Four new species of Microdochium (Microdochiaceae, Xylariales) from Hainan, China

151 Four new species of Microdochium (Microdochiaceae, Xylariales) from Hainan, China Yuxin Shang1, Qiyun Liu1, Zhaoxue Zhang1, Mengyuan Zhang1, Zixu Dong1, Duhua Li1, Yaling Wang1, Congcong Ai2, Xiuguo Zhang1, Jiwen Xia3, Zhuang Li1 1 Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Taian, 271018, China 2 College of Life Sciences, Shandong Normal University, Jinan 250358, China 3 College of Agriculture and Forestry, Linyi University, Linyi, Shandong, 276000, China Corresponding author: Zhuang Li ([email protected]) Copyright: © Yuxin Shang et al. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International – CC BY 4.0). Research Article Abstract Microdochium has been reported worldwide as a plant pathogen, endophyte, or saprotroph. This study utilized four genetic loci (ITS, LSU, rpb2, and tub2) along with morphological characteristics, employing maximum likelihood and Bayesian inference analyses to identify and establish the taxonomic status of four new species from two host plants (Bambusaceae sp. and Phragmites australis) in Hainan Province, China. We propose four new species, Microdochium australiana sp. nov., M. baishamenense sp. nov., M. bambusina sp. nov., and M. bambusarum sp. nov. We provide detailed descriptions and micrographs of these species and compare them with other Microdochium species. Key words: Microdochium, multigene phylogeny, new taxa, taxonomy Introduction Microdochium (Microdochiaceae, Xylariales) was established by Sydow with its type species M. phragmitis, a fungus from Germany inhabiting living leaves of Phragmites australis (Sydow 1924). It belongs to the family Microdochiaceae, proposed by Hernández-Restrepo et al. (2016), which also includes genera such as Idriella and Selenodriella. Members of this family are characterized by a sexual morph resembling Monographella, producing ascocarps and hyaline ascospores, while the asexual morph exhibits multibudded, sympodial, or annellated conidiophores that bear hyaline conidia of various shapes (e.g., cylindrical, fusiform, oval) without accessory structures; some species also form chlamydospores. Although the genus exhibits a cosmopolitan distribution, most records are from Europe and Asia, and approximately 71 species are currently listed in Index Fungorum (http://www.indexfungorum.org/; accessed 30 June 2025). Notably, the morphological characteristics of perithecia, asci, and ascospores show only subtle differences between Microdochium and Monographella (Amphisphaeriaceae), whereas their anamorphic features are highly similar, leading to frequent historical misclassification of Microdochium species within Monographella (Parkinson et al. 1981; Samuels and Hallett 1983; Von Arx 1984). Academic editor: Xin-Cun Wang Received: 31 August 2025 Accepted: 24 November 2025 Published: 15 November 2025 Citation: Shang Y, Liu Q, Zhang Z, Zhang M, Dong Z, Li D, Wang Y, Ai C, Zhang X, Xia J, Li Z (2025) Four new species of Microdochium (Microdochiaceae, Xylariales) from Hainan, China. MycoKeys 126: 151–170. https://doi.org/10.3897/ mycokeys.126.170451 MycoKeys 126: 151–170 (2025) DOI: 10.3897/mycokeys.126.170451 152 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species Accurate species delimitation within Microdochium remains challenging due to insufficient molecular data and morphological convergence among some taxa. Although traditional taxonomy relied heavily on morphological characteristics prior to the adoption of DNA-based phylogenetic analyses (Hibbett et al. 2011), reliable species identification now necessitates the integration of molecular sequence data with detailed morphological characterization. Microdochium species exist not only as plant pathogens but also frequently as endophytes and