<8) MycoKeys MycoKeys 120: 173-192 (2025) DOI: 10.3897/mycokeys.120.156619 Research Article New species of Vamsapriya (Vamsapriyaceae, Xylariales) and Neolinocarpon (Linocarpaceae, Chaetosphaeriales) from Yunnan, China Xingyu Luo’, Changtao Lu’*, Kamran Habib’, Yulin Ren'”®, Yilan Jin’, Wenmei Li’?, Pingzhu Lu’, Yinggian Kang», Xiangchun Shen'2®, Hind A. AlShwaiman®, Abdallah M._ Elgorban”®, Nalin N. Wijayawardene®, Yang Chen", Qirui Li'?© 1 State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University, University Town, Guian New District, Guiyang, Guizhou province, 561113, China 2 The High Efficacy Application of Natural Medicinal Resources Engineering Center of Guizhou Province (The Key Laboratory of Optimal Utilization of Natural Medicine Resources), School of Pharmaceutical Sciences, Guizhou Medical University, University Town, Guian New District, Guiyang, Guizhou province, 561113, China Anshun University Agricultural College, Anshun, 561000, China 4 Chongqing Three Gorges Medical College, Wanzhou Wanzhou District, Chongqing, 404100, China 5 Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, School of Basic Medical Sciences, Guizhou Medical University, Gui’an New District, Guiyang, 561113, China 6 Add Department of Botany and Microbiology, College of Science, King Saud University, PO. 2455, Riyadh 11451, Saudi Arabia 7 Center of Excellence in Biotechnology Research (CEBR), King Saud University, Riyadh, Saudi Arabia 8 Center for Yunnan Plateau Biological Resources Protection and Utilization, College of Biology and Food Engineering, Qujing Normal University, Qujing, Yunnan 655011, China Corresponding authors: Qirui Li (Iqrnd2008@163.com); Yang Chen (36498787@qq.com) OPEN Qaceess Academic editor: Nattawut Boonyuen Received: 22 April 2025 Accepted: 5 July 2025 Published: 29 July 2025 Citation: Luo X, Lu C, Habib K, Ren Y, Jin Y, Li W, Lu P Kang Y, Shen X, Al-Shwaiman HA, Elgorban AM, Wijayawardene NN, Chen Y, Li Q (2025) New species of Vamsapriya (Vamsapriyaceae, Xylariales) and Neolinocarpon (Linocarpaceae, Chaetosphaeriales) from Yunnan, China. MycoKeys 120: 173-192. https://doi.org/10.3897/ mycokeys.120.156619 Copyright: © Xingyu Luo 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 During a survey of ascomycetes in southwestern China, three new bambusicolous fungi were collected and identified as members of the genera Neolinocarpon and Vamsapriya. The newly identified species are designated Neolinocarpon bambusicola sp. nov., Vam- sapriya tongluobaensis sp. nov., and V. zhaotongensis sp. nov., delineated through an analysis of both morpho-anatomical characteristics and multi-locus phylogenetic anal- yses based on ITS, LSU, rpb2, and tub2 sequence data. The distinction of these species from their known counterparts was verified through comparisons of morphological char- acters and phylogenetic analyses. Comprehensive morphological descriptions, illustra- tions, and a phylogenetic tree showing the placement of the new taxa are provided. Key words: Ascomycetes, bambusicolous fungi, fungal systematics, new species Introduction The genus Vamsapriya was introduced by Gawas and Bhat (2005) to accom- modate synnematous hyphomycetes colonizing bamboo in India, with V. indi- ca designated as the type species. Dai et al. (2017) described the first sexual morphs of Vamsapriya and placed the genus within Xylariaceae, as accepted by Hyde et al. (2020). Later, multi-locus phylogenetic analyses by Sun et al. (2021) revealed that Vamsapriya, along with its closely related genera Diabolocovidia 173 Xingyu Luo et al.: Three species from Yunnan, China and Didymobotryum, formed a distinct monophyletic clade within Xylariales, clearly separated from the Xylariaceae. This finding led to the establishment of the new family Vamsapriyaceae, with Vamsapriya designated as the type genus. The asexual morph of Vamsapriya is characterized by colonies on natural sub- stratum that are black, mycelium immersed, septate, branched, conidiophores macronematous, synnematous, erect, dark brown, cylindrical, synnemata with cylindrical to clavate apical fertile part composed of compactly arranged con- idiophores; conidiogenous cells monotretic, integrated, terminal, brown, cylin- drical to clavate; conidia catenate, acrogenous, cylindrical, broadly fusiform, or obclavate, and brown to dark-brown conidia (Sun et al. 2021). The sexual morph of the genus is characterized by black perithecial, immersed ascomata, sub-glo- bose, ostiolate, surrounded by dark brown peridium, 8-spored short pedicellate asci, each with a J+ apical ring and fusiform to broadly fusiform, apiosporous ascospores, hyaline, pointed at both ends, with or without a mucilaginous sheath (Sun et al. 2021). Fourteen species of Vamsapriya have been document- ed worldwide, but only eight species of the genus are known to possess a Sexu- al morph (Dai et al. 2017; Sun et al. 2021; Samarakoon et al. 2022; Dissanayake et al. 2024; Liu et al. 2025). Species of the genus are predominantly found in tropical and subtropical regions, with records from China, Cuba, India, and Thai- land, with the majority of species reported from Thailand and China. Ecological- ly, Vamsapriya species are primarily saprobes on decaying bamboo culms (Sun et al. 2021; Samarakoon et al. 2022; Dissanayake et al. 2024; Liu et