Skip to main content
NCBI home page
MycoKeys logoLink to MycoKeys
. 2025 Nov 27;125:347–368. doi: 10.3897/mycokeys.125.174645

Morphological and phylogenetic analyses reveal three new species associated with Pueraria from Xizang Autonomous Region, China

Gui-Li Zhao 1,2, Yong-Zhong Lu 2,3, Xing-Juan Xiao 3, Ying Liu 1, Qiang Chen 1, Ning-Guo Liu 2,3,✉
PMCID: PMC12679286  PMID: 41356274

Abstract

During a survey of microfungi associated with Pueraria in the Xizang Autonomous Region, China, three saprobic taxa were isolated from the dead vines of Pueraria sp. Based on morphological characteristics and multi-gene phylogenetic analyses, the three taxa are identified as new species, viz., Hermatomyces nyingchiensis, H. xizangensis, and Rhytidhysteron xizangense. Morphological descriptions and illustrations of these new collections, along with a synopsis of Hermatomyces species, are provided.

Key words: 3 new taxa, Dothideomycetes , phylogeny, Pueraria-associated microfungi, taxonomy

Introduction

Species of Pueraria (Leguminosae) are widely distributed across Asia, North America, and South America (Wang et al. 2020). They exhibit strong colonization ability and ecological adaptability, enabling them to survive and thrive even in post-mining environments, thereby contributing to the restoration of degraded lands (Singhal 2009). Members of Pueraria contain numerous compounds, including puerarin, isoflavones, flavones, flavonols, coumestrols, and other related phytochemicals (Chen et al. 2018; Wang et al. 2020; Chen et al. 2021; Wang et al. 2021; Zhang et al. 2021). These compounds possess diverse pharmacological activities, including anti-alcoholism (Xiong et al. 2010), antioxidant (Zhang et al. 2017; Son et al. 2019), hepatoprotective (Xu et al. 2013), and antidiabetic effects (Shukla et al. 2017; Wang et al. 2017; Sun et al. 2019). Although previous studies have investigated the arbuscular mycorrhizal fungi associated with Pueraria (Guo et al. 2022), research on saprophytic fungi related to Pueraria remains limited. In this study, we aim to explore the diversity of fungi associated with Pueraria, with a focus on saprobic fungal communities.

Hermatomyces was established by Spegazzini (1910), with H. tucumanensis as the type species. It was initially placed within Lophiotremataceae (Doilom et al. 2016; Tibpromma et al. 2017). However, Hashimoto et al. (2017) excluded Hermatomyces from Lophiotremataceae and reinstated Hermatomycetaceae based on molecular evidence. Most species of Hermatomyces are known from their asexual morph, characterized by sporodochial conidiomata and dimorphic conidia (cylindrical and lenticular) (Doilom et al. 2016; Tibpromma et al. 2017; Koukol et al. 2018; Ren et al. 2021; Zhang et al. 2023; Liu et al. 2024). The sexual morph was recently described and is characterized by dark brown to black ascomata with a central ostiole, 8-spored, bitunicate asci, and hyaline, broadly fusiform, 1-septate ascospores (de Silva et al. 2022).

Rhytidhysteron was originally established by Spegazzini (1881) and was historically placed within Patellariaceae (Bezerra and Kimbrough 1982). However, molecular evidence provided by Boehm et al. (2009) demonstrated that Rhytidhysteron is not phylogenetically related to Patellariaceae but rather belongs to Hysteriaceae. Currently, 44 records of Rhytidhysteron are listed in Index Fungorum (http://www.indexfungorum.org, accessed 13 October 2025). Most species are known from their sexual morphs, whereas only four species have been recognized from their asexual morphs, which produce two types of conidia: aposphaeria-like and diplodia-like (Samuels and Müller 1979; Thambugala et al. 2016; Ren et al. 2022). The sexual morph of Rhytidhysteron is characterized by prominent, sizable, navicular ascomata, typically with perpendicular striations along the margins (Thambugala et al. 2016; de Silva et al. 2020; Wanasinghe et al. 2021; Ren et al. 2022; Du et al. 2023). The ascospores are pigmented, septate, and muriform to sub-muriform (Thambugala et al. 2016; Ren et al. 2022). Species within this genus play essential ecological roles, functioning as saprobes, endophytes, and mild pathogens on woody substrates in terrestrial and aquatic environments. Occasionally, some species have also been reported as human pathogens (Thambugala et al. 2016; Soto-Medina and Lücking 2017; de Silva et al. 2020; Wanasinghe et al. 2021; Ren et al. 2022; Du et al. 2023).

We aim to carry out a survey investigating fungal diversity associated with Pueraria sp. from the Xizang Autonomous Region, China. During this survey, three taxa were obtained. Based on morphological characterization, illustrations, and multi-gene phylogenetic analyses, Hermatomyces nyingchiensis sp. nov., H. xizangensis sp. nov., and Rhytidhysteron xizangense sp. nov. are described herein.

Materials and methods

Specimen collection, examination, and isolation

Specimens were collected from dead vines of Pueraria species in the Xizang Autonomous Region, China, and packed into plastic bags with recorded details such as collection date, locality, and host. The specimens were transported to the laboratory for examination and isolation. Morphological examination and single-spore isolations followed the methodologies described by Senanayake et al. (2020). Germinated spores were carefully transferred onto fresh potato dextrose agar (PDA) medium and incubated at room temperature. During this period, morphological characteristics of the colonies, such as color, form, and texture, were observed and recorded. Fungal colonies growing on the substrate were examined using Nikon SMZ 745 and SMZ 800N (Nikon, Tokyo, Japan) dissecting microscopes. Detailed microscopic observations and image capture were performed with a Nikon ECLIPSE microscope equipped with a Nikon DS-Ri2 digital camera. Pure cultures were deposited in the Guizhou Culture Collection (GZCC), Guiyang, China, and dried specimens were preserved in the Herbarium of Cryptogams, Kunming Institute of Botany, Academia Sinica (HAKS), Kunming, China. The new species were registered in Fungal Names (https://nmdc.cn/fungalnames/).

DNA extraction, PCR amplification, and sequencing

Molecular analyses were conducted following the protocols outlined by Dissanayake et al. (2020). Genomic DNA was extracted from freshly grown mycelia on PDA plates using the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech, Shanghai, China), according to the manufacturer’s instructions. Amplification of the large subunit ribosomal RNA region (LSU) was performed with the primers LR0R and LR5 (Vilgalys and Hester 1990); the internal transcribed spacer (ITS) region with ITS5 and ITS4 (White et al. 1990); the translation elongation factor 1-alpha (tef1-α) with EF1-983F and EF1-2218R (Rehner and Buckley 2005); the RNA polymerase II second largest subunit (rpb2) with fRPB2-5F and fRPB2-7cR (Liu et al. 1999); the β-tubulin (tub2) with primers T1 and T22 (O’Donnell and Cigelnik 1997); and the small subunit ribosomal RNA region (SSU) with NS1 and NS4 (White et al. 1990). Polymerase chain reaction (PCR) was conducted in a 25 μl volume containing 21 μl of 1.1 × T3 Super PCR Mix (Qingke Biotech, Chongqing, China), 1 μl of each primer, and 2 μl of DNA template. PCR amplification was initiated by denaturation at 98 °C for 2 min, followed by 40 cycles of 98 °C for 10 s, annealing at 55 °C for 1 min, and extension at 72 °C for 30 s. A final extension was performed at 72 °C for 2 min. Purification and sequencing of the PCR products were conducted by Tsingke Biotechnology Co., Ltd. (Beijing, China).

