(83 MycoKeys MycoKeys 112: 35-58 (2025) DOI: 10.3897/mycokeys.112.137098 Research Article Molecular phylogeny and morphology reveal four new wood-inhabiting fungi of Asterostroma and Radulomyces (Basidiomycota) from Southwestern China Junhong Dong™, Qiaohua Deng'™®, Minglan Chen’®, Daxiang Chen2®, Chungin Zhou®®, Changlin Zhao'*® &e wo NY — College of Forestry, Southwest Forestry University, Kunming 650224, China Tongbiguan Provincial Nature Reserve, Mangshi 678499, China Management and Conservation Bureau, Yunnan Wumeng Mountain National Nature Reserve, Zhaotong, 657000, China Yunnan Key Laboratory of Gastrodia and Fungal Symbiotic Biology, Zhaotong University, Zhaotong 657000, China Corresponding author: Changlin Zhao (fungi@swfu.edu.cn, fungichanglinz@163.com) OPEN Qaccess This article is part of: Exploring the Hidden Fungal Diversity: Biodiversity, Taxonomy, and Phylogeny of Saprobic Fungi Edited by Samantha C. Karunarathna, Danushka Sandaruwan Tennakoon, Ajay Kumar Gautam Academic editor: Samantha C. Karunarathna Received: 14 September 2024 Accepted: 3 December 2024 Published: 8 January 2025 Citation: Dong J, Deng Q, Chen M, Chen D, Zhou C, Zhao C (2025) Molecular phylogeny and morphology reveal four new wood- inhabiting fungi of Asterostroma and Radulomyces (Basidiomycota) from Southwestern China. MycoKeys 112: 35-98. https://dol.org/10.3897/ mycokeys.112.137098 Copyright: © Junhong Dong et al. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Abstract In the ecosystem, wood-inhabiting fungi play an indispensable role in wood degra- dation and the cycle of substances. They are regarded as the “key player” in the pro- cess of wood decomposition because of their ability to produce various enzymes that break down woody lignin, cellulose, and hemicellulose. In this study, four new wood-inhabiting fungal species, Asterostroma paramuscicola, Radulomyces bam- businus, R. fissuratus, and R. sinensis, were collected from southwestern China and were proposed based on the morphological and molecular evidence. Asterostroma paramuscicola is characterised by the felted-membranous to pellicular basidiomata with pinkish to slightly salmon-buff, a smooth hymenial surface, a monomitic hyphal system, and generative hyphae bearing simple-septate and subglobose, thin-walled, echinulate basidiospores measuring as 8-8.8 x 7-8 um. Radulomyces bambusinus is characterised by the resupinate basidiomata with pinkish-white to pink, a tuberculate hymenial surface, a monomitic hyphal system and generative hyphae bearing clamp connections, and subglobose, slightly thick-walled, smooth basidiospores measuring as 6-7.5 x 5.5-7.3 um. Radulomyces fissuratus is characterised by the coriaceous basidiomata with grey to grey-buff, a tuberculate hymenial surface, a monomitic hy- phal system and generative hyphae bearing clamp connections, and globose, slightly thick-walled, smooth basidiospores measuring as 7-9.5 x 6.5-8.5 um. Radulomyces sinensis is characterised by the coriaceous basidiomata with straw to cinnamon to ocherous, a tuberculate hymenial surface, a monomitic hyphal system and genera- tive hyphae bearing clamp connections, and broadly ellipsoid, slightly thick-walled, smooth basidiospores measuring as 7.5-9 x 6.2—7.5 um. Sequences of the internal transcribed spacer (ITS) and large subunit (nrLSU) markers of the studied samples were generated, and phylogenetic analyses were performed with maximum likelihood, maximum parsimony, and Bayesian inference methods. Phylogenetic analyses of ITS+nrLSU nuclear RNA gene regions showed that four new species were assigned to the genera Asterostroma and Radulomyces. The phylogenetic tree inferred from the ITS sequences revealed that A. paramuscicola was closely associated with A. macros- porum and A. muscicola. Based on the ITS sequences, the topology showed that Rad- ulomyces bambusinus was retrieved as a sister to R. zixishanensis. The taxon R. fis- suratus forms a monophyletic lineage. The other one species, R. sinensis, was closely associated with R. molaris and R. yunnanensis. 35 Junhong Dong et al.: Four new wood-inhabiting fungi Key words: Biodiversity, molecular systematics, new species, taxonomy, wood-decaying fungi, Yunnan Province Introduction The term “eukaryote” refers to cell structure and means that an organism's ge- netic information is housed inside a structure called a nucleus (Money 2016). Fungi are eukaryotic microorganisms that play key ecological roles as decom- posers, mutualists, or pathogens (Hyde et al. 2021). Fungi are a distinct, di- verse, and ecologically important branch of the tree of life (James et al. 2020). Classification of the fungi has proven challenging due to the small number of known as compared to estimated species and a lack of sequence data for many extant taxa (Hyde et al. 2021). The phylum Basidiomycota constitutes a major group of the kingdom fungi and is second in species numbers to the phylum Ascomycota (Wijayawardene et al. 2018). Wood-inhabiting fungi have important industrial, medicinal, edible, and economic values, and a small num- ber of them contain toxic metabolites (Niego et al. 2023). The genus Asterostroma Massee, belonging to the family Peniophoraceae (Russulales, Basidiomycota), is typified by A. apalum (Berk. & Broome) Massee, and it is characterised by the resupinate, effused, brittle, loosely adnate, mem- branaceous to pellic basidiomata; a cream to ochraceous, smooth hymenial surface; a dimitic hyphal system with simple-septate on generative hyphae and dextrinoid asterosetae; present gloeocystidia; utriform basidia with 4-sterig- mata and a basal simple septum; and subglobose to ellipsoid, smooth or tu- berculate, amyloid basidiospores (Massee 1889; Bernicchia and Gorjon 2010). Based on the MycoBank database (http://www.mycobank.org, accessed on 9 November 2024) and the Index Fungorum (http://www.indexfungorum.org, ac- cessed on 9 November 2024), the genus Asterostroma has registered 41 spe- cific and infraspecific names, and six have been recorded from China (Liu et al. 2017; Deng et al. 2024; Dong et al. 2024a; Zhou et al. 2024). The genus Radulomyces M.P. Christ., belonging to the family Radulomyceta- ceae (Agaricales, Basidiomycota), is typified by R. confluens (Fr.) M.P. Christ., and it is characterised by the resupinate, adnate, effused, ceraceous, hygropha- nous basidiomata, smooth, tuberculate, odontioid