Alanomyces indica Rohit Sharma, 2017

Sharma, Rohit, Kulkarni, Girish & Sonawane, Mahesh S., 2017, Alanomyces, a new genus of Aplosporellaceae based on four loci phylogeny, Phytotaxa 297 (2), pp. 168-178 : 172-174

Alanomyces indica Rohit Sharma , sp. nov. ( Figs. 1 , 2 )

MycoBank no.: MB 804155.

Etymology:—The epithet indica , is derived from the country of sample collection India.

Diagnosis:—Colony fast growing, initially greenish-black, later fuscous black- violaceous black. Pycnidia fuscous black, immersed to semi immersed, setose at base, multilocular. Peridium textura angularis dark brown outer layers of pycnidia. Conidiogenous cells hyaline. Chlamydospores dark brown.

Type:— INDIA. Madhya Pradesh, Anuppur (Amarkantak), 22.67 °N, 81.75 °E, from soil attached to fruiting body of an unidentified macrofungus, 23 December 2011, Ruby Sharma (holotype AMH 9513, culture ex-type MCC 1039, CBS 134264 = MMI00067) [GeneBank accession numbers—LSU: HF563623, ITS: HF563622, tef: AB872219, β tub: HF563625.

Saprobic on soil attached to the base of macrofungus stipe in mixed forest. Asexual morph: Pycnidia formed readily on Petri dish in 2–4 weeks, solitary or in groups, initial colony greenish-black VI’’’’’k (124), later turning to fuscous black VI’’’’’k (104)- violaceous black V’’’’k (114), 1600–2000 × 200–350 μm, immersed to semi-immersed, erumpent, multi locular, locules divided by pale brown III’’m (63) wall of 6–12 cell-layers thick (40–60 μm). Pycnidia in longitudinal section of dark brown IV‘’’m (84) textura globuloso-angularis, locule wall made of several layers of thick-walled, dark-brown IV‘’’m (84) textura angularis, becoming hyaline toward conidiogenous region. Exposed parts setose; setae hyaline to pale brown, smooth or verrucose, 100–200 × 10–12 μm. Conidiophores reduced to conidiogenous cells. Conidiogenous cells holoblastic, hyaline, cylindrical to doliiform, smooth-walled. Conidia not observed. Spermatia aseptate, hyaline, guttulate, smooth-walled, cylindrical, with rounded ends, 7.12–7.98 × 1.5–2.54 μm (mean =7.6–2.1 μm), L/W ratio = 3.45. Hyphae brown, 2–2.5 μm wide. Chlamydospore in chains of 2–5, brick brown III’’ (60) to sepia III’’ (63), thick-walled, 10–15 μm (mean =12 μm) wide.

Cultural characteristics—Colonies on PDA after 2-week in the dark: dark greenish black in the center, greenish olivaceous towards the margin, similar in reverse; later turning to fuscous black, aerial mycelium appressed. Colonies flat, showing zones with undulate edge, 35–40 mm diam after 2 d at 30 ± 0.2°C, reaching the edge of the Petri dish within 4 d.

Habitat:—Isolated from soil attached to the base of macrofungus stipe in mixed forest.

Distribution:— Amarkantak, District Anuppur, State of Madhya Pradesh, India.

Sexual morph:—Not known.

Additional culture examined:— INDIA, Madhya Pradesh, Anuppur (Amarkantak), from the same soil sample, 23 December 2011, Ruby Sharma (culture strain MMI00068).

Remarks:—The strain MCC 1039 is characterized by fast growing dark greenish black colony ( Figs. 1–2 ), long neck-like multilocular-papillate pycnidia and small <10 μm cylindrical, hyaline spermatia. The fungus was also tested for its temperature range on 5 different media. It shows optimum growth at 30 ± 0.2 °C (mesophilic in nature) with maximum tolerance up to 35 ± 0.2 °C and minimum tolerance up to 15 ± 0.2 °C. It did not grow at 40 ± 0.2 °C and 10 ± 0.2 °C ( Fig. 3 ). It showed maximum growth on PDA followed PCA and MEA. The fungus was also grown on dead wood, pinus needle (leaves) for approximately 90 d for the development of conidia but without success, only pycnidia and spermatia were formed ( Fig. 4).

The ITS sequence of 2 strains obtained during the study (designated as MMI00067 and MMI00068) are identical when aligned in MEGA5. The BLASTn result of ITS region (ITS– 5.8S –ITS2) show 90% similarity (query coverage: 100%) with Aplosporella artocarpi B0391 (KM006450) and Ap. prunicola CBS 121167 (KF766147). The alignment of combined dataset of 47 concatenaed sequences (ITS–LSU– tef– β tub) belonging to order Botryosphaeriales consisted of 2523 bases, out of which 1296 were conserved, 1176 were variable, 958 were parsimonious informative, and 212 were singleton. The alignment has been submitted to TreeBASE (ID 18163; www.treebase.org).

Figures

FIGURE 1. Fruiting structures of Alanomyces indica (MCC 1039) A–C: Culture of Alanomyces indica with several neck–like pycnidia. D: SEM of large pycnidia. Bars: A–C 2000 μm; D 200 μm.

FIGURE 2. Microscopic structures of Alanomyces indica (MCC 1039). A: Chlamydospores in chains. B: SEM of chlamydospores chain. C–D: DIC image of Spermatia at 1000 x. E: SEM of section of single locule of pycnidia. F: SEM of cylindrical spermatia. Bars: A 20 μm; B, C, D, E 10 μm; F 2 μm.

FIGURE 3. Temperature–growth relationship of Alanomyces indica (MCC 1039) on Malt Extract Agar (MEA), Czapek Dox Agar (CDA), Oat Meal Agar (OA), Potato Dextrose Agar (PDA), Potato Carrot Agar (PCA) (mm diam. of 7 d old colonies).