Additions to the Entoloma (Agaricales, Entolomataceae) from China: Description of five species with one new to science

89 Additions to the Entoloma (Agaricales, Entolomataceae) from China: Description of five species with one new to science Xu Liu1, Qiang Chen1, Quanju Xiang1, Yunfu Gu1, Xixi Han2,3,4, Rui Xing5,6, Ruilin Zhao2,7 , Maoqiang He1 1 College of Resource Sciences and Technology, Sichuan Agricultural University, Chengdu 611130, China 2 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China 3 Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand 4 School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand 5 Northwest Institute of Plateau Biology,Chinese Academy of Sciences, Xining,Qinghai, China 6 Qinghai Provincial Key Laboratory of Crop Molecular Breeding, Xining, Qinghai, China 7 College of Life Sciences, University of Chinese Academy of Sciences, Huairou District, Beijing, 100408, China Corresponding author: Maoqiang He ([email protected]) Copyright: © Xu Liu 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). Research Article Abstract Entoloma is one of the top three genera in species diversity of Agaricomycotina. In this study, five species from Qilian Mountain in China were documented. One new species E. griseopileum belonging to subg. Leptonia was introduced based on morphological and phylogenetic analyses. This new species is characterized by its gray basidiomes with fibrillose pileus and wood-decaying lifestyle. Four known species are E. glaucobasis, E. incanum, and E. verae in subg. Cyanula, and E. fuligineocinereum in subg. Nolanea. All five species are documented with descriptions, photo plates, and molecular data including ITS, LSU, and mtSSU sequences. Key words: Entolomataceae, new taxa, phylogeny, taxonomy Introduction Entoloma (Fr.) P. Kumm., typified by E. sinuatum (Bull.) P. Kumm., is one of the top three genera in species diversity of Agaricomycotina. It is estimated that there are more than 1800 species of Entoloma worldwide (He et al. 2019a). The general morphological characteristics of Entolomaare:basidiomes agaricoid, secotioid or gasteroid; agaricoid habit variable as tricholomatoid, mycenoid, collybioid, clitocyboid, omphalioid or pleurotoid; pink spore print or pinkish brown, lamellae almost free, adnexed to adnate, adnate-emarginate or adnate-decurrent, spores angular in all views which is entirely covered with facets delineated by ridges; clamp-connections often absent; cheilocystidia rare; pileipellis variable (cutis, trichoderm, hymeniderm) with diverse pigmentation(Noordeloos 1981; Largent 1986). Most species of this genus are saprotrophic, but there are also some mycorrhizal or weakly parasitic taxa. For example, members of sections Entoloma and Rhodopolia are ectomycorrhizal, and Rosaceae-associated members of section Nolanidea are considered weakly parasitic (Agerer and Waller 1993; Tedersoo et al. 2010). Academic editor: Thorsten Lumbsch Received: 19 June 2025 Accepted: 23 September 2025 Published: 10 October 2025 Citation: Liu X, Chen Q, Xiang Q, Gu Y, Han X, Xing R, Zhao R, He M (2025) Additions to the Entoloma (Agaricales, Entolomataceae) from China: Description of five species with one new to science. MycoKeys 123: 89–103. https://doi.org/10.3897/ mycokeys.123.162577 MycoKeys 123: 89–103 (2025) DOI: 10.3897/mycokeys.123.162577 90 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China A robust molecular-based taxonomic system for Entoloma is not established yet. Traditional taxonomic studies divided Entoloma into 13 subgenera based on