to raduloid hymenial surface, a monomitic hyphal system with clamp connections on generative hyphae, clavate, sinuous basidia with 4-sterigmata and a basal clamp connection, with abundant oil drops, and ellipsoid to globose, smooth or minutely ornamented (spore dimorphism occurs in some species), slightly thick-walled, acyanophi- lous basidiospores (Christiansen 1960; Bernicchia and Gorjon 2010). Based on the MycoBank database (http://www.mycobank.org, accessed on 9 November 2024) and the Index Fungorum (http://www.indexfungorum.org, accessed on 9 November 2024), the genus Radulomyces has registered 30 specific and in- fraspecific names, and three have been found from China (Dong et al. 2024b). Based on the morphological characteristics of asterosetae, the genus Aster- ostroma was placed in the family Lachnocladiaceae D.A. Reid (Parmasto 1971; Hallenberg and Eriksson 1985). Later, Asterostroma belonged to the russuloid lineage and was located in the family Peniophoraceae Lotsy in their phyloge- netic analysis of ITS+nrLSU (Larsson and Larsson 2003; Miller et al. 2006; Lars- MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 36 Junhong Dong et al.: Four new wood-inhabiting fungi son 2007; Liu et al. 2017). According to the morphological characteristics of the basidiospores in Asterostroma (Parmasto 1971; Boidin et al. 1997), this genus was divided into two subgenera: Austroasterostroma Parmasto (smooth and amyloid basidiospores) and Asterostroma ornamented and amyloid basid- iospores (Liu et al. 2017; Zhou et al. 2024). Recently, phylogenetic analyses on Asterostroma from China based on ITS+nrLSU sequences and morphology have identified three new species: A. rhizomorpharum H.M. Zhou & C.L. Zhao, A. roseoalbum J.H. Dong & C.L. Zhao and A. yunnanense Y.L. Deng & C.L. Zhao were described (Deng et al. 2024; Dong et al. 2024a; Zhou et al. 2024). The placement of two genera, Aphanobasidium Julich and Radulomyces, was previously located in the family Pterulaceae Corner by the phylogenetic reconstructions of corticioid taxa (Larsson et al. 2004; Larsson 2007). Phylo- genetically, Radulomyces belonged in the Pterulaceae (Agaricales) and was most closely related to Radulotubus Y.C. Dai, S.H. He & C.L. Zhao and Apha- nobasidium (Zhao et al. 2016). These three genera form a strongly supported clade sister to the Pterula-Deflexula-Pterulicium-Merulicium-Coronicium clade based on sequence analyses of ITSt+nrLSU (Zhao et al. 2016; Wang et al. 2018). Radulomycetaceae was characterized by the combination of resupinate basid- iomes, a mMonomitic hyphal system, and the absence of cystidia, in which the phylogenetic analyses strongly support the segregation of Radulomycetaceae from Pterulaceae (Leal-Dutra et al. 2020). Based on the phylogenetic and mor- phological analysis, Leal-Dutra et al. (2020) revealed that no members of the three genera (i.e., Aphanobasidium, Radulomyces, and Radulotubus) within this superclade were pteruloid (i.e., coralloid basidiomes with a dimitic hyphal sys- tem) in their morphology. They were distinct from nearly all the other members of Pterulaceae, clearly by morphology and phylogeny, and then consequently Leal-Dutra et al. (2020) proposed the new family Radulomycetaceae to accom- modate the three genera. Based on the ITS+nrLSU sequence and morpholog- ical characteristics, three new species, R. hydnoides J.H. Dong & C.L. Zhao, R. yunnanensis J.H. Dong & C.L. Zhao, and R. zixishanensis J.H. Dong & C.L. Zhao, were introduced from China (Dong et al. 2024b). During investigations on wood-inhabiting fungi in southwestern China, some specimens were collected. To clarify the placement and relationships of these specimens, we carried out a phylogenetic and taxonomic study on the genera Asterostroma and Radulomyces based on the ITS and ITS+nrLSU sequences. These specimens were assigned to the genera Asterostroma and Radulomy- ces. Therefore, four new species, Asterostroma paramuscola, R. bambusinus, R. fissuratus, and R. sinensis, are proposed, based on the morphological char- acteristics and phylogenetic analyses. Materials and methods Sample collection and herbarium specimen preparation The fresh fruiting bodies on the dead bamboo and fallen angiosperm branches were collected from Dehong, Lincang, Pu’er, and Zhaotong of Yunnan Province, China. The samples were photographed in situ, and fresh macroscopic details were recorded. Photographs were recorded by a Nikon D7100 camera. All the photos were focus-stacked using Helicon Focus software. Macroscopic details MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 37 Junhong Dong et al.: Four new wood-inhabiting fungi and collection information (Rathnayaka et al. 2024) were taken and transported to a field station where the fruit body was dried on an electronic food dryer at 45 °C (Hu et al. 2022). Once dried, the specimens were sealed in an envelope and zip-lock plastic bags and labeled. The dried specimens were deposited in the herbarium of the Southwest Forestry University (SWFC), Kunming, Yunnan Province, China. Morphology The macromorphological descriptions were based on field notes and photos captured in the field and lab. The color terminology follows Petersen (1996). The micromorphological data were obtained from the dried specimens after observation under a light microscope with a magnification of 10 x 100 oil. Sections were mounted in 5% KOH and 2% phloxine B (C,,,H,Br,Cl,Na,O.), and other reagents were also used, including cotton blue and Melzer’s reagent to observe micromorphology following Wu et al. (2022). To show the variation in spore sizes, 5% of measurements were excluded from each end of the range and shown in parentheses. At least thirty basidiospores from each specimen were measured. Stalks were excluded from basidia measurements, and the hi- lar appendage was excluded from basidiospore measurements. The following abbreviations are used: KOH = 5% potassium hydroxide water solution, CB- = acyanophilous, IKI- = both inamyloid and non-dextrinoid, L = mean spore length (arithmetic average for all spores), W = mean spore width (arithmetic average for all spores), Q = variation in the L/W ratios between the specimens studied, Q_ represented the average Q of basidiospores measured + standard deviation, and n = a/b (number of spores (a) measured from given number (b) of specimens). Molecular phylogeny