al. 2025). The genus Neolinocarpon was introduced by Hyde (1992a) to accommodate many Linocarpon-like species, with N. globosicarpum as the type species. The genus is characterized by a clypeus with a dense, blackened, shiny central papil- la; deeply immersed, oval to globose ascomata; 8-spored unitunicate asci, each with a reflective apical ring; and filiform, fasciculate ascospores containing re- fringent bands, with or without appendages (Hyde 1992a, b; Hyde et al. 1998; Vitoria et al. 2013; Konta et al. 2017). Neolinocarpon is morphologically similar to Linocarpon, particularly in the apical structure of the ascus and ascospore morphology, but differs in having deeply immersed ascomata that form below a slightly raised or flattened clypeus, with a refractive globose body beneath the ascus apical ring (Hyde 1992b). Nearly all species of Neolinocarpon are found on hosts in the Arecaceae, except for N. penniseti and N. phayaoense, which occur on hosts from Poaceae and Euphorbiaceae, respectively (Hyde 1992b; Hyde et al. 1998; Hyde and Alias 1999; Bhilabutra et al. 2006; Vitoria et al. 2013; Senwanna et al. 2018). As of March 10, 2025, Index Fungorum lists 13 species in this genus (www.indexfungorum.org). During a mycological investigation of ascomycetes associated with decaying bamboo in Yunnan Province, Southwestern China, we found three novel saprobic bambusicolous fungi belonging to the genera Vamsapriya and Neolinocarpon, each with distinct morpho-anatomy that does not fit with any of the previously de- scribed species within their respective genera. Saprobic fungi play an important role in the natural breakdown of organic materials and often harbor biotechnolog- ically relevant enzymes and secondary metabolites with potential applications in agriculture, medicine, and environmental management (Corbu et al. 2023). This study provides the basis for future investigations into these fungi, which may lead to valuable applied research and potential biotechnological innovations. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 174 Xingyu Luo et al.: Three species from Yunnan, China Materials and methods Sample collection and morphological study The specimens were collected during surveys conducted in Yunnan Prov- ince in China from August to November in 2024. All related habitat informa- tion was recorded. Photographs of the collected materials were taken using a Canon G15 camera (Canon Corporation, Tokyo, Japan). Materials were placed in paper bags and taken to the lab for morphological characteriza- tion and isolation. The specimens were placed in a ventilated area at room temperature for drying. All specimens were deposited at the Herbarium of Guizhou Medical University (GMB) and the Herbarium of Cryptogams, Her- barium of Kunming Institute of Botany, Chinese Academy of Sciences (KUN HKAS). Living cultures were deposited at the Guizhou Medical University Cul- ture Collection (GMBC). All scientific names of fungi follow the entries in MycoBank and Index Fungorum; hence, no authorities and year of publication are given in the text. Morphological characterization and isolation Macroscopic characteristics were examined under an Olympus SZ61 stereomi- croscope (Japan) and photographed with a Canon 700D digital camera (Can- on Inc., Tokyo, Japan). The morphological features were studied as described by Li et al. (2024a). The samples were mounted in water for microscopic ex- amination; Melzer’s reagent was added when necessary. The Tarosoft Image Framework (v.0.9.7) program and Adobe Photoshop CS6 software (Adobe Sys- tems, USA) were used for measuring and processing images. Axenic cultures were obtained from single spores or tissues as described by Senanayake et al. (2015). Pure cultures on potato dextrose agar (PDA) were isolated using single-ascospore isolation techniques (Liu et al. 2025). The cultures were in- cubated at 25-30 °C for 4-6 weeks, with weekly observations of growth and development (Senanayake et al. 2020). DNA extraction, PCR amplification, and sequencing Mycelium was scraped from pure culture plates using a sterilized scalpel and used for DNA extraction following the manufacturer's instructions for the BIO- MIGA fungus genomic DNA extraction kit. The DNA samples were kept at —20 °C. Internal transcribed spacer region (ITS), large subunit (LSU), ribosomal small subunit (SSU), the second largest subunit of RNA polymerase II (rpb2), and B-tubulin (tub2) were amplified by PCR with primers ITS1/ITS4 (White et al. 1990; Gardes and Bruns 1993), LROR/LR5 (Vilgalys and Hester 1990), NS1/NS4 (White et al. 1990), fRPB2-5F/fRPB2-7cR (Liu et al. 1999; Bischoff et al. 2006), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The components of a 25 uL PCR mixture were 9.5 uL of double-distilled water, 12.5 tL of PCR Mas- ter Mix, 1 uL of each primer, and 1 uL of template DNA. The PCR amplification was conducted as reported by Samarakoon et al. (2022). PCR products were checked through 1.5% agarose gel electrophoresis stained with Golden View and were sent to Sangon Co., China, for sequencing. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 175 Xingyu Luo et al.: Three species from Yunnan, China Sequence alignments and phylogenetic analyses All the obtained sequences were compared with known sequences in Gen- Bank using the BLAST algorithm for precise identification (https://www.ncbi. nlm.nih.gov) (Altschul et al. 1990). The reference sequences retrieved from open databases originated from recently published data (Liu et al. 2025) and the BLASTn results of close matches. Sequences were aligned using the MAFFT v.7.110 online program (Katoh et al. 2019) with default settings. The alignments were adjusted manually using BioEdit v.7.0.5.3 (Hall 1999) where necessary. Maximum likelihood (ML) analyses were implemented in RAx- ML v.8.2.12 using the GTRGAMMA substitution model with 1,000 bootstrap replicates (Stamatakis 2014). Phylogenetic analyses were also performed for Bayesian inference in MrBayes v.3.2.2 (Ronquist et al. 2012) online. The Markov chain Monte Carlo (MCMC) sampling in MrBayes v.3.2.2 was used to determine the posterior probabilities (PP). Six simultaneous Markov chains were run for 1,000,000 generations, and trees were sampled every 1,000" generation. The first 25% of the trees were discarded as burn-ins. The re- mainder was used to calculate the posterior probabilities (PPs) for individual branches. The phylogenetic tree was visualized in FIGTREE v.1.4.4 (Rambaut 2018). All analyses were run on the CIPRES Science Gateway v.3.3 web por- tal (Miller et al. 2010). All the obtained sequences were deposited in Gen- Bank (Tables 1, 2). Results Phylogenetic analyses 1 The aligned dataset of Vamsapriyaceae (Fig. 1), comprising 3,080 characters (ITS/LSU/tub2/rpb2), contains 29 strains, including the newly generated se- quences and outgroup taxa. Barrmaelia oxyacanthae and B. rhamnicola were selected as outgroup taxa. The tree topology from the ML analysis was sim- ilar to that from the BI analysis. The best-scoring RAXML tree is presented in Fig. 1. The sequences of our specimen, Vamsapriya zhaotongensis, formed a well-supported (ML/BI: 100/1.00) clade in a sister relationship with V. sichuan- ensis. The lineage branch of this clade contains the second collection, Vam- sapriya tongluobaensis, which shows a close relationship with V. mucosa and V. kailiensis. Although this clade lacks robust support, it remained stable in re- peated phylogenetic analyses. Phylogenetic analyses 2 The aligned dataset of Linocarpaceae comprised 2,454 characters (ITS/ LSU/SSU) after exclusion of ambiguously aligned regions and long gaps. Sordaria fimicola and Gelasinospora tetrasperma were selected as outgroup taxa. In our phylogram (Fig. 2), the sequences of our collection, Neolino- carpon bambusicola, nested within the phylogenetic branch of the genus Neolinocarpon, formed a separate clade outside a group comprised of N. phayaoense and N. rechedis, demonstrating its status as an independent species. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 176 Xingyu Luo et al.: Three species from Yunnan, China Vamsapriya breviconidiophora MFLUCC 14-0436 82/089!“ Vamsapriya khunkonensis MFLUCC 13-0497" “0921 ' Vamsapriya khunkonensis MELUCC 11-0475 ~ Vamsapriya yunnana KUMCC 18-00087 Vamsapriva aquatica DLUCC:970° Vamsapriya indica MFLUCC 12-0544? Vamsapriya indica MFLUCC 21-0088 100/1. Vamsapriva uniseptata GZCC 21-0892! Vamsapriya uniseptata MFLU23-0261 Vamsapriya yadongensis HKAS 134926 Vamsapriya chiangmaiensis MFLUCC 21-0065 9710.96, Vamsapriya bambusicola MFLUCC 11-0637 100/1|" Vamsapriya bambusicola MFLUCC 11-0477 Vamsapriya shiwandashanensis GMB5632! A -/0.90 e a Vamsapriya shiwandashanensis GMB5641 Sa eal Vamsapriya zhaotongensis GMB64017 = uae Vamsapriya zhaotongensis GMB6402 — = eed *., r ~ ~ Vamsapriya sichuanensis HKAS 1303047 100/1| Vamsapriya damingshanensis GMB5643 Vamsapriya damingshanensis GMB5627' 100/1| Vamsapriya kailiensis GMB6236! Vamsapriya kailiensis GMB6237 Vamsapriya mucosa MFLU 18-0103! sit ~ EJ 100/1| Vamsapriya tongluobaensis GMB6404" ‘\ Vamsapriya tongluobaensis GMB6405 “ j : | Vamsapriya clypeata GMB5630° 100/0.9 = PM pate) c=. ae — A & ) fe) pels) fq) i Vamsapriya clypeata GMB5642 100/1{~ Barrmaelia oxyacanthae CBS 142770 Barrmaelia rhamnicola CBS 142772 Outgroup 0.03 Figure 1. RAxML tree of Vamsapriya and its related taxa based on a combined LSU, ITS, rpb2, and tub2 sequences dataset. Bootstrap support values for maximum likelihood (MLB) greater than 75% and Bayesian posterior probabilities (BYPP) greater than 0.90 are displayed above the respective branches (ML/BI). The newly described species are marked in bold. 100/17 Neolinocarpon rachidis MFLUCC 15-0814b 1009/1" * Neolinocarpon rachidis MFLUCC 15-0814a zs 69/- Neolinocarpon rachidis MFLUCC 15-0332! = 82/086) Neolinocarpon phayaoense MFLUCC17-00073b (=) 6/3] 24 Neolinocarpon phayaoense MFLUCC 17-00074! ¢) Neolinocarpon phayaoense MFLUCC 17-00073a ——— . = too 100/1| Neolinocarpon bambusina eRe | = oe he =| Neolinocarpon bambusina GMB6408 ‘Se | ee 99 100/1| Neolinocarpon huaxiense GMB6204 so CO sie eh Neolinocarpon huaxiense GMB6203' (a 5 =" Sy S Neolinocarpon arengae MFLUCC 15-0323! oO 96/1 Linocarpon cocois MFLUCC 15-08127 Linocarpon arengae MFLUCC 15-0331 91/0.99 Ruzenia spermoides SMH4606 : 100/1 Synaptospora plumbea SMH3962 I * 100/1 a Helminthosphaeria clavariarum SMH4609! Helminthos- 100/1 Echinosphaeria canescens SMH4666! ; O Hilberina caudata SMH15427 phaeriaceae 99/1 Endophragmiella dimorphospora FMR 12150 96/0.9| Leptosporella gregaria SMH 42907 81/09! Lepiosporella gregaria SMH 4673 oon} |_J Leptosporella arengae MFLUCC 15-0330" Leptosporellaceae “— Leptosporella cocis MFLUCC 15-0816" Leptosporella bambusae MFLUCC 12-0846! 