Phylogenetic analyses

All newly generated sequences were checked using BioEdit v. 7.0.5.3 to assess sequence quality. Forward and reverse sequences were assembled using SeqMan v. 7.0.0 (DNASTAR, Madison, WI, USA) (Clewley 1995). Sequences used in phylogenetic analyses were obtained from GenBank (http://blast.ncbi.nlm.nih.gov/) based on BLASTn search results and relevant literature (Doilom et al. 2016; Tibpromma et al. 2016, 2017; Soto-Medina and Lücking 2017; Koukol et al. 2018; de Silva et al. 2020; Ren et al. 2021; Du et al. 2023; Zhang et al. 2023). Multiple sequence alignments were carried out using the online MAFFT version 7 (https://mafft.cbrc.jp/alignment/server/) (Katoh and Standley 2013). Single alignment trimming was conducted using trimAl v. 1.2 (Capella-Gutiérrez et al. 2009) with the gappyout option. Maximum Likelihood (ML) and Bayesian Inference (BI) analyses were conducted online via the CIPRES Science Gateway platform (https://www.phylo.org/portal2/home.action). The ML trees were constructed using RAxML-HPC v. 8 on XSEDE (version 8.2.12), employing the GTRGAMMA model and 1,000 bootstrap replicates. Posterior probabilities (PP) were estimated through BI analyses using the Markov chain Monte Carlo (MCMC) algorithm implemented in MrBayes on XSEDE (version 3.2.7a) (Ronquist et al. 2012). In Hermatomyces, the optimal models applied were: TNe+R2 for LSU, TIM2e+I+G4 for ITS, TN+F+R3 for tef1-α, TNe+G4 for rpb2, and K2P+I for tub2. In Rhytidhysteron, the optimal models were: TN+F+R2 for LSU, K2P+G4 for ITS, K2P+I for SSU, and TIM2e+I+G4 for tef1-α. Markov chains were run for 50,000,000 generations, with trees sampled every 100 generations. Phylogenetic trees were visualized using FigTree v. 1.4.4, and final figure layouts were edited using Adobe Illustrator CC 2019 (version 23.1.0; Adobe Systems, San Jose, CA, USA). The sequences generated in this study were submitted to GenBank, and their accession numbers are listed in Tables 1, 2.

Table 1.

Strains and sequence accession numbers included for analysis of Hermatomyces. The newly generated sequences are in bold. “T” indicates the type strains. NA indicates sequence unavailability.

Fungal Species Strain numbers Conidia type Country LSU ITS tef1-α rpb2 tub2 References
Anteaglonium parvulum MFLUCC 14-0823 - Thailand KU922917 NA KU922922 NA NA Jayasiri et al. (2016)
A. thailandicum MFLUCC 14-0816 - Thailand KU922909 NA KU922920 NA NA Jayasiri et al. (2016)
Hermatomyces amphisporus CBS 146613 2 USA LR812662 LR812662 LR812657 LR812668 LR812673 Delgado et al. (2020)
H. amphisporus CBS 146614 2 USA LR812666 LR812666 LR812660 LR812671 LR812676 Delgado et al. (2020)
H. anomianthi MFLUCC 21-0202T 1 (Sexual) Thailand OK655817 OL413437 OM117546 NA NA de Silva et al. (2022)
H. bauhiniae MFLUCC 16-0395T 2 Thailand MK443378 MK443382 MK443384 MK443386 NA Hyde et al. (2019)
H. bifurcatus CCF 5899 2 Panama NA LS398262 LS398416 LS398343 LS398441 Koukol et al. (2018)
H. bifurcatus CCF 5900T 2 Panama NA LS398263 LS398417 LS398344 LS398442 Koukol et al. (2018)
H. clematidis MFLUCC 17-2085T 2 Thailand MT214556 MT310603 MT394735 MT394684 NA Phukhamsakda et al. (2020)
H. constrictus CCF 5904T 2 Panama LS398264 LS398264 LS398418 LS398345 NA Koukol et al. (2018)
H. griseomarginatus PRC 7201 1 Benin NA PP779919 PP840395 PP840388 NA Koukol et al. (2025)
H. griseomarginatus PRC 7203T 1 Benin NA PP779918 PP840396 PP840389 NA Koukol et al. (2025)
H. hainanensis GZCC 23-0592T 2 China OR091329 OR098708 NA NA NA Zhang et al. (2023)
H. hongheensis KUNCC 23-13503T 1 China PP189897 PQ340461 PQ456948 PQ348601 NA Shen et al. (2024)
H. hongheensis KUNCC 23-14231T 1 China PP189898 PQ340462 PQ456949 NA NA Shen et al. (2024)
H. indicus (H. thailandicus) MFLUCC 14-1145 2 Thailand KU764694 KU144922 KU872756 KU712490 NA Doilom et al. (2016)
H. indicus (H. thailandicus) MFLUCC 14-1143T 2 Thailand KU764692 KU144920 KU872754 KU712488 NA Doilom et al. (2016)
H. iriomotensis KH 361T 2 Japan LC194367 LC194483 LC194394 LC194449 NA Hashimoto et al. (2017)
H. jinghaensis HKAS 112167T 2 China MW989519 MW989495 MZ042642 NA NA Ren et al. (2021)
H. krabiensis MFLUCC 16-0249T 2 Thailand KX525742 KX525750 KX525758 KX525754 NA Tibpromma et al. (2016)
H. krabiensis (H. chiangmaiensis) MFLUCC 16-2817 2 Thailand KY559394 NA NA NA NA Tibpromma et al. (2016)
H. maharashtraense NFCCI 4879T 1 India MZ099917 MZ147016 MZ130659 MZ130660 NA Wijayawardene et al. (2021)
H. maharashtraense NFCCI 4880 1 India MZ147042 MZ147019 MZ130661 MZ130662 NA Wijayawardene et al. (2021)
H. megasporus CCF 5897 2 Panama NA LS398265 LS398419 LS398346 LS398444 Koukol et al. (2018)
H. megasporus CCF 5898T 2 Panama NA LS398266 LS398420 NA LS398445 Koukol et al. (2018)
H. nabanheensis KUMCC 16-0149T 2 China KY766059 KY766058 KY766061 NA NA Hyde et al. (2017)
H. nujiangensis HKAS 144367 1 China PV264903 PV264897 PV261935 PV261929 NA Yang et al. (2025)
H. nujiangensis HKAS 144368 1 China PV264904 PV264898 PV261936 PV261930 NA Yang et al. (2025)
H. nyingchiensis GZCC 25-0849T 2 China PX413305 PX561052 PX436849 PX436845 PX436853 This study
H. pyriformis CGMCC 3.27462T 1 China PP491962 PP491964 PP505452 PP505454 NA Du et al. (2024)
H. pyriformis UESTCC 23.0441 1 China PP491963 PP491965 PP505453 PP505455 NA Du et al. (2024)
H. reticulatus CCF 5893 1 Panama LS398267 LS398267 LS398421 LS398347 LS398446 Koukol et al. (2018)
H. reticulatus (H. subiculosus) MFLUCC 15-0843 1 Thailand KX259523 KX259521 KX259527 KX259529 NA Hyde et al. (2016)
H. sphaericoides CCF 5908T 1 Panama LS398273 LS398273 LS398427 LS398352 LS398450 Koukol et al. (2018)
H. sphaericoides CCF 5895 1 Panama LS398270 LS398270 LS398424 LS398350 LS398447 Koukol et al. (2018)
H. sphaericus PMA 116080 1 Panama LS398281 LS398281 LS398431 LS398356 LS398454 Koukol et al. (2018)
H. sphaericus PRC 4104 1 Panama NA LS398278 LS398430 LS398355 LS398453 Koukol et al. (2018)
H. sphaericus MFLUCC 21-0036 1 Thailand MW989516 MW989492 MZ042639 MZ042636 MZ042643 Ren et al. (2021)
H. sphaericus (H. biconisporus) KUMCC 17-0183 2 China MH260296 MH275063 MH412771 MH412755 NA Tibpromma et al. (2018)
H. sphaericus (H. chromolaenae) MFLUCC 16-2818 1 Thailand KY559393 NA NA NA NA Tibpromma et al. (2017)
H. sphaericus (H. pandanicola) MFLUCC 16-0251 2 Thailand KX525743 KX525751 KX525759 KX525755 NA Tibpromma et al. (2016)
H. sphaericus (H. saikhuensis) MFLUCC 16-0266 1 Thailand KX525740 KX525748 KX525756 KX525752 NA Tibpromma et al. (2016)
H. sphaericus (H. tectonae) MFLUCC 14-1141 2 Thailand KU764696 KU144918 KU872758 NA NA Doilom et al. (2016)
H. sphaericus (H. tectonae) MFLUCC 14-1140 2 Thailand KU764695 KU144917 KU872757 KU712486 NA Doilom et al. (2016)
H. trangensis BCC 80742 1 Thailand KY790601 KY790599 KY790607 KY790605 NA Nuankaew et al. (2019)
H. trangensis BCC 80741T 1 Thailand KY790600 KY790598 KY790606 KY790604 NA Nuankaew et al. (2019)
H. tucumanensis CCF 5912 2 Panama LS398288 LS398288 LS398435 LS398360 LS398458 Koukol et al. (2018)
H. tucumanensis CCF 5915 2 Panama LS398290 LS398290 LS398437 LS398362 NA Koukol et al. (2018)
H. turbinatus MFLUCC 21-0038T 2 Thailand MW989518 MW989494 MZ042641 MZ042638 MZ042645 Ren et al. (2021)
H. verrucosus CCF 5903T 1 Panama LS398292 LS398292 LS398439 LS398364 LS398462 Koukol et al. (2018)
H. verrucosus CCF 5892 1 Panama LS398291 LS398291 LS398438 LS398363 LS398461 Koukol et al. (2018)
H. xizangensis GZCC 25-0850T 1 China PX413303 PX561050 PX436847 PX436843 NA This study
H. xizangensis GZCC 25-0853 1 China PX413304 PX561051 PX436848 PX436844 NA This study
Open in a new tab

Table 2.

