Four new species of Entoloma subgen. Cubospora (Entolomataceae, Agaricales) from Yunnan Province, China

331 Four new species of Entoloma subgen. Cubospora (Entolomataceae, Agaricales) from Yunnan Province, China Ze-Wei Liu1,2,3 , Jing Li4, Zhu L. Yang1,2 , Yang-Yang Cui1,2 1 Key Laboratory of Phytochemistry and Natural Medicines, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China 2 Yunnan Key Laboratory for Fungal Diversity and Green Development, Kunming 650201, China 3 University of Chinese Academy of Sciences, Beijing 100049, China 4 Faculty of Geography, Yunnan Normal University, Kunming 650500, China Corresponding authors: Yang-Yang Cui ([email protected]); Zhu L. Yang ([email protected]) Copyright: © Ze-Wei 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 Species of Entoloma are exceptionally diverse and widely distributed, with cuboid-spored members occurring mainly in subtropical to tropical regions. These cuboid-spored taxa are primarily classified into two subgenera, E. subg. Cuboeccilia and E. subg. Cubospora. During surveys conducted in Yunnan Province, a representative subtropical region of China, four new species belonging to E. subg. Cubospora were identified through multigene phylogenetic analyses (ITS, LSU, rpb2, and tef1) combined with detailed morphological observations. Entoloma acutiflavum is distinguished by its yellow basidiomata, papillate pileus, heterogeneous lamellar edge, and the presence of clamp connections. Entoloma bichromum can be recognized by a centrally papillate pileus, serrate pileal and lamellar margins, and clavate to cylindrical pleurocystidia that are flexuous, furcate, or occasionally ventricose. Entoloma guttuliferum is characterized by its yellowish-white to white papillate pileus, clavate to elongated clavate terminal cells in both the pileipellis and stipitipellis, and pigmented cystidia, lamellar trama, pileipellis, and stipitipellis. Entoloma rufosquamulosum is identified by its reddish-brown to grayish-violet squamose pileus, with reddish-brown pigments present in the upper hyphal layer and terminal cells of the pileipellis. Detailed descriptions, line drawings, habitat photographs, SEM micrographs, and comparisons with morphologically and phylogenetically related taxa are provided. A key to the species of Entoloma subg. Cubospora from China is also included. Key words: Cuboid spores, new taxa, phylogeny, subtropical, taxonomy Introduction Species of Entoloma are globally distributed and abundant in both temperate and tropical regions, with some species even extending into alpine and frigid zones (Largent 1977, 1994; Romagnesi and Gilles 1979; Horak 1980, 2008; Baroni 1981; Noordeloos 1988, 1992, 2004; Gates and Noordeloos 2007; Noordeloos and Hausknecht 2007; Gates et al. 2009; Noordeloos and Gates 2009, 2012; He 2012; Karstedt et al. 2024). To date, more than 2,000 species have been described. Despite their remarkable morphological diversity, all Entoloma species produce pink spore prints, and their basidiospores are angular in all orientations (Co-David et al. 2009; Noordeloos et al. 2018). Academic editor: Bao-Kai Cui Received: 9 September 2025 Accepted: 24 October 2025 Published: 11 November 2025 Citation: Liu Z-W, Li J, Yang ZL, Cui Y-Y (2025) Four new species of Entoloma subgen. Cubospora (Entolomataceae, Agaricales) from Yunnan Province, China. MycoKeys 124: 331–356. https://doi. org/10.3897/mycokeys.124.171541 MycoKeys 124: 331–356 (2025) DOI: 10.3897/mycokeys.124.171541 332 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Among angular-spored Entoloma species, those with cuboid basidiospores are relatively easy to recognize. Historically, however, taxa with this character were inconsistently classified into different subgenera or sections based solely on morphology—such as sect. Staurospora within subg. Nolanea or subg. Inocephalus (Noordeloos 1992; Largent 1994). To clarify their phylogenetic positions, Karstedt et al. (2019) conducted extensive sampling and demonstrated that these taxa form two distinct subgenera, subg. Cuboeccilia and subg. Cubospora, as supported by both molecular data and morphological features of basidiomata and cystidia. In recent decades, numerous cuboid-spored species have been discovered across tropical and subtropical regions. To date, approximately 190 such species have been reported worldwide, with only a few occurring in temperate zones (Horak 1976, 1977; Romagnesi and Gilles 1979; He et al. 2015a; Karstedt et al. 2019, 2024; Reschke et al. 2022a; Morozova and Pham 2023; Chen et al. 2024; Sato et al. 2025). In China, several new cuboid-spored species have been described from subtropical provinces such as Guangdong, Fujian, and Jiangxi (He et al. 2015a; Chen et al. 2024). Through field explorations in Yunnan Province, we discovered four novel species belonging to E. subg. Cubospora. These taxa exhibit distinct morphological characteristics and phylogenetic differentiation from all previously known species, as demonstrated by both morphological and multilocus analyses. Materials and methods Morphological studies Specimens collected between 2010 and 2023 in Yunnan Province, China, were deposited in the Herbarium of the Kunming Institute of Botany, Chinese Academy of Sciences (KUN-HKAS). Macroscopic characters were recorded and photographed from fresh basidiomata in the field. Color designations followed Kornerup and Wanscher (1978). Microscopic characters were examined under a ZEISS Axio Scope A1 light microscope. Sections of dried material were rehydrated in 5% KOH or H2O and stained with 1% (w/v) aqueous Congo red when necessary. Melzer’s reagent was used to test for spore amyloidity (Clémençon et al. 2004; Horak 2005). Line drawings were prepared freehand under a phase-contrast objective (1000×). At least 20 basidiospores were measured, and data are presented as (a/b/c) (d)e–f–g(h) × (i)j–k–l(m) μm [Q = (n)o–p(q), Q = r ± s]. Here, a–c indicates that a basidiospores were measured from b basidiomata of c specimens. Parameters d and h represent the minimum and maximum (5% extreme) values of length; e–g denote the central 90% range. The same applies to the width parameters (i–m) and the Q values (n–q). The mean length (f), width (k), and Q ratio (r), together with the standard deviation of Q (s), are also provided (Liu et al. 2022; Feng