumbbell-shape, 18–25 × 11–15 µm, hyaline, forming on surface or below agar; aerial mycelium sparse to dense, gray. Conidiophores semi-macronematous, solitary to caespitose (forming rather loose sporodochia), short, constricted, arising vertically or laterally from hyphae, pale to dark brown, smooth, thick-walled, branched; phialides round-bottom flask or bottle-shape, up to 20 µm long, 3–4 µm width at base, discrete to integrated, terminal or intercalary, pale brown, thick-walled, with cylindrical collarettes of 8–10 × 2 µm; conidia aseptate, cylindrical oblong to fusiform, abundantly aggregated in slimy heads, 4–5×1 µm, hyaline, thin-walled.

Figure 5. 

Neobelonopsis cinnabarina (TNS-F-86682, holotype) A fresh apothecia on the decaying culm of Miscanthus sinensis B immature apothecium protruding from the stroma C dried apothecia D one month old colony on PDA E texture of stroma (in LA) F vertical section of the apothecium (in LA) G yellowish reflective vacuoles in fresh paraphyses (in water) H yellowish vacuoles in outermost cells of fresh ectal excipulum (in water) I immature ascus with yellowish cytoplasm (in water) J ascospores with yellowish cytoplasm (in water) K apothecium that dissolved magenta pigments when immersed in 3% KOH L outermost cells that turned magenta (in 3% KOH) M fresh paraphyses that turned magenta, note that the tips have a darker color (in 3% KOH) N ascospores (in CB/LA) O asci with ascospores (in CB/LA) P paraphyses with wide, blunt head (in CB/LA) Q blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) R crozier at the base of ascus (in CB/LA); S collapse conidiophores with slimy conidial drops on CMA (arrows) T hyphae with oblong-shape crystals on CMA U conidia (in water) V clusters of conidiophores (in water) W, X discrete conidiophores (in water). Scale bars: 1 mm (A, S); 0.1 mm (B); 0.5 mm (C); 50 μm (F–H, M, T, V); 20 μm (E); 10 μm (I, J, L, N–R, U, W, X).

TNS-F-86690 and 86692, Yuzawa Town, Minami-uonuma County, Niigata Pref., 31 October 2021, on decaying culms of Miscanthus sinensis; TNS-F-86701, Daigenta Lake, Yuzawa Town, Minami-uonuma County, Niigata Pref., 31 October 2021, on decaying culms of M. sinensis, culture NBRC 115669; TNS-F-86704 and 86716 (culture NBRC 115670), Toukamachi City, Niigata Pref., 31 October 2021, on decaying culms of M. sinensis.

Neobelonopsis cinnabarina is easily distinguished from other species by the reddish orange disc, slightly clavate paraphyses, and strong magenta pigment release of apothecia in KOH. In particular, the brilliant color of disc of this fungus is a rare feature in mollisioid fungi, except for Mollisia purpurea Rhem and M. russea (Schmid-Heckel) Baral. These two species share several characters with N. cinnabarina, such as dark scuta [N. russea has “dunkelbraunen Schild” sensu Schmid-Heckel (1988)], bright orange vacuoles in paraphyses that become intensely magenta (red violet) in KOH, ocher to brown receptacle, asci arising from croziers, aseptate ascospores, and monocot host (Rhem 1907; Schmid-Heckel 1988; Baral and Marson 2005; Richter and Baral 2008). As the tips of fresh paraphyses turn dark magenta in 3% KOH (Fig. 5M), this phenomenon is suggested to be a vital reaction as the pigments diffuse uniformly in the paraphyses after heat drying. Richter and Baral (2008) also described the same reaction in M. russea. These features imply a close relationship among N. cinnabarina, M. purpurea, and M. russea. However, M. russea has no subiculum, ascospores are shorter (11–16 × 2.5–3.5 µm) than N. cinnabarina, and paraphyses are not clavate. Mollisia purpurea also differs from N. cinnabarina in having crystals in medullary excipulum and shorter ascospores (12–14 × 2.5–3 µm) than N. cinnabarina. Genetic comparison among these species could not be conducted as M. purpurea and M. russea lack available DNA sequences.

Neobelonopsis cinnabarina produces conidiophores only on CMA (Fig. 5S), and conidia mostly germinate (Fig. 5U). The asexual stage of N. cinnabarina is unique in loose sporodochia (Fig. 5V), longer collarettes, and oblong conidia (Figs 5U–X, 14C).

Named after two-celled ascospores.

Resembles Neobelonopsis bicolor, but distinguishable by sparse, minute guttles in living/dead ascospores, shorter asci, and wider host range including woody bamboos.

TNS-F-13501, Yakushima Island, Kagoshima Pref., Japan, 19 October 2005, on decaying culms of woody bamboos, ex-holotype culture NBRC 115354.