Strains and sequence accession numbers included for analysis of Rhytidhysteron. The newly generated sequences are in bold. “T” indicates the type strains. NA indicates sequence unavailability.

Fungal Species Strain numbers LSU ITS tef1-α SSU
Gloniopsis calami MFLUCC 15-0739 NG_059715 KX669036 KX671965 KX669034
G. praelonga CBS 112415 FJ161173 NA FJ161090 FJ161134
Rhytidhysteron bannaense KUMCC 21-0483 OP526409 OP526399 OP572200 OP526395
R. bannaense KUMCC 21-0482T OP526408 OP526398 OP572199 OP526395
R. bruguierae KUMCC 21-0484 OP482285 OP494090 OP572207 OP482277
R. bruguierae MFLU 18-0571T MN017833 NA MN077056 MN017901
R. bruguierae MFLUCC 17-1515 MN632452 MN632457 MN635661 MN632463
R. bruguierae MFLUCC 17-1511 MN632454 MN632459 NA MN632465
R. bruguierae MFLUCC 17-1502 MN632453 MN632458 MN635662 MN632464
R. bruguierae MFLUCC 17-1509 MN632455 MN632460 NA MN632466
R. camporesii KUMCC 21-0488 OP482286 OP494091 OP572208 OP482278
R. camporesii HKAS 104277T MN429072 MN429069 MN442087 NA
R. chromolaenae MFLUCC 17-1516T MN632456 MN632461 MN635663 MN632467
R. coffeae KUMCC 21-0492 OP526406 OP605963 OP572201 OP526412
R. coffeae KUMCC 21-0489T OP526407 OP605964 OP572202 OP526413
R. cozumelense A. Cobos-Villagrán951 MW939459 MZ056797 MZ457338 NA
R. cozumelense T. Raymundo 7321 MW939460 MZ056798 MZ457339 NA
R. erioi MFLU 16-0584T MN429071 MN429068 MN442086 NA
R. esperanzae T. Raymundo 6579 MZ477203 MZ056795 MZ457336 NA
R. esperanzae R. Valenzuela 17206 MZ477204 MZ056796 MZ457337 NA
R. hongheense KUMCC 21-0487 OP482287 OP494092 OP572209 OP482279
R. hongheense KUMCC 20-0222 MW264193 MW264214 MW256815 MW264223
R. hongheense HKAS112348 MW541820 MW541824 MW556132 MW541831
R. hongheense HKAS112349 MW541821 MW541825 MW556133 MW541832
R. hysterinum EB 0351 GU397350 NA GU397340 NA
R. hysterinum CBS 316.71 MH871912 MH860141 NA NA
R. ligustrum SICAUCC 20-0004T MT062446 MT062850 MT075600 MT062451
R. ligustrum SICAUCC 19-0007 MN956789 MN956777 MT027603 MN956798
R. magnoliae KUMCC 21-0478 OP482288 OP494093 OP572210 OP482280
R. magnoliae MFLUCC 18-0719T MN989384 MN989383 MN997309 MN989382
R. mangrovei MFLU 18-1894T MK357777 MK425188 MK450030 NA
R. mengziense KUMCC 21-0490T OP526396 OP526402 OP572203 OP526414
R. mengziense KUMCC 21-0491 OP526397 OP526403 OP572204 OP526415
R. mesophilum A. Trejo 74 MW939461 MZ056799 MZ457340 NA
R. mesophilum A. Cobos-Villagrán 1800 MW939462 MZ056800 MZ457341 NA
R. mexicanum RV17107.1T MT626028 MT626026 NA NA
R. mexicanum RV17107.2 MT626029 MT626027 NA NA
R. neorufulum KUMCC 21-0480 OP482290 OP494095 OP572212 OP482282
R. neorufulum MFLUCC 21-0035 MZ346015 MZ346025 MZ356249 MZ346020
R. neorufulum MFLUCC 13-0216T KU377566 KU377561 KU510400 KU377571
R. neorufulum GKM 361A GQ221893 NA NA GU296192
R. neorufulum HUEFS 192194 KF914915 NA NA NA
R. neorufulum MFLUCC 12-0528 KJ418117 KJ418118 NA KJ418119
R. neorufulum CBS 306.38 FJ469672 NA GU349031 GU296191
R. neorufulum MFLUCC 12-0011 KJ418109 KJ206287 NA KJ418110
R. neorufulum MFLUCC 12-0567 KJ526126 KJ546124 NA KJ546129
R. neorufulum MFLUCC 12-0569 KJ526128 KJ546126 NA KJ546131
R. neorufulum EB 0381 GU397351 NA NA GU397366
R. opuntiae GKM 1190 GQ221892 NA GU397341 NA
R. rufulum MFLUCC 14-0577T KU377565 KU377560 KU510399 KU377570
R. rufulum EB 0384 GU397354 NA NA GU397368
R. rufulum EB 0382 GU397352 NA NA NA
R. rufulum EB 0383 GU397353 NA NA GU397367
R. rufulum MFLUCC 12-0013 KJ418111 KJ418112 NA KJ418113
R. sichuanense SICAUCC 19-0005T MN956787 MN956775 MT027601 MN956796
R. sichuanense SICAUCC 19-0006 MN956788 MN956776 MT027602 MN956797
R. sichuanense SICAUCC 20-0005 MT062447 MT062851 MT075601 MT062452
R. subrufulum SICAUCC 19-0011T MN956793 MN956781 MT027607 MN956802
R. subrufulum SICAUCC 19-0010 MN956792 MN956780 MT027606 MN956801
R. subrufulum SICAUCC 20-0003 MT062445 MT062849 MT075599 MT062450
R. tectonae KUMCC 21-0479 OP482291 OP494096 OP572213 OP482283
R. tectonae MFLUCC 21-0037 MZ346013 MZ346023 MZ356247 MZ346018
R. tectonae MFLUCC 21-0034 MZ346014 MZ346024 MZ356248 MZ346019
R. tectonae MFLUCC 13-0710T KU764698 KU144936 KU872760 KU712457
R. thailandicum KUMCC 21-0493 OP482292 OP494097 OP572214 OP482284
R. thailandicum MFLUCC 14-0503T KU377564 KU377559 KU497490 KU377569
R. thailandicum MFLUCC 12-0530 KJ526125 KJ546123 NA KJ546128
R. thailandicum MFLU17-0788 MT093472 MT093733 NA MT093495
R. thailandicum MFLUCC 13-0051 MN509434 MN509433 MN509435 NA
R. xiaokongense KUMCC 20-0158 MZ346011 MZ346021 MZ356245 MZ346016
R. xiaokongense KUMCC 20-0160T MZ346012 NR_185637 MZ356246 NG_242421
R. xizangense GZCC 25-0851T PX413306 PX561053 PX425314 PX413301
R. xizangense GZCC 25-0852 PX413307 NA PX436846 PX413302
R. yunnanense KUMCC 21-0485T OP526404 OP526410 OP572205 OP526400
R. yunnanense KUMCC 21-0486 OP526405 OP526411 OP572206 OP526401
Open in a new tab

Taxonomy and results

Phylogenetic analyses of Hermatomyces

The concatenated dataset includes 53 strains, including the new collections. The best-scoring RAxML tree is shown in Fig. 1, with a final likelihood value of −15404.961825. The alignment contains a total of 3,809 characters (with gaps included), distributed as follows: ITS (1–503), LSU (504–1,320), rpb2 (1,321–2,325), tef1-α (2,326–3,249), and tub2 (3,250–3,809). The tree was rooted with Anteaglonium parvulum (MFLUCC 14-0823) and A. thailandicum (MFLUCC 14-0816). The alignment matrix exhibited 1,110 distinct patterns, with undetermined characters or gaps constituting 24.90% of the total dataset. Phylogenetic trees derived from ML and BI analyses exhibited fundamentally comparable topologies.