et al. 2025). Basidiospores were further examined using a Zeiss Sigma 300 scanning electron microscope (Oberkochen, Germany). DNA extraction, PCR amplification, and DNA sequencing Genomic DNA was extracted from dried basidiomata using the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech, Shanghai, China). Four nuclear loci were amplified: the internal transcribed spacer (ITS), the large subunit 333 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China ribosomal RNA (LSU), RNA polymerase II subunit 2 (rpb2), and translation elongation factor 1-alpha (tef1). PCR reactions (25 μL) contained 12.5 μL of 2× Taq Master Mix, 8.5 μL of nuclease-free water, 2 μL of primers, and 2 μL of DNA template. Primer pairs were ITS1F/ITS4 and LR0R/LR5 for ITS and LSU (Vilgalys and Hester 1990; Gardes and Bruns 1993), rpb2-i6f/rpb2-RhoR1 for rpb2, and EF1-983ClitoF1/EF1-1953ClitoR1 or EF1-983ClitoF4/EF1-1953ClitoR4 for tef1 (Kluting et al. 2014; Zhong 2019; Jian 2020). PCR conditions were as follows: for ITS, initial denaturation at 94 °C for 3 min; 35 cycles of 94 °C for 30 s, 53 °C for 40 s, and 72 °C for 1 min; and a final extension at 72 °C for 8 min. For LSU, the protocol was 94 °C for 3 min; 35 cycles of 94 °C for 30 s, 48 °C for 40 s, and 72 °C for 90 s; and a final extension at 72 °C for 8 min. Protocols for rpb2 and tef1 followed Zhong (2019) and Jian (2020), respectively. All sequencing and cloning were conducted by Sangon Biotech (Shanghai, China). Alignment and phylogenetic analysis Sequences of Entoloma subg. Cubospora were downloaded from GenBank and combined with sequences newly generated in this study. Three representative species from E. subg. Nolanea were selected as outgroups. The four gene regions (ITS, LSU, rpb2, and tef1) were aligned separately using MAFFT v7 (Katoh and Standley 2013), manually adjusted in BioEdit v7.2.5 (Hall 1999; Thompson et al. 1997), and concatenated into a single matrix. MODELTEST v2.3 was used to determine the best-fit substitution model for each gene based on the Akaike Information Criterion (Posada and Crandall 1998). Bayesian inference (BI) was performed using MrBayes implemented in PhyloSuite v1.2.3, with two independent runs of 60 million generations, sampling every 10,000 generations. The analysis was terminated when the average standard deviation of split frequencies dropped below 0.01, and the first 25% of trees were discarded as burn-in using the “sump” and “sumt” commands (Zhang et al. 2020). Maximum likelihood (ML) analysis was conducted using RAxMLGUI v2.0.5 with 1,000 bootstrap replicates under the GTRGAMMAI model (Edler et al. 2021). Results Phylogenetic relationships The combined dataset included 358 sequences representing 154 specimens and 68 species, comprising 53 newly generated and 305 retrieved from GenBank (Table 1). The concatenated alignment consisted of 3,254 characters. The best-fit models were GTR+I+G for ITS and LSU, SYM+I+G for rpb2, and HKY+I+G for tef1. After 60 million generations, the BI analysis reached convergence (average split frequency < 0.01). The BI and ML topologies were highly congruent; therefore, the BI tree is presented (Fig. 1). Entoloma subg. Cubospora formed a robust monophyletic clade (BS/Bayesian posterior probability [BPP] = 100/1.00). The four new taxa—E. acutiflavum, E. bichromum, E. guttuliferum, and E. rufosquamulosum—each formed well-supported, independent lineages within E. subg. Cubospora. No closely related species were identified for E. bichromum, E. guttuliferum, or E. rufosquamulosum, whereas E. acutiflavum was recovered as sister to E. submurrayi. 334 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Figure 1. Bayesian inference analysis of Entoloma subg. Cubospora based on ITS, LSU, rpb2, and tef1 sequence data. E. atropapillatum, E. cetratum, and E. sericeum serve as outgroups. Bootstrap values (BS) from maximum likelihood ≥ 75 and Bayesian posterior probabilities (BPP) ≥ 0.95 are shown on each branch (BS/BPP). The new species are marked in bold. E. dragonosporum FK2019 Brazil E. subcycneum HFJAU4738 Fujian China E. dragonosporum MC4600 Brazil E. cervinum FK0940 Brazil E. acutipallidum FK1893 Brazil E. azureoviride FK1123 Brazil E. luteobrunneum FK1693 Brazil holotype E. smurfetti FK1741 Brazil holotype E. smurfetti FK1709 Brazil E. smurfetti FK1717 Brazil E. gatesianum ACM498 Brazil holotype E. gatesianum ACM499 Brazil E. manausense ACM500 Brazil E. manausense FK2083 Brazil E. altissimum LE262945 Vietnam E. quadratum WU21098 France E. virescens MCA2479 Australia E. virescens MEL: 2379813 E. virescens DLL9972 Australia E. mengsongense HKAS 90774 Yunnan China holotype E. rufosquamulosum HKAS 69226 Yunnan China sp. nov. E. rufosquamulosum HKAS 130186 Yunnan China holotype sp. nov. E. rufosquamulosum HKAS 150125 Yunnan China sp. nov. E. rufosquamulosum HKAS 143019 Yunnan China sp. nov. E. rufosquamulosum HKAS 136639 Yunnan China sp. nov. E. plicatum DLL10083 Australia E. plicatum DLL9691 Australia E. plicatum DLL10091 Australia E. carneum LE262912 Vietnam E. carneum LE262954 Vietnam E. acutoconicum ZTMyc42856 Papua New Guinea E. phlebophyllum HFJAU4261 Fujian China holotype E. phlebophyllum HFJAU4263 Fujian China E. pallidoflavum LE262934 Vietnam E. laccarioides GDGM 26298 Guangdong China holotype E. luteum GDGM 27698 Hainan China E. tomentosum HFJAU5159 Fujian China holotype E. tomentosum HFJAU5160 Fujian China E. bichromum HKAS 150131 Yunnan China sp. nov. E. bichromum HKAS 150132 Yunnan China holotype sp. nov. E. sp. DLL9679 Australia E. sp. DLL9823 Australia E. neotropicale FK2016 Brazil E. neotropicale FK2130 Brazil holotype E. kermesinum KYO-HC59 Japan E. kermesinum TNS-F-82986 Japan E. kermesinum TNS-F-83014 Japan holotype E. quadratum PAN241 Panama E. albidoquadratum P. Manimohan 667 India holotype E. dragorufescens FK2102 Brazil holotype E. dragorufescens FK2116 Brazil E. dragoluteum FK2120 Brazil E. luteolamellatum MCA1480 Guyana holotype E. luteolamellatum 11RMT109 Brazil E. luteolamellatum FK1866 Brazil E. dennisii 8263 TJB USA E. caxiuanense FK1871 Brazil holotype Inocephalus squamulosus MCA1867 E. capes FK2096 Brazil holotype E. borbonicum WU21097 France holotype E. petchii GDGM 27696 Hainan China E. petchii HKAS 56716 Yunnan China E. petchii HKAS 122493 Yunnan China E. acutipallidum 11RMT078 Brazil E. dennisii CP47/04 Brazil