Apothecia superficial, without subiculum and scuta, 0.1–0.2 mm high, with blackish green (C100M100Y80–100K30) to black receptacle; disc 0.5–1 mm diam., white to bluish gray (C60M30–40Y20K60) when fresh, shrunk to 0.3–0.7 mm diam., cream (Y20K10) or olive (C40M40Y60–100K10) when dried. Ectal excipulum 30–50 µm thick at base, 20–25 µm thick at the upper flank to margin; cortical cells obovoid to clavate, (10–)12–15 × 7.5–10 µm at base, becoming slender and closely packed at the upper flank to margin, containing refractive vacuoles at the protruding cells when mounted fresh in water. Medullary excipulum 25–38 µm thick, frequently dichotomously branched, radially spreading toward the upper flask. Asci (50–)52–60(–65) × 5–7.5 µm, arising from croziers, with MLZ + apical pore. Ascospores 10–14(–16) × 2.5–3.5 µm, ellipsoid to fusiform, with subacute to acute extremes, frequently constrict at the septum, (0–)1–2-septate, hyaline, containing scattered small guttules. Paraphyses (45–)53–65 × 2.5–3(–4) µm, simple, (1–)2–3-septate, containing long refractive vacuoles in the apical cells and first 2–3 lower cells. Colony of NBRC 115354 on PDA flat, entire, dense, cottony to felted, dark brown (C60M80Y80–100K10) to beige (C10M20Y20–40K10) at the center, becoming pale brown toward to the edge, same colors at the reverse side, without soluble pigment and crystals; aerial mycelium sparse to dense, white to beige. Conidiophores solitary to occasionally aggregated on aerial hyphae, semi-macronematous, short, arising vertically or laterally from hyphae, pale to dark brown, smooth, thick-walled, sometimes branched 2–3 times, constricted at the septa, 2–3 µm width; phialides ampulliform, up to 15 µm long, 3.5 µm width at base, discrete or integrated, terminal or intercalary, hyaline to pale brown, thick-walled, with cylindrical to wide funnel-shape collarettes of 4.5–7.5 × 3 µm; conidia aseptate, subspherical to ellipsoid, abundantly aggregated in slimy head, 1.5–1.8 µm diam., hyaline, thin-walled.

Figure 6. 

Neobelonopsis didymospora (TNS-F-13501, holotype) A fresh apothecia on decaying Bamboo culm B dried apothecia on decaying Bamboo culm C fresh apothecia on decaying unidentified wood (TNS-F-86670) D dried apothecia on decaying unidentified wood (TNS-F-86670) E vertical section of the apothecium (in LA) F reflective vacuoles in fresh marginal cells (in water) G vertical section at the basal apothecium (in lactic acid) H ascospores (in CB/LA) I asci with ascospores (in CB/LA) J paraphyses (in CB/LA) K fresh paraphyses with long refractive vacuoles (in water) L blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) M crozier at the base of ascus (in CB/LA) N three months old colony on PDA O clusters of conidiophores (in CB/LA) P–S conidiophores with conidia (in CB/LA). Scale bars: 1 mm (A–D); 50 μm (E); 20 μm (O), 10 μm (F–M, P–S).

TNS-F-13509, Yakushima Island, Kagoshima Pref., 19 October 2005, on decaying wood of Elaeocarpus japonicus, culture NBRC 115651; TNS-F-86178, Shishizuka Pond, Tsuchiura City, Ibaraki Pref., 29 October 2018, on decaying wood of Albizia julibrissin, culture NBRC 115657; TNS-F-88720, Shirokanedai, Meguro Ward, Tokyo, 6 July 2018, on dead stem of Trachycarpus fortunei; TNS-F-86661 and TNS-F-86652, Yugashima, Izu City, Shizuoka Pref., 15 October 2021 on decaying culms of woody bamboos and unidentified wood, respectively; TNS-F-86670, Kawazu City, Kamo County, Shizuoka Pref., 16 October 2021, on unidentified decaying wood; TNS-F-86718, Mt. Katsuu, Nago City, Okinawa Pref., 27 October 2021, on decaying wood of Alnus sp.

Neobelonopsis didymospora forms apothecia in autumn (October–December) and has a wide host range, but limited to woody plants, including woody bamboo. Neobelonopsis didymospora forms its asexual stage only on CMA (Fig. 6O, S). This fungus is superficially similar to N. bicolor, but differs in fewer guttules in the cytoplasm.

Based on a BLAST search of the GenBank database, the closest hits to the ITS sequences of N. didymospora were three sequences of Mollisia sp. from New Zealand collected from the dead frond of Rhopalostylis sapida [MG195516; Identities=553/554 (99.8%), no gaps], fallen unidentified wood [MG195517; Identities=551/554 (99.5%), one gap], and fallen wood of Coriaria arborea [MG195518; Identities=511/511 (100%), no gaps]. The presence of these sequence data suggests that distribution of N. didymospora is not limited in Japan, but also in New Zealand.

Named after its small ascospores.

Characterized by narrow, aseptate ascospores.

TNS-F-18068, Yuzawa Town, Minami-uonuma County, Niigata Pref., Japan, 18 July 2006, on decaying culms of Sasa sp., ex-holotype culture NBRC 115567.