The CTAB rapid plant genome extraction kit-DN14 (Aidlab Biotechnologies Co., Ltd., Beijing, China) was used to extract genomic DNA from the dried speci- mens according to the manufacturer's instructions. The nuclear ribosomal in- ternal transcribed spacer (ITS) region was amplified with ITS5 and ITS4 primers (White et al. 1990). The nuclear large subunit (nrLSU) region was amplified with the LROR and LR7 primer pair (Vilgalys and Hester 1990; Rehner and Samuels 1994). The PCR procedure for ITS was as follows: initial denaturation at 95 °C for 3 min, followed by 35 cycles at 94 °C for 40 s, 58 °C for 45 s, and 72 °C for 1 min, and a final extension of 72 °C for 10 min. The PCR procedure for nrLSU was as follows: initial denaturation at 94 °C for 1 min, followed by 35 cycles at 94 °C for 30 s, 48 °C for 1 min, and 72 °C for 1.5 min, and a final extension of 72 °C for 10 min. The PCR products were purified and sequenced at Kunming Tsingke Biological Technology Limited Company (Yunnan Province, PR. China). The newly generated sequences were deposited in NCBI GenBank (Table 1). The sequences were aligned in MAFFT v. 7 (Katoh et al. 2019) using the G-INS-i strategy. The alignment was adjusted manually using AliView v. 1.27 (Larsson 2014). The dataset was aligned first, and then the sequences of ITS+nrLSU were combined with Mesquite v. 3.51. The combined ITS+nrLSU se- quences and ITS datasets were used to infer the position of the new species MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 39 Junhong Dong et al.: Four new wood-inhabiting fungi Table 1. List of species, specimens, and GenBank accession numbers of sequences used in this study. [New species is shown in bold; * type material]. GenBank Accession No. Species Name Sample No. ITS arLSU Country References Aphanobasidium pseudotsugae HHB-822 GU187509 | GU187567 USA Larsson (2007) Aphanobasidium pseudotsugae UC 2023153 KP814353 | AY586696 | Sweden Larsson (2007) Asterostroma andinum He 20120921-17 a KY 263874 China Liu et al. (2017) Asterostroma andinum HHB-8546-sp = AF518600 USA Hibbett and Binder (2002) Asterostroma bambusicola He 4128 KY263864 = Thailand Liu et al. (2017) Asterostroma bambusicola He 4132 KY263865 | KY263871 | Thailand Liu et al. (2017) Asterostroma cervicolor He 2314 KY263860 | KY263868 | China Liu et al. (2017) Asterostroma cervicolor He 4020 KY263859 | KY263869 | China Liu et al. (2017) Asterostroma cervicolor KHL 9239 AF506408 | AF506408 | Puerto Larsson and Larsson (2003) Rico Asterostroma cervicolor TMI 21362 AB439560 7 Japan Larsson and Larsson (2003) Asterostroma laxum EL 33-99 AF506410 | AF506410 | Estonia Larsson and Larsson (2003) Asterostroma macrosporum TMI 25696 AB439544 = Japan Suhara et al. (2010b) Asterostroma macrosporum TMI 25697 AB439545 = Japan Suhara et al. (2010b) Asterostroma medium HFRG_EJ220212_2_FRDBI | 0Q133615 = UK Deng et al. (2024) 23891920 Asterostroma medium HFRG_EJ210127_2FRDBI | OL828779 a UK Deng et al. (2024) 18772203 Asterostroma muscicola He 4106 KY263861 | KY263873 | Thailand Liu et al. (2017) Asterostroma muscicola He 20121104-1 KY263862 | KY263872 China Liu et al. (2017) Asterostroma muscicola TUMH 10017 AB439552 = Japan Suhara et al. (2010b) Asterostroma ochroleucum HB 9/89 = AF323737 | Germany Wagner (2001) Asterostroma paramuscicola CLZhao 8594 PP392895 PQ306584 | China Present study Asterostroma rhizomorpharum CLZhao 31212 OR672732 | OR879302 | China Zhou et al. (2024) Asterostroma rhizomorpharum CLZhao 31216 OR672733 = China Zhou et al. (2024) Asterostroma vararioides He 4136 KY263866 = Thailand Liu et al. (2017) Asterostroma vararioides He 4140 KY263867 | KY263870 | Thailand Liu et al. (2017) Asterostroma yunnanense CLZhao 22781 ORO48809 | OR506285 | China Deng et al. (2024) Asterostroma yunnanense CLZhao 22786 ORO048811 OR506286 | _ China Deng et al. (2024) Confertobasidium olivaceoalbum FP 90196 AF511648 | AF511648 USA Larsson and Larsson (2003) Dichostereum durum FG 1985 AF506429 | AF506429 | France Larsson and Larsson (2003) Dichostereum effuscatum GG 930915 AF506390 | AF506390 | France Larsson and Larsson (2003) Gloiothele lactescens EL 8-98 AF506453 AF506453 | Sweden | Larsson and Larsson (2003) Gloiothele lamellosa KHL 11031 AF506454 | AF506454 USA Larsson and Larsson (2003) Merulicium fusisporum Hjm s.n. EU118647 | EU118647 | Sweden Larsson (2007) Peniophora cinerea NH 9808/1788 AF506424 | AF506424 |_ Spain Larsson and Larsson (2003) Peniophora incarnata NH 10271/1909 AF506425 AF506425 | Denmark | Larsson and Larsson (2003) Pterula echo AFTOL-ID711 DQ494693 | AY629315 USA Larsson and Larsson (2003) Radulomyces bambusinus CLZhao 35383 * PQ306582 | PQ306589 China Present study Radulomyces bambusinus CLZhao 35384 PQ306583 | PQ306590 = China Present study Radulomyces confluens Cui 5977 KU535661 | KU535669 China Wang et al. (2018) Radulomyces confluens He 2224 KU535662 | KU535670 | China Wang et al. (2018) Radulomyces copelandii Dai 15061 KU535664 | KU535672 | China Wang et al. (2018) Radulomyces copelandii Wu 9606-5 KU535663 | KU535671 China Wang et al. (2018) Radulomyces fissuratus CLZhao 29670 * PQ306579 | PQ306586 | China Present study Radulomyces fissuratus CLZhao 29695 PQ306580 | PQ306587 | China Present study Radulomyces fissuratus CLZhao 29713 PQ306581 | PQ306588 | China Present study Radulomyces hydnoides CLZhao 21632 ORO96184 | OR449914 | China Dong et al. (2024b) Radulomyces hydnoides CLZhao 21668 ORO96185 | OR449915 |_ China Dong et al. (2024b) MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 39 Junhong Dong et al.: Four new wood-inhabiting fungi Species Name Radulomyces molaris Radulomyces molaris Radulomyces paumanokensis Radulomyces rickii Radulomyces rickii Radulomyces sinensis Radulomyces yunnanensis Radulomyces yunnanensis Radulomyces zixishanensis Radulotubus resupinatus Radulotubus resupinatus Scytinostroma portentosum Vararia gallica Vararia ochroleuca Vesiculomyces citrinus GenBank Accession No. Sample No. Country References ITS nrLSU ARAN-Fungi 2003 - MT232311| Spain Olariaga et al. (2020) ML0499 AY463459 | AY586705 | Sweden Larsson et al. (2004) IMG 5985-16 MG050100 | MG050110| = Spain Wang et al. (2018) JK 951007 7 AY586706 | Sweden Larsson et al. (2004) G1066 = MK278543 | Hungary Varga et al. (2019) CLZhao 25667 * PQ306578 | PQ306585 China Present study CLZhao 1262 ORO96191 | OR449917 China Dong et al. (2024b) CLZhao 7364 