100/1 7910.99 Chloridium lignicola CBS 143.547 sai Dictyochaeta fuegiana FMR 13126 100/1 Zanclospora iberica FMR 11584! ° Chaetosphaeria chlorotunicata SMH1565* Chaetosphaeriaceae Exserticlava vasiformis TAMA 450 ~_ _Umbrinosphaeria caesariata CBS 102664 100/1 100/1| Neolinocarpon globosicarpum HKUCC 2983 7 Neolinocarpon enshiense HKUCC 2983 Linocarpaceae Gelasinospora tetrasperma CBS 178.33" eri funicola CBS 508.50 Outgroup 0.04 Figure 2. Phylogenetic tree based on combined partial SSU, LSU, and ITS sequences showing the relationship of one new species of Neolinocarpon from Yunnan Province with other species. Numbers at the branches indicate support values (RAx- ML-BS/BI-PP) above 60%/0.9. Ex-type materials are marked with “T.” Materials in bold type are those analyzed in this study. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 17 Xingyu Luo et al.: Three species from Yunnan, China Table 1. Information and GenBank accession numbers about species used for the phylogenetic tree (Fig. 1). Species Barrmaelia oxyacanthae Barrmaelia rhamnicola Vamsapriya aquatica Vamsapriya bambusicola Vamsapriya breviconidiophora Vamsapriya chiangmaiensis Vamsapriya clypeata Vamsapriya damingshanensis Vamsapriya zhaotongensis Vamsapriya indica Vamsapriya kailiensis Vamsapriya khunkonensis Vamsapriya mucosa Vamsapriya shiwandashanensis Vamsapriya tongluobaensis Vamsapriya sichuanensis Vamsapriya uniseptata Vamsapriya yadongensis Vamsapriya yunnana Voucher / information CBS 142770 CBS 142772" DLUCC:970" MFLUCC 11-0477 MFLUCC 11-0637 MFLUCC 14-0436 MFLUCC 21-0065 GMB 5630° GMB 5642 GMB 5627' GMB 5643 GMB6401 GMB6402 MFLUCC 12-0544" MFLUCC 21-0088 GMB6237 GMB6236' MFLUCC 13-0497° MFLUCC 11-0475 MFLU 18-0103" GMB 5632" GMB 5641 GMB6404 GMB6405 HKAS 1303047 GZCC 21-0892" MFLU23-0261 HKAS 134926 KUMCC 18-0008" Host / Substrate Salix caprea Rhamnus alpina bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo bamboo wood bamboo ITS MF488988 MF488991 MZ420740 KM462835 KU940159 MF621584 MZ613171 PQ884676 PQ884677 PQ884680 PQ884681 PV637143 PV6437144 KM462839 MZ613172 PQ874043 PQ874042 KM462830 MW240620 MW240622 PQ884678 PQ884679 PV637141 PV637142 PP584757 MZ613173 OR259106 PQ350312 MG833874 LSU NA NA NA KM462836 KU863147 MF621588 MZ613168 PQ885388 PQ885389 PQ885392 PQ885393 PV637137 PV637138 KM462840 MZ613169 PQ860492 PQ860491 KM462831 MW240549 MW240551 PQ885390 PQ885391 PV637135 PV637136 PP584830 MZ613170 OR259087 PQ350314 MG833873 GenBank Accession Number rpb2 MF488997 MF489000 NA KM462834 KU940182 NA NA NA NA NA NA PV642220 PV642221 KM462841 OK560921 NA NA KM462829 KM462829 MW658614 PQ893613 PQ893614 PV642218 PV642219 PP993507 NA NA PQ563198 MG833875 tub MF489016 MF489019 NA KM462833 NA NA NA PQ893585 PQ893586 PQ893589 PQ893590 NA NA KM462838 NA PQ864006 PQ864005 KM462828 KM462828 MW775580 PQ893587 PQ893588 PV642222 PV642223 NA NA OR269618 PQ563196 NA Reference Voglmayr et al. (2018) Voglmayr et al. (2018) Bao et al. (2021) Dai et al. (2014) Dai et al. (2017) Direct Submission Sun et al. (2021) Liu et al. (2025) Liu et al. (2025 Liu et al. (2025) Liu et al. (2025) This study This study Dai et al. (2014) Sun et al. (2021) Habib et al. (2025) Habib et al. (2025) Dai et al. (2014) Dai et al. (2014) Samarakoon et al (2022) Liu et al. (2025) Liu et al. (2025) This study This study Dissanayake et al (2024) Sun et al. (2021) Sun et al. (2021) Wang et al. (2024) Jiang et al. (2022) Notes: Type specimens are marked with T; “NA” indicates no sequence available in GenBank; newly generated sequences are indicated in bold. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 Taxonomy Vamsapriya zhaotongensis X. Y. Luo, K. Habib & Q. R. Li, sp. nov. MycoBank No: 859145 Fig. 3 Etymology. The specific epithet “zhaotongensis” refers to the location, Zha- otong City, where the holotype specimen was collected. Type. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'20.40'N, 104°5'12.09"E), altitude: 1315 m, on dead culms of bamboo, Jun 2024, collected by Xing Yu Luo (holotype: GMB6401; isotype: KUN-HKAS 146990; ex-type living culture: GMBC64071). Description. Saprobic on dead bamboo culm. Sexual morph: Ascomata 560-675 x 720-800 um (x = 606 x 761 um, n = 5), immersed, visible as black, circular dots, solitary, scattered, in cross-section subglobose, with mostly flat- tened base. Ostioles centric, ostiolar canal periphysate. Peridium 12-17 um (x = 14.1 um, n = 10) wide, two-layered, outer layer comprising dark brown, 178 Xingyu Luo et al.: Three species from Yunnan, China Table 2. Information and GenBank accession numbers of species used in phylogenetic tree (Fig. 2). Species Chaetosphaeria chlorotunicata Chloridium lignicola Dictyochaeta fuegiana Echinosphaeria canescens Endophragmiella dimorphospora Exserticlava vasiformis Gelasinospora tetrasperma Helminthosphaeria clavariarum Hilberina caudata Leptosporella arengae Leptosporella bambusae Leptosporella cocis Leptosporella gregaria Linocarpon arengae Linocarpon cocois Neolinocarpon arengae Neolinocarpon bambusicola Neolinocarpon enshiense Neolinocarpon globosicarpum Neolinocarpon huaxiense Neolinocarpon phayaoense Neolinocarpon rachidis Ruzenia spermoides Sordaria fimicola