Apothecia developed from scuta. Scuta superficial, scattered to gregarious, flat discoid, dark brown (C60M80Y80–100K60) to black, 125–450 µm diam., textura epidermoidea. Apothecia 0.1–0.2 mm high, with grayish brown (C10–30M30–40Y60K60) receptacle; disc 0.3–1.5 mm diam., cream (Y10–30K10) when dried. Ectal excipulum 25–35 µm thick at base, 15–25 µm thick at the upper flask to margin; cortical cells hemispherical to obpyriform, 12–15(–17) × 7.5–11 µm at base, becoming smaller and hyaline at the upper flask to margin. Medullary excipulum, 25–50 µm thick. Asci (40–)45–55(–63) × 3.7–5 µm, arising from croziers, with MLZ + apical pore. Ascospores (7.5–)9.5–12.5(–16) × 2–2.5 µm, cylindrical to subcylindrical-fusoid-clavate with rounded extremes, aseptate, without guttules. Paraphyses (47–)52–62(–67.5) × 2–3.5(–4) µm, simple, rarely branched, (1–)2–3-septate. Subiculum thinly developed at the surface of substrates, sparse overall, shiny brown; subicular hyphae straight to undulate, frequently forming monilioid cells at the tip of the hyphae, 3–5 µm diam. with 0.5–1 µm thick-walls, perpendicularly branched. Colony of NBRC 115567 on PDA entire, flat to winkled at the center, floccose to felted, gray (K50–70) from the surface, darker from the reverse, without soluble pigment and crystals; aerial mycelium sparse to dense, white to pale gray. Conidiophores solitary to occasionally aggregated, semi-macronematous, short, arising vertically or laterally from fascicular hyphae, pale to dark brown, smooth, thick-walled, sometimes branched, constricted at the septa, 2.5–3 µm width; phialides ampulliform, up to 15 µm long, 3–4 µm width at base, discrete or integrated, terminal or intercalary, thick-walled, with cylindrical to long funnel-shape collarettes; collarettes of 6–8 × 2–3 µm, dark brown, occasionally covered with granules; conidia aseptate, cylindrical oblong to fusiform, abundantly aggregated in slimy heads, 4–5 × 1–1.5 µm, hyaline, thin-walled.

Figure 7. 

Neobelonopsis microspora (TNS-F-18068, holotype) A dried apothecia on the decaying culm of Sasa sp. B immature apothecia protruding from the stromata (arrows) C vertical section of the apothecium (in LA) D ascospores (in CB/LA) E asci with ascospores (in CB/LA) F paraphyses (in CB/LA) G blue-stained apical pore of asci (in Melzer’s solution after 3% KOH pretreatment) H, I croziers at the base of asci (in CB/LA) J conidiophores with pigmented collarets K conidiophores. Scale bars: 0.5 mm (A, B); 50 μm (C); 10 μm (D–K).

TNS-F-16804, Sugadaira Montane Research Center, Ueda City, Nagano Pref., 7 July 2007, on unidentified fallen branches, culture NBRC 115653; TNS-F-17105, Nozori Lake, Kuni Village, Agatsuma County, Gunma Pref., 15 May 2004, on decaying culms of Sasa sp., culture NBRC 115650; TNS-F-86453, Shiromine, Shiroyama City, Ishikawa Pref., 18 June 2021, on decaying culms of Sasa palmata, culture NBRC 115660; TNS-F-86584, Kawakami Town, Noboribetsu City, Hokkaido, 2 August 2021, on decaying culms of Sasa kurilensis, culture NBRC 115662.

The minimum length of the ascospores of N. microspora is the shortest in Neobelonopsis but its maximum length is overlapped with the other species. This fungus resembles B. eriophori Raitv. in macroscopic appearance of apothecia and in having short, aseptate ascospores (16–19 × 3–3.5 µm), but ascospores of B. eriophori become uniseptate at maturity while that of N. microspora remain aseptate (Raitviir 2003),

Neobelonopsis microspora produces conidiophores only on CMA, and the conidia germinate frequently (Figs 7J, K, 14E). The asexual morphology of N. microspora is very similar to that of N. cinnabarina, with long collarets and oblong conidia, except conidiophores do not form a sporodochium.

Named after the abundant number of guttules in the ascospores.

Resembles N. acutata, but distinguishable by more sparsely formed conidiophores, longer asci, and longer ascospores with rounded extremes (vs. more acute in N. acutata).

TNS-F-86402, Sugadaira Research Station, Mountain Science Center, Ueda City, Nagano Pref., 5 June 2021, on decaying culms of Sasa kurilensis, ex-holotype culture NBRC 115371.

Apothecia developed from scuta. Scuta superficial, scattered to gregarious, flat discoid, approximately 0.2 mm diam., blackish brown (C80M100Y80–100K60), textura epidermoidea, consisting of thick-walled cells. Apothecia 0.2–0.4 mm high, with dark brown (C80M80Y80–100K60) receptacle; disc 0.5–1.6 mm diam., white to pale gray (K10) when fresh, shrunk to 0.4–1.3 mm diam., pale yellow (Y10) when dried. Ectal excipulum 37–50 µm thick at base, 25–35 µm thick at the upper flank to margin; cortical cells hemispherical to short clavate, 12–17 × 9–10(–12) µm at base, becoming slender and closely packed toward the upper flank to margin. Medullary excipulum 37–87 µm thick. Asci (63–)78–98(–105) × 5–8 µm, arising from croziers, with MLZ + apical pore. Ascospores (12–)17–26(–27.5) × 2.5–3.5 µm, long ellipsoid to fusiform with rounded extremes, (1–)3-septate, containing abundant guttules. Paraphyses (62–)74–90(–100) × 2.5–3 µm, simple, with long apical cell. Subiculum sparsely developed, covering the surface of substrates in patches, shiny brown; subicular hyphae straight to curved, usually constricted at septum, fascicular, 3–5 µm width with 0.5–1 µm thick-walls, septate every 15–25(–50) µm, branched at right angle, covered by gelatinous substance. Colony of NBRC 115371 on PDA flat to slightly winkled, entire to undulate, floccose to woolly, grayish brown (C20–30M40Y40K60) from the surface, forming indistinct section observed clearer from the reverse, without soluble pigment and crystals; aerial mycelium moderately abundant at the center, sparse at the edge, pale gray (K10–30) to white. Conidiophores semi-macronematous, short, arising vertically from aerial hyphae, pale to dark brown, smooth, thick-walled, constricted, occasionally loosely branched; phialides cylindrical to ampulliform, up to 16 µm long, 3 µm width at base, discrete, arranged terminal or intercalary, pale brown, thick-walled, with cylindrical funnel-shape collarettes of 4.5–6.5 × 2–3 µm; conidia aseptate, spherical to subspherical, abundantly aggregated in slimy heads, 1.5–1.7 µm diam., hyaline, thin-walled.