Figure 1.

Figure 1.

Open in a new tab

The ML tree based on a combined dataset of LSU, ITS, tef1-α, rpb2 and tub2 sequences. Bootstrap support values for ML greater than 75% and PP greater than 0.95 are marked near each node as ML/PP. Newly generated isolates are highlighted in bold red.

In the phylogenetic tree (Fig. 1), our strain of Hermatomyces nyingchiensis (GZCC 25-0849) formed a distinct clade that was sister to H. hannanensis (GZCC 23-0592) with 87% ML and 1 PP support. Our two strains of Hermatomyces xizangensis (GZCC 25-0850 and GZCC 25-0853) grouped together, forming a distinct clade sister to H. nujiangensis (HKAS 144367 and HKAS 144368) with 100% ML and 1 PP support.

. Hermatomyces nyingchiensis

G.L. Zhao, N.G. Liu & Y.Z. Lu sp. nov.

B9972C58-C34B-552F-A6E7-1A6F71857BC8

Fungal Names: FN 573085

Fig. 2

Figure 2.

Figure 2.

Open in a new tab

Hermatomyces nyingchiensis (HKAS 149901, holotype). a, b. Colonies on natural substrates; c. Conidial mass; d. Subicular hyphae; e–h. Lenticular conidia (side view); i. Conidiophore with conidium; j, k. Cylindrical conidia; l–o. Lenticular conidia (front view); p. Colony on PDA (left from above, right from below). Scale bars: 50 μm (c); 20 μm (d–o).

Etymology.

Referring to the location Nyingchi City, where the holotype was collected.

Holotype.

HKAS 149901.

Description.

Saprobic on dead vine of Pueraria sp. in a terrestrial habitat. Sexual morph Undetermined. Asexual morph Hyphomycetous. Colonies on natural substrates superficial, effuse, scattered, comprising a greenish brown sterile mycelial outer region, and a dark brown to blackish brown center. Mycelium partly immersed and partly superficial, consisting of pale to brown, branched hyphae. Conidiophores micronematous, mononematous, geniculate, hyaline to pale brown, septate, thick-walled. Conidiogenous cells monoblastic, integrated, terminal, hyaline to pale brown. Conidia dimorphic, (1) cylindrical conidia 36–42 × 14–17 µm (x̄ = 38 × 15 µm, n = 15), hyaline to subhyaline, frequently exhibiting a distinct dark brown pigment, which either extends from the top downward or at the conidial rim, straight or broadly curved, phragmoseptate or muriform, with oblique septa occasionally present, constricted at septa, composed of two columns arising from one or two basal cells. (2) lenticular conidia 30–40 × 23–29 µm (x̄ = 35 × 27 µm, n = 30), 16–21 µm thick (x̄ = 18.5 µm, n = 10), in front view, subglobose to ellipsoidal, or slightly irregular, peripheral cells pale brown to brown, forming a distinct ring, constricted at the septa, central cells dark brown, in side view, rugby-shaped, oval, two distinct adpressed halves can be recognized, each half arranged by 6–10 cells, end cells subhyaline to pale brown, middle cells dark brown.

Culture characteristics.

Conidial germination was observed within 24 hours incubated on PDA medium at 28 °C. Colony reached a diameter of 1.9 cm after 19 days at room temperature, circular, dense, convex at the center, margin entire, pale brown on the front, dark brown in reverse.

Material examined.

China, Xizang Autonomous Region, Nyingchi City, Bomi County, (30°3'16"N, 95°12'20"E, 2320 m), on dead vine of Pueraria sp., 26 July 2024, G.L. Zhao, DDQGF14A (HKAS 149901, holotype), DDQGF14B (HKAS 149903, isotype) , ex-type culture GZCC 25-0849.

Notes.

Hermatomyces nyingchiensis resembles those Hermatomyces species that possess dimorphic conidia (Tibpromma et al. 2016; Hashimoto et al. 2017; Koukol et al. 2018; Ren et al. 2021; Zhang et al. 2023). In the phylogenetic tree (Fig. 1), our isolate (GZCC 25-0849) formed a distinct clade and was sister to H. hainanensis (GZCC 23-0592) with 87% ML-BS and 1 PP support (Fig. 1). However, the two conidial types of H. nyingchiensis are smaller than those of H. hainanensis: (1) cylindrical conidia, 36–42 × 14–17 µm vs. 51–67 × 16–24 μm; and (2) lenticular conidia, 30–40 × 23–29 µm vs. 44–52 × 29–39 µm (Zhang et al. 2023). Therefore, based on phylogenetic analyses and morphological characteristics, H. nyingchiensis is identified as a new species.

. Hermatomyces xizangensis

G.L. Zhao, N.G. Liu & Y.Z. Lu sp. nov.

399F781F-8D84-5A02-A30A-A7CD700C0FD9

Fungal Names: FN 573046

Fig. 3

Figure 3.

Figure 3.

Open in a new tab

Hermatomyces xizangensis (HKAS 149902, holotype). a, b. Colonies on natural substrates; c. Subicular hyphae; d–l. Conidiophores and conidia; m. Germinated conidium; n, o. Colony on PDA (n. From above; o. From below). Scale bars: 50 μm (d); 20 μm (e–m).

Etymology.

Referring to the location Xizang Autonomous Region, where the fungus was collected.

Holotype.

HKAS 149902.

Description.

Saprobic on dead vine of Pueraria sp. in a terrestrial habitat. Sexual morph Undetermined. Asexual morph Hyphomycetous. Colonies on natural substrate sporodochial, superficial, scattered, effuse, consisting of a velvety, sparse, brown sterile mycelial outer zone enclosing a brown to black, glistening, abundantly sporulating center, conidia not easily dislodged when disturbed. Mycelium mostly immersed and consisting of branched, smooth, septate, brown hyphae. Conidiophores micronematous, mononematous, curved, unbranched, light brown to brown thick-walled. Conidiogenous cells monoblastic, integrated, terminal, subcylindrical, hyaline to brown. Conidia 26–48 × 17–39 µm (x̄ = 36 × 27 µm, n = 30), 13–20 µm thick (x̄ = 15.5 µm, n = 15), monomorphic, lenticular, dry muriform, smooth, in front view, ellipsoidal, peripheral cells pale brown to brown, forming a distinct ring, constricted at the septa, central cells dark brown to blackish brown, in side view, rugby-shaped, oval, two distinct adpressed halves can be recognized, each half arranged by 8–10 cells, end cells subhyaline to pale brown, middle cells brown to dark brown.

Culture characteristics.

Conidia germinated within 24 hours on PDA medium at 28 °C. The colony reached 1.8 cm diam after 13 days at room temperature, circular, dense, bulge at middle with entire margin, reverse brown, with a lighter coloration at the margins and a darker pigmentation toward the center.

Material examined.

China, Xizang Autonomous Region, Nyingchi City, Bomi County, (30°3'16"N, 95°12'20"E, 2320 m), on dead vines of Pueraria sp., 26 July 2024, G.L. Zhao, DDQGF12A (HKAS 149902, holotype), ex-type culture GZCC 25-0850; ibid., DDQGF12B (HKAS 149904, isotype), ex-isotype culture GZCC 25-0853.

Notes.

Based on multi-gene phylogenetic analysis, our collections of Hermatomyces xizangensis (GZCC 25-0850 and GZCC 25-0853) formed an independent clade, sister to H. nujiangensis (HKAS 144367 and HKAS 144368) with 100% ML and 1 PP support (Fig. 1). Morphologically, conidia of H. xizangensis consist of 8–10 cells per half in side view, whereas those of H. nujiangensis have 15–22 cells (Yang et al. 2025). Furthermore, three Hermatomyces species, viz., H. dimorphus, H. uniseriatus, and H. truncatus, for which molecular data are lacking, are characterized by dimorphic conidia, including both lenticular and cylindrical types (Rao and de Hoog 1986; Leão-Ferreira et al. 2013; Koukol and Delgado 2019). In contrast, H. xizangensis produces only lenticular conidia. Therefore, based on molecular evidence and morphological comparisons, H. xizangensis is identified as a new species.