ORON96192 | OR449918 China Dong et al. (2024b) CLZhao 21127 ON033887 | OR449922 China Dong et al. (2024b) Cui 8383 KU535660 | KU535668 China Zhao et al. (2016) Cui 8462 KU535657 | KU535665 China Zhao et al. (2016) EL 11-99 AF506470 AF506470 | Sweden | Larsson and Larsson (2003) CBS 656.81 = AF323742 | France Wagner (2001) CBS 465.61 = AF323743 | France Wagner (2001) EL 53-97 AF506486 AF506486 | Sweden | Larsson and Larsson (2003) and related species. The sequences of Confertobasidium olivaceoalbum (Bour- dot & Galzin) Jiilich were retrieved from GenBank and used as outgroup taxa in the ITS+nrLSU analysis (Fig. 1) in the family Peniophoraceae; Scytinostroma portentosum (Berk. & M.A. Curtis) Donk was selected as the outgroup taxon for the ITS analysis (Fig. 2) in the genus Asterostroma (Deng et al. 2024). The sequences of Pterula echo D.J. McLaughlin & E.G. McLaughlin and Merulicium fusisporum (Romell) J. Erikss. & Ryvarden were selected as the outgroup taxon for the ITS+nrLSU analysis (Fig. 3) of the family Radulomycetaceae; Radulotu- bus resupinatus Y.C. Dai, S.H. He & C.L. Zhao was selected as outgroup taxa in the ITS analysis (Fig. 4) in the genus Radulomyces (Zhao et al. 2016). Maximum parsimony (MP), maximum likelihood (ML), and Bayesian infer- ence (BI) analyses were applied to the combined three datasets following a previous study (Zhao and Wu 2017), and the tree construction procedure was performed in PAUP* v. 4.0610 (Swofford 2002). All characters were equally weighted, and gaps were treated as missing data. Trees were inferred using the heuristic search option with TBR branch swapping and 1000 random sequence additions. Max trees were set to 5000, branches of zero length were collapsed, and all parsimonious trees were saved. Clade robustness was assessed using bootstrap (BT) analysis with 1000 replicates (Felsenstein 1985). Descriptive tree statistics, tree length (TL), consistency index (Cl), retention index (RI), res- caled consistency index (RC), and the homoplasy index (HI) were calculated for each maximum parsimonious tree generated. The multiple sequence align- ment was also analysed using Maximum Likelihood (ML) in RAxML-HPC2 on XSEDE v. 8.2.8 with default parameters (Miller et al. 2012). Branch support (BS) for ML analysis was determined by 1,000 bootstrap replicates. jModelTest v. 2 (Darriba et al. 2012) was used to determine the best-fit evo- lutionary model for each data set for Bayesian inference (BI), which was per- formed using MrBayes 3.2.7a (Ronquist et al. 2012). The first one-fourth of all generations was discarded as burn-in. The majority rule consensus tree of all remaining trees was calculated. Branches were considered as significant- ly supported if they received maximum likelihood bootstrap value (BS) = 70%, maximum parsimony bootstrap value (BT) = 70%, or Bayesian posterior proba- bilities (BPP) = 0.95. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 A0 Junhong Dong et al.: Four new wood-inhabiting fungi - /100/1.00 [ Asterostroma medium HFRG_EJ220212_2 FRDBI 23891920 UK 81/52/1.00 Asterostroma medium HFRG_EJ210127_2FRDBI 18772203. UK 100/89/1.00 [~ Asterostroma bambusicola He 4132 Thailand Asterostroma bambusicola He 4128 Thailand 99/100/1.00 ; ; - /98/ - “™.- Asterostroma muscicola He 4106 Thailand 93/95/1.00 Asterostroma muscicola He 20121104-1 China 88/62/).98 Asterostroma paramuscicola CLZhao 8594 China 100/97/1.00. We Asterostroma macrosporum TMI 25696 Japan 91/98/ - Asterostroma macrosporum TMI 25697 Japan -/-/1.00 Asterostroma ochroleucum HB 9/89 Germany 100/1100/1.00 Asterostroma rhizomorpharum CLZhao 31216 China 96/55/0.98 Asterostroma rhizomorpharum CLZhao 31212 China 100/99/1.00 IPL VUIOSOLIISP | Asterostroma yunnanense CLZhao 22781 China Asterostroma yunnanense CLZhao 22786 China 99/64) - Asterostroma cervicolor TMI 21362 Japan -/98/1.00} Asterostroma cervicolor He 4020 China Asterostroma cervicolor He 2314 China 97/99/1.00 : Asterostroma cervicolor KHL 9239 Puerto Rico 100/100/1.00 Asterostroma vararioides He 4136 Thailand Asterostroma vararioides He 4140 ‘Thailand Laevispora clade ENC Asterostroma laxum EL 33-99 — Estonia 100/100/1.00 Gloiothele lactescens EL8-98 Sweden 81/75/0.99 Gloiothele lamellosa KHL 11031 USA | Gloiothele a TNE Vesiculomyces citrinus EL 53-97 Sweden | Vesiculomyces Scytinostroma portentosum EL 11-99 Sweden | Scytinostroma is -/~/1.00 Asterostroma andinum HHB-8546-sp USA 100/99/1.00 © Asterostroma andinum He 20120921-17 China 00F - Dichostereum « m Fungi Gallici 1985 France A, andinum clade Peniophora incarnata NH 10271/1909 Denmark ' 99/100/1.00 Peniophora cinerea NH 9808/1788 Spain | Peniophora Confertobasidium olivaceoalbum FP 90196 USA | Outgroup ;———! 50 Figure 1. Maximum parsimony strict consensus tree illustrating the phylogeny of Asterostroma and related genera in the family Peniophoraceae based on ITS+nrLSU sequences. Branches are labelled with maximum likelihood bootstrap value > 70%, parsimony bootstrap value = 50%, and Bayesian posterior probabilities = 0.95. Results Molecular phylogeny The aligned dataset comprised 34 specimens representing 23 species. Four Markov chains were run for two runs from random starting trees, each for one million generations for the combined ITSt+nrLSU (Fig. 1) data set with trees and parameters sampled every 1000 generations. The dataset had an aligned length of 2206 characters, of which 1453 characters are constant, 276 are vari- able and parsimony uninformative, and 477 are informative. Maximum parsi- mony analysis yielded nine equally parsimonious trees (TL = 1735, Cl = 0.6317, HI = 0.3683, RI = 0.6751, and RC = 0.4265). The best model for the ITS+nrL- SU dataset, estimated and applied in the Bayesian analysis, was GTR+I+G. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 1 Junhong Dong et al.: Four new wood-inhabiting fungi 100/100/1.00 f Asterostroma bambusicola He 4128 Thailand 83/93/1.00 Asterostroma bambusicola He 4132 Thailand Asterostroma Asterostroma medium HFRG_EJ220212 2 FRDBI 23891920 UK 100/100/1.00 4sterostroma medium HFRG_EJ210127_2FRDBI 18772203 UK 89/97/0.99 [— Asterostroma muscicola He 4106 Thailand 95/97/1.00 Asterostroma muscicola TUMH 10017 — Japan Asterostroma paramuscicola CLZhao 8594 China 75/57/0.99 75/63/0.98 Asterostroma macrosporum TMI 25696 Japan 87/92/ - 100/100/1.00! 