TNS-F-18023, Shirakamisanchi, Aomori Pref., 24 May 2006, on decaying culms of Sasa sp.; TNS-F-39229, Mt. Tsukuba, Tsukuba City, Ibaraki Pref., 22 April 2011, on decaying culms of Sasa sp.; TNS-F-54941, Omama Town, Midori City, Gunma Pref., 9 May 2018, on decaying culms of Sasa sp.; TNS-F-61278, Mt. Tsukuba, Tsukuba City, Ibaraki Pref., 16 April 2014, on fallen cupules of Fagus crenata; TNS-F-61280, Hakone Town, Ashigara-shimo County, Kanagawa Pref., 23 May 2014, on fallen cupules of F. crenata Blume, culture NBRC 115655; TNS-F-81133, Sugadaira Research Station, Montane Science Center, Ueda City, Nagano Pref., June 2017, on decaying culms of Sasa kurilensis; TNS-F-86224, Sekimoto Town, Kita-ibaraki City, Ibaraki Pref., 3 June 2020, on dead branches on living Stephanandra incisa, culture NBRC 115365; TNS-F-86426, Mt. Amari, Asahi Town, Nirasaki City, Yamanashi Pref., 14 June 2021, on decaying culms of Sasa sp.; TNS-F-86465, Nagataki Town, Noumi City, Ishikawa Pref., 18 June 2021, on decaying culms of Sasa palmata, culture NBRC 115661.

Figure 8. 

Neobelonopsis multiguttata (TNS-F-86402, holotype) A fresh apothecia on decaying culm of Sasa kurilensis B dried apothecia C stromata with sparse subiculum D vertical section of apothecium (in LA) E ascospores (in CB/LA) F asci with ascospores (in CB/LA) G paraphyses (in CB/LA) H blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) I croziers at the base of asci (in CB/LA) J texture of stroma (in LA) K subicular hyphae (in lactic acid) L three months old colony on PDA M conidia (in CB/LA) N–R conidiophores, arrows in N indicated conidiogenous cells (in CB/LA). Scale bars: 0.5 mm (A–C); 50 μm (D, K); 20 μm (N), 10 μm (E–J, M, O–R).

Neobelonopsis multiguttata has a wide host range, such as Sasa spp., Fagus crenata, and Stephanandra incisa, and occurs on various substrates, such as culms, branches, and cupules. Neobelonopsis multiguttata was found in spring and its morphology overlaps with N. bicolor in the dimensions of apothecia and paraphyses. However, the ITS sequence similarity with N. acutata is only 93.8%. Further, the two species form phylogenetically distinct clades (Fig. 1). The conidiophores of N. multiguttata on CMA are discrete (Figs 8N–R, 14F), rather than aggregated as in N. acutata (Fig. 3N, O).

Based on a BLAST search for the ITS sequences of Neobelonopsis multiguttata in the GenBank database, the closest hit was Ascomycota sp. (MK842071), isolated from the needles and roots of pine trees in South Korea [Identities=531/531 (100%), no gaps]. The endophytic isolate was recognized as Mollisia sp. by Rim et al. (2021). This result suggests that N. multiguttata has an endophytic phase as part of its life cycle.

Named after rounded apices of ascospores.

Differs from N. acutata and N. multiguttata, which share 3-septate ascospores, by shorter ascospores with obtuse extremes and occurring only on woody substrates.

TNS-F-15602, Iryuda, Odawara City, Kanagawa Pref., 12 April 2007, on decaying wood of Aucuba japonica Thunb. var. japonica, ex-holotype culture NBRC 115381.