Phylogenetic analyses of Rhytidhysteron

The final ML tree of Rhytidhysteron exhibited a topological pattern congruent with that reported by Du et al. (2023). The tree was constructed using a concatenated dataset composed of four gene regions: ITS, LSU, SSU, and tef1-α, totaling 3,280 aligned characters (including gaps). The partitioning scheme was as follows: ITS (1–579), LSU (580–1,433), SSU (1,434–2,427), and tef1-α (2,428–3,280). Gloniopsis calami (MFLUCC 15-0739) and G. praelonga (CBS 112415) were selected as outgroup taxa. The ML analysis of the concatenated matrix yielded the best-scoring tree (Fig. 4) with a final optimization likelihood score of −11534.338749. The matrix had 831 distinct alignment patterns, and undetermined characters or gaps accounted for 20.42% of the total.

Figure 4.

Figure 4.

Open in a new tab

Phylogenetic tree from ML analysis based on the combined ITS, LSU, SSU, and tef1-α sequence datasets. Bootstrap support values for ML greater than 75% and PP greater than 0.95 are marked near each node as ML/PP. Newly generated isolates are highlighted in bold red.

Within Rhytidhysteron, our new taxon, R. xizangense constitutes a distinct clade and is sister to R. hysterinum (EB 0351 and CBS 316.71).

. Rhytidhysteron xizangense

G.L. Zhao, N.G. Liu & Y.Z. Lu sp. nov.

B4CD6DCC-3A19-5648-845B-0340E8E466B9

Fungal Names: FN 573045

Figs 5, 6

Figure 5.

Figure 5.

Open in a new tab

Rhytidhysteron xizangense (HKAS 149907, holotype). a, b. Appearance of hysterothecia on the host; c. Vertical section through hysterothecium; d. Exciple; e. Pseudoparaphyses; f. Epithecium mounted in water; g, h. Pedicel of asci; i–k. Asci; l–q. Ascospores; r, s. Colony on PDA medium. Scale bars: 100 μm (c); 50 μm (d); 20 μm (e–k); 10 μm (l–q).

Figure 6.

Figure 6.

Open in a new tab

Rhytidhysteron xizangense (HKAS 149907, holotype) on PDA. a, b. Obverse and reverse colony on PDA; c. Conidiomata on PDA; d, e. Vertical section through conidiomata; f, h. Conidiomatal wall; g, h. Conidiogenous cells and conidia; i–l. Conidia. Scale bars: 100 μm (d, e); 20 μm (f, h); 10 μm (g); 5 μm (i–l).

Etymology.

Referring to the Xizang Autonomous Region, where the holotype was collected.

Holotype.

HKAS 149907.

Description.

Saprobic on dead vine of Pueraria sp. in a terrestrial habitat. Sexual morph Ascomata 1432–1818 µm long × 780–1265 µm wide × 648–1100 µm high (x̄ = 1622 × 959 × 822 µm, n = 5), scattered to gregarious, semi-immersed to superficial, hysterothecial, bilabiate, closed, black, perpendicular striae. Exciple 150–220 µm (x̄ = 170 µm, n = 5) wide, consisting of dark brown, thick-walled cells arranged in a textura angularis structure, outer layer brown to dark brown, inner layer hyaline. Hamathecium 2–3 μm (x̄ = 2.5, n = 10) wide at the base, 3.5–6.5 μm (x̄ = 4.5, n = 10) wide at swollen tips, dense, brown at the swollen apex, hyaline at the base, septate, branched, pseudoparaphyses, forming a brown epithecium above the asci, swollen with dense septa at the apex. Asci 199–275 × 13–18 µm (x̄ = 224 × 16 µm, n = 20), 8-spored, bitunicate, clavate to cylindrical, short pedicel, rounded at the apex, with an ocular chamber. Ascospores 23–30 × 9–14 µm (x̄ = 26.5 × 11 µm, n = 30), uni-seriate, slightly overlapping, 1-septate, hyaline when immature, brown to dark brown when mature, fusiform to ellipsoidal, slightly pointed at both ends, thick-walled. Asexual morph Sporulation in PDA media, coelomycetous. Conidiomata 260–519 µm high × 391–467 µm wide (x̄ = 411 × 424 µm, n = 10), superficial, subglobose, solitary or aggregated, dark brown to black, thick-walled. Conidiomata wall 63–288 µm (x̄ = 124 µm, n = 15) wide, consisting of brown, thick-walled cells arranged in a textura angularis structure. Conidia 2.5–4.5 µm diam. (x̄ = 3.0 µm, n = 40), globose, hyaline, smooth-walled.

Culture characteristics.

Conidia germinated within 24 hours on PDA medium at 28 °C. After one month of incubation at room temperature, colonies reached a diameter of 3.35 cm. Colonies velvety, subrounded, with wavy margins, yellow in the center, brown in the middle, with light yellow at the edge.

Material examined.

China, Xizang Autonomous Region, Nyingchi City, Bomi County (29°53'11"N, 95°42'9"E, 2720 m), on dead vine of Pueraria sp., 26 July 2024, G.L. Zhao, CGGF7A (holotype, HKAS 149907), ex-type living culture GZCC 25-0851; ibid., CGGF7B (isotype, HKAS 149908), ex-isotype living culture GZCC 25-0852.

Notes.

Rhytidhysteron xizangense is similar to R. neohysterinum in the shapes of ascomata, asci, and ascospores (Cobos-Villagrán et al. 2020). However, in the phylogenetic tree (Fig. 4), R. xizangense (GZCC 25-0851 and GZCC 25-0852) formed an independent clade and is sister to R. hysterinum (EB 0351 and CBS 316.71). BLAST analyses of the ITS and LSU sequences revealed that our strain (GZCC 25-0851) shares 92% (36/439, including 8 gaps) and 97% (21/786, including 0 gap) similarity with R. hysterinum (CBS 316.71), respectively. Morphologically, Rhytidhysteron xizangense differs from R. hysterinum by its larger ascomata (780–1265 µm vs. 500 µm) (Thambugala et al. 2016; Ren et al. 2022; Du et al. 2023). Therefore, based on molecular data and morphological comparisons, we introduce Rhytidhysteron xizangense as a new species.

Discussion

In this study, three novel species, namely Hermatomyces nyingchiensis, H. xizangensis, and Rhytidhysteron xizangense, are introduced. These collections were isolated from decaying vines of Pueraria sp. in the Xizang Autonomous Region. The findings of this study further clarify the species diversity within Hermatomyces and Rhytidhysteron while also supplementing the distribution information of fungi associated with Pueraria hosts.

Among the 26 recognized species of Hermatomyces (Table 1), 15 species exhibit conidial dimorphism, such as H. amphisporus, H. bifurcatus, H. hainanensis, H. nabanheensis, and H. turbinatus (Hyde et al. 2017; Koukol et al. 2018; Delgado et al. 2020; Ren et al. 2021; Zhang et al. 2023). Nine species produce exclusively monomorphic conidia, viz., H. griseomarginatus, H. hongheensis, H. maharashtraense, H. pyriformis, H. reticulatus, H. sphaericoides, H. trangensis, H. verrucosus, and H. xizangensis (Koukol et al. 2018, 2025; Nuankaew et al. 2019; Wijayawardene et al. 2021; Du et al. 2024; Shen et al. 2024). One species, H. sphaericus, is known to exhibit either monomorphic or dimorphic conidia (Doilom et al. 2016; Tibpromma et al. 2016, 2017, 2018; Koukol et al. 2018; Ren et al. 2021), whereas H. anomianthi is known solely from its sexual morph (de Silva et al. 2022). Phylogenetic analysis indicates that conidial morphology (monomorphic vs. dimorphic) does not exhibit a clear phylogenetic clustering pattern. For instance, H. megasporus (dimorphic conidia) and H. reticulatus (monomorphic conidia) cluster in clade 2 (Fig. 1) (Hyde et al. 2016; Koukol et al. 2018), while H. jinghaensis (dimorphic conidia) and H. trangensis (monomorphic conidia) also group together in clade 3 (Fig. 1) (Nuankaew et al. 2019; Ren et al. 2021). Furthermore, conidial type shows no significant correlation with biogeographical distribution. Although both H. hongheensis and H. nabanheensis were collected from Yunnan, China, the former produces monomorphic conidia, whereas the latter develops dimorphic conidia (Hyde et al. 2017; Shen et al. 2024). To elucidate the underlying mechanisms governing the formation of dimorphic conidia and monomorphic conidia in Hermatomyces, broader sampling is essential across diverse geographical regions, ecological niches, and developmental stages. Future research integrating comparative genomics, transcriptomics, and ecological niche modeling will be crucial for elucidating the evolutionary and ecological drivers of conidial dimorphism within the genus.