4sterostroma macrosporum TMI 25697 Japan Asterostroma cervicolor He 2314 China EOD NOOBS Asterostroma cervicolor TMI 21362 Japan sate Asterostroma cervicolor He 4020 China Wihne 98/80/-|| Asterostroma yunnanense CLZhao 22781 China 79/95/-| Asterostroma yunnanense CLZhao 22786 China Asterostroma rhizomorpharum CLZhao 31212 China 100/100/1.00! Asterostroma rhizomorpharum CLZhao 31216 China 100/100/1.00 f 4sterostroma vararioides He 4140 Thailand 100/89/ - Asterostroma vararioides He 4136 Thailand Asterostroma laxum EL 33-99 — Estonia Scytinostroma portentosum EL 11-99 Sweden I 10 Figure 2. Maximum parsimony strict consensus tree illustrating the phylogeny of Asterostroma paramuscicola and re- lated species in the genus Asterostroma based on ITS sequences. Branches are labelled with maximum likelihood boot- strap value = 70%, parsimony bootstrap value = 50%, and Bayesian posterior probabilities = 0.95. Both Bayesian analysis and ML analysis resulted in a similar topology to MP analysis, with an average standard deviation of split frequencies = 0.004237 (BI), and the effective sample size (ESS) for Bayesian analysis across the two runs is double the average ESS (avg. ESS) = 505. The aligned dataset comprised 20 specimens representing 11 species. Four Markov chains were run for two runs from random starting trees, each for 0.5 million generations for the ITS (Fig. 2) data set with trees and param- eters sampled every 1000 generations. The dataset had an aligned length of 620 characters, of which 384 characters are constant, 51 are variable and par- simony uninformative, and 185 are informative. Maximum parsimony analy- sis yielded one equally parsimonious tree (TL = 412, Cl = 0.7694, HI = 0.2306, RI = 0.8450, and RC = 0.6502). The best model for the ITS dataset, estimated and applied in the Bayesian analysis, was HKY+G. Both Bayesian analysis and ML analysis resulted in a similar topology to MP analysis with an aver- age standard deviation of split frequencies = 0.004683 (BI), and the effective sample size (ESS) for Bayesian analysis across the two runs is double the average ESS (avg. ESS) = 435. The aligned dataset comprised 26 specimens representing 15 species. Four Markov chains were run for two runs from random starting trees, each for one million generations for the combined ITS+nrLSU (Fig. 3) data set with trees and parameters sampled every 1000 generations. The dataset had an aligned length of 2115 characters, of which 1699 characters are MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 42 Junhong Dong et al.: Four new wood-inhabiting fungi - /100/1.00 t——] 20 100/100/1.00 Aphanobasidium pseudotsugae UC 2023153 Sweden Pterula echo AFTOL-ID711 USA Radulomyces fissuratus CLZhao 29695 — China 98/96/1.00 | Radulomyces fissuratus CLZhao 29713 China 92/76/1.00}] *Radulomyces fissuratus CLZhao 29670 China Radulomyces rickii JK 951007 Sweden 98/80/\- Radulomyces rickii G1066 Hungary 100(100/1.00 [— Radulomyces bambusinus CLZhao 35384 China Radulomyces bambusinus CLZhao 35383 China Radulomyces sinensis CLZhao 25667 China 76/60/1.00 Radulomyces confluens Cui 5977 — China Radulomyces confluens He 2224 China Radulomyces copelandii Wu 9606-5 China 79/5A/ - 94/90/1.00 Radulomyces copelandii Dai 15061 China Radulomyces molaris ML0499_ Sweden Radulomyces molaris ARAN-Fungi 2003 Spain saatumojnpoy Radulomyces yunnanensis CLZhao 1262 China 88/78/1.00 & Radulomyces yunnanensis CLZhao 7364 China Radulomyces zixishanensis CLZhao 21127 China -/-/1.00 | Radulomyces hydnoides CLZhao 21632 China 84/92/ - Radulomyces hydnoides CLZhao 21668 China -/-/0.97 ‘ Radulomyces paumanokensis IMG 5985-16 Spain 100/100/1.00 | Radulotubus resupinatus Cui 8383 China Radulotubus Radulotubus resupinatus Cui 8462 China Aphanobasidium pseudotsugae HHB-822 USA | 4p hanobasidium Merulicium fusisporum Hjms.n. Sweden | Outgroup Figure 3. Maximum parsimony strict consensus tree illustrating the phylogeny of Radulomyces and related genera in the family Radulomycetaceae based on ITS+nrLSU sequences. Branches are labelled with maximum likelihood bootstrap value = 70%, parsimony bootstrap value = 50%, and Bayesian posterior probabilities = 0.95. constant, 132 are variable and parsimony uninformative, and 284 are in- formative. Maximum parsimony analysis yielded 143 equally parsimonious trees (TL = 711, Cl = 0.7496, HI = 0.2504, RI = 0.7623, and RC = 0.5715). The best model for the ITS+nrLSU dataset, estimated and applied in the Bayesian analysis, was GTR+I+G. Both Bayesian analysis and ML analysis resulted in a similar topology to MP analysis, with an average standard de- viation of split frequencies = 0.008051 (BI), and the effective sample size (ESS) for Bayesian analysis across the two runs is double the average ESS (avg. ESS) = 535.5. The aligned dataset comprised 21 specimens representing 12 species. Four Markov chains were run for two runs from random starting trees, each for 0.5 million generations for the ITS (Fig. 4) data set with trees and param- eters sampled every 1,000 generations. The dataset had an aligned length of 671 characters, of which 444 characters are constant, 74 are variable and parsimony uninformative, and 153 are informative. Maximum parsimo- ny analysis yielded ten equally parsimonious trees (TL = 373, Cl = 0.7587, MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 13 Junhong Dong et al.: Four new wood-inhabiting fungi 100/100/1.00 | Radulomyces bambusinus CLZhao 35383 Radulomyces bambusinus CLZhao 35384 Radulomyces zixishanensis CLZhao 21127 China Radulomyces 100/97/1.00 — Radulomyces yunnanensis CLZhao 1262 China -/10/0.96| ! Radulomyces yunnanensis CLZhao 7364 China ee Radulomyces molaris 5364 Italy Radulomyces sinensis CLZhao 25667 China Radulomyces copelandii Dai 15061 China ‘| Radulomyces copelandii Wu 9606-5 China Radulomyces confluens He 2224 China Radulomyces confluens Cui 5977 — China - /62/ - /97/ - 100/100/1.00 | Radulomyces hydnoides CLZhao 21632 China - /81/- Radulomyces hydnoides CLZhao 21668 China 86/85/0.96 Radulomyces paumanokensis LE-BIN 4691 Russia 100/100/1.00! Radulomyces paumanokensis IMG 5985-16 USA - /100/ - 95/64/ - | Radulomyces fissuratus CLZhao 29670 China Radulomyces fissuratus CLZhao 29713 China 99/100/1.00 : Radulomyces fissuratus CLZhao 29695 China Radulomyces notabilis UC2023084 USA Radulotubus resupinatus Cui 8383 China Radulotubus resupinatus Cui 8462 China — 10 Figure 4. Maximum parsimony strict consensus tree illustrating