Apothecia superficial without subiculum and scuta, 0.2–0.3 mm high, with blackish brown (C80M80–100Y80–100K60) receptacle; disc 0.5–1.5 mm diam., white to pale gray when fresh, often turned grayish blue (C30–40M10Y10K30 or C40M20Y20K30) when moist, shrunk to 0.3–1 mm diam., pale yellow (Y20–30) or buff (M10Y30–40) when dried. Ectal excipulum 37–63 µm thick at base, 25–35 µm thick at the upper flank to margin; cortical cells hemispherical to short clavate, (10–)12–18 × (7–)8–12 µm at base, becoming slender and closely packed toward the upper flank to margin, containing refractive vacuoles at the protruding cells when mounted fresh in water. Medullary excipulum 60–75 µm thick, frequently dichotomously branched toward the margin. Asci (52–)56–78(–98) × 6–8.5(–10) µm, arising from croziers, with MLZ + apical pore. Ascospores (8–)13–17(–20) × 2.5–3.5 µm, subcylindrical with obtuse to subacute extremes, (1–)3-septate, containing small guttules. Paraphyses (40–)47–63(–70) × 2.5–3 µm, simple, (1–)2–3-septate, containing long refractive vacuoles at the apical cells when mounted fresh in water. Colony of NBRC 115381 on PDA entire, convex with abundant aerial hyphae, woolly to hairy, dark beige (M10Y20K30) from the surface, forming indistinct section and zonation observed clearer from the reverse, without soluble pigment and crystals; aerial mycelium abundant, membranous in the center, becoming densely fascicular, beige (C10–20M30Y30K10) to white. Conidiophores aggregated in inconspicuous clusters on aerial hyphae, (semi-)macronematous, constricted, arising vertically or laterally from hyphae, pale to dark brown, smooth, thick-walled, frequently branched; phialides ampulliform to lageniform with determinate collarettes, up to 15 µm long, approximately 3 µm width at base, discrete to integrated, terminal or intercalary, pale brown, thick-walled, with cylindrical to wide funnel-shape collarettes of 4–6.5 × 2–3 µm; conidia aseptate, spherical to subspherical, abundantly aggregated in slimy heads, 2–2.5 µm diam., hyaline, thin-walled.

Figure 9. 

Neobelonopsis obtusa (TNS-F-15602, holotype) A fresh apothecia on decaying unidentified wood B fresh apothecia during moist conditions C dried apothecia D vertical section of the apothecium (in LA) E refractive vacuoles in fresh marginal cells (in water) F vertical section at the basal apothecium (in LA) G ascospores (in CB/LA) H asci with ascospores (in CB/LA) I paraphyses (in CB/LA) J fresh paraphyses with long refractive vacuoles (in water) K blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) L croziers at the base of asci (in CB/LA) M three months old colony on PDA N conidiophores with conidia (in water). Scale bars: 1 mm (A, C); 0.5 mm (B); 50 μm (D); 10 μm (E–L, N).

TNS-F-44017, Yoyogi, Shibuya Ward, Tokyo, 8 November 2011, on unidentified decaying wood, culture NBRC 115654; TNS-F-54934, Omama Town, Midori City, Gunma Pref., 21 April 2018, on unidentified decaying wood, culture NBRC 115656; TNS-F-86359, Mt. Yamizo, Daigo City, Kuji County, Ibaraki Pref., 24 May 2021, on decaying wood of Lindera sp., culture NBRC 115659; TNS-F-86638, Ikaho, Shibukawa Town, Gunma Pref., 5 October, 2021, on decaying wood of Quercus sp.; TNS-F-86658, Yugashima, Izu City, Shizuoka Pref., 15 October 2021, on decaying wood of Cornus controversa, culture NBRC 115664; TNS-F-86668, Kawazu City, Kamo County, Shizuoka Pref., 15 October 2021, on decaying wood of Morus australis.

The ectal excipulum consisting of closely packed brownish cells of Neobelonopsis obtusa is similar to that of N. didymospora. However, the two species can be easily distinguished by the stable number of septa of ascospores (3-septate vs. 1-septate). Neobelonopsis obtusa forms an asexual stage on CMA (Figs 9N, 14G) which closely resembles that of N. acutata in dendroid (irregularly branched) conidiophores (Figs 3N–O, 14A).

Named after the frequently branched paraphyses.

Characterized by multi-septate, frequently 1–3 times branched paraphyses and long ascospore with 0–3 septum.

TNS-F-86030, Daimyoujin Fall, Ueda City, Nagano Pref., 6 August 2018, on decaying culms of Sasa sp., ex-holotype culture NBRC 115362.

Apothecia developed from scuta. Scuta superficial, scattered to gregarious, flat discoid, 140–185 mm diam., dark brown (C40–60M80Y80K60), consisting of closely packed brown cells and hyphae with thick-walls. Apothecia 0.1–0.2 mm high, with dark brown (C60M80Y80–100K60) receptacle; disc 0.1–1.5 mm diam., cream (Y10–30K10) when dried. Ectal excipulum 37.5–45 µm thick at base, 25–37 µm thick at the upper flank to margin; cortical cells pyriform to short clavate, paler toward to margin, 11–14(–16) × 7–10 µm at base, becoming slender and smaller toward margin. Medullary excipulum 25–63 µm thick. Asci (63–)74–88(–98) × 5–7.5 µm, arising from croziers, with MLZ + apical pore. Ascospores (12–)16–22(–25) × 2.5–3 μm, long subcylindrical to fusiform, with subacute extremes, 0–3-septate, sparsely containing guttules. Paraphyses (60–)65–77(–85) × 2–2.5 µm, frequently branching 1–3 times at the middle cells, multi-septate. Subiculum covering the surface of substrates in patches, sparse to moderately abundant around the scuta and apothecia, shiny brown; subicular hyphae straight or gently curved, sometimes forming fascicules with 2–3 hyphae, approximately 5 µm diam. with 0.5–1 µm thick-walls, septate every 20–50 µm, perpendicularly branched, covered by gelatinous substance. Colony of NBRC 115362 on PDA undulate, flat, floccose to cottony, sepia (C30–60M100Y60–80K60) from the surface and near the center, paler toward the margin, forming an indistinct section, darker from the reverse, without soluble pigment and crystals; aerial mycelium sparse to moderately abundant at the center, white to beige.