Supplementary Material

XML Treatment for Hermatomyces nyingchiensis
XML Treatment for Hermatomyces xizangensis
XML Treatment for Rhytidhysteron xizangense

Acknowledgments

We would like to thank Yuan-Pin Xiao, Dan-Feng Bao, Xue-Yan Chen, and Li-Juan Luo for their help with this study.

Citation

Zhao G-L, Lu Y-Z, Xiao X-J, Liu Y, Chen Q, Liu N-G (2025) Morphological and phylogenetic analyses reveal three new species associated with Pueraria from Xizang Autonomous Region, China. MycoKeys 125: 347–268. https://doi.org/10.3897/mycokeys.125.174645

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.

Funding

This study was supported by the Guizhou Institute of Technology 2024 Annual Academic Seedling Cultivation and Innovation Exploration Project (No. 2024XSXM001, 2024XSXM010), the Science and Technology Foundation of Guizhou Province (Qian Ke He Pingtai ZSYS [2025] 029), the Guizhou Provincial Basic Research Program (Natural Science) (Qiankehe Jichu ZK [2023] yiban 132), and the Guizhou Science and Technology Plan Project (Qiankehe Support [2023] Generally 295).

Author contributions

Morphological data, photo plates, and phylogenetic analyses were completed by Gui-Li Zhao. Writing – original draft: Gui-Li Zhao. Writing – review and editing: all co-authors.

Author ORCIDs

Gui-Li Zhao https://orcid.org/0000-0001-8370-4844

Yong-Zhong Lu https://orcid.org/0000-0002-1033-5782

Xing-Juan Xiao https://orcid.org/0009-0003-8769-4534

Ying Liu https://orcid.org/0009-0003-4850-4929

Qiang Chen https://orcid.org/0009-0004-2035-6562

Ning-Guo Liu https://orcid.org/0000-0002-9169-2350

Data availability

All of the data that support the findings of this study are available in the main text.

