Calycina montana (W.Y. Zhuang & H.D. Zheng) Lestari & Chethana, 2022

Lestari, Anis S. & Thilini Chethana, K. W., 2022, Morpho-phylogenetic insights reveal Bisporella montana as Calycina montana comb. nov. (Pezizellaceae, Helotiales), Phytotaxa 558 (2), pp. 185-202 : 190-193

Calycina montana (W.Y. Zhuang & H.D. Zheng) Lestari & Chethana , comb. nov. ( FIGURE 2–4 )

Basionym: Bisporella montana W.Y. Zhuang & H.D. Zheng, Zhuang, Zheng & Ren, Mycosystema 36(4): 407 (2017)

Index Fungorum number: IF 570309, Facesoffungi number: FoF 10812

Holotype: HMAS 275566

Saprobic on decayed woods covered with mosses. Sexual morph: Apothecia 1.1–1.6 × 0.6–1.4 mm, gregarious, arising singly or in clusters, amber colored, stipitate. Stipe 0.6–1.2 x 0.2–0.4 mm in dried condition. Receptacle discoid, bright amber color to hyaline. Margin hyaline with brown outer layer at the flank. Disc concave. Ectal excipulum 55–68 µm (= 59.8 µm, n = 10) at the flank near the stipe, composed of gelatinized thick-walled, hyaline cells of textura porrecta. Medullary excipulum 36–80 µm (= 57.7 µm, n = 10) at the flank near the stipe, distinctively separate from ectal excipulum, composed of thin-walled, hyaline cells of textura oblita, hypha with refractive guttules. Hymenium 141– 180 µm (= 168.9 µm, n = 15), amber colored. Paraphyses 1.8–2.5 µm in diam. (= 2.1 µm, n = 15) at the terminal, filiform, septate, branched at the 1/3 length from the base, guttules. Asci 101–118 × 4.7–6.3 µm (= 108.6 × 5.5 µm, n = 15), unitunicate, 8-spored, cylindrical and subclavate at the top, rounded (in alive state) to conical apex (dead state), J+ (eu-amyloid, in Melzer’s with and without KOH treatment), long substipitate base, arising from croziers. Ascospores 7.1–9.2 × 4.1–5 µm (= 8 × 4.4 µm, n = 20), ellipsoid, 0-1 septate, partially biseriate to uniseriate, hyaline, rounded at both ends with guttules. Asexual morph: Hypomycetous, on the lower flank and receptacle of the apothecia’s surface. Conidiogenous cells 12–19 × 3.7–5.1 (= 14.3 × 4.3 µm, n = 10) µm, phialidic, black to dark brown. Conidiophore 1.5–3.7 × 1.6–2.4 (= 3 × 1.9, n = 5) µm, black to dark brown. Conidia 3.2–3.8 × 1.7–2 (= 3.5 × 1.9 µm, n = 2) µm ellipsoid, aseptate, hyaline, smooth.

Material examined: Thailand, Chiang Mai Province , Chomthong District , Doi Inthanon , Kew Mae Pan nature trail, 20 October 2021, Anis S. Lestari, KMP3 21 ( MFLU 22-0055 ), KMP3 41 ( MFLU 22-0056 ) ( Figure 2 & 3 ); CHINA, Yunnan, Shangrila, Pingbian, Daweishan on rotten hard wood, 5 September 1999, X.Q. Zhang & D.Z. Ren, ( HMAS 275566 , holotype) ( Figure 4 ) .

GenBank numbers: MFLU 22-0055: ITS – ON156524 , LSU – ON156523 ; MFLU 22-0056: ITS – ON176689 , LSU – ON325438

Notes: In the current study, Bisporella montana ( HMAS 275566, holotype) and our specimens clustered under Calycina , as Calycina montana . Calycina montana ( HMAS 275566), previously known as Bisporella montana , was discovered in a mountainous area in China as a saprobe on woody substrates ( Zhuang et al. 2017). The characters of both specimens, especially the amber-colored apothecia and gelatinized textura porrecta cells of ectal excipulum, fit the generic concept of Calycina ( Baral 1993) . Another character which further confirms the combination is the appearance of a calycina-like apical ring shown clearly in our collection ( Figure 3j ) and the type specimen ( Figure 4m ). Based on phylogenetic analyses using ITS and LSU sequence data, our collection clusters with the type strain with high statistical support of 99% ML and 0.99 BI ( Figure 1 ). Across the 466 base pairs of ITS (+ 5.8S) gene of our Calycina montana strains ( MFLU 22-0055 and MFLU 22-0056) and the ex-type HMAS 275566, only one base pair difference (0.22 %) was observed.

Figures

FIGURE 1. The phylogram of combined ITS and LSU sequence data for genera in Pezizellaceae. Maximum likelihood bootstrap values greater than 70% and posterior probability values greater than 0.95 given near the nodes. The new geographical record highlighted in red and type strains are in bold. The tree is rooted to Chlorencoelia torta (JAC 14068, KUSF 52256 and ICMP 21732).

FIGURE 2. Calycina montana (MFLU 22-0055, new geographical record).a Rotten wood substrates covered with mosses. b–c Apothecia on the substrate. d Close-up of an apothecium sectioning. e Close-up of hymenium at the margin. f Phialide cells on lower flank surface. g–h Conidiogenous cells bearing conidiospore i–j A conidiogenous cell (j Mounted in Congo red). Scale bars: b = 950 μm, c = 470 μm, d = 315 μm, e = 141 μm, f = 40 μm, g = 17 μm, h–j = 13 μm.

FIGURE 3. Calycina montana (MFLU 22-0055, new geographical record). a Close up of excipulum cells. b Ectal excipulum cells (b mounted in Congo Red). c Medullary excipulum cells. d–e Filiform paraphyses with refractive guttules. f–g Asci (mounted in Congo red). h Asci (alive state). i Close up of crozier. j Tip of the ascus (J+, in Melzerr’s agent with KOH treatment). k–m Ascospores (m mounted in Congo red). Scale bars: a = 40 μm, b–c = 25 μm, d–h = 14 μm, i = 10 μm, j–m = 4 μm.

FIGURE 4. Calycina montana (HMAS 275566, holotype). a A herbarium package. b An apothecium on the woody substrate. c–d Cross sections of an apothecium (d mounted in Congo red). e Close up of hymenium at the margin. f Close up of ectal excipulum cells. g–h Filiform paraphyses (dead state). i–l Asci (k mounted in Melzer’s agent, l mounted in Congo red). m Tip of the ascus (J+, in Melzer’s agent with KOH treatment). n Close up of crozier at the base of an immature ascus. o–p Ascospores (o mounted in Congo red). Scale bars: b = 500 μm, c–d = 380 μm, e = 43 μm, f = 25 μm, g–h = 23 μm, i–l = 19 μm, m = 4 μm, n = 8 μm, m, o–p = 4 μm.