morphological characteristics, such as basidiome forms, pileipellis, basidiospores, cystidia, and clamp connections (Noordeloos and Gates 2012a; Kaygusuz et al. 2024). Another opinion treated these subgenera as genera (Aime et al. 2010, Karstedt et al. 2020; Niveiro et al. 2021). Entoloma constitutes the dominant species diversity within Entolomataceae. The first comprehensive molecular phylogenetic study of Entolomataceae indicated there are two main clades of Entoloma, basal and crown clades. The basal clade is equal to Prunuloides clade and crown clade is composed of Rhodopoloid clade, Inocephalus-Cyanula clade, and Nolanea-Claudopus. Subgenus Leptonia is found to be polyphyletic and distributed in two different subclades in the crown clade (Co-David et al. 2009). In the past decades, most taxonomic studies of Entoloma are focused on American and European areas while species diversity of Entoloma in China is relatively poorly known. In recent years, species diversity of Entoloma in China has been explored.There are many new species of Entoloma introduced from China based on morphological and molecular data (He et al. 2012, 2013, 2015, 2019b; Chen et al. 2024, 2025). The Qilian Mountains, located between Qinghai and Gansu provinces in northwest China, is a critical biodiversity corridor connecting northern and southern China. During our macrofungi diversity surveys in Qilian Mountains, several notable samples of Entoloma were collected. In this study, based on morphological and multigene phylogenetic analyses, we identified and documented five Entoloma species including one new to science in subgenus Leptonia. Materials and methods Morphological examination Specimens were collected during the rainy season (July to August). Photographs were taken immediately in the field. Basidiomes were wrapped in aluminium foil or put in plastic boxes separately. Macro morphological characteristics and habit information were recorded when specimens were fresh. Every specimen was dried in an electrical food drier at 40 °C, then kept in a plastic ziplock bag and deposited in Herbarium MycologicumAcademiaeSinicae (HMAS). Anatomical and cytological characteristics were observed using dry specimens under the light microscope (SOPTOP EX33). Observed characteristics included basidiospores, basidia, cystidia and pileipellis. At least 20 measurements were made for each characteristic. Measurements were analyzed and recorded as X= the mean of length by width ± SD, Q = the quotient of basidiospore length to width, and Qm = the mean of Q values ± SD. All the protocols of morphological study followed Largent’s methodology (Largent 1986). DNA extraction and PCR Genomic DNA of each specimen was extracted by abroad-spectrum plant Rapid Genomic DNA Kitfollowed the manufacturer’s protocol. Three genes were applied in the phylogenetic analyses viz. internal transcribed spacer (ITS), large ribosomal subunit (LSU), and mitochondrial ribosomal small subunit (mtSSU). The primers for each gene are ITS4/ITS5, LR5/LROR, and NS1/NS4, respectively. 