Neobelonopsis Itagaki & Hosoya, 2023

Itagaki, Hiyori & Hosoya, Tsuyoshi, 2023, A new genus Neobelonopsis and two new species of Trichobelonium (Helotiales, Ascomycota) discovered mainly from poaceous grasses native to Asia in Japan, MycoKeys 99, pp. 45-85 : 45

Neobelonopsis Itagaki & Hosoya gen. nov.

Figs 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14

Etymology.

Refers to the morphological similarity with the genus, Belonopsis .

Diagnosis.

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.

Type species.

Neobelonopsis multiguttata Itagaki & Hosoya.

Description.

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.

Figures

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).

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).

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).

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).

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).

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).

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).

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).

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).

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).

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.

Figure 14. Asexual morph and hyphal structures of the Neobelonopsis and Trichobelonium species A conidiophores of N. acutata B monilioid hyphae of N. bicolor C conidiophores with conidia of N. cinnabarina D conidiophores with conidia of N. didymospora E conidiophores with conidia of N. microspora F conidiophores with conidia of N. multiguttata G conidiophores with conidia of N. obtusa H conidiophores with conidia of T. albobarbatum I hyphal mass with melanized appressorium-like structure of T. miscanthi. Scale bars: 10 μm.