Synaptospora plumbea Umbrinosphaeria caesariata Zanclospora iberica Voucher / information SMH 1565" CBS 143.547 FMR_13126 SMH 4666" FMR_12150 TAMA 450 CBS 178.337 SMH 46097 SMH 15427 MFLUCC 15-0330° MFLUCC 12-0846° MFLUCC 15-0816' SMH 4290' SMH 4673 MFLUCC 15-0331' MFLUCC 15-0812' MFLUCC 15-0323" GMB6407 GMB6408 HKUCC2983 HKUCC2983 GMB6204 GMB6203' MFLUCC 17-00073a MFLUCC 17-00073b MFLUCC 17-00074" MFLUCC 15-0332" MFLUCC 15-0814a MFLUCC 15-0814b SMH4606 CBS 508.50 SMH 3962 CBS 102664 FMR_115847 Host / Substrate decorticated wood soil leaves dead branches soil rotten wood soil soil rotten wood Arenga pinnata bamboo cocoies rotten wood rotten wood Arenga pinnata Cocos nucifera Arenga pinnata bamboo bamboo dead petiole decaying intertidal fronds bamboo bamboo Hevea brasiliensis Hevea brasiliensis Hevea brasiliensis Arenga pinnata Cocos nucifera Cocos nucifera rotten wood wood wood decayed wood wood GenBank Accession Number LSU AF466064 MH868806 KY853500 KF765605 KY853502 AB753846 DQ470980 AY346283 KF765615 MG272246 KU863122 NA AY346290 HM171287 MG272247 MG272248 MG272249 PV637139 PV637140 DQ810221 DQ810224 PQ860477 PQ860476 MG581933 MG581934 MG581935 MG272250 MK106353 MK106354 AY436422 AY681160 KF765621 AF261069 KY853544 SSU NA NA NA NA HF937351 NA DQ471032 NA NA NA NA MG366595 NA NA MG272252 MG272253 MG366597 PV637147 PV637148 NA DQ810258 PQ860578 PQ860577 MG581936 MG581937 MG581938 MG366598 MK106367 MK106368 NA NA NA NA ITS NA MH857273 KY853440 NA KY853442 NA NA NA NA MG272255 KU940134 MG272256 NA NA NA MG272257 MG272258 PV637145 PV637146 NA NA PQ874031 PQ874030 NA NA NA NA MK106342 NA NA AY681188 NA NA Reference Miller and Huhndorf (2004) Vu et al. (2019) Spegazzini (1923) Miller and Huhndorf (2004) Hernandez-Restrepo et al. (2017) Lin et al. (2019) Cai et al. (2006) Huhndorf et al. (2004) Miller et al. (2014) Konta et al. (2017) Dai et al. (2017) Dai et al. (2017) Huhndorf et al. (2004) Huhndorf and Miller (2011) Konta et al. (2017) Konta et al. (2017) Konta et al. (2017) This study This study Bahl (2006) Hyde and Alias (1999) Habib et al. (2025) Habib et al. (2025) Senwanna et al. (2018) Senwanna et al. (2018) Senwanna et al. (2018) Konta et al. (2017) Konta et al. (2017) Konta et al. (2017) Miller and Huhndorf (2004) Niranjan and Sarma (2021) Miller et al. (2014) Réblova and Winka (2001) NG_070789 | KY853480 | Herndandez-Restrepo et al. (2017) Notes: Type specimens are marked with T; “NA” indicates no sequence available in GenBank; newly generated sequences are indicated in bold. thick-walled cells of textura angularis, inner layer composed of hyaline, thin- walled cells of textura angularis. Paraphyses 1.7-4.2 um (x =3.3 um, n = 30) wide, longer than asci, numerous, guttulate. Asci 125-180 x 8-14.5 um (x = 151.1 x 10.7 um, n = 25), 8-spored, unitunicate, cylindrical, short pedicellate, with a 1.6—-2.7 x 3.0-4.2 um (x = 2.1 x 3.4 um, n = 30), trapezoid apical ring, J+ in Melzer’s reagent, apex rounded. Ascospores 17.5—21 x 5.4-8.2 um (x = 19.5 x 6.7 um, n = 30), uniseriate, hyaline, ellipsoidal to broadly fusiform, rounded at both ends, apiosporous; smaller cell 4.4—5.1 um (x = 4.7 um, n = 30) long, larger cell 13-16 um (x = 14.8 um, n = 30) length, usually with large guttules, lacking mucilaginous sheath. Asexual morph: Undetermined. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 179 Xingyu Luo et al.: Three species from Yunnan, China Figure 3. Vamsapriya zhaotongensis (GMB6401, holotype). A. Type material; B. Ascomata on the host surface; C. Cross-section of ascoma; D, E. Longitudinal sections of ascomata; F. Peridium; G. Paraphyses; H. A J+ subapical ring bluing in Melzer’'s reagent; I, J. Ascospores; K-N. Asci. Scale bars: 1 mm (B-D); 100 um (E); 10 um (F-N). MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 180 Xingyu Luo et al.: Three species from Yunnan, China Culture characters. Ascospores cultured on PDA medium at 27 °C for 4-5 weeks, colony diameter 4—4.5 cm, circular, cottony, slightly raised in center, with a distinct margin. White from above; pale brown from below, white in center. Paratype. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'14.07"N, 104°5'20.06'E), altitude: 1253 m, on bamboo, Jun 2024, collected by Xingyu Luo (GMB6402; GMBC6402). Notes. Phylogenetically, Vamsapriya zhaotongensis is closely related to V. sichuanensis. In the BLAST search, the closest match for the ITS sequence of Vamsapriya zhaotongensis was V. sichuanensis (HKAS 130304) with 95.59% similarity, followed by V. damingshanensis (GMB5627) with 89.84% similarity. The LSU sequences of V. zhaotongensis showed 99.51% similar- ity to V. sichuanensis (HKAS 130304) and 98.39% to V. damingshanensis (GMB5627). The rpb2 sequences showed 99.59% similarity to V. sichuanensis (HKAS 130304). Morphologically, V. sichuanensis can be easily distinguished from V. zhaotongensis in having ascospores surrounded by a mucilaginous sheath. Moreover, the V. sichuanensis has smaller stromata (250-400 x 350- 500 um vs. 560-675 x 720-800 um) and smaller asci (100-160 x 7-10 um vs. 125.9-180.9 x 8.0-14.3 um). Vamsapriya damingshanensis differs from V. zhaotongensis in having larger ascospores (21-25 x 7-12.5 um vs. 17.5- 21 x 5.4-8.2 um), with a large cell (16.4-19 um long vs. 13-16 um long) and basal cell (5-8.5 um long vs. 4.4-5.1 um long), and ascospores of V. daming- shanensis surrounded by a thin gelatinous mucilaginous sheath vs. lacking in V. zhaotongensis. Vamsapriya tongluobaensis X.Y. Luo, K. Habib & Q.R. Li, sp. nov. MycoBank No: 859144 Fig. 4 Etymology. The specific epithet “tongluobaensis” refers to the geographical location, Tongluo Dam National Forest Park, where the holotype specimen was collected. Type. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'18.26'N, 104°5'20.28"E), altitude: 1306 m, on dead culms of bamboo, Jun 2024, collected by Xingyu Luo (holotype: GMB6404; iso- type: KUN-HKAS 146991; ex-type living culture: GMBC6404). Description. Saprobic on dead bamboo culms. Sexual morph: Ascoma- ta 600-875 x 440-610 um (x = 791 x 550 um, n = 5), immersed, visible as black dots, with clypeus-solitary, scattered, in cross-section globose to subglo- bose. Ostioles centric, raised, ostiolar canal periphysate. Peridium 46-133 um (x = 83.7 um, n= 10) wide, two-layered, outer layer comprising dark brown, thick- walled cells of textura angularis, inner layer composed of hyaline, thin-walled cells of textura angularis. Paraphyses 2.8-6.2 um (x = 4.3 um, n = 30) wide, long, septate, constricted at septa, guttulate. Asci 115.5-170 x 7.6-11.5 um (x = 146.5 x 8.9 um, n = 25), 8-spored, unitunicate, cylindrical, short pedicellate, with a 2-3.4 x 1.5-2.3 um (x = 3.1 x 1.9 um, n = 30), trapezoid apical ring, J+ in Melzer’s reagent. Ascospores 15-24 x 4.4-6.8 um (x = 19.3 x 5.9 um, n = 30), L/W 3.3, uniseriate, hyaline, ellipsoidal, apiosporous; smaller cell 2.8-5.1 um (x = 3.7 um, n = 30) long, conical; larger cell 13.4-21 um (x = 15.6 um, n = 30) MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 181 Xingyu Luo et al.: Three species from Yunnan, China Figure 4. Vamsapriya tongluobaensis (GMB6404, holotype). A. Type material; B. Ascomata on the host surface; C. Cross-section of ascoma; D, E. Longitudinal sections of ascomata; F. Peridium; G. A J+ subapical ring bluing in Melzer’s reagent; H. Paraphyses; I-K. Ascospores; L-N. Asci. Scale bars: 0.5 mm (B-D); 100 um (E); 10 um (F-N). MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 182 Xingyu Luo et al.: Three species from Yunnan, China length, usually with large guttules, lacking mucilaginous sheath. Asexual morph: Undetermined. Culture characters. Ascospores cultured on PDA medium at 27 °C for 3-4 weeks, colony diameter 4—5 cm, circular, cottony surface, slightly raised in center, with a distinct margin. White from above; yellow-white to pale brown from below. Paratype. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'22.16"N, 104°5'10.06'E), altitude: 1411 m, on dead bamboo culms, Jun 2024, collected by Xingyu Luo (GMB6405; paratype living culture: GMBC6405). Note. Phylogenetically, Vamsapriya tongluobaensis is closely related to V. mu- cosa. In the BLAST search, the closest match of the ITS sequences of V. ton- gluobaensis showed 89.37% similarity to V. mucosa MFLU 18-0103, followed by V. kailiensis GMB6236 with 85.90% similarity. Its LSU sequences demonstrated 98.23% and 97.66% similarity to V. mucosa (MFLU 18-0103) and V. kailiensis (GMB6236), respectively, while its rp>b2 sequences showed 91.64% similarity to V. mucosa (MFLU 18-0103). Morphologically, V. mucosa differs in having small- er ascomata (260-300 x 320-380 um vs. 600-875 x 440-610 um), small- er asci (80-95 x 5-7.5 um vs. 115.5-170 x 7.6-11.5 of V. tongluobaensis), smaller ascospores (10-14 x 3.5-4.5 um vs. 15-24 x 4.4-6.8 um of V. ton- gluobaensis), and ascospores surrounded by a 4-10 um wide mucilaginous sheath vs. lacking in V. tongluobaensis (Samarakoon et al. 2022). Morphologically, Vamsapriya zhaotongensis resembles V. tongluobaensis in the lack of a mucilaginous sheath around the ascospores. However, V. zha- otongensis differs in having shorter ascospores (17.5-21 x 5.4-8.2 um vs. 15-24 x 4.4-6.8 um of V. tongluobaensis) with a ratio of basal to apical cell in V. zhaotongensis of 3.1, while in V. tongluobaensis, it is 4.2. Additionally, the ITS sequences of Vamsapriya tongluobaensis show 90.94% similarity to V. zhaoton- gensis. LSU sequences show 98.17% similarity. Based on these morphological differences and phylogeny, we introduce V. tongluobaensis as a new species. Neolinocarpon bambusicola X.Y. Luo, K. Habib & Q.R. Li, sp. nov. MycoBank No: 859146 Figs'5 Etymology. The specific epithet “bambusicola’” refers to the host genus Bambu- sa (Poaceae) on which the holotype specimen was collected. Type. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'14.48'"N, 104°5'22.40"E), altitude: 1273 m, on dead culms of bamboo, Jun 2024, collected by Xingyu Luo (holotype: GMB6407; iso- type: KUN-HKAS 146992; ex-type living culture: GMBC6407). Description. Saprobic on dead bamboo culm. Sexual morph: Ascomata 410-690 x 380-745 um (x = 519 x 576 um, n = 5), immersed, perithecial, soli- tary, superficial, globose to subglobose, black, clypeus slightly raised. Ostioles papillate, central, black. Peridium 19-55 um wide (x = 40 um, n = 10), outer cells merging with host epidermal cells, composed of dark brown to black cells of textura angularis. Paraphyses 2.5—4 um wide (x = 3 um, n = 20), numerous, hyaline, hypha-like, long-cylindrical, unbranched, septate, longer than asci. Asci 142-230 x 10-15 um (x = 176.8 x 13.3 um, n = 25), 