References

  1. Bezerra JL, Kimbrough JW. (1982) Culture and cytological development of Rhytidhysterium rufulum on citrus. Canadian Journal of Botany 60(5): 568–579. 10.1139/b82-078 [DOI] [Google Scholar]
  2. Boehm EW, Schoch CL, Spatafora JW. (2009) On the evolution of the Hysteriaceae and Mytilinidiaceae (Pleosporomycetidae, Dothideomycetes, Ascomycota) using four nuclear genes. Mycological Research 113: 461–479. 10.1016/j.mycres.2008.12.001 [DOI] [PubMed] [Google Scholar]
  3. Capella-Gutiérrez S, Silla-Martinez JM, Gabaldon T. (2009) TrimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15): 1972–1973. 10.1093/bioinformatics/btp348 [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen X, Yu J, Shi J. (2018) Management of diabetes mellitus with puerarin, a natural isoflavone from Pueraria lobata. The American Journal of Chinese Medicine 46(8): 1771–1789. 10.1142/S0192415X18500891 [DOI] [PubMed] [Google Scholar]
  5. Chen F, Chen ZQ, Wang H, Zhu JJ. (2021) Puerarin pretreatment inhibits myocardial apoptosis and improves cardiac function in rats after acute myocardial infarction through the PI3K/Akt signaling pathway. Advances in Clinical and Experimental Medicine 30(3): 255–261. 10.17219/acem/131754 [DOI] [PubMed] [Google Scholar]
  6. Clewley JP. (1995) Macintosh sequence analysis software – DNAStar’s LaserGene. Molecular Biotechnology 3: 221–224. 10.1007/BF02789332 [DOI] [PubMed] [Google Scholar]
  7. Cobos-Villagrán A, Hernández-Rodríguez C, Valenzuela R, Villa-Tanaca L, Calvillo-Medina RP, Mateo-Cid LE, MartínezPineda M, Raymundo T. (2020) El género Rhytidhysteron (Dothideomycetes, Ascomycota) en México. Acta Botánica Mexicana 127: e1675. 10.21829/abm127.2020.1675 [DOI]
  8. de Silva NI, Tennakoon DS, Thambugala KM, Karunarathna SC, Lumyong S, Hyde KD. (2020) Morphology and multigene phylogeny reveal a new species and a new record of Rhytidhysteron (Dothideomycetes, Ascomycota) from China. Asian Journal of Mycology 3(1): 295–306. 10.5943/ajom/3/1/4 [DOI] [Google Scholar]
  9. de Silva NI, Hyde KD, Lumyong S, Phillips AJL, Bhat DJ, Maharachchikumbura SSN, Thambugala KM, Tennakoon DS, Suwannarach N, Karunarathna SC. (2022) Morphology, phylogeny, host association and geography of fungi associated with plants of Annonaceae, Apocynaceae and Magnoliaceae. Mycosphere: Journal of Fungal Biology 13(1): 955–1076. 10.5943/mycosphere/13/1/12 [DOI] [Google Scholar]
  10. Delgado G, Koukol O, Heredia G, Piepenbring M. (2020) Texas microfungi: Hermatomyces amphisporus (Pleosporales, Dothideomycetes) revisited. Czech Mycology 72(1): 95–107. 10.33585/cmy.72107 [DOI] [Google Scholar]
  11. Dissanayake AJ, Bhunjun CS, Maharachchikumbura SSN, Liu JK. (2020) Applied aspects of methods to infer phylogenetic relationships amongst fungi. Mycosphere: Journal of Fungal Biology 11: 2652–2676. 10.5943/mycosphere/11/1/18 [DOI] [Google Scholar]
  12. Doilom M, Dissanayake AJ, Wanasinghe DN, Boonmee S, Liu JK, Bhat DJ, Taylor JE, Bahkali AH, McKenzie EHC, Hyde KD. (2016) Microfungi on Tectona grandis (teak) in Northern Thailand. Fungal Diversity 82: 107–182. 10.1007/s13225-016-0368-7 [DOI] [Google Scholar]
  13. Du TY, Dai DQ, Mapook A, Lu L, Stephenson SL, Suwannarach N, Tibpromma S. (2023) Additions to Rhytidhysteron (Hysteriales, Dothideomycetes) in China. Journal of Fungi 9(2): e148. 10.3390/jof9020148 [DOI] [PMC free article] [PubMed]
  14. Du HZ, Liu NG, Cheewangkoon R, Liu JK. (2024) Hermatomyces pyriformis sp. nov., a novel dematiaceous hyphomycete (Hermatomycetaceae, Pleosporales) associated with medicinal plants in Yunnan Province, China. Phytotaxa 650(1): 103–115. 10.11646/phytotaxa.650.1.9 [DOI] [Google Scholar]
  15. Guo Y, Bei Q, Dzomeku BM, Martin K, Rasche F. (2022) Genetic diversity and community composition of arbuscular mycorrhizal fungi associated with root and rhizosphere soil of the pioneer plant Pueraria phaseoloides. iMeta 1(4): e51. 10.1002/imt2.51 [DOI] [PMC free article] [PubMed]
  16. Hashimoto A, Matsumura M, Hirayama K, Tanaka K. (2017) Revision of Lophiotremataceae (Pleosporales, Dothideomycetes): Aquasubmersaceae, Cryptocoryneaceae, and Hermatomycetaceae fam. nov. Persoonia 39: 51–73. 10.3767/persoonia.2017.39.03 [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hyde KD, Hongsanan S, Jeewon R, Bhat DJ, McKenzie EHC, Jones EBG, Phookamsak R, Ariyawansa HA, Boonmee S, Zhao Q, Abdel-Aziz FA, Abdel-Wahab MA, Bannmai S, Chomnunti P, Cui BK, Daranagama DA, Das K, Dayarathne MC, de Silva NI, Dissanayake AJ, Doilom M, Ekanayaka AH, Gibertoni TB, Góes-Neto A, Huang SK, Jayasiri SC, Jayawardena RS, Konta S, Lee HB, Li WJ, Lin CG, Liu JK, Lu YZ, Luo ZL, Manawasinghe IS, Manimohan P, Mapook A, Niskanen T, Norphanphoun C, Papizadeh M, Perera RH, Phukhamsakda C, Richter C, de A. Santiago ALCM, Drechsler-Santos ER, Senanayake IC, Tanaka K, Tennakoon TMDS, Thambugala KM, Tian Q, Tibpromma S, Thongbai B, Vizzini A, Wanasinghe DN, Wijayawardene NN, Wu HX, Yang J, Zeng XY, Zhang H, Zhang JF, Bulgakov TS, Camporesi E, Bahkali AH, Amoozegar MA, Araujo-Neta LSA, Ammirati JF, Baghela A, Bhatt RP, Bojantchev D, Buyck B, Silva GAS, de Lima CLF, de Oliveira RJV, de Souza CAF, Dai YC, Dima B, Duong TTD, Ercole E, Mafalda-Freire F, Ghosh A, Hashimoto A, Kamolhan S, Kang JC, Karunarathna SC, Kirk PM, Kytövuori I, Lantieri A, Liimatainen K, Liu ZY, Liu XZ, Lücking R, Medardi G, Mortimer PE, Nguyen TTTN, Promputtha I, Raj KNA, Reck MA, Lumyong S, Shahzadeh-Fazel SA, Stadler M, Soudi MR, Su HY, Takahashi T, Tangthirasunun N, Uniyal P, Wang Y, Wen TC, Xu JC, Zhang ZK, Zhao YC, Zhou JL, Zhu L. (2016) Fungal diversity notes 367–490: taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 80: 1–270. 10.1007/s13225-016-0373-x [DOI] [Google Scholar]
  18. Hyde KD, Norphanphoun C, Abreu VP, Bazzicalupo A, Thilini Chethana KW, Clericuzio M, Dayarathne MC, Dissanayake AJ, Ekanayaka AH, Zhao RL, Mortimer PE. (2017) Fungal diversity notes 603–708: Taxonomic and phylogenetic notes on genera and species. Fungal Diversity 87: 1–235. 10.1007/s13225-017-0391-3 [DOI] [Google Scholar]
  19. Hyde K, Tennakoon D, Jeewon R, Bhat DJ, Maharachchikumbura S, Rossi W, Leonardi M, Lee H, Mun HY, Houbraken J, Nguyễn T, Jeon S, Frisvad J, Wanasinghe D, Lücking R, Aptroot A, Cáceres M, Karunarathna S, Hongsanan S, Doilom M. (2019) Fungal diversity notes 1036–1150: Taxonomic and phylogenetic contributions on genera and species of fungal taxa. Fungal Diversity 96: 1–242. 10.1007/s13225-019-00429-2 [DOI] [Google Scholar]
  20. Jayasiri SC, Jones EBG, Kang JC, Promputtha I, Bahkali AH, Hyde KD. (2016) A new species of genus Anteaglonium (Anteagloniaceae, Pleosporales) with its asexual morph. Phytotaxa 263(3): 233–244. 10.11646/phytotaxa.263.3.4 [DOI] [Google Scholar]
  21. Katoh K, Standley D. (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772–780. 10.1093/molbev/mst010 [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Koukol O, Delgado G. (2019) Do not forget Africa-revision of fungarium collections at Kew revealed a new species of Hermatomyces (Hermatomycetaceae, Pleosporales). Nova Hedwigia 109(3/4): 413–423. 10.1127/nova_hedwigia/2019/0559 [DOI]
  23. Koukol O, Delgado G, Hofmann TA, Piepenbring M. (2018) Panama, a hot spot for Hermatomyces (Hermatomycetaceae, Pleosporales) with five new species, and a critical synopsis of the genus. IMA Fungus 9(1): 107–141. 10.5598/imafungus.2018.09.01.08 [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koukol O, Delgado G, Tabé A, Yorou NS. (2025) The genus Hermatomyces in Benin, with the description of H. griseomarginatus, sp. nov. Mycologia 117(2): 286–296. 10.1080/00275514.2024.2433367 [DOI] [PubMed] [Google Scholar]
  25. Leão-Ferreira S, Gusmao L, Castañeda-Ruiz R. (2013) Conidial fungi from the semi-arid Caatinga biome of Brazil. Three new species and new records. Nova Hedwigia 96: 479–494. 10.1127/0029-5035/2013/0084 [DOI] [Google Scholar]
  26. Liu YJ, Whelen S, Hall BD. (1999) Phylogenetic relationships among ascomycetes: Evidence from an RNA polymerse II subunit. Molecular Biology and Evolution 16: 1799–1808. 10.1093/oxfordjournals.molbev.a026092 [DOI] [PubMed] [Google Scholar]
  27. Liu NG, Hyde KD, Sun YR, Bhat DJ, Jones EBG, Jumpathong J, Lin CG, Lu YZ, Yang J, Liu LL, Liu ZY, Liu JK. (2024) Notes, outline, taxonomy and phylogeny of brown-spored hyphomycetes. Fungal Diversity 129(1): 1–281. 10.1007/s13225-024-00539-6 [DOI] [Google Scholar]
  28. Nuankaew S, Suetrong S, Wutikhun T, Pinruan U. (2019) Hermatomyces trangensis sp. nov., a new dematiaceous hyphomycete (Hermatomycetaceae, Pleosporales) on sugar palm in Thailand. Phytotaxa 391(5): 277–288. 10.11646/phytotaxa.391.5.1 [DOI] [Google Scholar]
  29. O’Donnell K, Cigelnik E. (1997) Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution 7: 103–116. 10.1006/mpev.1996.0376 [DOI] [PubMed] [Google Scholar]
  30. Phukhamsakda C, McKenzie EHC, Phillips AJL, Gareth Jones EB, Jayarama Bhat D, Stadler M, Bhunjun CS, Wanasinghe DN, Thongbai B, Camporesi E, Ertz D, Jayawardena RS, Perera RH, Ekanayake AH, Tibpromma S, Doilom M, Xu JC, Hyde KD. (2020) Microfungi associated with Clematis (Ranunculaceae) with an integrated approach to delimiting species boundaries. Fungal Diversity 102: 1–203. 10.1007/s13225-020-00448-4 [DOI] [Google Scholar]