E. atropapillatum FK0898 Brazil holotype E. cetratum LE311888 Sweden neotype E. sericeum KaiR237 Germany E. sericeum VHAs03/02 E. dragoluteum FK2131 Brazil holotype E. rufosquamulosum HKAS 150126 Yunnan China sp. nov. E. rufosquamulosum HKAS 150124 Yunnan China sp. nov. E. canoconicum PDD75649 New Zealand E. canoconicum ZTMyc42850 New Zealand E. luteum 6562 TJB USA E. luteum 7771 TJB USA E. aurantiovirescens KaiR623 Panama holotype E. aurantiovirescens PAN419 Panama E. murrayi SDR NAMA 2017-160 USA E. quadratum S.D. Russell ONT iNaturalist 136495142 USA E. quadratum iNAT: 16890676 USA E. luteum ACAD21101F Canada E. luteum ACAD21151F Canada E. quadratum 7794 TJB USA E. quadratum 8214 TJB USA E. quadratum EQ7695 USA E. voltavelhense FK1694 Brazil holotype E. voltavelhense FK2118 Brazil E. murrayi 8210 TJB USA E. murrayi VHAs0202 E. murrayi ECO-TA-HO 7874 Mexico E. quadratum GDGM 28953 Jiangxi China E. quadratum HFJAU4223 Fujian China E. quadratum HFJAU5173 Fujian China E. quadratum HFJAU4265 Fujian China E. quadratum HFJAU5179 Fujian China E. submurrayi HFJAU1050 Jiangxi China E. submurrayi HFJAU3587 Fujian China holotype E. acutiflavum HKAS 150127 Yunnan China sp. nov. E. acutiflavum HKAS 150128 Yunnan China holotype sp. nov. E. murrayi HKAS 52597 Yunnan China E. album KA12-1260 Korea E. murrayi QI 1001 Liaoning China E. murrayi QI 1002 Liaoning China E. quadratum HFJAU2527 Zhejiang China E. quadratum HFJAU2612 Zhejiang China E. quadratum LE254355 Russia E. flavescens TNS-F-82996 Japan E. flavescens TNS-F-82990 Japan E. flavescens TNS-F-82995 Japan holotype E. quadratum LE253781 Russia E. excavatum HFJAU2013 Zhejiang China holotype E. excavatum HFJAU4774 Zhejiang China E. murrayi MHHNU 30602 Hunan China E. rufomarginatum HFJAU1933 Zhejiang China holotype E. rufomarginatum HFJAU4070 Zhejiang China E. lacticolor HFJAU3736 Fujian China holotype E. lacticolor HFJAU3737 Fujian China E. guttuliferum HKAS 107829 Yunnan China sp. nov. E. guttuliferum HKAS 150130 Yunnan China sp. nov. E. semilanceatum NS2283 Cameroon E. guttuliferum HKAS 150129 Yunnan China holotype sp. nov. E. album KA12-1315 Korea E. hochstetteri TL2570 New Zealand E. hochstetteri TL2573 New Zealand E. hochstetteri ZTMyc42838 New Zealand E. hochstetteri ZTMyc42841 New Zealand E. canoconicum ZTMyc42846 New Zealand E. latericolor ZTMyc42832 New Zealand E. cervinum FK1770 Brazil Inocephalus “argenteus” MCA1475 E. subcycneum HFJAU3124 Fujian China holotype E. cycneum LE F-343654 Vietnam holotype E. cycneum LE F-343655 Vietnam E. peristerinum LE F-343650 Vietnam E. peristerinum LE F-343653 Vietnam holotype E. mocamboense FK1899 Brazil holotype E. caribaeum FK1790 Brazil E. tenue FK1922 Brazil E. procerum ME Noordeloos 2004070 Australia E. procerum PDD75517 New Zealand E. procerum ZTMyc42821 New Zealand E. amazonicum 11RMT126 Brazil E. amazonicum FK1815 Brazil holotype E. vinososquamulosum FK1745 Brazil holotype E. paulense FK0821 Brazil E. paulense FK1151 Brazil holotype 100/1 --/-- --/-- --/-- --/0.995 --/0.995 100/1 100/1 100/1 --/1 --/-- --/-- --/-- 100/1 --/-- 100/1 --/-- 89/1 90/0.998 100/1 100/-- --/0.998 --/-- --/0.956 90/-- 99/0.993 95/0.989 96/-- --/-- 100/0.994 100/1 100/1 100/1 100/1 96/1 97/1 --/-- --/-- 92/1 100/1 100/1 99/1 100/1 100/1 E. arenicola FK1811 Brazil holotype E. arenicola FK2089 Brazil E. arenicola NMJ195 Brazil E. kovalenkoi LE312529 Vietnam holotype Rhodophyllus lactifluus 8753 TJB USA Rhodophyllus lactifluus TB7962 E. albogracile ZTMyc42855 Papua New Guinea --/-- 100/1 100/1 --/-- --/-- --/-- --/-- 98/-- --/-- 98/1 100/1 94/-- 100/1 --/-- --/0.965 97/0.983 --/-- --/-- --/-- --/-- --/-- 98/1 99/1 100/1 97/0.996 99/-- 95/-- 0.05 --/0.973 91/0.999 --/-- 82/0.944 95/-- 75/-- 76/0.957 --/-- --/-- --/-- --/-- --/-- --/-- --/-- --/-- --/0.959 --/-- 79/9.992 100/1 98/1 --/-- --/-- --/-- --/-- --/-- 84/-- 95/-- 100/-- 100/-- 100/-- 97/-- 100/0.951 --/-- --/-- --/-- --/-- 100/0.988 --/-- 83/0.986 99/-- 100/1 --/-- --/-- 100/0.993 --/-- 99/-- --/-- --/-- --/0.991 --/-- --/-- --/-- 76/0.971 98/1 88/0.959 89/-- 98/0.999 100/1 98/1 86/1 --/-- --/-- 99/-- 88/-- --/-- 100/-- --/-- --/-- --/-- --/-- --/-- --/-- 100/1 100/0.999 --/-- 100/1 --/-- 100/1 99/0.929 --/-- 335 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Table 1. Specimens used in phylogenetic analysis and GenBank accession numbers. No. Species Specimen voucher GenBank accession numbers Locality Reference ITS LSU rpb2 tef1 1. Entoloma acutiflavum HKAS 150127 PX269826 PX269836 PX255550 PX255527 China in this study 2. E. acutiflavum HKAS 150128, holotype PX269825 PX269837 PX255549 PX255526 China in this study 3. E. acutipallidum 11RMT078 –MW624792 MW624730 –Brazil Karstedt et al. 2024 4. E. acutipallidum FK1893 –MG018325 –MH190147 Brazil Karstedt et al. 2019 5. E. acutoconicum ZTMyc42856 –MW624791 – – Papua New Guinea Karstedt et al. 2024 6. E. albidoquadratum P. Manimohan 667, holotype –GQ289151 GQ289223 –India Co-David et al. 2009 7. E. albogracile ZTMyc42855 –MH190207 – – Papua New Guinea Karstedt et al. 2019 8. E. album KA12-1260 KR673507 –––Korea Kim et al. 2015 9. E. album KA12-1315 KR673546 –––Korea Kim et al. 2015 10. E. altissimum LE262945 MF476912 MW624793 MW624731 –Vietnam Karstedt et al. 2024 11. E. amazonicum 11RMT126 –MW624794 MW624732 MW624839 Brazil Karstedt et al. 2024 12. E. amazonicum FK1815, holotype –MW624795 – – Brazil Karstedt et al. 2024 13. E. arenicola FK1811, holotype –MW624796 MW624733 –Brazil Karstedt et al. 2024 14. E. arenicola FK2089 –MW624797 MW624734 MW624840 Brazil Karstedt et al. 2024 15. E. arenicola NMJ195 –MW624799 MW624735 MW624841 Brazil Karstedt et al. 2024 16. E. atropapillatum FK0898, holotype KF679354 KF738940 MH190107 MH190137 Brazil Karstedt et al. 2019, 2020 17. E. aurantiovirescens KaiR623, holotype MZ611665 –––Panama Reschke et al. 2022a 18. E. aurantiovirescens PAN419 MZ611691 –––Panama Reschke et al. 2022a 19. E. azureoviride FK1123 –MW624830 MW624752 MW624855 Brazil Karstedt et al. 2024 20. E. bichromum HKAS 150131 PX269814 PX269835 PX255539 PX255531 China in this study 21. E. bichromum HKAS 150132, holotype PX269813 PX269834 PX255538 PX255530 China in this study 22. E. borbonicum WU21097, holotype –MH190198 MH190131 MH190166 France Karstedt et al. 2019 23. E. canoconicum PDD75649 –MW624800 – – New Zealand Karstedt et al. 2024 24. E. canoconicum ZTMyc42846 –MW624801 – – New Zealand Karstedt et al. 2024 25. E. canoconicum ZTMyc42850 –MW624802 MW624736 –New Zealand Karstedt et al. 2024 26. E. capes FK2096, holotype –MW624803 MW624737 –Brazil Karstedt et al. 2024 27. E. caribaeum FK1790 –MH190214 MH190114 MH190146 Brazil Karstedt et al. 2019 28. E. carneum LE262912 –MH190181 MH190119 MH190152 Vietnam Karstedt et al. 2019 29. E. carneum LE262954 –MH190184 MH190121 –Vietnam Karstedt et al. 2019 30. E. caxiuanense FK1871, holotype –MW624804 – – Brazil Karstedt et al. 2024 31. E. cervinum FK0940, holotype – – MG018332 MH190138 Brazil Karstedt et al. 2019 32. E. cervinum FK1770 –MW624805 – – Brazil Karstedt et al. 2024 33. E. cetratum LE311888, neotype OL338280 –OL405215 OL405538 Sweden Reschke et al. 2022b 34. E. cycneum LE F-343654, holotype OQ779461 OQ804518 –OQ779183 Vietnam Morozova and Pham 2023 35. E. cycneum LE F-343655 OQ779463 OQ804519 –OQ779182 Vietnam Morozova and Pham 2023 36. E. dennisii 8263 TJB –MH190195 MH190128 MH190164 USA Karstedt et al. 2019 37. E. dennisii CP47/04 –MH190210 – – Brazil Karstedt et al. 2019 38. E. dragoluteum FK2120 –MW624807 MW624739 MW624842 Brazil Karstedt et al. 2024 39. E. dragoluteum FK2131, holotype –MW624806 MW624738 –Brazil Karstedt et al. 2024 40. E. dragonosporm FK2019 –MH190179 MG018336 MH190150 Brazil Karstedt et al. 2019 41. E. dragonosporm MC4600 –MH190186 MH190122 MH190156 Brazil Karstedt et al. 2019 336 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China No. Species Specimen voucher GenBank accession numbers Locality Reference ITS LSU rpb2 tef1 42. E. dragorufescens FK2102, holotype –MW624810 MW624740 MW624843 Brazil Karstedt et al. 2024 43. E. dragorufescens FK2116 –MW624811 MW624741 –Brazil Karstedt et al. 2024 44. E. excavatum HFJAU2013, holotype PP796416 PP789602 – – China Chen et al. 2024 45. E. excavatum HFJAU4774 PP796431 PP789614 – – China Chen et al. 2024 46. E. flavescens TNS-F-82990 ITS1:LC786658 ITS2:LC786709 KF723693 KF723647 –Japan Sato et al. 2025 47. E. flavescens TNS-F-82995, holotype ITS1:LC786663 ITS2:LC786714 KF723691 KF723645 –Japan Sato et al. 2025 48. E. flavescens TNS-F-82996 ITS1:LC786664 ITS2:LC786715 KF723692 KF723646 –Japan Sato et al. 2025 49. E. gatesianum ACM498, holotype –MW624812 MW624742 MW624844 Brazil Karstedt et al. 2024 50. E. gatesianum ACM499 –MW624813 – – Brazil Karstedt et al. 2024 51. E. guttuliferum HKAS 107829 PX269816 PX269840 PX255541 –China in this study 52. E. guttuliferum HKAS 150129, holotype PX269815 PX269838 PX255540 PX255528 China in this study 53. E. guttuliferum HKAS 150130 PX269817 PX269839 PX255542 PX255529 China in this study 54. E. hochstetteri TL2570 KP191939 KP191755 – – New Zealand Unpublished 55. E. hochstetteri TL2573 KP191941 KP191758 – – New Zealand Unpublished 56. E. hochstetteri ZTMyc42838 –MW624814 – – New Zealand Karstedt et al. 2024 57. E. hochstetteri ZTMyc42841 –OP836300 – – New Zealand Karstedt et al. 2024 58. E. kermesinum KYO-HC59 ITS1:LC786696 ITS2:LC786749 –––Japan Sato et al. 2025 59. E. kermesinum TNS-F-82986 ITS1:LC786654 ITS2:LC786705 –––Japan Sato et al. 2025 60. E. kermesinum TNS-F-83014, holotype ITS1:LC786677 ITS2:LC786729 –––Japan Sato et al. 2025 61. E. kovalenkoi LE312529, holotype OK257210 OK257207 –OK256169 Vietnam Crous et al. 2021 62. E. laccarioides GDGM 26298 –JQ993091 – – China He et al. 2013 63. E. lacticolor HFJAU3736, holotype OR683793 OR687490 OR738710 OR699451 China Chen et al. 2024 64. E. lacticolor HFJAU3737 OR683794 OR687491 OR738711 OR699452 China Chen et al. 2024 65. E. latericolor ZTMyc42832 –MW624815 –MW624845 New Zealand Karstedt et al. 2024 66. E. luteobrunneum FK1693, holotype –MW624816 –MW624846 Brazil Karstedt et al. 2024 67. E. luteolamellatum 11RMT109 –MH190170 MH190105 –Brazil Karstedt et al. 2019 68. E. luteolamellatum FK1866 –MW624817 MW624743 –Brazil Karstedt et al. 2024 69. E. luteolamellatum MCA1480 holotype –MH190213 MH190135 MG702644 Guyana Karstedt et al. 2019 70. E. luteum 6562 TJB –KR869944 – – USA Largent et al. 2016 71. E. luteum 7771 TJB –MH190212 MH190125 MH190161 USA Karstedt et al. 2019 72. E. luteum ACAD21101F OM716995 –––Canada Unpublished 73. E. luteum ACAD21151F ON412813 –––Canada Unpublished 74. E. luteum GDGM 27698 JQ281486 JQ320121 – – China He et al. 2012, 2013 75. E. manausense ACM500 –MW624818 MW624744 MW624847 Brazil Karstedt et al. 2024 76. E. manausense FK2083 –MW624819 MW624745 –Brazil Karstedt et al. 2024 77. E. mengsongense HKAS90774, holotype KU131556 –––China Ediriweera et al. 2017 78. E. mocamboense FK1899, holotype –MW624820 –MW624848 Brazil Karstedt et al. 2024 79. E. murrayi 8210 TJB –MH190193 MH190127 –USA Karstedt et al. 2019 80. E. murrayi ECO-TA-HO 7874 MF156254 –––Mexico Hernández 2017 81. E. murrayi HKAS 52597 –KJ648469 – – China He 2012 82. E. murrayi MHHNU 30602 MK250917 – – –China Chen and Zhang 2019 337 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China No. Species Specimen voucher GenBank accession numbers Locality Reference ITS LSU rpb2 tef1 83. E. murrayi QI 1002 KJ658968 JQ993089 JQ993082 –China He et al. 2013 84. E. murrayi QI 1001 KJ658967 JQ993090 JQ993081 –China He et al. 2013 85. E. murrayi SDR NAMA 2017-160 MK575459 –––USA Unpublished 86. E. murrayi VHAs0202 –GU384620 GU384637 –Baroni et al. 2011 87. E. neotropicale FK2016 –MW624822 MW624746 MW624850 Brazil Karstedt et al. 2024 88. E. neotropicale FK2130, holotype –MW624825 MW624748 MW624853 Brazil Karstedt et al. 2024 89. E. pallidoflavum LE262934 OQ779469 MH190183 MH259314 MH190155 Vietnam Karstedt et al. 2019 90. E. paulense FK0821 –MW624826 MW624749 –Brazil Karstedt et al. 2024 91. E. paulense FK1151, holotype –MW624827 MW624750 –Brazil Karstedt et al. 2024 92. E. peristerinum LE F-343650 OQ779467 OQ804524 –OQ779186 Vietnam Morozova and Pham 2023 93. E. peristerinum LE F-343653, holotype OQ779466 OQ804522 –OQ779188 Vietnam Morozova and Pham 2023 94. E. petchii GDGM 27696 –JX992853 – – China He 2012 95. E. petchii HKAS 56716 JQ281485 JQ320120 – – China He et al. 2012 96. E. petchii HKAS 122493 ON794324 –––China Unpublished 97. E. phlebophyllum HFJAU4261, holotype OR827447 OR825714 OR827308 OR827307 China Chen et al. 2024 98. E. phlebophyllum HFJAU4263 OR827448 –––China Chen et al. 2024 99. E. plicatum DLL9691 –JQ624610 JQ624617 –Australia Largent et al. 2013 100. E. plicatum DLL10083 –JQ624612 JQ624619 