8-spored, unitunicate, cy- MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 183 Xingyu Luo et al.: Three species from Yunnan, China Figure 5. Neolinocarpon bambusicola (GMB6407; holotype) A. Type material; B. Ostioles; C. Cross-section of ascoma; D, E. Longitudinal sections of ascomata; F. Peridium; G-J. Ascospores; K. Paraphyses; L. Asci. Scale bars: 1 mm (C, D); 100 um (E); 10 um (F-L). MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 Xingyu Luo et al.: Three species from Yunnan, China lindrical, short-pedicellate, with a J-, wedge-shaped, subapical ring. Ascospores 106-144 x 3-5 um (x = 130 x 4.5 um, n = 30), filiform, straight or curved, hya- line, aseptate, containing numerous refringent septum-like bands up to 50, ends rounded, lacking polar mucilaginous appendages, smooth-walled. Asexual morph: Undetermined. Culture characteristics. Ascospores germinating on PDA within 24 h. Col- onies on PDA reached 50 mm in diameter after one month at 27 °C, colonies circular, edge entire, velvety to woolly, from above white, below yellow-white to light brown. Paratype. CHINA * Yunnan Province, Zhaotong City, Shuifu County, Tongluo Dam National Forest Park (28°26'20.44'"N, 104°5'16.28"E), altitude: 1098 m, on bamboo, Jun 2024, collected by Xingyu Luo (GMB6408; paratype living culture: GMBC6408). Note. Phylogenetic analysis (Fig. 2) based on ITS, LSU, and SSU sequences revealed Neolinocarpon bambusicola to be closely related to N. phayaoense. In the BLAST search, the closest match for the LSU sequence of N. bambusi- cola was N. phayaoense (MFLUCC 17-00073a) with 93.2% similarity, followed by N. arengae (MFLUCC 15-0323) with 91.5% similarity. The SSU sequence of N. bambusicola showed 98.88% similarity to N. phayaoense (MFLUCC 17- 00073a) and 99.29% similarity to N. arengae (MFLUCC 15-0323). Morphologically, Neolinocarpon bambusicola resembles N. phayaoense in having filiform ascospores lacking mucilaginous appendages. However, N. pha- yaoense can be distinguished from N. bambusicola by its host Hevea brasilien- sis in Euphorbiaceae vs. bamboo, smaller ascomata with 1-3 locules (250- 550 x 120-300 um vs. 410-690 x 380-745 um), longer asci (115-130 um vs. 142-230 um), and smaller ascospores (77-92 x 2-5 um, x = 85 x 4 um vs. 106-144 x 3-5 um, X = 130 x 4.5 um) (Senwanna et al. 2018). Morphologically, Neolinocarpon bambusicola resembles N. arengae in ascal size and the lack of mucilaginous appendages on ascospores. However, the latter differs in host preference, being reported on dead leaflets of Arenga pin- nata (a palm, Arecaceae), and in having slightly smaller ascospores (114-134 x 3-4 um), whereas N. bambusicola is found on dead bamboo culms (Poaceae) and has slightly larger ascospores (106-144 x 3-5 um). Although both spe- cies occur on monocotyledonous hosts, the two species are phylogenetically distinct. Based on these morphological differences and phylogenetic evidence, we introduce N. bambusicola as a new species (Konta et al. 2017). Discussion China possesses the richest bamboo resources in the world, accounting for about one-third of the global bamboo forest area (Dlamini et al. 2022). This ecological dominance is reflected in its rich fungal diversity and hosts over one-third of the world’s documented bambusicolous Ascomycota (Jiang et al. 2022). Up to 2022, 174 bambusicolous ascomycete taxa had been document- ed in Southwest China, representing approximately 34% of China’s total known bambusicolous Ascomycota (Jiang et al. 2022). Since then, about 150 new bambusicolous ascomycete species have been described from China, mostly from southwestern regions (Liang et al. 2023; Wu et al. 2023; Yu et al. 2023; Dissanayake et al. 2024; Habib et al. 2024; Li et al. 2024a, b; Shen et al. 2024; MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 185 Xingyu Luo et al.: Three species from Yunnan, China Yu et al. 2024a, b; Zhou et al. 2024; Liu et al. 2025). This remarkable increase highlights the hyperdiversity of bambusicolous fungi in China, and Yang et al. (2024) reported that bamboo forests in southwestern China are still unexplored for bamboo-associated fungi, with many species yet to be discovered. This study contributes to the knowledge of bambusicolous fungal diversity in southern China through the description of three novel species assigned to the genera Vamsapriya and Neolinocarpon. All species of Vamsapriya have been re- ported on bamboo, except V. yadongensis, whose host remains unidentified. The two new species described in this study were also found on bamboo, suggesting that the genus may exhibit a strong host specificity. Globally, 14 species of Vam- sapriya have been documented, and among these, eight species are known to possess a sexual morph (Dai et al. 2017; Sun et al. 2021; Samarakoon et al. 2022; Dissanayake et al. 2024; Liu et al. 2025). In this study, we added another Vamsap- riya species from its sexual morph. The species in this genus with sexual morphs can be divided into two sections: those with a mucilaginous sheath on the asco- spore and those without. Among the previously known species, V. clypeata, V. kail- iensis, and V. shiwandashanensis lack a mucilaginous sheath, whereas V. bam- busicola, V. chiangmaiensis, V. damingshanensis, V. mucosa, and V. sichuanensis are characterized by its presence. The species of the genus can be