saprophytes, and they are routinely isolated from a diverse range of plant hosts (Von Arx 1987; Glynn et al. 2005; Jewell and Hsiang 2013; Mandyam et al. 2013; Hiruma et al. 2018; Liang et al. 2019; Lu et al. 2023; Zhang et al. 2023). As pathogenic agents, they primarily affect grasses and cereal crops, causing serious diseases in economically important plants and resulting in yield reductions that lead to substantial economic losses (Gao et al. 2022). In southern China, this genus has been identified as the causal agent of tar spot disease on vetiver grass (Chrysopogon zizanioides) (Lu et al. 2023). The disease initially presents as scattered or aggregated black ascomatal stromata on the leaf surface, which are embedded within the leaf tissues (Lu et al. 2023). Microdochium species can also colonize plant tissues as endophytes. Many endophytic fungi inhabit plant tissues without causing harm, engaging in mutualistic relationships with their hosts (Rashid et al. 2021). The host provides nutrients, while endophytes enhance plant defense against pests and pathogens and stimulate immune responses (Rashid et al. 2021). Additionally, they suppress pathogenic fungi and promote plant growth (Ali et al. 2024). Certain species within the genus Microdochium are also recognized as saprophytes and soil inhabitants (Gao et al. 2022). Saprophytic fungi obtain nutrients from dead organic matter and play a crucial role as decomposers in ecosystems (Saldajeno et al. 2008). With the advancement of sequencing technologies, phylogenetic studies have transitioned from early single-gene analyses to more comprehensive approaches (Glynn et al. 2005; Jewell and Hsiang 2013; Hiruma et al. 2018; Liang et al. 2019; Liu et al. 2020; Zhang et al. 2023). Hong et al. (2008) employed the ITS sequence to preliminarily examine the phylogenetic relationships among three Microdochium species and their type species. Barr (1990) had previously used the LSU sequence to classify the related genus Monographella within the Subploasporaceae, a conclusion later revised by Lumbsch and Huhndorf (2010), who reassigned it to the Amphisphaeriaceae. We identified four novel species of Microdochium from specimens collected in Hainan Province, China. Two species were isolated from Bambusaceae and the other two from Phragmites australis. Morphological characteristics and molecular sequence data for these species are provided, with taxonomic implications discussed below. Materials and methods Morphological study Specimens were collected in 2024 from Hainan Province, China, and transported to the laboratory for analysis. Multiple fungal strains were typically obtained from a single specimen, from which pure cultures were subsequently acquired using both single-spore isolation and tissue isolation techniques (Zhang et al. 2025). 153 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species Tissue fragments (5 × 5 mm) were excised from the edges of leaf lesions and surface-sterilized by sequential immersion in 75% ethanol for 60 s, followed by rinsing in sterile distilled water for 45 s, then immersion in 5% sodium hypochlorite for 45 s, and repeated times in sterile distilled water. The sterilized leaf fragments were blotted dry with sterile paper towels and placed on potato dextrose agar (PDA). All plates were incubated in a biochemical incubator at 25 °C for 3–4 days, after which hyphae were picked from the colony margins and transferred onto new PDA plates. Pure cultures on PDA were incubated at 25 °C for 14 days, with images captured on days 7 and 14 using a Sony Alpha 6400 L digital camera (Sony Group Corporation, Tokyo, Japan). Microscopic examination of fungal structures was performed using an Olympus SZ61 stereomicroscope and an Olympus BX43 compound microscope (Olympus Corporation, Tokyo, Japan), which was