MG702626 Australia Largent et al. 2013; Karstedt et al. 2019 101. E. plicatum DLL10091 –JQ624613 JQ624620 –Australia Largent et al. 2013 102. E. procerum ME Noordeloos 2004070 –GQ289183 GQ289254 –Australia Co-David et al. 2009 103. E. procerum PDD75517 –MH190189 – – New Zealand Karstedt et al. 2019 104. E. procerum ZTMyc42821 –MH190201 –MH190167 New Zealand Karstedt et al. 2019 105. E. quadratum 7794 TJB –MH190192 MH190126 –USA Karstedt et al. 2019 106. E. quadratum 8214 TJB –MH190194 –MH190162 USA Karstedt et al. 2019 107. E. quadratum EQ7695 –AF261303 – – USA Moncalvo et al. 2002 108. E. quadratum GDGM 28953 –KJ648471 KP226183 –China He et al. 2015a, b 109. E. quadratum HFJAU2527 PP796418 PP789604 PP873244 PP873227 China Chen et al. 2024 110. E. quadratum HFJAU2612 PP796419 PP789605 –PP873228 China Chen et al. 2024 111. E. quadratum HFJAU4223 PP796428 PP789611 PP873249 PP873233 China Chen et al. 2024 112. E. quadratum HFJAU4265 PP796429 PP789612 PP873250 PP873234 China Chen et al. 2024 113. E. quadratum HFJAU5173 PP796437 PP789620 PP873256 PP873240 China Chen et al. 2024 114. E. quadratum HFJAU5179 PP796438 PP789621 PP873257 PP873241 China Chen et al. 2024 115. E. quadratum iNAT:16890676 ON366783 –––USA Unpublished 116. E. quadratum LE253781 –MH190180 MH190118 –Russia Karstedt et al. 2019 117. E. quadratum LE254355 KC898452 KC898504 – – Russia Morozova et al. 2014 118. E. quadratum PAN241 MZ611690 –––Panama Reschke et al. 2022b 119. E. quadratum S.D. Russell ONT iNaturalist 136495142 OP749675 –––USA Unpublished 120. E. quadratum WU21098 –MW624828 – – France Karstedt et al. 2024 121. E. rufomarginatum HFJAU1933, holotype PP796415 PP789601 – – China Chen et al. 2024 122. E. rufomarginatum HFJAU4070 PP883966 PP789608 – – China Chen et al. 2024 123. E. rufosquamulosum HKAS 69226 PX269824 PX269831 – – China in this study 124. E. rufosquamulosum HKAS 130186, holotype PX269818 PX269828 PX255543 PX255532 China in this study 125. E. rufosquamulosum HKAS 136639 PX269822 PX269833 PX255547 PX255536 China in this study 338 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China No. Species Specimen voucher GenBank accession numbers Locality Reference ITS LSU rpb2 tef1 126. E. rufosquamulosum HKAS 143019 PX269823 PX269832 PX255548 PX255537 China in this study 127. E. rufosquamulosum HKAS 150124 PX269821 PX269830 PX255546 PX255535 China in this study 128. E. rufosquamulosum HKAS 150125 PX269820 PX269827 PX255545 PX255534 China in this study 129. E. rufosquamulosum HKAS 150126 PX269819 PX269829 PX255544 PX255533 China in this study 130. E. semilanceatum NS2283 MN069544 –––Cameroon Largent et al. 2020 131. E. sericeum KaiR237 OL338118 OL338542 OL405220 –Germany Reschke et al. 2022b 132. E. sericeum VHAs03/02 DQ367430 DQ367423 DQ367435 DQ367428 Hofstetter et al. 2014 133. E. smurfetti FK1709 –MW624831 – – Brazil Karstedt et al. 2024 134. E. smurfetti FK1717 –MW624832 – – Brazil Karstedt et al. 2024 135. E. smurfetti FK1741, holotype –MW624829 MW624751 MW624854 Brazil Karstedt et al. 2024 136. E. sp. DLL9679 OR083035 KR233854 KR233915 –Australia Unpublished 137. E. sp. DLL9823 –KR233861 KR233920 –Australia Unpublished 138. E. subcycneum HFJAU3124, holotype PP796420 –PP873245 PP873229 China Chen et al. 2024 139. E. subcycneum HFJAU4738 PP796430 PP789613 – – China Chen et al. 2024 140. E. submurrayi HFJAU1050 MN622719 –––China Chen et al. 2024 141. E. submurrayi HFJAU3587, holotype PP796423 PP789606 –PP873230 China Chen et al. 2024 142. E. tenue FK1922 –MH190176 MH190115 –Brazil Karstedt et al. 2019 143. E. tomentosum HFJAU5159, holotype PP796434 PP789617 PP873253 PP873237 China Chen et al. 2024 144. E. tomentosum HFJAU5160 PP796435 PP789618 PP873254 PP873238 China Chen et al. 2024 145. E. vinososquamulosum FK1745, holotype –MW624833 – – Brazil Karstedt et al. 2024 146. E. virescens DLL9972 –KR869937 KR869957 MG702628 Australia Karstedt et al. 2019; Largent et al. 2016 147. E. virescens MCA2479 –GU384622 GU384640 MG702629 Australia Karstedt et al. 2019 148. E. virescens MEL:2379813 MF977981 ––– Unpublished 149. E. voltavelhense FK1694, holotype –MW624834 MW624753 MW624856 Brazil Karstedt et al. 2024 150. E. voltavelhense FK2118 –MW624835 MW624754 –Brazil Karstedt et al. 2024 151. Inocephalus “argenteus” MCA1475 –GU384619 GU384636 –Baroni et al. 2011 152. I. squamulosus MCA1867 –GU384621 GU384638 –Baroni et al. 2011 153. Rhodophyllus lactifluus (GB as I. lactifluus) 8753 TJB –MH190196 MH190129 MH190165 USA Karstedt et al. 2019 154. R. lactifluus TB7962 –AF261304 – – Moncalvo et al. 2002 Taxonomy Entoloma acutiflavum Z.W. Liu, Y.Y. Cui & Zhu L. Yang, sp. nov. MycoBank No. 860479 Figs 2–4 Diagnosis. Pileus greyish yellow, with acuminate papilla at center. Basidiospores cuboid. Lamellar edge heterogeneous. Cheilocystidia elongated clavate, subcylindrical elongated clavate, or subcylindrical. Pileipellis with intracellular and incrusted pigment. Clamp connections present. Holotype. China • Yunnan Province: Baoshan, Longyang District, 25.2996°N, 98.7855°E, elev. 2200 m, in the litter layer of mixed broadleaf-conifer forest, 7 August 2022, Jin Li 085 (HKAS 150128). 339 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Etymology. Refers to the yellowish pileus with acuminate papilla at center. Description. Basidioma small to medium-sized. Pileus 28–42 mm in diam., conical, with acuminate papilla at center; greyish beige (4C2), light yellow (3A4) to greyish yellow (3C3, 4C5), margin concolorous or paler to yellowish white (2A2); striate sometimes shallowly sulcate, beige (4C3) or concolorous with pileus, towards the center up to 1/3 to 4/5 diam.; surface dry, smooth, occasionally hygrophanous, margin undulate; context thin. Lamellae sinuate, slightly crowded, 2–3 mm wide, with 1–3 tiers of lamellulae, narrowly ventricose, blond (4C4) or brownish orange (5C4), edge concolorous. Stipe 46–115 × 4–6 mm, central, cylindrical, usually tapering upwards, hollow; apex yellowish white (1A2) or greyish yellow (2C4, 3C3), sometimes downwards darker to beige (4C3); slightly with longitudinal or oblique striae, with white fibrils or smooth, base slightly swollen and with white mycelium. Odor and taste not observed. Basidiospores [69/2/2] (9.3)9.6–10.9–12.3(12.9) × (8.9)9.2–10.4– 12.1(12.7) μm [Q = 1.00–1.12(1.20), Q = 1.05 ± 0.04], cuboid, most with 4 angles in