Neophyllachora religiosa Literatus, Haituk & Cheew., 2023

Literatus, Irish C. E., Maharachchikumbura, Sajeewa S. N., Withee, Patchareeya, Haituk, Sukanya, Tamakaew, Nisachon, Nguanhom, Jeerapa, Monkhung, Sararat, Thiyagaraja, Vinodhini & Cheewangkoon, Ratchadawan, 2023, Taxonomic and phylogenetic appraisal of a new holomorphic Neophyllachora species from Chiang Mai, Thailand, Phytotaxa 600 (5), pp. 259-271 : 264-266

Neophyllachora religiosa Literatus, Haituk & Cheew. , sp. nov.

Index Fungorum number: IF900393

Etymology:— The epithet “ religiosa ” refers to the epithet name of the host plant “ Ficus religiosa ”, from which the holotype was collected.

Holotype:— MFLU 23-0258

Associated with living leaves of Ficus religiosa . Pseudostromata elongated, irregular, discrete, sparse, coalescent, glabrous, shiny, intraepidermal to subepidermal, epiphyllous, multilocular, occasionally amphigenous, often or rarely covering the leaf surface. Sexual morph: Ascomata 83–172 × 127–349 µm (x̄ = 127.5 × 238 µm, n = 20), perithecial, globose to subglobose, solitary or aggregated, ostiolate. Ostiole conspicuous. Peridium 21–30 µm thick, dark brown to black, lateral part wider than basal part, compactly arranged strongly melanized cells. Paraphyses 1–2 µm wide, filiform, numerous, persistent, septate, unbranched, longer than asci. Asci 59–140 × 14–25 µm (x̄ = 88 × 20 µm, n = 30), 8-spored, J-, unitunicate, persistent, cylindrical to fusiform, short-pedicellate, walls uniform in thickness but not specially thickened at the apex and without visible apical structures. Ascospores 8–12 × 5–9 µm (x̄ = 13 × 7 µm, n = 30), uniseriate to biseriate, overlapping, hyaline to olivaceous, globose to elliptical, 1-2 guttules, with a central concave depression, covered by a mucilaginous sheath, irregularly thickened sheath, 1–4.5 µm thickness, widely thickened at the lateral. Asexual morph: Spermacial or Conidial. Conidiomata bearing conidiogenous cells. Conidiogenous cells holoblastic. Conidia 3.5–6 × 2–4 µm (x̄ = 4.75 × 3 µm, n = 30), ellipsoidal, hyaline, rarely septate. Spermatial locules ampulliform. Spermatiogenous cells produce spermatia. Spermatia 7–16 × 0.9–1.4 µm (x̄ = 11.5 × 1.15 µm, n = 30), botuliform or falciform, narrowly rounded at both ends, curved, aseptate, hyaline, smooth-walled.

Material examined— THAILAND, Chiang Mai Province, on living leaves of Ficus religiosa (Moraceae) , (19 December 2022 ( MFLU 23-0252), 18 November 2022 ( MFLU 23-0253), 1 December 2022 ( MFLU 23-0254), 22 October 2022 ( MFLU 23-0255), 27 November 2022 ( MFLU 23-0256), 3 September 2022 ( MFLU 23-0257), 6 September 2022 ( MFLU 23-0258)), Irish C. E. Literatus & Ratchadawan Cheewangkoon ( MFLU 23-0258; Holotype, CRC-H196; Isotype) ( Fig. 3 , 4 , 5 ).

GenBank Accession Numbers for ITS — MFLU 23-0252; OQ821004, MFLU 23-0253; OQ821005, MFLU 23- 0254; OQ821006, MFLU 23-0255; OQ821007, MFLU 23-0256; OQ821008, MFLU 23-0257; OQ821009, MFLU 23-0258; OQ821010.