equipped with a BioHDA20c color digital camera (FluoCa Scientific, Shanghai, China) for documentation and image capture of fungal structures. All fungal strains were preserved in sterile 10% glycerol at 4 °C for further studies. Specimens were deposited in the Herbarium of the Department of Plant Pathology, Shandong Agricultural University (HSAUP). Live cultures were stored in the Shandong Agricultural University Culture Collection (SAUCC) and the China General Microbiological Culture Collection Center (CGMCC). Taxonomic information for the new taxa has been submitted to MycoBank (http://www.mycobank.org/, accessed 30 June 2025). The abbreviation of the genus name used in this study is as follows: M. = Microdochium. DNA extraction, PCR amplification, and sequencing Genomic DNA was extracted from fungal mycelia scraped from colonies grown on PDA medium using the cetyltrimethylammonium bromide (CTAB) method (Guo et al. 2000; Zhang ang Wang 2020). Four gene regions (ITS, LSU, rpb2, and tub2) were amplified using the primer pairs listed in Table 1. PCR amplifications were performed in a 25 μL reaction volume using an Eppendorf Master Thermocycler (Hamburg, Germany), containing 12.5 μL of 2 × Hieff Canace® Plus PCR Master Mix (Cat. No. 10154ES03, Yeasen Biotechnology, Shanghai, China), 1 μL each of forward and reverse primers (10 μM, TsingKe, Qingdao, China), 1 μL of template genomic DNA (approximately 10 ng/μL), and double-distilled water to adjust the final volume. PCR products were separated by 1% agarose gel electrophoresis, stained with GelRed, and visualized under ultraviolet light to confirm bands of the expected size. Amplicons were then purified using a gel extraction kit (Cat. AE0101-C; Shandong Sparkjade Biotechnology Co., Ltd., Jinan, China). Bidirectional sequencing was performed by Youkang Company Limited (Zhejiang, China). Consensus sequences were assembled using MEGA v. 7.0 (Kumar et al. 2016). All sequences generated in this study have been deposited in GenBank, with accession numbers provided in Table 2. Phylogenetic analyses In this study, 32 newly generated sequences were submitted to the NCBI GenBank database, and available reference sequences of Microdochium species were retrieved from GenBank for phylogenetic analysis. Sequences newly generated in this study were aligned alongside relevant sequences obtained from GenBank via the online MAFFT 7 tool implemented with the Auto strategy (Katoh et al. 2019; 154 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species Table 2. GenBank accession numbers of the taxa used in the phylogenetic reconstruction. Species Culture accession GenBank accession numbers Reference ITS LSU tub2 rpb2 Microdochium albescens CBS 243.83 KP858994 KP858930 KP859057 KP859103 (Hernández-Restrepo et al. 2016) M. albescens CBS 291.79 KP858996 KP858932 KP859059 KP859105 (Hernández-Restrepo et al. 2016) M. australiana SAUCC 6340-2-6* = CGMCC 3.28622 PQ807110 PV609100 PV686755 PV975978 This study M. australiana SAUCC 8723-2 PQ807111 PV609101 PV686756 PV975979 This study M. bambusae SAUCC 1862-1* OR702567 OR702576 OR715791 OR715785 (Zhang et al. 2023) M. bambusae SAUCC 1866-1 OR702568 OR702577 OR715792 OR715786 (Zhang et al. 2023) M. bambusina SAUCC 7531-3* = CGMCC 3.28623 PQ807108 PV609098 PV686753 PV975976 This study M. bambusina SAUCC 7638-2 PQ807109 PV609099 PV686754 PV975977 This study M. bambusarum SAUCC 7611-3* = CGMCC 3.28624 PQ807112 PV609102 PV686757 PV975980 This study M. bambusarum SAUCC 6699-4 PQ807113 PV609103 PV686758 PV975981 This study M. baishamenense SAUCC 8129-1* = CGMCC 3.28625 PQ807114 PV609104 PV686759 PV975982 This study M. baishamenense SAUCC 7263-1 PQ807115 PV609105 PV686760 PV975983 This study M. bolleyi CBS 540.92 KP859010 KP858946 KP859073 KP859119 (Hernández-Restrepo et al. 2016) M. chrysanthemoides