side-view, rarely with 5 angles, lacking elongated angles. Basidia 45–79 × 11–22 μm, clavate, 4-spored, colorless. Lamellar edge heterogeneous. Cheilocystidia 28–89 × 7–18 μm, elongated clavate or subcylindrical, occasionally ventricose, hyaline or with pale yellowish green droplet-like content, thin-walled. Pleurocystidia absent. Lamellar trama regular, made up of cylindrical hyphae 3–12 μm diam., smooth, colorless. Pileipellis a cutis composed of cylindrical and fusiform hyphae, 2–16 μm wide, thin-walled, with yellowish brown intracellular pigment, also sometimes incrusting. Stipitipellis composed of longitudinally arranged, cylindrical hyphae 3–16 μm wide; terminal cells clavate to elongated clavate, 37–76 × 7–12 μm; both hyphae and terminal cells in stipitipellis with colorless intracellular content. Clamp connections present in all tissue. Refractive hyphae sometimes occur in lamellar trama. Habitat. Solitary or scattered in the litter layer of mixed broadleaf-conifer forests. Known distribution. Yunnan Province, China. Additional material examined. China • Yunnan Province: Baoshan, Longyang District, 25.2996°N, 98.7855°E, elev. 2200 m, in the litter layer of mixed broadleaf-conifer forest, 7 August 2022, Xue-Ping Fan 502 (HKAS 150127). Notes. Entoloma acutiflavum is similar to E. submurrayi and E. murrayi morphologically; meanwhile, E. acutiflavum and E. submurrayi form a close and stable clade in the phylogenetic tree. Compared to E. submurrayi reported from subtropical regions of China, E. acutiflavum differs in its dry pileus with brownish and orangish tinges, white fibrils on the stipe, heterogeneous lamellar edge, and encrusted pigment on the pileipellis (Chen et al. 2024). Entoloma murrayi, a widespread species, can be distinguished by its absence of clamp connections and sterile lamellar edge (Selby et al. 1859; Horak 1976; He 2012). In addition, E. neotropicale and E. flavescens have yellow basidiomata that are similar to those of E. acutiflavum. However, E. neotropicale has an appressed-fibrillose pileus with a more obtuse papilla and indistinct striations on the pileus margin (Karstedt et al. 2024). Entoloma flavescens possesses a shiny pileus partially covered with whitish silky scales, shorter cheilocystidia, and an almost hyaline pileipellis (Sato et al. 2025). 346 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Among them, E. lacticolor is also similar to E. guttuliferum morphologically, but the former lacks pleurocystidia and has a sterile lamellar edge. Except for E. lacticolor, the other four species can be easily distinguished by the color of their basidiomata (Selby et al. 1859; Horak 1976; He 2012; Chen et al. 2024). Entoloma albogracile and E. dennisii share similarities with E. guttuliferum in Figure 6. Microscopic features of Entoloma guttuliferum (HKAS 150129, holotype). a. Basidiospores; b. Pleurocystidia; c. Basidia; d. Cheilocystidia; e. Pileipellis; f. Stipitipellis. Scale bars: 10 μm (a–d); 20 μm (e, f). 347 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China pileus color and shape. However, E. albogracile differs in having pink to porphyry-pink lamellae, pigmentless cheilocystidia and pileipellis, and the absence of clamp connections. Entoloma dennisii has smaller spores (5.5–7 μm) and pigmentless cheilocystidia (Horak 1976). In addition, E. album and E. caribaeum also resemble E. guttuliferum, although the color of their pilei tends to be white. Compared to the new species, they lack pleurocystidia. Furthermore, the hyphae in the pileipellis of E. album lack contents. Entoloma caribaeum has larger spores with elongated angles (15–18 × 15–20 μm) and 2-spored basidia (Karstedt et al. 2024; Sato et al. 2025). Entoloma rufosquamulosum Z.W. Liu, Y.Y. Cui & Zhu L. Yang, sp. nov. MycoBank No. 860482 Figs 2, 3, 7 Diagnosis. Pileus reddish with densely reddish brown or greyish violet squamae at center, becoming sparse towards margin. Basidiospores cuboid. Upper hyphae and terminal cells in pileipellis with reddish brown intracellular pigment. Holotype. China • Yunnan Province: Nujiang Lisu Autonomous Prefecture, Fugong County, elev. 1750 m, in the litter layer of evergreen broad-leaved forest, 14 August 2023, Zhu L. Yang 7052 (HKAS 130186). Etymology. Refers to the pileus with densely reddish squamae. Description. Basidioma small. Pileus 10–30 mm in diam., broadly conical to campanulate; violet brown (10E7) when young, becoming dull lilac (16C3), greyish magenta (13E4), or reddish brown (8E6) at center with age, disc paler to greyish lilac (16C2), greyish brown (8D3), reddish brown (8D4), or greyish ruby (12C3); striations none in most of specimens, occasionally present but indistinct, towards the center up to 1/3 to 3/4 diam.; surface dry, with densely reddish brown (8E7), greyish ruby (12E5) or greyish violet (18E5) squamae at center, becoming radial and gradually sparse towards margin, not hygrophanous, margin undulate, occasionally rimose and uplifted. Lamellae adnate to sinuate, crowded, 1–3 mm wide, with 1–3 tiers of lamellulae, narrowly ventricose, white to yellowish white (2A2), with pinkish tinged at age, edge serrate and concolorous. Stipe 22–114 × 3–6 mm, central, cylindrical, hollow, usually tapering upwards; greyish orange (5B3), reddish white (9A2), or greyish brown (8D3), sometimes with reddish brown (8D4) bruises; nearly smooth, sometimes with tiny granular squamules, with longitudinal or oblique striae, base slightly swollen and with white tomentum. Odor and taste not observed. Basidiospores [110/4/4] (9.0)9.4–10.3–11.2(11.8) × (8.3)8.9–9.7– 10.6(11.0) μm [Q = 1.00–1.14(1.18), Q = 1.06 ± 0.04], cuboid, most with 4 angles in side-view, rarely with 5 angles, lacking elongated angles. Basidia 37–59 × 12–18 μm, clavate, 4-spored, colorless. Lamellar edge heterogeneous. Cheilocystidia 21–98 × 6–13 μm, clavate to elongated clavate, sometimes flexuous or furcate, with pale yellowish green droplet-like content or hyaline, thin-walled. Pleurocystidia absent. Lamellar trama regular, made up of cylindrical hyphae 3–15 μm diam., smooth, thin-walled, sometimes with droplet-like content. Pileipellis a cutis composed of cylindrical hyphae 3–20 μm wide, thin-walled; terminal cells clavate or fusiform, 27–111 × 6–17 μm; both upper hyphae and terminal cells in pileipellis with reddish brown intracellular