further dif- ferentiated by the size of ascomata and asci and the shape and size of their as- cospores. The two newly recognized species of Vamsapriya do not possess a mucilaginous sheath. Among the species lacking a sheath on the ascospores, V. kailiensis possesses the smallest ascospores in the genus (<14 um), whereas V. shiwandashanensis has the largest (20-29 x 5.5-7.5 um). Vamsapriya clypeata can be confused with V. zhaotongensis because of their similar ascospore size (18.5-21 x 7.2-8.9 um) and shape. However, V. zhaotongensis differs in having slightly smaller asci (115.5-170 x 7.6-11.5 um vs. 129-181 x 8.2-9.6 um in V. clypeata) and is phylogenetically distantly related. Vamsapriya tongluobaensis can be easily distinguished from V. clypeata by its larger ascospores (15-24 x 4.4—6.8 um) pointed at both ends, vs. rounded ends in V. clypeata (Liu et al. 2025). Nearly all previously known species of Neolinocarpon have been found on hosts in the Arecaceae, indicating a strong host preference within this genus. The exceptions are N. penniseti, which occurs on the dead stems of Pennisetum purpureum (Poaceae), and N. phayaoense, found on branches of Hevea brasilien- sis (Euphorbiaceae) (Hyde 1992b; Hyde et al. 1998; Hyde and Alias 1999; Bhila- butra et al. 2006; Vitoria et al. 2013; Senwanna et al. 2018). Within the Arecaceae, Neolinocarpon species are reported on different plants and are associated with various plant parts. Neolinocarpon australiense is associated with dead rattan of Calamus moti, while N. attaleae occurs on dead rachises of Attalea funifera. Simi- larly, N. calami and N. enshiense inhabit dead petioles of Calamus conirostris and Trachycarpus fortunei, respectively. Neolinocarpon globosicarpum and N. nypico- la are reported on decaying intertidal fronds and intertidal petioles of Nypa fruti- cans. Neolinocarpon inconspicuum and N. nonappendiculatum both occur on Ar- chontophoenix alexandrae but occupy different tissues: dead rachises and dead petioles, respectively (Hyde 1992b; Hyde et al. 1998; Hyde and Alias 1999; Vitoria et al. 2013). Recently, anew species, N. huaxiense, has been reported from China, isolated from bamboo (Habib et al. 2025). This study expands the known ecolog- ical range of the genus by reporting N. bambusicola on dead bamboo (Poaceae) culms, marking the third documented occurrence of Neolinocarpon on Poaceae. MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 186 Xingyu Luo et al.: Three species from Yunnan, China The genus Neolinocarpon and its type species, Neolinocarpon globosicarpum, were introduced by the renowned mycologist K.D. Hyde, who also established several other species within the genus, including Neolinocarpon arengae S. Konta & K.D. Hyde, Neolinocarpon phayaoense Senwanna & K.D. Hyde, and Neolinocar- pon rachidis S. Konta & K.D. Hyde. The holotype sequences of these latter species have been verified and form a well-supported monophyletic clade, within which our newly described species also clusters. However, phylogenetic analyses re- veal inconsistencies within the genus, particularly concerning N. globosicarpnum and N. enshiense, whose available sequences, derived from non-type specimens, cluster distantly from the main Neolinocarpon clade and do not group within the familial boundaries, rendering the genus polyphyletic. This phylogenetic incon- gruence highlights a systematic issue within Neolinocarpon that can only be re- solved by sequencing the holotype of N. globosicarpum. A detailed molecular study, including the re-examination and sequencing of existing type specimens, is urgently needed to clarify the true phylogenetic placement of these taxa. Until this is addressed, the delimitation of Neolinocarpon remains provisional, and the phylogenetic positions of N. globosicarpum and N. enshiense should be inter- preted with caution rather than being excluded from the genus. These findings not only highlight the vast, yet-to-be-explored fungal diversity in China’s bamboo ecosystems but also pave the way for further research to uncover additional species and clarify their ecological roles. Additional information Conflict of interest The authors have declared that no competing interests exist. Ethical statement No ethical statement was reported. Use of Al No use of Al was reported. Funding This research was supported by the National Natural Science Foundation of China (31960005, 32000009) and the Guizhou Medical University High-Level Talent Launch Fund Project (2023-058). The authors extend their appreciation to the Ongoing Research Funding Program (ORF-2025-500), King Saud University, Riyadh, Saudi Arabia. Author contributions All authors have contributed equally. Author ORCIDs Kamran Habib © https://orcid.org/0000-0003-2572-0306 Yulin Ren © https://orcid.org/0009-0003-9063-425X Xiangchun Shen ® https://orcid.org/0000-0002-4333-9106 Abdallah M. Elgorban © https://orcid.org/0000-0003-3664-7853 Nalin N. Wijayawardene © https://orcid.org/0000-0003-0522-5498 Qirui Li © https://orcid.org/0000-0001-8735-2890 MycoKeys 120: 173-192 (2025), DOI: 10.3897/mycokeys.120.156619 187 Xingyu Luo et al.: Three species from Yunnan, China Data availability All of the data that support the findings of this study are available in the main text. 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