LC5363* KU746690 KU746736 KU746781 KY883244 (Zhang et al. 2017) M. chrysanthemoides LC5466 KU746689 KU746735 KU746782 KY883245 (Zhang et al. 2017) M. chrysopogonis GDMCC 3.683 MT988022 MT988024 MW002441 MW002444 (Lu et al. 2023) M. chuxiongense YFCC 8794 OK5861616 OK586160 OK556901 OK584019 (Tang et al. 2022) M. citrinidiscum CBS 109067* KP859003 KP858939 KP859066 KP859112 (Hernández-Restrepo et al. 2016) M. colombiense CBS 624.94* KP858999 KP858935 KP859062 KP859108 (Hernández-Restrepo et al. 2016) M. dawsoniorum BRIP 65649* MK966337 ON394569 - - (Crous et al. 2020) M. dawsoniorum BRIP 67439a MN492650 OM333563 -ON624208 (Crous et al. 2020) M. fisheri CBS 242.90 KP859015 KP858951 KP859079 KP859124 (Hernández-Restrepo et al. 2016) M. graminearum CGMCC 3.23524 OP103965 OP104015 OP242835 OP236026 (Gao et al. 2022) M. graminearum CGMCC 3.23525* OP103966 OP104016 OP236029 OP236027 (Gao et al. 2022) M. graminis ZJE01778* PP111928 PP111935 PP112588 PP112593 (Yan and Zhang 2024) M. graminis B13 HQ696038 - - - (Shen et al. 2014) M. graminis PE110 JX875927 - - - (Shen et al. 2012) M. gongcheniae YNE01155 PP111925 PP111932 PP112585 - (Yan and Zhang 2024) M. gongcheniae YNE01164* PP111926 PP111933 PP112586 - (Yan and Zhang 2024) M. hainanense SAUCC 210781* OM956295 OM959323 OM981146 OM981153 (Liu et al. 2022) M. hainanense SAUCC 210782 OM956296 OM959324 OM981147 OM981154 (Liu et al. 2022) M. hongkuii YNE00384 PP111922 PP111929 PP112582 PP112589 (Yan and Zhang 2024) M. hongkuii YNE00483* PP111923 PP111930 PP112583 - (Yan and Zhang 2024) M. hongkuii N115 MK304137 - - - From NCBI M. indocalami SAUCC 1016* MT199884 MT199878 MT435653 MT510550 (Huang et al. 2020) M. insulare BRIP 75114a OQ917075 OQ892168 -OQ889560 (Tan and Shivas 2023a) M. lycopodinum CBS 109397 KP859004 KP858940 KP859067 KP859113 (Hernández-Restrepo et al. 2016) M. lycopodinum CBS 109398 KP859005 KP858941 KP859068 KP859114 (Hernández-Restrepo et al. 2016) M. lycopodinum CBS 125585* KP859016 KP858952 KP859080 KP859125 (Hernández-Restrepo et al. 2016) Table 1. Gene loci and corresponding PCR primers and programs used in this study. LocusPCR primers Sequence (5’ – 3’) PCR cycles References ITS ITS5 GGA AGT AAA AGT CGT AAC AAG G (94 °C: 30 s, 55 °C: 30 s, 72 °C: 45 s) × 29 cycles (White et al. 1990) ITS4 TCC TCC GCT TAT TGA TAT GC LSU LR0R GTA CCC GCT GAA CTT AAG C (94 °C: 30 s, 48 °C: 50 s, 72 °C: 1 min 30 s) × 35 cycles (Vilgalys and Hester 1990) LR5 TCC TGA GGG AAA CTT CG rpb2 RPB2-5F2 GGG GWG AYC AGA AGA AGG C (94 °C: 45 s, 60 °C: 45 s, 72 °C: 2 min) × 5 cycles, (94 °C: 45 s, 54 °C: 45 s, 72 °C: 2 min) × 30 cycles (Liu et al. 1999; Sung et al. 2007) RPB2-7CR CCC ATR GCT TGY TTR CCC AT tub2 Btub526-F CGA GCG YAT GAG YGT YTA CTT (95 °C: 30 s, 56 °C: 30 s, 72 °C: 45 s) × 35 cycles (Jewell and Hsiang 2013) Btub1332-R TCA TGT TCT TGG GGT CGA A 155 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species Species Culture accession GenBank accession numbers Reference ITS LSU tub2 rpb2 M. maculosum COAD 3358* OK966954 OK966953 -OL310501 (Crous et al. 2021a, b) M. majus CBS 741.79 KP859001 KP858937 KP859064 KP859110 (Hernández-Restrepo et al. 2016) M. miscanthi SAUCC 211092* OM956214 OM957532 OM981141 OM981148 (Liu et al. 2022) M. miscanthi SAUCC 211093 OM956215 OM957533 OM981142 OM981149 (Liu et al. 2022) M. musae CBS 143499 MH107894 MH107941 MH108040 - (Crous et al. 2018) M. musae CBS 143500* MH107895 MH107942 -MH108003 (Crous et al. 2018) M. nannuoshanense SAUCC 2450-1* OR702569 OR702578 OR715793 OR715787 (Zhang et al. 2023) M. nannuoshanense SAUCC 2450-3 OR702570 OR702579 OR715794 OR715788 (Zhang et al. 2023) M. neoqueenslandicum CBS 108926* KP859002 KP858938 KP859065 KP859111 (Hernández-Restrepo et al. 2016) M. neoqueenslandicum CBS 445.95 KP858997 KP858933 KP859060 KP859106 (Hernández-Restrepo et al. 2016) M. nivale