Figure 10. 

Neobelonopsis ramosa (TNS-F-86030, holotype) A dried apothecia on the decaying culm of Sasa sp. B stromata with sparse subiculum C vertical section of the apothecium (in LA) D asci (in CB/LA) E paraphyses (in CB/LA) F ascospores (in CB/LA) G blue-stained apical pore of asci (in Melzer’s solution after 3% KOH pretreatment) H croziers at the base of asci (in CB/LA) I subicular hyphae (in LA) J texture of stroma (in LA) K three months old colony on PDA. Scale bars: 0.5 mm (A); 0.25 mm (B); 50 μm (C, I); 20 μm (J); 10 μm (D–H).

Neobelonopsis ramosa is morphologically distinguished from other Neobelonopsis species by its frequently branching paraphyses (Figs 10E, 12). Neobelonopsis microspora also have branched paraphyses (mostly branched once), but differ from N. ramosa in ascospore with a stable number of septum (1- or 0-sepatate, respectively). Belonopsis pamparum Speg. resembles N. ramosa in habitat (apothecia on culm of poaceous grass, Aristida) and having frequently branched paraphyses, but differs in having larger ascospores (30–35 × 3–4 μm) with 5–7-pseudosepta (Spegazzini 1909). No asexual stage was observed in colonies of NBRC 115362 on artificial media.

Named after the anchoring hyphae between the cortical cells of receptacle and subiculum, which resembles a white beard (albo and barbata in Latin, respectively).

Resembles T. kneiffii, but distinguished by its larger ascospores.

TNS-F-86430, Sawara Pond, Asahi Town, Nirasaki City, Yamanashi Pref., 14 June 2021, on decaying poaceous grass culm lying on the wet ground close to the pond, ex-holotype culture NBRC 115568.

Apothecia developed scuta. Scuta superficial, scattered to gregarious, flat to protruded discoid, 125–375 µm diam., blackish brown (C80M100Y80–100K60), textura epidermoidea. Apothecia sessile, globose to pulvinate when immature, discoid to saucer-shape when mature, flat to concave when fresh, doliiform to pulvinate when dried, 0.1–0.3 mm high, with brown (C40–80M80Y100K30) receptacle; disc 0.5–1.5 mm diam., entire to undulate, without hairs at margin, waxy, yellow (Y30–60) when fresh, shrunk to 0.2–1 mm diam., pulverulent, yellowish orange (M10–40Y80–100) when dried, turned to brown (C30–60M80Y80–100) with senescence. Ectal excipulum 30–40 µm thick at base, 20–30 µm thick at the upper flank to margin, textura globulosa and angularis, composed of 2–4 layers of brown thick-walled cells; cortical cells hemispherical, 10–15(–17) × 6–10(–12) µm, ending up in cylindrical clavate cells, thick-walled, paler toward the margin; anchoring hyphae connecting the cortical cells of the flank and subiculum, radially extending from apothecium, 2.5–3 µm width, septate every 20–35 µm, thin-walled, hyaline, becoming conspicuous when apothecia dried. Medullary excipulum 100–150 µm thick, textura intricata to prismatica, hyaline, containing crystals below giving a rough texture, composed of loosely interwoven thin-walled hyphae which is frequently dichotomously branching. Asci (75–)85–100(–107) × 12–16(–20) µm, cylindrical-clavate to saccate, 8-spored, arising from croziers, containing yellowish oil globules in cytoplasm that disappear when mature, with a thick-walled conical apex; apex MLZ+ with or without 3% KOH pretreatment. Ascospores (25–)30–35(–38) × 4.5–6 µm, fusiform-clavate, with rounded or subacute extremes, straight to sigmoid curved, thin-walled, (0–)3-septate, sometimes constricted at the septum, hyaline, with numerous guttules. Paraphyses 85–100(–115) × 2.5–4.5 µm, occasionally branching at base, cylindrical, often becoming slightly wider toward the apex, 2–3-septate, thin-walled, hyaline, containing fragmented refractive vacuoles when mounted fresh in water. Subiculum covering the surface of substrates in patches, sparse to especially abundant around the apothecia and scuta, shiny dark brown, consisting of 1–3 layers of closely packed subicular hyphae; subicular hyphae 2–5 µm diam., thick-walled, brown. Colony of NBRC 115568 on PDA entire to partially filamentous at the margin, flat to slightly convex with aerial hyphae, cottony to woolly, agate (C10–30M60Y60) to amber (C10–40M100Y60K60) from the surface, appearing maroon (C10–40M100Y60K60) from reverse, with apricot (M20–40Y60) soluble pigment uniformly diffuse in agar; crystals aggregating plate-like or small clusters, acicular, moderately abundant on colony surface and surrounding agar, 0.1–0.3 mm across, pale yellow (Y10–20); aerial mycelium especially abundant in the center and edge, blush pink (M20–30Y20); mycelium containing guttules, pale to dark brown, thick-walled, sometimes covered with exudates. Conidiophores (semi-)macronematous, arising vertically or laterally from hyphae, pale to dark brown, smooth, containing oil globules in the hyphal cell, constricted at the septum, thick-walled, occasionally 2–3 series of branches, 2–3 µm width; phialides cylindrical to ampulliform, up to 10 µm long, 2.5–4 µm width, discrete to integrated, terminal, pale brown, thick-walled, with short cylindrical or wide funnel-shape collarettes of 2.5–4 × 3 µm at the upper edge, hyaline to pale brown, thin-walled; conidia aseptate, ellipsoid, abundantly aggregated near the collarettes, 2–3 × 1 µm, hyaline, thin-walled.