91 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China The PCR programs are followed describe in He et al. (2017). The PCR products were sequenced in Sangon Biotech (Shanghai) Co., Ltd. Both directions were sequenced to ensure accuracy of each gene. Phylogenetic analyses Information of reference and new generated sequences were listed in Suppl. material 1. New generated sequences were checked in Geneious Prime v.2025.0.3. Alignments were made using Muscle for each gene separately (Edgar 2004), then adjusted by hand to remove the ambiguous region. The dataset was made for genus Entoloma. The Entolomadataset includes 72ITS, 53LSU, and 43mtSSU. Maximum likelihood (ML) analysis and bootstrap values calculation were performed in raxmlGUI 1.5b1 with GTRGAMMA model with 1000 replicates (Silvestro and Michalak 2012). ModelFinder v2.2.0 (Kalyaanamoorthy et al. 2017) was used to select the best-fit partition model (Edge-linked) using AIC criterion. In the Entoloma dataset, Best-fit model according to AIC are GTR+F+I+G4 for ITS, GTR+F+I+G4 for LSU, and GTR+F+I+G4 for mtSSU. Bayesian Inference (BI) analysis was performed by MrBayes 3.1.2. Ten million generations were run for six Markov chains and sampled every 1000th generation. Burn-ins was determined in Tracer v1.6 with effective sample sizes (ESS) higher than 200 (http://tree.bio.ed.ac.uk/software/ tracer). Remaining trees were used to calculate Bayesian posterior probabilities (PP). Maximum likelihood (ML) analysis and bootstrap values calculation were performed in raxmlGUI 1.5b1 with GTRGAMMA model with 1000 replicates (Silvestro and Michalak 2012). Phylogenetic trees were presented in Fig. 1. Phylogenetic results The ML tree of Entoloma is presented in Fig. 1. There are 75 samples involved representing 48 species of Entoloma and one outgroup species Clitopilus hirneolus. These 48 species represent twelve subgenera of Entoloma. Samples collected from Qilian Mountains distributed in three subgenera, viz. subg. Cyanula, subg. Nolanea, subg. Leptonia. In the Cyanula clade, three samples from Qilian Mountains clustered in E. incanum, E. verae,and E. glaucobasis clades with full bootstrap values. Four samples clustered in E. fuligineocinereum clade in subg. Nolanea with full bootstrap and PP values. Three samples clustered in Entoloma and sister to the E. tallingiorum with the statistical values PP/BS = 1.0/99.Three Qinghai samples clustered together and sister to the clade composed by E. austriacum, E. alnetorum,and E. tjallingiorum with the statistical values PP/BS = 1.0/100. Taxonomy Entoloma fuligineocinereum Mešić & Tkalčec, Phytotaxa 289(3): 297 (2016) Fig. 2 Basionym. Nolanea latifolia Kauffman, Pap. Mich. Acad. Sci. 5: 140, 1926 [1925]. Description. Pileus is 23.0–32.2 mm in diameter, convex or broadly convex, occasionally umbonate on the disc, margin entire, occasionally straight; dry orhygrophanous, smooth, background gray or dark brown. Lamellae up to 3mm broad, adnate or slightly sinuate, light brown, brown, getting dark near the 92 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China E.lepidissimum PRM755801 Czech Republic T 0.07 E.violaceozonatum L275 Estonia T E.subaraneosum GDGM 28823 China T E.tjallingiorum UPS:BOT:F-016378 Sweden T E.violaceovillosum 645 India Kerala E.serrulatum LE254361 Russia Caucasus E.fuligineocinereum OMD LiNat 230202861 Canada Ontario E.fuligineocinereum iNAT 18337020 USA Washington E.kruticianum LE311767 Russia Europeanpart E.placidum UPS:BOT:F-121714 Sweden E.atricolor LE295001 Vietnam E.incanum ACD0182 USA Devil sLake E.percoelestinum LE254390 Spain T E.turbidum MEN200351 Slovakia E. fuligieneocinereum ZRL20234471 China Qinghai E.fuligineocinereum OMDLK CananMO 342838 USA Washington E.conferendum MEN200330 Slovakia E.glaucobasis FA4320 France Savoie E. verae ZRL20235682 China Qinghai E.verae KaiR 990 Germany Clitopilus hirneolus MEN199956 Italy E.bidupense LE262935 Vietnam T E.incanum FLAS F 69723 USA Kentucky E.atricolor LE295003 Vietnam E.bidupense LE262941 Vietnam E.chytrophilum L855 Spain Canary Islands T E.indutoides