CBS 116205* KP859008 KP858944 KP859071 KP859117 (Hernández-Restrepo et al. 2016) M. nivale CBS 288.50 MH856626 MH868135 - - (Vu et al. 2019) M. novae-zelandiae CBS 143847* LT990655 LT990627 LT990608 LT990641 (Marin-Felix et al. 2019) M. novae-zelandiae CPC 29693 LT990656 LT990628 LT990609 LT990642 (Marin-Felix et al. 2019) M. paspali CBS 138620* KJ569513 -KJ569518 - (Zhang et al. 2015) M. paspali QH-BA-48 KJ569510 -KJ569515 - (Zhang et al. 2015) M. phyllosaprophyticum SAUCC 3583-1* OR702571 OR702580 OR715795 OR715789 (Zhang et al. 2023) M. phyllosaprophyticum SAUCC 3583-6 OR702572 OR702581 OR715796 OR715790 (Zhang et al. 2023) M. phragmitis CBS 285.71* KP859013 KP858949 KP859077 KP859122 (Hernández-Restrepo et al. 2016) M. phragmitis CBS 423.78 KP859012 KP858948 KP859076 KP859121 (Hernández-Restrepo et al. 2016) M. poae LC12114* MH740898 -MH740914 MH740906 (Liang et al. 2019) M. ratticaudae BRIP 68298* MW481661 MW481666 -MW626890 (Crous et al. 2021a, b) M. rhopalostylidis CBS 145125* MK442592 MK442532 -MK442667 (Crous et al. 2019) M. salmonicolor KCTC 56427 NR173378 MK836108 - - (Das et al. 2020) M. seminicola KAS1516 KP859025 KP858961 KP859088 KP859134 (Hernández-Restrepo et al. 2016) M. seminicola KAS3576 KP859038 KP858974 KP859101 KP859147 (Hernández-Restrepo et al. 2016) M. shilinense CGMCC 3.23531* OP103972 OP104022 OP242834 - (Gao et al. 2022) M. sichuanense KUNCC23-13008* OQ616510 OQ616434 -OQ623473 (Dissanayake et al. 2023) M. sinense SAUCC 211097* OM956289 OM959225 OM981144 OM981151 (Liu et al. 2022) M. sinense SAUCC 211098 OM956290 OM959226 OM981145 OM981152 (Liu et al. 2022) M. sorghi CBS 691.96 KP859000 KP858936 KP859063 KP859109 (Hernández-Restrepo et al. 2016) Microdochium sp. YNE01043 PP111924 PP111931 PP112584 PP112591 (Yan and Zhang 2024) Microdochium sp. YNE01771 PP111927 PP111934 PP112587 PP112592 (Yan and Zhang 2024) Microdochium sp. ZJ40 KJ572190 - - - From NCBI M. streetiae BRIP 74742a* OR947072 OR947079 - - (Tan and Shivas 2023b) M. streetiae BRIP 74752a OR947071 OR947078 - - (Tan and Shivas 2023b) M. tainanense CBS 269.76* KP859009 KP858945 KP859072 KP859118 (Hernández-Restrepo et al. 2016) M. tainanense CBS 270.76 KP858995 KP858931 KP859058 KP859104 (Hernández-Restrepo et al. 2016) M. trichocladiopsis CBS 623.77* KP858998 KP858934 KP859061 KP859107 (Vu et al. 2019) M. triticicola RR 241 AJ748691 ---(Kwasna and Bateman 2007) M. yunnanense SAUCC 1011* MT199881 MT199875 MT435650 MT510547 (Huang et al. 2020) M. yunnanense SAUCC 1015 MT199883 MT199877 MT435652 MT510549 (Huang et al. 2020) Peglionia verticiclada CBS 127654* ON400763 ON400815 -ON399352 (Hernández-Restrepo et al. 2022) Selenodriella brasiliana CBS 140227* ON400769 ON400821 -ON399356 (Hernández-Restrepo et al. 2022) Selenodriella cubensis CBS 683.96* KP859053 KP858990 - - (Hernández-Restrepo et al. 2016) Selenodriella fertilis CBS 772.83 KP859055 KP858992 - - (Hernández-Restrepo et al. 2016) Selenodriella fertilis CPC 16273 ON400771 ON400823 -ON399358 (Crous et al. 2019) Muscodor fengyangensis CGMCC 2862* HM034856 HM034859 HM034843 HM034849 (Zhang et al. 2010) Muscodor thailandicus MFLUCC 17-2669* MK762707 MK762714 MK776960 MK791283 (Samarakoon et al. 2020) Notes: Species established in this study are shown in bold. Those marked “*” in the table are represented as ex-type or ex-epitype strains. “-’’: Not available. http://mafft.cbrc.jp/alignment/server/), followed by manual refinement using BioEdit (Hall 2006). To ascertain the species affiliation of the isolates, initial phylogenetic analyses were carried out for each locus individually, which was subsequently followed by a concatenated phylogenetic analysis incorporating all four loci (ITS, LSU, rpb2, and tub2). Phylogenetic analyses were conducted on both the 156 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species