Figure 11. 

Trichobelonium albobarbatum (TNS-F-86430, holotype) A fresh apothecia on the decaying culm of unidentified grass (Poaceae) B dried apothecium with hyaline connective hyphae C protruding immature apothecia from the stromata D vertical section of the apothecium (in LA) E crystals in medullary excipulum (in LA) F connecting hyphae between ectal excipulum and subiculum (in LA) G texture of the stroma (in LA) H ascospores (in CB/LA) I ascus with mature ascospores (in CB/LA) J ascus with immature ascospores (in CB/LA) K immature ascus containing yellowish oil globes in the cytoplasm (in CB/LA) L paraphysis (in CB/LA) M rehydrated paraphysis with fragmented refractive vacuoles (in water) N blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) O croziers at the base of asci (in CB/LA) P three months old colony on PDA Q yellow crystals formed on hyphae R acicular crystals (in water) S–V conidiophores with conidia (in CB/LA); W hyphae with exudates (in CB/LA). Scale bars: 0.5 mm (A, Q); 0.25 mm (B, C); 50 μm (D–F); 20 μm (R); 10 μm (G, H–O, S–W).

The yellowish color of the hymenium is due to the oil globules in immature asci (Fig. 11D, K). The oil globules gradually disappear as the ascospores mature (Fig.) . Trichobelonium albobarbatum forms conidiophores immersed in agar, especially at the bottom of the Petri dish (Figs 11S–V, 14H). Both T. albobarbatum and T. kneiffii have well-developed dark brown subiculum, white anchoring hyphae, yellow hymenium, abundant crystals in excipulum, long asci (approximately 80–100 µm length), and 1–3-septate ascospores (Schröter 1908). However, T. albobarbatum has wider asci (vs. 5–6 µm width) and larger ascospores (vs. 16–18 × 2–2.5 µm) than T. kneiffii.

Named after the genus of its host, Miscanthus.

Characterized by 5-septate ascospores and sparse subiculum

TNS-F-17835, Sugadaira Montane Research Center, Ueda City, Nagano Pref., 17 September 2005, on decaying culm of Miscanthus sinensis, ex-holotype culture NBRC 115566.

Apothecia developed from scuta. Scuta superficial, scattered to gregarious, flat discoid, 145–180 µm diam., dark brown (C80M100Y80K60), textura epidermoidea, gradually becoming textura porrecta and connecting to subiculum. Apothecia sessile, globose to pulvinate when immature, discoid to saucer-shape when mature, flat to slightly convex when fresh, 0.2 mm high, with brown (C30–60M60Y80–100K60) receptacle; disc 0.5–1.5 mm diam., entire to slightly undulate, without hairs at margin, waxy, white to pale yellow (Y10–30) when fresh, shrunk to 0.3–1 mm diam., pulverulent, cream (Y20–40K10) when dried. Ectal excipulum 25–35 µm thick at base, approximately 25 µm thick at the upper flank to margin, textura globulosa and angularis, composed of 2–4 layers of brown thick-walled cells, not gelatinized, without crystals or exudates; cortical cells in middle to lower flank pyriform to clavate, with protruded cells, 16–21(–23) × 5–7 µm, containing with refractive vacuoles at margin when mounted fresh in water; anchoring hyphae connecting the cortical calls of the flank and subiculum, radially extending from apothecium, 2.5–3 µm width, thin-walled, brown. Medullary excipulum 40–75 µm thick, textura intricata to prismatica, hyaline, composed of loosely interwoven thin-walled hyphae which is frequently dichotomously branching. Asci (77–)79–85(–90) × 12.5–15(–17.5) µm, cylindrical-clavate to saccate, 8-spored, arising from croziers, containing hyaline oil globules in cytoplasm that disappear when mature, with a thick-walled conical apex; apex MLZ+ with or without 3% KOH pretreatment. Ascospores (32–)37–47(–57.5) × 4.5–5.5 µm, long fusiform, with acute extremes, curved to sigmoid, occasionally constricted, thin-walled, often 5-septate, hyaline, containing large or abundant minute guttles. Paraphyses (70–)83–105(–115) × 2.5–3(–4) µm, simple, occasionally branching at base, cylindrical, often becoming slightly wider toward the apex, 2–3-septate, thin-walled, hyaline, (2–)3-septate, containing long refractive vacuoles when mounted fresh in water. Subiculum thinly developed the surface of substrates in patches, sparse to especially abundant around the mature apothecia, shiny brown; subicular hyphae straight to curved, usually swelling in a globose, 3–5 µm diam. with 0.5–1 µm thick-walls, septate every 15–30 µm, perpendicularly branched. Colony of NBRC 115566 on PDA, flat to slightly convex with aerial hyphae, cottony, grayish orange (C0–20M60Y60K10) from the surface, appearing cinnamon (C20–40M80Y100K30) from reverse side, with apricot (M20–40Y60) soluble pigment uniformly diffuse in agar; aerial mycelium dense, white to pale yellow (Y10–20); crystals aggregating plate-like or small clusters, acicular, moderately abundant on colony surface and surrounding agar, 0.1–0.5 mm across, pale yellow. Conidiophores macronematous to mononematous, arising from subicular hyphae, straight, pale to dark brown, thick-walled, smooth, 2–3 µm width; phialides ampulliform, up to 15 µm long, 2.5–4 µm width, integrated, arranged penicillately, pale brown, thick-walled, with cylindrical to wide funnel-shape collarettes of 3.5–5 × 2–3 µm; conidia aseptate, spherical to subspherical, 2–2.5 µm diam., hyaline, thin-walled.