pigment, lower 348 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China hyphae with colorless intracellular content. Stipitipellis composed of longitudinally arranged, cylindrical hyphae 3–17 μm wide; terminal cells sparse, elongated cylindrical, 44–68 × 9–13 μm; both hyphae and terminal cells in stipitipellis with colorless intracellular content. Clamp connections present in all tissue. Refractive hyphae occasionally occur in lamellar trama and pileipellis. Habitat. Solitary or scattered in the litter layer of evergreen broad-leaved forest, coniferous forest dominated by Keteleeria fortunei, or mixed broadleaf-conifer forests. Known distribution. Yunnan Province, China. Additional materials examined. China • Yunnan Province: Tengchong, Zhonghe Town, elev. 1970 m, in the litter layer in coniferous forest dominated by Keteleeria fortunei, 13 August 2010, Yan-Jia Hao 242 (HKAS 69226); • Mingguang Town, 25.7456°N, 98.5594°E, elev. 2037 m, in the litter layer of broadleaf Figure 7. Microscopic features of Entoloma rufosquamulosum (HKAS 130186, holotype). a. Basidiospores; b. Basidia; c. Cheilocystidia; d. Pileipellis; e. Stipitipellis. Scale bars: 10 μm (a–c); 20 μm (d, e). 349 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China forest, 28 July 2022, Peng-Cheng Yuan 917 (HKAS 143019); • Diantan Town, 25.2936°N, 98.4506°E, elev. 2170–2220 m, in the litter layer of broadleaf-conifer forest, 27 July 2022, Xue-Ping Fan 316 (HKAS 136639); • Baoshan, Longyang District, 25.2996°N, 98.7855°E, elev. 2200 m, in the litter layer of broadleaf-conifer forest, 7 August 2022, Xue-Ping Fan 540 (HKAS 150126); • Longling County, 24.7789°N, 98.7931°E, elev. 1950 m, in the litter layer of broadleaf-conifer forest, 29 August 2022, Xue-Ping Fan 769 (HKAS 150125); • Longling County, 24.9478°N, 98.6108°E, elev. 2050 m, in the litter layer of broadleaf-conifer forest, 1 September 2022, Xue-Ping Fan 931 (HKAS 150124). Notes. Entoloma rufosquamulosum is characterized by its pileus with reddish or violet squamules and cuboid spores. E. rufosquamulosum forms a stable clade with E. bichromum, E. carneum, E. pallidoflavum, E. phlebophyllum, E. plicatum, and E. tomentosum in the phylogenetic tree. Among them, E. phlebophyllum resembles E. rufosquamulosum when young, but the former has a pileus with a depressed center at maturity, a sterile lamellar edge, and a pileipellis with encrusted pigment (Chen et al. 2024). Entoloma carneum has been reported to have a pale carneous pileus with a red center and radiating fibers, but it differs greatly from E. rufosquamulosum in microstructure, including smaller spores, the presence of pleurocystidia, and the absence of clamp connections (Bi et al. 1986). Entoloma bichromum, E. pallidoflavum, E. plicatum, and E. tomentosum can be easily distinguished from E. rufosquamulosum by the distinctly different colors of the pileus and stipe (Horak 1976; Largent et al. 2013; Chen et al. 2024). In addition, three species from Brazil—E. arenicola, E. vinososquamulosum, and E. voltavelhense—are similar to E. rufosquamulosum macroscopically. However, they have smaller spores or straw-yellow intracellular pigment in the pileipellis (Karstedt et al. 2024). Key to species of Entoloma subg. Cubospora from China 1 Pileus yellow, orange or brown .....................................................................2 – Pileus other colored ....................................................................................12 2 Pileus with obvious acute papilla at center .................................................3 – Pileus obtuse at the apex or depressed at center ....................................... 8 3 Lamellae edge with red-brown underlined .........Entoloma rufomarginatum – Lamellae edge without red-brown underlined .............................................4 4 Hyphae of pileipellis hyaline .........................................................................5 – Hyphae of pileipellis with pigment ...............................................................6 5 Clamps absent ................................................. .................. Entoloma murrayi – Clamps present ............................................................ Entoloma flavescens 6 Pileus rimose-fibrillose and orange .............................Entoloma quadratum – Pileus smooth and yellow .............................................................................7 7 Lamellae edge heterogeneous .................................. Entoloma acutiflavum – Lamellae edge sterile .................................................. Entoloma submurrayi 8 Pileus and stipe densely covered with scales ...................Entoloma petchii – Pileus and stipe glabrous, slightly fibrillose or partially pruinose ..............9 9 Spores less than 7.5 μm length ..................................................................10 – Spores more than 7.5 μm length ................................................................11 10 Cheilocystidia present ..........................................Entoloma cuboidosporum – Cheilocystidia absent .................................................Entoloma conspicuum 350 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China 11 Lamellae bright yellow, adnexed to sinuate ................Entoloma excavatum – Lamellae white to pale salmon, adnate to almost free .....Entoloma luteum 12 Pileus white to beige ...................................................................................13 – Pileus other colored ....................................................................................19 13 Lamellae short decurrent; cheilocystidia absent .......................................... ........................................................................Entoloma pseudogriseoalbum – Lamellae not short decurrent; cheilocystidia present ...............................14 14 Pleurocystidia present ................................................................................15 – Pleurocystidia absent .................................................................................. 