individual and concatenated alignments of ITS, LSU, rpb2, and tub2 sequences using maximum likelihood (ML) and Bayesian inference (BI) algorithms. Prior to BI analysis, the optimal evolutionary model for each partition was determined using MrModelTest v. 2.3 based on the Akaike Information Criterion (AIC), and the selected models were integrated into the analytical framework. ML and BI analyses were performed via either the CIPRES Science Gateway portal (https://www. phylo.org/, accessed on 30 June 2025) or offline software: ML was run using RAxML-HPC2 on XSEDE v. 8.2.12, and BI was conducted using MrBayes v. 3.2.7a on Linux with 64 threads. ML analysis used default parameters and was run with the GTR+G+I substitution model for 1,000 rapid bootstrap replicates, while BI utilized a rapid bootstrapping algorithm combined with an automatic stop feature. Finally, the resulting phylogenetic trees were visualized using FigTree v. 1.4.4 (http://tree.bio.ed.ac.uk/software/figtree, accessed on 30 June 2025) or ITOL: Interactive Tree of Life (https://itol.embl.de/, accessed on 30 June 2025), and final tree layouts were refined using Adobe Illustrator CC 2019. Results Phylogenetic analyses A phylogenetic analysis of Microdochium strains included a total of 80 sequences, with Muscodor fengyangensis (CGMCC 2862) and Muscodor thailandicus (MFLUCC 17-2669) as outgroups and sequences of Selenodriella and Peglionia as sister groups. The final concatenated alignment comprised 3,070 characters, viz. 1–681 (ITS), 682–1,515 (LSU), 1,516–2,369 (rpb2), and 2,370–3,274 (tub2). The final maximum likelihood (ML) log likelihood was 26959.183878. The matrix contained 1,300 distinct alignment patterns, with 27.86% of characters or gaps remaining undetermined. Estimated base frequencies were A = 0.219963, C = 0.292052, G = 0.257504, and T = 0.230481; substitution rates were AC = 1.126641, AG = 4.443240, AT = 1.324086, CG = 0.757689, CT = 6.915959, and GT = 1.000000. The alignment contained 1,304 unique site patterns (ITS: 315, LSU: 148, rpb2: 430, tub2: 411). The topology of the ML tree was congruent with that of the BI tree; thus, only the ML tree is shown (Fig. 1). The present study identified four novel species, viz. Microdochium australiana sp. nov., M. baishamenense sp. nov., M. bambusina sp. nov., and M. bambusarum sp. nov. Taxonomy Microdochium australiana Y.X. Shang, Z. Li & X.G. Zhang, sp. nov. MycoBank No: 857092 Fig. 2 Etymology. Referring to the species name of the host plant, Phragmites australis. Holotype. HSAUP 6340-2-6. Description. On leaves of Phragmites australis, Mycelia superficial and immersed, 2.2–2.9 µm wide, branched, membranous, hyaline. Conidiophores straight or slightly curved, aseptate, aggregated in the aerial mycelium, often reduced to conidiogenous cells borne directly from the hyphae. Conidiogenous cells terminal or intercalary, transparent, smooth, 157 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species 0.05 Microdochium triticicola RR 241 Microdochium nannuoshanense SAUCC 2450-3 Microdochium bolleyi CBS 540.92 Microdochium gongcheniae YNE01164* SAUCC 7263-1* Selenodriella cubensis CBS 683.96* Microdochium chuxiongense YFCC 8794 Microdochium albescens CBS 243.83 Microdochium streetiae BRIP 74742a* Microdochium novae-zelandiae CPC 29693 Muscodor thailandicus MFLUCC 17-2669* Microdochium paspali CBS 138620* Microdochium tainanense CBS 269.76 * Microdochium hainanense SAUCC 210781 * Microdochium rhopalostylidis CBS 145125 * Microdochium hongkuii YNE00483* Microdochium miscanthi SAUCC 211092* Microdochium poae LC12114 * Microdochium salmonicolor KCTC 56427 Microdochium seminicola CBS KAS3576* Microdochium dawsoniorum BRIP 65649* Microdochium bambusae SAUU 1866-1 Microdochium sp.YNE01043 Microdochium citrinidiscum CBS 109067* Microdochium shilinense CGMCC 