Figure 12. 

Trichobelonium miscanthi (TNS-F-17835, holotype) A fresh apothecia on the decaying culm of Miscanthus sinensis B dried apothecia with stromata (arrows) C brown connective hyphae extending from the base of fresh apothecium D texture of stroma (in LA) E conidiophore (in LA) F vertical section of the apothecium (in LA) G fresh outermost cells of ectal excipulum with refractive vacuoles (in water) H ascospores (in CB/LA) I fresh ascospores with droplets (in water) J blue-stained apical pore of ascus (in Melzer’s solution after 3% KOH pretreatment) K croziers at the base of ascus (in CB/LA) L long refractive vacuoles in fresh paraphysis (in water) M branching paraphysis (in CB/LA) N immature ascus containing oil globes in the cytoplasm (in CB/LA) O ascus with mature ascospores (in CB/LA) P immature ascus (in water) Q ascus with ascospores (in water) R one month old colony on PDA S pale yellow crystals formed on the edge of the colony T acicular crystals (in water) U hyphal mass attached to the bottom of the Petri dish V–X swollen cells with melanized appressorium-like structure. Scale bars: 1 mm (S); 0.5 mm (A, B); 0.25 mm (C); 50 μm (F, U); 20 μm (G, T); 10 μm (D, E, H–Q, V–X).

TNS-F-30037, Hachimantai City, Iwate Pref., 12 October 2009 on decaying culm of Miscanthus sinensis, culture NBRC 115652; TNS-F-81751, Kiritappu Wetland, Hamanaka City, Akkeshi County, Hokkaido, 29 August 2019, on decaying culm of Phragmites australis; TNS-F-86581, Higashi Ward, Sapporo City, Hokkaido, 13 August 2021, on decaying culm of Ph. australis; TNS-F-86672 (culture NBRC 115667) and 86695, Yuzawa Town, Minami-uonuma County, Niigata Pref., 17 and 31 October 2021 (respectively), on decaying culm of M. sinensis; TNS-F-86700, Daigenta Lake, Yuzawa Town, Minami-uonuma County, Niigata Pref., 31 October 2021, on decaying culm of M. sinensis, culture NBRC 115668; TNS-F-86715, Toukamachi City, Niigata Pref., 31 October 2021, on decaying culm of M. sinensis.

Trichobelonium miscanthi occurs with Neobelonopsis cinnabarina as they share the same host, Mollisia sinensis, and fruiting season (autumn). Brown phialides (Fig. 12E) and spherical conidia, regarded as asexual stage of T. miscanthi, were observed to accompany subiculum, but we could not induce conidial reproduction under culture.

From the reverse of the two months old colony of T. miscanthi on CMA, clumps of dark cells strongly attached to the bottom of the Petri dish (Fig. 12U) were observed. The clumps are composed of swollen cells with melanized ring. The swollen cell is usually obovoid to pyriform, sometimes lobed or hyphoid, 10–15 × 6–10 µm, arising vertically from hyphae, thick-walled, and containing abundant guttles (Figs 12V–X, 14I). The brown ring structure has an outer diameter of 8–10 µm and inner diameter of 2–3 µm, and is formed at the cell and Petri dish interface. Very similar hyphal structures were reported by Aebi (1972) in the culture of T. kneiffii, but its function is unknown. The clumps of dark cells of Phialocephala bamuru P.T.W. Wong & C. Dong, known as plant pathogen, are interpreted as appressoria with infected pegs (Wong et al. 2015). Although this structure may be appressorium, direct observation of the mycelium of T. miscanthi on the host epidermis and inoculation experiments are needed to clarify whether the clamps of T. miscanthi function as an appressorium during the infection process.

Figure 13. 

Line-drawings of the Neobelonopsis and Trichobelonium species I vertical section of the apothecium showing the marginal structure of ectal excipulum, medullary excipulum, and hymenium II asci III paraphyses IV ascospores. Scale bars: 10 μm.

Trichobelonium miscanthi resembles T. albobarbatum in remarkable oil globules in immature asci, anchoring hyphae, and saccate form of asci. Although T. miscanthi lacks crystals in the excipulum, the culture produced abundant acicular crystals on PDA (Fig. 12R, S, T).

Mollisia diesbachiana Tanney & Seifert, Studies in Mycology 95: 331, 2020.