Notes — The fungus formed solitary to gregarious black spots, mainly on the upper surface of the host. The new species shows a close phylogenetic association with N. fici , and both species are reported in the same host genus ( Ficus ) but in different localities ( Thailand vs. Taiwan). Morphologically, N. religiosa differs from N. fici in the dimensions of asci (55–185 × 11–26 vs. 90–100 × 15–19 µm) and the thickness of the peridium (15–40 vs. 20–25 µm), and further differs in the ascospore characteristics. The new species possesses ascospores with guttules and are covered with thick gelatinous sheath, whereas N. fici lacks sheath in the ascospores. N. religiosa differs from the type N. myrciae in the dimension of pseudostromata (2–3 vs. 3–6 µm), thinner paraphyses (1.5–2.5 vs. 2.5–4.5 µm), shape of asci (fusoid vs. cylindrical to fusiform), shape of ascospores (lunate vs. globose to elliptical), presence of sheath in the ascospores, arrangement of ascospores (1-2 seriate vs. biseriate to multiseriate), color of ascospores (hyaline to olivaceous vs. hyaline), host ( Ficus religiosa vs. Myrcia sp. ) and the distribution ( Thailand vs. Brazil) but shares similar characteristics in the size of ascomata, asci and coelomycetes asexual state. N. religiosa shares similar ascospores characteristics to N. cerradensis in elliptic-oblong shape ascospores with gelatinous sheath. However, N. religiosa differs in the size of ascospores (8–15 × 5–12 vs. 15–22 × 6–9 µm), thickened gelatinous sheath with hyaline to olivaceous color ascospores, while N. cerradensis has thin walled and hyaline ascospores ( Table 1 ). Neophyllachora religiosa mainly differs from the extant species in the ascospores characteristics, which has a thick gelatinous sheath that is irregularly thickened, irregularly guttulate at the immature stage and large guttules present at maturity. The shape ranges from globose to elliptical, and a central concave depression present in the globose-shaped ascospores. In this study, all the newly generated sequences clustered together and formed a sister clade to N. fici with high bootstrap support (100%) ( Fig. 2 ). The ITS region comprised 588 aligned characters, including gaps. The base pair comparison between N. fici and the newly obtained sequences shows more than 2% differences in the ITS region without the gap. The base pair comparison among the newly generated sequences revealed a minor difference in the ITS region. Thus, we assumed that the newly generated sequences obtained in this study are from the same species.

Figures

FIGURE 2. RAxML tree based on analyses of ITS sequence data. Bootstrap support values for ML equal or greater than 70% and Bayesian posterior probabilities (BP) equal or greater than 0.95 are given as ML/BP above the nodes. The tree is rooted to Telimena bicincta (MM-108), T. canafistulae (MM-13) and T. leeae (TH549).

FIGURE 3. Neophyllachora religiosa (MFLU 23-0254, MFLU 23-0256).a. Tarspot on living leaves.b. Upper leaf surface with symptoms. c. Lower leaf surface with symptoms.d. Pseudostroma e. Horizontal section through pseudostroma.f, g.Vertical section through ascomata. h. Vertical section through peridium. i. Asci and paraphyses. j. Paraphyses. k–n. Asci (In ‘l’, arrow showing central concave depression; In ‘n’, arrows showing guttules). o–p. Ascospores (In ‘p’ arrows showing mucilaginous sheath). u–q. Germinated ascospores. Scale bars: f, g =30 µm, h = 10 µm, i = 50 µm, j = 5 µm, k–n = 20 µm, o–q = 10 µm.

FIGURE 4. Neophyllachora religiosa (MFLU 23-0252, MFLU 23-0253, MFLU 23-0258). a. Tar spot on living leaves. b. Upper leaf surface with symptoms. c. Lower leaf surface with symptoms. d. Pseudostroma. e. Horizontal section through pseudostroma. f. Vertical section through pseudostroma.g. Verticalsection through ascomata.h.Vertical section through peridium.i, j.Asci (In i & marrows showing central concave depression, In ‘n’, arrows showing gattules). k. Paraphyses. l–n. Asci. o–p. Ascospores (arrows showing mucilaginous sheath). q. Germinated ascospores. Scale bars: f, g =30 µm, h–k = 10 µm, l–n = 20 µm, o–q = 10 µm.

FIGURE 5. Neophyllachora religiosa (MFLU 23-0255, MFLU 23-0257). a. Vertical section through spermatogonia showing spermatia bearing cells and spermatia.b. Spermatia. c. Conidiomata. d. Conidiogenous cells (arrows showing conidiogenous cells), e. Conidia. Scale bars: f–j = 20 µm, k–q = 10 µm