LE254354 Russia Leningrad Region E. fuligieneocinereum ZRL20234535 China Qinghai E.nitidum MEN200324 Slovakia E.alnetorum G-00111402 Switzerland T E.cetratum LE235480 Russia Leningrad Region E. griseopileum ZRL20230583 China Qinghai T E.callidermum GENT Malaysia E.atricolor LE295000 Vietnam T E.quadratum LE254355 Russia Far East E.fuligineocinereum O18044 Mongolia E.griseocyaneum LE254351 Russia Caucasus E.bulakhae LE253787 Russia Far East E.altissimum LE262945 Vietnam E. griseopileum ZRL20230545 China Qinghai E.verae HMAS293479 China E. glaucobasis ZRL20234505 China Qinghai E.bulakhae LE312416 Russia Far East E.dichroides LE F-343682 Vietnam E.clypeatum MEN198302 The Netherlands E.incanum HKAS49360 China E.sinuatum 8181-FTUR F inland E.undatum LE312417 Russia Novgorod Region E.atricolor LE295002 Vietnam E.tadungense LE F-343681 Vietnam E. incanum ZRL20234451 China Qinghai E.chalybeum LE254353 Russia Leningrad Region E.araneosum MEN200314 Belgium E.violaceoserrulatum JV8329F E.incanum OMD L iNat 172316324 USA Virginia E.callichroum ZT 71/58 Switzerland T E. griseopileum ZRL20230575 China Qinghai E.dichroum LE.227472 Russia European part E.sublacteisporum LIP-JVG-070823T Spain E.venustum E17/10 Germany T E.eugenei LE253771 Russia Primorsky Territory T E.euchroum LE262995 Russia Caucasus E. fuligieneocinereum ZRL20230695 China Qinghai E. fuligieneocinereum ZRL20230456 China Qinghai E.incanum LE312503 Sweden Ostersund T E.austriacum LE254343 Russia Far East E.glaucobasis Roth 16-6-95 Germany E.abortivum LE312419 Russia Far East E.lampropus UPS:BOT:F-176490 Sweden E.verrnum LE312418 Russia Novgorod Region E.allochromum 29-07-1973 The Netherlands T E.verae LE227613 Russia Samara Region T E.coelestinum LE258103 Russia Ural 1.0/96 -/75 -/100 0.9/100 0.9/100 -/100 -/92 1.0/100 1.0/99 -84 1.0/100 -/60 1.0/100 -/55 1.0/100 1.0/100 1.0/100 1.0/97 1.0/99 -/99 1.0/100 -/92 -/76 1.0/100 1.0/100 0.9/100 -/53 1.0/100 -/93 -/73 0.9/100 1.0/75 1.0/100 -/79 -/79 1.0/98 1.0/99 1.0/85 1.0/100 0.9/91 -/100 1.0/97 -/93 1.0/100 1.0/100 1.0/100 1.0/96 1.0/100 1.0/100 -/75 -/92 1.0/99 Subg. Cyanula Outgroup Subg. Calliderma Subg. Entolocybe Subg. Entoloma Subg. Pouzarella Subg. Leptonia Subg. Nolanea Subg. Griseorubida Subg. Inocephalus Subg. Violaceozonata Subg. Claudopus Figure 1. Maximum likelihood tree of Entoloma based on ITS, LSU, and mtSSU genes, rooted with Clitopilus hirneolus. Bayesian posterior probability (PP) values ≥ 90% or Bootstrap support (BS) values ≥ 50% are indicated at the internodes (PP/BS). New samples are in bold.The type strains cited indicated by a bold “T”. pileus edge, eroded,veined. Stipe 72.4–80.2 × 3.2–3.8 mm, concolor with the pileus, paler toward base, cylindrical,equal, surface dry, smooth, silky, occasionally with the white fibrils or punctate near the pileus. 93 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China Figure 2. Entoloma fuligineocinereum. a, b. Basidiomes in the field; c–g. Basidia; h, i. Basidiospores; j. Pileipellis hyphae; k, l. Lamellae edge. Scale bars: 5 mm (a, b); 5 μm (c–g); 10 μm (h–I); 40 μm (j); 5 μm (k, l). 