  31. Rao V, de Hoog GS. (1986) New or critical hyphomycetes from India. Studies in Mycology 28: 1–84. [Google Scholar]
  32. Rehner SA, Buckley E. (2005) A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: Evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97: 84–98. 10.1080/15572536.2006.11832842 [DOI] [PubMed] [Google Scholar]
  33. Ren GC, Wanasinghe DN, Monkai J, Mortimer PE, Hyde KD, Xu JC, Pang A, Gui H. (2021) Novel saprobic Hermatomyces species (Hermatomycetaceae, Pleosporales) from China (Yunnan Province) and Thailand. MycoKeys 82: 57–79. 10.3897/mycokeys.82.67973 [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ren GC, Wanasinghe DN, Jeewon R, Monkai J, Mortimer PE, Hyde KD, Xu JC, Gui H. (2022) Taxonomy and phylogeny of the novel Rhytidhysteron-like collections in the Greater Mekong Subregion. MycoKeys 86: 65–85. 10.3897/mycokeys.86.70668 [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP. (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542. 10.1093/sysbio/sys029 [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Samuels GJ, Müller E. (1979) Life-history studies of Brazilian Ascomycetes. 7. Rhytidhysteron rufulum and the genus Eutryblidiella. Sydowia 32: 277–292. [Google Scholar]
  37. Senanayake IC, Rathnayaka AR, Marasinghe DS, Calabon MS, Gentekaki E, Lee HB, Hurdeal VG, Pem D, Dissanayake LS, Wijesinghe SN, Bundhun D, Nguyen TT, Goonasekara ID, Abeywickrama PD, Bhunjun CS, Jayawardena RS, Wanasinghe DN, Jeewon R, Bhat DJ, Xiang MM. (2020) Morphological approaches in studying fungi: Collection, examination, isolation, sporulation and preservation. Mycosphere: Journal of Fungal Biology 11(1): 2678–2754. 10.5943/mycosphere/11/1/20 [DOI] [Google Scholar]
  38. Shen HW, Luo ZL, Bao DF, Luan S, Bhat DJ, Boonmee S, Wang WP, Su XJ, Li YX, Al-Otibi F, Lu YZ, Yang LQ, Hyde KD. (2024) Lignicolous freshwater fungi from China IV: Morphology and phylogeny reveal new species of Pleosporales from plateau lakes in Yunnan Province, China. Mycosphere: Journal of Fungal Biology 15(1): 6439–6524. 10.5943/mycosphere/15/1/28 [DOI] [Google Scholar]
  39. Shukla R, Pandey N, Banerjee S, Tripathi YB. (2017) Effect of extract of Pueraria tuberosa on expression of hypoxia inducible factor-1α and vascular endothelial growth factor in kidney of diabetic rats. Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie 93: 276–285. 10.1016/j.biopha.2017.06.045 [DOI] [PubMed] [Google Scholar]
  40. Singhal RK. (2009) Mining and the environment: From ore to metal. International Journal of Mining, Reclamation and Environment 23(4): e241. 10.1080/17480930903429794 [DOI]
  41. Son E, Yoon JM, An BJ, Lee YM, Cha J, Chi GY, Kim DS. (2019) Comparison among activities and isoflavonoids from Pueraria thunbergiana aerial parts and root. Molecules 24(5): e912. 10.3390/molecules24050912 [DOI] [PMC free article] [PubMed]
  42. Soto-Medina E, Lücking R. (2017) A new species of Rhytidhysteron (Ascomycota: Patellariaceae) from Colombia, with a provisional working key to known species in the world Artículo original. Revista de la Academia Colombiana de Ciencias Exactas, Físicas y Naturales 41(158): 59–63. 10.18257/raccefyn.423 [DOI] [Google Scholar]
  43. Spegazzini C. (1881) Fungi argentini additis nonnullis brasiliensibus montevideensibusque. Pugillus quartus (Continuacion). Anales de la Sociedad Científica Argentina 12: 174–189. [Google Scholar]
  44. Spegazzini C. (1910) Mycetes argentinenses (Series V). Anales del Museo Nacional de Historia Natural Buenos Aires 20: 329–467. [Google Scholar]
  45. Sun R, Deng X, Zhang D, Xie F, Wang D, Wang J, Tavallaie MS, Jiang F, Fu L. (2019) Anti-diabetic potential of Pueraria lobata root extract through promoting insulin signaling by PTP1B inhibition. Bioorganic Chemistry 87: 12–15. 10.1016/j.bioorg.2019.02.046 [DOI] [PubMed] [Google Scholar]
  46. Thambugala KM, Hyde KD, Eungwanichayapant PD, Romero AI, Liu ZY. (2016) Additions to the genus Rhytidhysteron in Hysteriaceae. Cryptogamie. Mycologie 37(1): 99–116. 10.7872/crym/v37.iss1.2016.99 [DOI] [Google Scholar]
  47. Tibpromma S, Bhat DJ, Doilom M, Lumyong S, Nontachaiyapoom S, Yang JB, Hyde KD. (2016) Three new Hermatomyces species (Lophiotremataceae) on Pandanus odorifer from Southern Thailand. Phytotaxa 275(2): 127–139. 10.11646/phytotaxa.275.2.4 [DOI] [Google Scholar]
  48. Tibpromma S, Hyde KD, Jeewon R, Maharachchikumbura SSN, Liu JK, Bhat DJ, Jones EBG, McKenzie EHC, Nontachaiyapoom S, Wen TC, Karunarathna SC. (2017) Fungal diversity notes 491–602: Taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 83: 1–261. 10.1007/s13225-017-0378-0 [DOI] [Google Scholar]
  49. Tibpromma S, Hyde KD, McKenzie EHC, Bhat DJ, Phillips AJL, Wanasinghe DN, Samarakoon MC, Jayawardena RS, Dissanayake AJ, Tennakoon DS, Doilom M, Phookamsak R, Tang AMC, Xu JC, Mortimer PE, Promputtha I, Maharachchikumbura SSN, Khan S, Karunarathna SC. (2018) Fungal diversity notes 840–928: Micro-fungi associated with Pandanaceae. Fungal Diversity 93: 1–160. 10.1007/s13225-018-0408-6 [DOI] [Google Scholar]
  50. Vilgalys R, Hester M. (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246. 10.1128/jb.172.8.4238-4246.1990 [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wanasinghe DN, Mortimer PE, Xu J. (2021) Insight into the systematics of microfungi colonizing dead woody twigs of Dodonaea viscosa in Honghe (China). Journal of Fungi 7(3): e180. 10.3390/jof7030180 [DOI] [PMC free article] [PubMed]
  52. Wang XL, Jiao FR, Yu M, Lin LB, Xiao J, Zhang Q, Wang L, Duan DZ, Xie G. (2017) Constituents with potent α-glucosidase inhibitory activity from Pueraria lobata (Willd.) ohwi. Bioorganic & Medicinal Chemistry Letters 27(9): 1993–1998. 10.1016/j.bmcl.2017.03.013 [DOI] [PubMed] [Google Scholar]
  53. Wang S, Zhang S, Wang S, Gao P, Dai L. (2020) A comprehensive review on Pueraria: Insights on its chemistry and medicinal value. Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie 131: e110734. 10.1016/j.biopha.2020.110734 [DOI] [PubMed]
  54. Wang HX, Zeng MS, Ye Y, Liu JY, Xu PP. (2021) Antiviral activity of puerarin as potent inhibitor of influenza virus neuraminidase. Phytotherapy Research: PTR 35(1): 324–336. 10.1002/ptr.6803 [DOI] [PubMed] [Google Scholar]
  55. White T, Bruns T, Lee S, Taylor J. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: A Guide to Methods and Applications 18: 315–322. 10.1016/B978-0-12-372180-8.50042-1 [DOI] [Google Scholar]
  56. Wijayawardene NN, Phillips AJL, Tibpromma S, Dai DQ, Selbmann L, Monteiro JS, Aptroot A, Flakus A, Rajeshkumar KC, Coleine C, Pereira DS, Fan X, Zhang L, Maharachchikumbura SSN, Souza MF, Kukwa M, Suwannarach N, Rodriguez-Flakus P, Ashtekar N, Dauner L, Tang LZ, Jin XC, Karunarathna SC. (2021) Looking for the undiscovered asexual taxa: Case studies from lesser studied life modes and habitats. Mycosphere: Journal of Fungal Biology 12(1): 1290–1333. 10.5943/mycosphere/12/1/17 [DOI] [Google Scholar]
  57. Xiong Y, Yang Y, Yang J, Chai H, Li Y, Yang J, Jia Z, Wang Z. (2010) Tectoridin, an isoflavone glycoside from the flower of Pueraria lobata, prevents acute ethanol-induced liver steatosis in mice. Toxicology 276(1): 64–72. 10.1016/j.tox.2010.07.007 [DOI] [PubMed] [Google Scholar]
  58. Xu L, Zheng N, He Q, Li R, Zhang K, Liang T. (2013) Puerarin, isolated from Pueraria lobata (Willd.), protects against hepatotoxicity via specific inhibition of the TGFbeta1/Smad signaling pathway, thereby leading to anti-fibrotic effect. Phytomedicine: International Journal of Phytotherapy and Phytopharmacology 20(13): 1172–1179. 10.1016/j.phymed.2013.06.008 [DOI] [PubMed] [Google Scholar]
  59. Yang YY, Li TS, Bhunjun CS, Thiyagaraja V, Bhat DJ, He SC, Zhao Q, Al-Otibi F, Kakumyan P, Hyde KD. (2025) Two novel species of Hermatomyces (Dothideomycetes) from Fagales in China. Mycological Progress. 10.1007/s11557-025-02105-z [DOI]
  60. Zhang B, Li W, Dong M. (2017) Flavonoids of kudzu root fermented by eurtotium cristatum protected rat pheochromocytoma line 12 (PC12) cells against H2O2-Induced apoptosis. International Journal of Molecular Sciences 18(12): e2754. 10.3390/ijms18122754 [DOI] [PMC free article] [PubMed]
  61. Zhang L, Fang DY, Liu C, Zhao DY, Wang YJ, Chen WN, Cui Y, Guo JF, Cong PW, Feng XF, Zhang YT. (2021) Mechanism of puerarin reversing SH-SY5Y cell injury induced by Aβ_(1-42) based on proteomics. Zhongguo Zhongyao Zazhi 46(14): 3650–3659. 10.19540/j.cnki.cjcmm.20210201.403 [DOI] [PubMed] [Google Scholar]
  62. Zhang YZ, Chen Q, Ma J, Lu YZ, Chen H, Liu NG. (2023) Morphological and multi-gene phylogenetic analyses reveal five new hyphomycetes from freshwater habitats. Frontiers in Microbiology 14: e1253239. 10.3389/fmicb.2023.1253239 [DOI] [PMC free article] [PubMed]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

XML Treatment for Hermatomyces nyingchiensis
mycokeys.125.174645-treatment1.xml (18.4KB, xml)
XML Treatment for Hermatomyces xizangensis
mycokeys.125.174645-treatment2.xml (16.2KB, xml)
XML Treatment for Rhytidhysteron xizangense
mycokeys.125.174645-treatment3.xml (20.5KB, xml)

Data Availability Statement

All of the data that support the findings of this study are available in the main text.

Articles from MycoKeys are provided here courtesy of Pensoft Publishers

RESOURCES