the phylogeny of three new and related species in the genus Radulomyces based on ITS sequences. Branches are labelled with maximum likelihood bootstrap value = 70%, parsimony bootstrap value = 50%, and Bayesian posterior probabilities = 0.95. HI = 0.2413, RI = 0.8109, and RC = 0.6153). The best model for the ITS data- set, estimated and applied in the Bayesian analysis, was HKY+G. Both Bayes- ian analysis and ML analysis resulted in a similar topology to MP analysis, China China with an average standard deviation of split frequencies = 0.006832 (BI), and the effective sample size (ESS) for Bayesian analysis across the two runs is double the average ESS (avg. ESS) = 346.5. The phylogram, based on the combined ITS+nrLSU sequences (Fig. 1) anal- ysis, showed that the new species, Asterostroma paramuscicola, was assigned to the genus Asterostroma within the family Peniophoraceae. The phylogenetic tree, based on ITS sequences (Fig. 2), revealed that A. paramuscicola was close- ly associated with A. macrosporum N. Maek. & Suhara. and A. muscicola (Berk. & M.A. Curtis) Massee. The phylogram, based on the combined ITS+nrLSU se- quences (Fig. 3) analysis, showed that three new species, Radulomyces bam- businus, R. fissuratus, and R. sinensis, were assigned to the genus Radulomyces within the family Radulomycetaceae. The phylogenetic tree, based on ITS se- quences (Fig. 4), revealed that R. bambusinus was retrieved as a sister to R. zix- ishanensis. The taxon R. fissuratus forms a monophyletic lineage. The other species, R. sinensis, was closely associated with R. molaris (Chaillet ex Fr.) M.P. Christ. and R. yunnanensis J.H. Dong & C.L. Zhao. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 Junhong Dong et al.: Four new wood-inhabiting fungi Taxonomy Asterostroma paramuscicola J.H. Dong & C.L. Zhao, sp. nov. MycoBank No: 855659 Figs 5A, 6, 7 Holotype. CHINA * Yunnan Province, Pu’er, Jingdong County, Taizhong Town, Ai- laoshan Ecological Station, GPS coordinates 24°31'N, 101°02E, altitude 2400 masl., a dead angiosperm tree, leg. C.L. Zhao, 24 August 2018, CLZhao 8594 (SWFC). Etymology. paramuscicola (Lat.): referring to its close phylogenetic relation- ship with A. muscicola. Basidiomata. Annual, resupinate, felted-membranous to pellicular, soft, without odour and taste when fresh, becoming coriaceous upon drying, up to 10 cm long, 5 cm wide, 50-150 um thick. Hymenial surface smooth, pinkish when fresh, turning to pinkish to slightly salmon-buff upon drying, rhizomor- phic. Sterile margin thin, cream to buff, fimbriate, up to 1 mm wide. Hyphal system. Dimitic, generative hyphae scattered, simple-septate, col- orless, thin-walled, occasionally branched, 2—3.5 um in diameter, IKI-, CB-, tissues unchanged in KOH. Asterosetae in subiculum abundant, predominant, yellowish brown, thick-walled, regularly star-shaped, 4-6 um in diameter, weakly dextrinoid, rays up to 75 um long, with acute tips, CB-, and tissues unchanged in KOH. 20 um 20 um Figure 5. Sections of hymenium of Asterostroma and Radulomyces A Asterostroma paramuscicola (holotype, CLZhao 8594) B Radulomyces bambusinus (holotype, CLZhao 35383) C Radulomyces fissuratus (holotype, CLZhao 26970) D Radulomyces sinensis (holotype, CLZhao 25667). 10 x 100 Oil. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 45 Junhong Dong et al.: Four new wood-inhabiting fungi 1 cm 3mm PAE o. Figure 6. Basidiomata of Asterostroma paramuscicola in general and detailed views (holotype, CLZhao 8594). Hymenium. Asterohyphidia in hymenium similar to asterosetae in subiculum, but smaller and less regularly shaped, 2—3.5 um in diameter, rays up to 25 um long, usually bifurcated at tips. Gloeocystidia subcylindrical to fusiform, thin-walled, with a basal simple septum, 45-83.5 x 9-21.5 um; cystidioles absent. Basidia subcylindrical, colorless, with four sterigmata and a basal simple septum, 47.5—58 x 7.5-9.5 um; basidioles dominant, similar to basidia in shape, but slightly smaller. Basidiospores. Subglobose, with a distinct apiculus, spines conical, 2-3 um long, colorless, thin-walled, echinulate, amyloid, with one guttula, CB-, (7.5- )8-8.8(-9) x (6.6-)7-8(-8.5) um, L = 8.39 um, W = 7.65 um, Q = 1.02-1.22, Q_ = 1.10 + 0.06 (n = 30/1). Radulomyces bambusinus J.H. Dong & C.L. Zhao, sp. nov. MycoBank No: 855660 Figs 5B, 8, 9 Holotype. CHINA * Yunnan Province, Zhaotong, Daguan County, Wumengshan Na- tional Nature Reserve, GPS coordinates 27°46'N, 103°52'E, altitude 2200 m asl., on the dead bamboo, leg. C.L. Zhao, 6 November 2023, CLZhao 35383 (SWFC). Etymology. bambusinus (Lat.): referring to the type species growing on bamboo. Basidiomata. Annual, resupinate, adnate, soft membranous, without odour or taste, becoming coriaceous upon drying, up to 5 cm long, 2 cm wide, 50- 100 um thick. Hymenial surface tuberculate, cream to pinkish when fresh, MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 AG Junhong Dong et al.: Four new wood-inhabiting fungi Figure 7. Microscopic structures of Asterostroma paramuscicola (holotype, CLZhao 8594) A basidiospores B basidia C basidioles D gloeocystidia E generative hyphae F asterohyphidia from hymenium G asterosetae from subiculum. Scale bars: 10 um (A-G). turning to pinkish-white to pink upon drying. Sterile margin cream to slightly pinkish, thinning out, up to 1 mm wide. Hyphal system. Monomitic, generative hyphae with clamp connections, col- orless, thin-walled, branched, interwoven, 2.5-3.5 um in diameter; IKI-, CB-, tissues unchanged in KOH. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 47 Junhong Dong et al.: Four new wood-inhabiting fungi lcm Figure 8. Basidiomata of Radulomyces bambusinus in general and detailed views A, B CLZhao 35383 (holotype) C, D CLZhao 35384. 