94 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China Basidiospores 8.6–11.6 × 8.1–9.8 μm [x¯ =9.6 ± 0.7 × 8.7 ± 0.4 μm, Q = 1.0– 1.2, Qm = 1.1 ± 0.1, n = 20], subglobose-angular, asymmetrical, with 5–7 side. Basidia 40.7–47.9 × 11.2–13.2 μm, 4-spored. Cheilocystidia absent. Pileipellis a cutis with a transition to a plagiotrichoderm and a trichoderm of cylindrical to slightly inflated hyphae14.2–28.9 μm wide, with lightmembranous brown pigment. Clamp connections absent. Lamellae edge is fertile. Notes. Our samples clustered with samples from Northern America and Mongolia of E. fuligineocinereum in the phylogenetic tree (Fig. 1). Morphologically, the morphological characteristics of our samples differs slightly from the previous descriptions. Basidiomes color of our samples is paler than those described in Largent (1971). In addition, our Chinese samples present slightly larger basidiospores compared with the holotype (7.5–9 × 7–8.5 μm, x¯ = 8.3 × 7.7 μm, Largent 1971). Specimen examined. China • Qinghai province, Qilian Mountain National Park, Sigou, Donghaikou, 9 July 2023, MQ He, ZRL20230456 (HMAS300882) • Qinghai Province, Haibei Tibetan Autonomous Prefecture, Qilian County, Binggoulinhai, 15 July 2023, MQ He, ZRL20230695 (HMAS300880) • Qinghai Province, Tongren City, ZamaoKasuhu, 23 August 2023, MQ He, ZRL20234471 (HMAS300879) • Qinghai Province, Huangnan Tibetan Autonomous Prefecture, Maixiu Forest Farm, 22 August 2023, MQ He, ZRL20234535 (HMAS300881). Entoloma glaucobasis Huijsman ex Noordel., Persoonia 12(4): 260 (1985) Fig. 3 Description. Pileus 29–33.4 mm in diameter, campanulate, occasionally with a broad umbo, disc depressed or deeply indented, margin entire, straight, slightly uplifted; surfacedry, background gray or light brown, covered with brown fibrillose scales in triangular shape, scales denser and darker at the disc. Lamellae up to 4 mm broad, adnate to decurrent, subdistant, white, light pink, entire, veined. Stipe 48.1–59.6 × 2.3–2.5 cm, light brown, concolor with the pileus, surface dry, smooth, base white with hypha. Basidiospores 9.3–12.0 × 6.7–9.2 μm [x¯ =10.7 ± 0.7 × 8.1 ± 0.6 μm, Q = 1.2–1.6, Qm = 1.3 ± 0.1, n = 20], heterodiametric, with 5–7 angles in sideview. Basidia 28.5–38.6 × 9.8–13.6 μm, 4-spored. Cheilocystidia basidia-like, 13.2–19.8 × 3.7–7.4 μm.Pileipellis a cutis with a transition to a plagiotrichoderm and a trichoderm of cylindrical to slightly inflated hyphae 8.2–17.9 μm wide, hyaline, membranous light brown. Clamp connections absent. Lamellae edge is heterogeneous. Notes. A sample from Qilian Mountain clustered two European samples of E. glaucobasisin the phylogenetic tree (Fig. 1). This species is originally described from Germany. Our sample generally coincides with the original description of E. glaucobasis. Difference is that the cheilocystidia is basidia-like in our sample while is clavate or vesicular according to the original description (Noordeloos 1985). Specimen examined. China • Qinghai Province, Huangnan Tibetan Autonomous Prefecture, Maixiu Forest Farm, 22 August 2023, MQ He, ZRL20234505 (HMAS300878). 95 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China Entoloma griseopileum M.Q. He & X. Liu, sp. nov. Fungal Names: FN 572836 Fig. 4 Etymology. the epithet “griseo” means gray, the name refers to the grayish pileus. Holotype. China • Qinghai Province Mutual Aid County, Beishan National Forest Park,Langshidang, 10 July 2023, MQ He,ZRL20230583 (HMAS300884). Diagnosis. This species is characterized by its gray basidiomes and fibrillose pileus. Original description. Pileus is 26–32 mm in diameter, broadly convex, umbonate on the disc, margin slightly decurved when young, getting straight or slightly uplifted when mature; surface dry, fibrillose, background gray, fibrils light brown, denser and darker on the disc. Lamellae up to 3 mm broad, adnate or slightly sinuate, eroded, white, slightly pink, getting brown or dark brown near the pileus edge. Stipe 33.53–42.20 × 3.2–3.5 mm, concolor with the pileus, darker toward base, cylindrical,equal,surface dry, smooth, silky, occasionally with the same fibrils as on pileus. Figure 3. Entoloma glaucobasis. a. Basidiomes in the field; b. Pileipellis hyphae; c–f. Basidiospores; g–j. Basidia; k–i.Cheilocystidia. Scale bars: 5 mm (a); 10 μm (b); 5 μm (c–f); 5 μm (g–j); 5 μm (k, l). 