3.23531 * Microdochium phragmitis CBS 285.71 * SAUCC 7638-2* SAUCC 8723-2* Microdochium sinense SAUCC 211097* Microdochium hongkuii N115 Microdochium miscanthi SAUCC 211093 Microdochium chrysopogonis GDMCC 3.683 Microdochium trichocladiopsis CBS 623.77 * Microdochium indocalami SAUCC 1016 * Microdochium albescens CBS 291.79 Microdochium lycopodinum CBS 109397 Microdochium nannuoshanense SAUCC 2450-1* Microdochium graminis PE110 Microdochium lycopodinum CBS 125585 * Microdochium sorghi CBS 691.96 Microdochium lycopodinum CBS 109398 Microdochium sp. ZJ40 SAUCC 7531-3 Microdochium bambusae SAUU 1862-1 Selenodriella fertilis CBS 772.83 SAUCC 6699-4 Microdochium yunnanense SAUCC1015 Microdochium chrysanthemoides LC5363* Selenodriella brasiliana CBS 140227* Microdochium majus CBS 741.79 Microdochium gongcheniae YNE01155 Microdochium streetiae BRIP 74752a Microdochium musae CBS 143500 * Microdochium seminicola KAS1516 Microdochium novae-zelandiae CBS 143847 * Microdochium maculosum COAD 3358 * Microdochium sp. YNE01771 Microdochium tainanense CBS 270.76 Microdochium yunnanense SAUCC1011* SAUCC 7611-3* Microdochium graminearum CGMCC 3.23525* Microdochium neoqueenslandicum CBS 445.95 SAUCC 8129-1 Microdochium chrysanthemoides LC5466 Microdochium nivale CBS 116205* Peglionia verticiclada CBS 127654* Muscodor fengyangensis CGMCC 2862* Microdochium hainanense SAUCC 210782 Microdochium colombiense CBS 624.94* Selenodriella fertilis CPC 16273 SAUCC 6340-2-6 Microdochium sichuanense KUNCC23-13008 * Microdochium phragmitis CBS 423.78 Microdochium dawsoniorum BRIP 67439a Microdochium insulare BRIP 75114a Microdochium neoqueenslandicum CBS 108926* Microdochium fisheri CBS 242.90 Microdochium ratticaudae BRIP 68298 * Microdochium phyllosaprophyticum SAUCC 3583-6 Microdochium paspali QH-BA-48 Microdochium graminis ZJE01778* Microdochium phyllosaprophyticum SAUCC 3583-1* Microdochium graminearum CGMCC 3.23524 Microdochium sinense SAUCC 211098 Microdochium nivale CBS 288.50 Microdochium musae CBS 143499 Microdochium graminis B13 35 96/1 74/0.94 77/0.99 100/1 98/1 100/1 100/1 35/0.76 99/1 76/0.92 100/1 29/0.5 83/0.73 100/1 42/0.99 93/0.86 30/0.66 100/1 100/0.71 100/1 94/1 100/1 40/0.99 66/0.63 89/0.86 100/1 50/1 100/1 100/1 68/0.86 100/1 54/1 100/1 100/1 63/0.99 98/0.99 100/1 90/0.99 100/0.99 100/1 100/1 100/1 100/1 100/1 100/1 97/1 99/1 98/1 76/0.99 97/0.9 92/0.99 100/1 100/1 95/1 100/1 100/1 91/1 100/1 94/1 54/0.72 70/1 100/0.92 67/0.98 100/1 29/0.99 97/1 94/1 100/1 Microdochium hongkuii YNE00384 99/1 100/1 23/0.57 77/0.99 61/0.98 100/1 33/0.86 60/0.72 49/0.74 97/1 91/0.99 45/0.76 80/0.82 Microdochium australiana sp.nov. Microdochium baishamenense sp.nov. Microdochium bambusarum sp.nov. Microdochium bambusina sp.nov. Figure 1. Maximum likelihood inference tree based on a combined dataset of analyzed ITS, LSU, rpb2, and tub2 sequences. The Bayesian inference posterior probability (left, BIPP ≥ 0.90) and the maximum likelihood bootstrap value (right, MLBV ≥ 75%) are shown as BIPP/MLBV above the nodes. Those marked “*” in the tree are represented as ex-type or ex-epitype strains. Strains isolated in this study are indicated in red. The scale bar at the bottom indicates 0.07 substitutions per site. To enhance the visual appeal of the evolutionary tree layout, certain branches are shortened by two diagonal lines (“//”) with the number of times. 