6 G6 6 sow HERE &: © © , | ZL SSS RES Figure 9. Microscopic structures of Radulomyces bambusinus (holotype, CLZhao 35383) A basidiospores B basidia C basidioles D part of the vertical section of hymenium. Scale bars: 10 um (A-D). MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 48 Junhong Dong et al.: Four new wood-inhabiting fungi Hymenium. Cystidia and cystidioles absent. Basidia clavate to barrelled, with 4 sterigmata and a basal clamp connection, occasionally constricted in the middle, 23.5-40.5 x 7.5-10.5 um; basidioles dominant, similar to basidia in shape, but slightly smaller. Basidiospores. Subglobose, slightly thick-walled, smooth, colorless, CB+, (5.7-)6-7.5(-8) x 5.5-7.3(-7.5) um, L = 6.76 pm, W = 6.43 um, Q = 1.01-1.18, Q_ = 1.05 + 0.05 (n = 60/2). Additional specimen (paratype) examined. CHINA * Yunnan Province, Zha- otong, Daguan County, Wumengshan National Nature Reserve, GPS coordi- nates 27°46'N, 103°52'E, altitude 2200 m asl., on the dead bamboo, leg. C.L. Zhao, 6 November 2023, CLZhao 35384 (SWFC). Radulomyces fissuratus J.H. Dong & C.L. Zhao, sp. nov. MycoBank No: 855661 Figss6, 10811 Holotype. CHINA * Yunnan Province, Dehong, Yingjiang County, Tongbiguan Provin- cial Nature Reserve, GPS coordinates 23°48'N, 97°38 'E, altitude 1000 m asl., on the fallen branch of angiosperm, leg. C.L. Zhao, 17 July 2023, CLZhao 29670 (SWFC). mm Figure 10. Basidiomata of Radulomyces fissuratus in general and detailed views A, B, CLZhao 29670 (holotype) C, D CLZhao 29713 E, F CLZhao 29695. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 49 Junhong Dong et al.: Four new wood-inhabiting fungi +34 ’ Go © = SESE 0 STS = = = B[V in NO Me Figure 11. Microscopic structures of Radulomyces fissuratus (holotype, CLZhao 29670) A basidiospores B basidia es = SH Pei See C basidioles D part of the vertical section of hymenium. Scale bars: 10 um (A-D). dl Dial B D Etymology. fissuratus (Lat.): referring to the cracked hymenial surface of the type specimen. Basidiomata. Annual, resupinate, adnate, membranaceous, without odour or taste, becoming hard coriaceous upon drying, up to 30 cm long, 2 cm wide, 50- 150 um thick. Hymenial surface tuberculate, cream when fresh, turning grey to grey-buff upon drying. Sterile margin cream, thinning out, up to 1 mm wide. Hyphal system. Monomitic, generative hyphae with clamp connections, col- orless, thin-walled, branched, interwoven, 2-3.5 um in diameter; IKI-, CB-, tis- sues unchanged in KOH. Hymenium. Cystidia and cystidioles absent. Basidia clavate to barrelled, with 4 sterigmata and a basal clamp connection, 21.5-32.5 x 8.5-10.5 um; basidioles dominant, similar to basidia in shape, but slightly smaller. Basidiospores. Globose, slightly thick-walled, smooth, colorless, CB+, (6.5- )7-9(-9.5) x (6.2-)6.5-8.5(-8.8) um, L = 8.07 um, W = 7.76 um, Q = 1.01-1.06, Q_ = 1.04 + 0.03 (n = 90/3). MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 50 Junhong Dong et al.: Four new wood-inhabiting fungi Additional specimens (paratypes) examined. CHINA * Yunnan Province, Dehong, Yingjiang County, Tongbiguan Provincial Nature Reserve, GPS coor- dinates 23°48'N, 97°38'E, altitude 1000 m asl., on the fallen branch of angio- sperm, leg. C.L. Zhao, 17 July 2023, CLZhao 29695; CLZhao 29713 (SWFC). Radulomyces sinensis J.H. Dong & C.L. Zhao, sp. nov. MycoBank No: 855662 Figs 5D, 12, 13 Holotype. CHINA * Yunnan Province, Lincang, Fengging County, Yaojie Town, Xingyu- an Village, GPS coordinates 24°58'N, 99°92'E, altitude 1600 m asl., on the fallen branch of angiosperm, leg. C.L. Zhao, 22 October 2022, CLZhao 25667 (SWFC). Figure 12. Basidiomata of Radulomyces sinensis in general and detailed views (holotype, CLZhao 25667). MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 51 Junhong Dong et al.: Four new wood-inhabiting fungi © O Figure 13. Microscopic structures of Radulomyces sinensis (holotype, CLZhao 25667) A basidiospores B basidia C basidioles D part of the vertical section of hymenium. Scale bars: 10 um (A-D). Etymology. sinensis (Lat.): referring to the locality (China) of the type specimen. Basidiomata. Annual, resupinate, adnate, soft coriaceous, without odour or taste, becoming hard coriaceous upon drying, up to 20 cm long, 2 cm wide, 100-150 um thick. Hymenial surface tuberculate, buff to slightly straw when fresh, turning to straw to cinnamon to ocherous upon drying. Sterile margin cream to slightly straw, abrupt, up to 1 mm wide. Hyphal system. Monomitic, generative hyphae with clamp connections, col- orless, thin-walled, branched, interwoven, 2.5-3.5 um in diameter; IKI-, CB-, tissues unchanged in KOH. Hymenium. Cystidia and cystidioles absent. Basidia clavate, with 4 sterigmata anda basal clamp connection, with a median constriction, 35-41.5 x 7.5-9.5 um; basidioles dominant, similar to basidia in shape, but slightly smaller. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 59 Junhong Dong et al.: Four new wood-inhabiting fungi Basidiospores. Broadly ellipsoid, slightly thick-walled, smooth, colorless, some with guttulae, CB+, 7.5-9(-9.5) x (5.8-)6.2-7.5(-7.7) um, L = 8.42 um, W = 6.88 um, Q = 1.10-1.33, Q_ = 1.23 + 0.08 (n = 30/1). Discussion In the present study, four new species, Asterostroma paramuscicola, Radulomy- ces bambusinus, R. fissuratus, and R. sinensis, are described based on phyloge- netic analyses and morphological characteristics. Asterostroma is a monophyletic genus in our phylogenetic analysis with low statistical support, in contrast to the previous study (Liu et al. 2017; Zhou et al. 2024). Seven species with ornamented basidiospores formed the section Asteros- troma as A. bambusicola S.L. Liu & S.H. He, A. cervicolor (Berk. & M.A. Curtis) Mas- see, A. macrosporum, A. medium Bres., A. muscicola, A. ochroleucum, A. paramus- cicola, A. rhizomorpharum, and A. yunnanense, while three species with smooth basidiospores formed two clades as the A. andinum Pat. clade (only A. andinum) and sect. Laevispora (A. laxum Bres. and A. vararioides S.L. Liu & S.H. He). Phylogenetically, the phylogram based on the combined ITS+nrLSU sequenc- es (Fig. 1) analysis showed that the new species Asterostroma paramuscicola was assigned to the genus Asterostroma within the family Peniophoraceae. The phylogenetic tree, based on ITS sequences (Fig. 2), revealed that Asteros- troma paramuscicola was closely associated with A. macrosporum and A. mus- cicola. However, the morphological characteristics of A. macrosporum differ from Asterostroma paramuscicola by having an ochreous to fulvous hymenial surface (Suhara et al. 2010b). The species A. muscicola differentiates from A. paramuscicola by having a salmon hymenial surface, smaller basidia (18-24 x 5-6 um), and shorter basidiospores (6-8 x 5.5—7.5 um; Boidin et al. 1997). The phylogram, based on the combined ITS+nrLSU sequences (Fig. 3) anal- ysis, showed that three new species, Radulomyces bambusinus, R. fissuratus, and R. sinensis, were assigned to the genus Radulomyces within the family Radulomycetaceae. The phylogenetic tree, based on ITS