96 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China Figure 4. Entoloma griseopileum. a–d. Basidiomes in the field; e–j. Basidiospores; k–o. Basidia; p. Pileipellis hyphae; q–r.Lamellae edge. Scale bars: 5 mm (a–d); 2 μm (e–j); 5 μm (k–o); 10 μm (p); 5 μm (q–r). 97 MycoKeys 123: 89–103 (2025), DOI: 10.3897/mycokeys.123.162577 Xu Liu et al.: Additions to the Entoloma from China Basidiospores 6.8–10.1 × 5.9–7.7 μm [x¯ =8.9 ± 0.7 × 6.7 ± 0.4 μm, Q = 1.0– 1.6, Qm = 1.3 ± 0.1, n = 20], heterodiametric, with 5–7 angles in side-view. Basidia 31.8–40.6 × 9.3–13.8 μm, 4-spored. Cheilocystidia absent. Pileipellis a cutis with a transition to a plagiotrichoderm and a trichoderm of cylindrical to slightly inflated hyphae 4.6–11.1 μm wide, with membranous brown pigment. Clamp connections absent. Lamellae edge is fertile. Habitat. Wood-inhabiting. Notes. Morphologically, E. griseopileum resembles E. bisporigerum in the field which also can be found in northern China (Li et al. 2015). Both species have grayish small basidiomes (pileus smaller than 4cm in diameter). But under microscope, E. bisporigerum can be distinguished from E. griseopileum by its larger basidiospores (11.5–13.5 × 7.5–9 μm). Furthermore, E. bisporigerum grows on soil, while E.griseopileum grows on wood. Other specimens examined. China • Qinghai Province, Beishan National Forest Park, Langshidang, 10 July 2023, MQ He, ZRL20230545 (HMAS300885), ZRL20230575 (HMAS300883). Entoloma incanum (Fr.) Hesler,Beih. Nova Hedwigia 23: 147 (1967) Fig. 5 Description. Pileus is 22.9–26.0 mm in diameter, broadly convex, disc depressed to deeply indented, margin entire;surfacesmooth or slightly with white fibrils, brown, yellowish brown, darker at the disc, getting paler or yellower towards the edge. Lamellae up to 3 mm broad, adnate to decurrent, subdistant, white, light yellow,entire,veined. Stipe 53.6 × 6.6 mm, yellow, or light greenish yellow, hollow,compressed, surface dry, smooth, base white with hypha. Basidiospores 9.8–13.6 × 6.3–8.4 μm [x¯ =11.6 ± 1.1 × 7.4 ± 0.6 μm, Q = 1.3–2.0, Qm = 1.6 ± 0.2, n = 20], heterodiametric, with 6–8 angles in side-view. Basidia 40.3–60.8 × 10.2–15.9 μm, 4-spored. Cheilocystidia absent. Pileipellis a cutis with a transition to a plagiotrichoderm and a trichoderm of cylindrical to slightly inflated hyphae 6.0–15.5 μm wide, with membranous light brown pigment. Clamp connections absent. Lamellae edge is fertile. Notes. Our Chinese samples clustered with northern America samples of E. incanum in the phylogenetic tree with full statistical values (Fig. 1). Entoloma incanum is a remarkable species in the field due to its greenish-brown or yellowish-brown basidiomes with a bright stipe in green or yellow. Our samples present a relatively browner pileus than American samples (Kuo 2013). Specimen examined. China • Qinghai Province, Tongren City, Zhamao Village, 23 August 2023, MQ He, ZRL20234451 (HMAS300876). Entoloma verae O.V. Morozova, Noordel., Reschke, F. Salzmann & Dima, Persoonia 47: 307 (2021) Fig. 6 Description. Pileus 21.4 mm in diameter, plane, disc deeply indented, margin entire, surfacedry, with distinct radial dark brown fibrils on pale background,sometimes cracked when dry or old, pileus edge dark brown. Lamellae