158 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species cylindric-clavate, 6.2–7.3 × 1.9–3.5 µm. Conidia solitary, cylindrical, ampulliform, 8.2–10.1 × 3.0–4.9 µm, multi-guttulate, 0–2-septate, apex rounded, base usually flattened. Sporodochia and chlamydospores not observed. Culture characteristics. Cultures incubated on PDA at 25 °C in darkness, reaching 48–50 mm diam, had a growth rate of 6.9–7.1 mm/day after 7 days. The center has obvious milky white aerial mycelium bulge, and the edge grayish, mycelium fluffy, back fawn. Material examined. China • Hainan Province, Jianfengling National Forest Park, on leaves of Phragmites australis, 26 June 2024, Y.X. Shang, (HSAUP 6340-2-6), ex-holotype culture SAUCC 6340-2-6 = CGMCC 3.28622; Ibid., (HSAUP 8723-2, paratype), living culture SAUCC 8723-2. Figure 2. Microdochium australiana (CGMCC 3.28622, ex-holotype culture). a. A leaf of Phragmites australis; b, c. Colonies on PDA from above and below after 7 days; d. Colony overview; e, f. Conidiogenous cells and conidia; g, h. Conidia. Scale bars: 10 μm (e–h). 159 MycoKeys 126: 151–170 (2025), DOI: 10.3897/mycokeys.126.170451 Yuxin Shang et al.: We propose four new species Notes. Microdochium australiana is closely related to M. miscanthi (SAUCC211092 and SAUCC211093) based on DNA sequence data in BLAST searches and phylogenetic analysis (Fig. 1). However, M. australiana differs from M. miscanthi by 28 nucleotides (1/541 in ITS, 1/866 in LSU, 13/730 in tub2, and 13/665 in rpb2). In morphology, they are distinguished by different hosts (Phragmites australis vs. Miscanthus sinensis), and M. australiana colonies on PDA exhibit a prominent, milky white aerial mycelium bulge, with a central pale orange region and a grayish edge. In contrast, M. miscanthi colonies are overall white, featuring a central dark-green plaque covered by white mycelia. Mycelial width (2.2–2.9 µm) in M. australiana vs. (1.5–2.3 µm) in M. miscanthi. Conidia in M. australiana differ from those in M. miscanthi (cylindrical, ampulliform vs. transparent, spindle-to-rod-shaped) (Liu et al. 2022). Therefore, we establish this fungus as M. australiana sp. nov. Microdochium baishamenense Y.X. Shang, Z. Li & X.G. Zhang, sp. nov. MycoBank No: 857094 Fig. 3 Etymology. The epithet “baishamenense” is named after Baishamen Park, where the fungus was collected. Holotype. HSAUP 8129-1. Type. China, Hainan Province, Baishamen Park, on leaves of Bambusaceae sp., 24 June 2024, Y.X. Shang, (HSAUP 8129-1), ex-holotype culture SAUCC 8129-1 = CGMCC 3.28625. Description. On leaves of Bambusaceae sp. Mycelium superficial and immersed; hyphae hyaline, branched, septate. Conidiophores slightly differentiated, bifurcate, hyaline, smooth. Conidiogenous cells terminal, sympodial, denticulate, cylindrical, 19–60 × 1.5–2 μm, hyaline, smooth. Conidia soli tary, dry, fusiform, obovoid, subpyriform, to clavate, 7–12 × 3–4 μm, 0–1-septate, hyaline, tapering to a subtruncate hilum; hilum unpigmented. Chlamydospores not observed. Culture characteristics. Cultures incubated on PDA at 25 °C in darkness, reaching 46–51 mm diam, had a growth rate of 6.5–7.2 mm/day after 7 days. The center has milky white, mycelia lush and the edge mycelium was sparse, pale brown, back light yellow. Material examined. China • Hainan Province, Baishamen Park, on leaves of Bambusaceae sp., 24 June 2024, Y.X. Shang, (HSAUP 8129-1), ex-holotype culture SAUCC 8129-1 = CGMCC 3.28625; Ibid., (HSAUP 7263-1, paratype), living culture SAUCC 7263-1. Notes. Phylogenetic analysis showed that Microdochium baishamenense formed an independent clade, where it shows a relationship with M. fisheri (CBS 242.90). M. baishamenense differs from M. fisheri (CBS 242.90) by its production of shorter conidiogenous cells (4.5–7.3 × 1.5–2.4 µm vs. 19–60 × 1.5–2 μm) and distinct morphological features. Specifically, the center of M. baishamenense colonies is milky white, with lush mycelia; the mycelia at the edge are sparse and pale brown, and the colony reverse is light yellow. In contrast, M. fisheri (CBS 242.90) exhibits a salmon-colored center, a peach-colored periphery, and an entire margin. 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The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/mycokeys.126.170451.suppl1