sequences (Fig. 4), revealed that Radulomyces bambusinus was retrieved as a sister to R. Zixis- hanensis. The taxon R. fissuratus forms a monophyletic lineage. The other one species, R. sinensis, was closely associated with R. molaris and R. yunnanen- sis. However, the morphological characteristics of R. zixishanensis differ from R. bambusinus by having a cream to slightly brown, smooth hymenial surface and ellipsoid, thin-walled, basidiospores (7-8.8 x 5.5-6.8 um; Dong et al. 2024b). The species R. molaris differs from R. sinensis by having a yellowish to cream, hydnoid hymenial surface (Bernicchia and Gorjén 2010). The spe- cies R. yunnanensis can be distinguished from R. sinensis by having a cream, smooth hymenial surface, shorter basidia (24-35 x 7-11 um), and thin-walled basidiospores (8.2—9.5 x 5.5—7 um; Dong et al. 2024b). Asterostroma paramuscicola, A. rhizomorpharum, and A. yunnanense are all described from China. However, A. rhizomorpharum can be distinguished from A. paramuscicola in its cream to buff, cracked hymenial surface, shorter basidia (30-45 x 5.5-8 um), and smaller basidiospores (5.5—6.8 x 4.6—5.9 um; Zhou et al. 2024). The species A. yunnanense can be distinguished fromA. paramuscicola in its cream to salmon-buff hymenial surface, smaller basidia (31-38 x 4—5 um), and smaller basidiospores (4.5—6 x 4—5 um; Deng et al. 2024). MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 53 Junhong Dong et al.: Four new wood-inhabiting fungi Morphologically, Asterostroma paramuscicola resembles A. bambusicola, A. boninense Suhara & N. Maek., and A. vararioides in sharing a fimbriate ba- sidiomata and subglobose to globose basidiospores. However, A. bambusicola differs from A. paramuscicola by having a brownish yellow, grayish brown, light brown, to brownish-red hymenial surface, shorter basidia (30-45 x 5.5-8 um; Liu et al. 2017). The species A. boninense differentiates from A. paramusci- cola by having the buff to partly ochreous hymenial surface and shorter ba- sidiospores (5.5-7.5 x 5-7.2 um; Suhara et al. 2010a). The species A. varari- oides can be distinguished from A. paramuscicola in its grayish brown, light brown, to dark brown hymenial surface and smooth, smaller basidiospores (6-7 x 5.5-6 um; Liu et al. 2017). Radulomyces bambusinus shares similarities with R. yunnanensis and R. zix- ishanensis in having a cracked hymenial surface. However, R. yunnanensis can be distinguished from R. bambusinus by its smooth hymenial surface, ellipsoid, thin-walled, shorter basidiospores (8.2—9.5 x 5.5-7 um; Dong et al. 2024b). The species R. zixishanensis differentiates from R. bambusinus by having the smooth hymenial surface, ellipsoid, thin-walled basidiospores (7- 8.8 x 5.5-6.8 um; Dong et al. 2024b). Radulomyces fissuratus resembles R. copelandii (Pat.) Hjortstam & Spooner, R. hydnoides, and R. paumanokensis J. Horman, Nakasone & B. Ortiz in sharing subglobose to globose, slightly thick-walled basidiospores. However, R. cope- landii differs from R. fissuratus due to its white hymenial surface, smaller basid- ia (29-35 x 6-7 um), and basidiospores (6.4-7 x 5.4-6.2 um; Ginns and Mill- man 2011). The species R. hydnoides can be distinguished from R. fissuratus by its hydnoid hymenial surface and shorter basidia (21-34 x 8.5-12.5 um; Dong et al. 2024b). The species R. paumanokensis differentiates from R. fissuratus by having a hydnoid hymenial surface, smaller basidia (25-31 x 5-7.5 um), and shorter basidiospores (5.8-6.9 x 5.2—6.4 um; Wang et al. 2018). Radulomyces sinensis shares similarities with R. arborifer Malysheva & Zmi- tr., R. molaris, and R. zixishanensis in having ellipsoid basidiospores. However, R. arborifer differs from R. sinensis due to its dendroid or coralloid hymenial surface (Malysheva 2006). The species R. molaris can be distinguished from R. sinensis by its hydnoid hymenial surface (Bernicchia and Gorjon 2010). The species R. zixishanensis differentiates from R. sinensis by having the cream to slightly brown, smooth hymenial surface and thin-walled basidiospores (7- 8.8 x 5.5-6.8 ym; Dong et al. 2024b). In recent years, the wood-inhabiting fungi have been an extensively studied group of Basidiomycota, which includes a number of poroid, smooth, gran- dinoid, odontioid, and hydnoid basidiomata in China (Liu et al. 2023; Mao et al. 2023; Zhao et al. 2023; Dong et al. 2024b; Wang et al. 2024; Zhao et al. 2024). This paper enriches our knowledge of fungal diversity in China. We an- ticipate that more undescribed wood-inhabiting fungi taxa will be discovered throughout China after extensive collection combined with morphological and molecular analyses. Additional information Conflict of interest The authors have declared that no competing interests exist. MycoKeys 112: 35-58 (2025), DOI: 10.3897/mycokeys.112.137098 54 Junhong Dong et al.: Four new wood-inhabiting fungi Ethical statement No ethical statement was reported. Funding The research was supported by the National Natural Science Foundation of China (Project Nos. 32170004, U2102220), the High-level Talents Program of Yunnan Prov- ince (YNQR-QNRC-2018-111), the Research Project of Key Laboratory of Forest Disas- ter Warning and Control in Universities of Yunnan Province (ZKJS-S-202208), and the Research Project of Yunnan Key Laboratory of Gastrodia and Fungal Symbiotic Biology (TMKF2023A03). Author contributions Conceptualization, C.Z.; methodology, C.Z. and J.D.; software, C.Z., J.D., Q.D., and M.C.; validation, C.Z. and J.D.; formal analysis, C.Z., J.D., and Q.D.; investigation, C.Z., J.D., D.C., and C.Z.; resources, C.Z. and J.D.; writing—original draft preparation, C.Z., J.D., Q.D., and M.C.; writing—review and editing, C.Z. and J.D.; visualization, C.Z. and J.D.; supervision, C.Z.; project administration, C.Z.; funding acquisition, C.Z. All authors have read and agreed to the published version of the manuscript. Author ORCIDs Junhong Dong ® hittps://orcid.org/0000-0001-8740-0805 Qiaohua Deng ® https://orcid.org/0009-0002-4683-2702 Minglan Chen © https://orcid.org/0009-0006-7890-2604 Daxiang Chen © https://orcid.org/0009-0001-2309-1991 Chungin Zhou © https://orcid.org/0009-0004-8117-1537 Changlin Zhao ® https://orcid.org/0000-0002-8668-1075 Data availability All of the data that support the findings of this study are available in the main text. 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