16 15 Pileus plano-convex, depressed at center with age ...... Entoloma laccarioides – Pileus conical to campanulate, with papilla at center................................... .................................................................................... Entoloma guttuliferum 16 Spores with elongated angles .............................................Entoloma album – Spores without elongated angles ...............................................................17 17 Lamellae edge heterogeneous .....................Entoloma pseudotomentosum – Lamellae edge sterile ..................................................................................18 18 Pileus with obvious acute papilla at the center and more than 15 mm in diam.; hyphae of pileipellis without pigment .................Entoloma lacticolor – Pileus without obvious acute papilla at the center and less than 15 mm in diam.; hyphae of pileipellis with pale yellow encrusting pigment ................ ....................................................................................Entoloma subcycneum 19 Pileus red to purple .....................................................................................20 – Pileus blue to green .....................................................................................23 20 Pileus smooth .............................................................. Entoloma bichromum – Pileus squamulose or fibrillose ..................................................................21 21 Clamps absent ..................................................................Entoloma carneum – Clamps present ...........................................................................................22 22 Pileus depressed at center; lamellae edge sterile .....Entoloma phlebophyllum – Pileus not depressed at center; lamellae edge heterogeneous ................... ...........................................................................Entoloma rufosquamulosum 23 Hyphae of pileipellis with bluish intracellular pigment .............................24 – Hyphae of pileipellis with pale yellowish or stramineous intracellular pigment .............................................................................................................25 24 Pileus non-striate and fibrillose; lamellae crowded; spores smaller (7– 10.5 μm); cheilocystidia longer and narrower, cylindrical to clavate ........... .......................................................................................Entoloma altissimum – Pileus slightly striate and glabrous to weakly furfuraceous; lamellae distant to subdistant; spores larger (8–12.5 μm); cheilocystidia shorter and wider, broadly clavate .............................................Entoloma subaltissimum 25 Spores less than 8 μm length ...............................Entoloma mengsongense – Spores more than 8 μm length ...................................... Entoloma virescens Discussion Entoloma subg. Cubospora was established by Karstedt et al. (2019) to accommodate species characterized by cuboid basidiospores; mycenoid, collybioid, or tricholomatoid basidiomata; cylindrical-clavate or clavate cheilocystidia; and a trichodermal to cutis-like pileipellis. Later, Karstedt et al. (2024) expanded 351 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China the subgenus with numerous new taxa from Brazil and proposed seven subclades based on LSU, mtSSU, rpb2, and tef1 data, although two subclades were weakly supported. Chen et al. (2024) subsequently described seven additional species of E. subg. Cubospora from subtropical regions of China based on LSU, rpb2, and tef1 analyses. Incorporating these taxa, the present study confirms the monophyly and morphological coherence of E. subg. Cubospora as defined by Karstedt et al. (2019). Our phylogenetic results also lend partial support to the subdivision proposed by Karstedt et al. (2024), though subclade /Murrayi exhibited weak support (BS/BPP = 37/0.535). Based on pileus coloration, the presence of cuboid basidiospores with non-elongated angles, and phylogenetic placement, the four newly described species fit within subclade /Murrayi. However, E. acutiflavum, E. bichromum, and E. guttuliferum possess smooth to nearly smooth pilei, differing from the appressed-fibrillose pileus typical of that group (Karstedt et al. 2024). Further taxonomic resolution and subclade delimitation will require additional sampling and data. Many specimens exhibiting yellow, orange, or white basidiomata have been historically identified as E. murrayi, E. quadratum, or E. album. However, these names likely encompass multiple undescribed species. For instance, E. submurrayi (Chen et al. 2024) and E. acutiflavum (this study) were both collected from subtropical China, highlighting the region’s underexplored fungal diversity. Although the view that cuboid-spored species are primarily distributed in tropical to subtropical regions is widely accepted (Horak 1976, 1977; Romagnesi and Gilles 1979; He et al. 2015a; Reschke et al. 2022a; Chen et al. 2024), recent findings challenge this opinion. Sato et al. (2025) described temperate-zone taxa morphologically similar to E. murrayi and E. quadratum, supporting the view that cuboid-spored species also occur in temperate regions. This observation aligns with early records such as E. luteum from northwestern North America (Peck 1902), further emphasizing the ecological diversity of this distinctive lineage. Acknowledgements We extend our sincere gratitude to X.P. Fan, P.C. Yuan, Y.J. Hao, J. Li, F.F. Liu, and J.Y. Tang (Kunming Institute of Botany, Chinese Academy of Sciences) for their invaluable assistance during fieldwork. We also thank Mr. Zhi-jia Gu (Platform for Plant Multi-dimensional Imaging and Diversity Analysis, Kunming Institute of Botany, CAS) for the SEM technical support. Additional information Conflict of interest The authors have declared that no competing interests exist. Ethical statement No ethical statement was reported. Use of AI No use of AI was reported. 352 MycoKeys 124: 331–356 (2025), DOI: 10.3897/mycokeys.124.171541 Ze-Wei Liu et al.: Four new species of Entoloma subgen. Cubospora from Yunnan Province, China Funding This study was supported by the Yunnan Fundamental Research Projects (202501CF070090), the Yunnan Revitalization Talent Support Program “Young Talent” Project (XDYC-QNRC-2022-0509), the Caiyun Postdoctoral Program, and the “Talent Introduction Program” of the Kunming Institute of Botany, Chinese Academy of Sciences. 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