Biodiversity Data Journal 11: e97439 OO) doi: 10.3897/BDJ.11.e97439 open access Taxonomy & Inventories A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) from freshwater habitats in China Xin-Yi Yan*S!, Jun-En Huang", Hai-Yan Song*, Yang GaoS!*, Hai-Jing Hul*8, Zhi-Jun Zhail, Jun-Qing Yan§, Guang-Hua Huot, Dian-Ming HuS $ Jiangxi Key Laboratory for Conservation and Utilization of Fungal Resources, Jiangxi Agricultural University, Nanchang, China § Jiangxi Agricultural University, Nanchang, China | Bioengineering and Technological Research Centre for Edible and Medicinal Fungi, Jiangxi Agricultural University, Nanchang, China | Chinese Academy of Sciences, Beijing, China # Key Laboratory of Crop Physiology, Ecology and Genetic Breeding, (Jiangxi Agricultural University), Ministry of Education of the P.R., Nanchang, China Corresponding author: Dian-Ming Hu (hudianming1@163.com) Academic editor: Christian Wurzbacher Received: 11 Nov 2022 | Accepted: 04 Apr 2023 | Published: 28 Apr 2023 Citation: Yan X-Y, Huang J-E, Song H-Y, Gao Y, Hu H-J, Zhai Z-J, Yan J-Q, Huo G-H, Hu D-M (2023) A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) from freshwater habitats in China. Biodiversity Data Journal 11: e€97439. https://doi.org/10.3897/BDJ.11.e€97439 Abstract Background Freshwater fungi refer to the fungi that depend on the freshwater habitats for the whole life cycle or part of their life cycle. In this context, a new aquatic hyphomycete was isolated from decaying wood in a freshwater habitat in Jiangxi Province, China. New information Dictyochaeta jiangxiensis sp. nov., a new aquatic hyphomycete, is characterised by its unbranched, septate, base-fertile conidiophores with multisepta and single phialide at the © Yan X et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 2 Yan X et al apex, brown, sterile seta, monophialidic, subcylindrical conidiogenous cells narrowing below the funnel-shaped collarette, hyaline, unicellular, thin-walled, smooth, guttulate, falcate to subclavate conidia narrowly rounded at both ends with hair-like appendages. Phylogenetically, the new species Dictyochaeta jiangxiensis clustered together with Dictyochaeta brevis MFLU 19-0216 in a well-supported clade, but formed a separate branch. In order to better define the taxonomic status of the new species, a phylogenetic tree of most closely-related taxa in Chaetosphaeriaceae was established, based on multi- locus sequences (ITS and LSU). The novel species is described and illustrated. Newly- generated molecular data of Dictyochaeta Jiangxiensis is also provided. Keywords dematiaceous hyphomycete, new species, taxonomy, phylogeny Introduction Spegazzini (1923) established the genus Dictyochaeta with D. fuegiana as type species, which was isolated from fallen leaves of Nothofagus betuloides (Mirb.) Oerst. Gamundi et al. (1977) and Godeas et al. (1977) verified the holotype and redescribed it. Later, Reblova (2004) re-examined the type species and gave more detailed description about the genus Dictyochaeta, as two-layer conidiophores, the upper layer setiform, when sterile, monophialidic or rarely polyphialidic, the lower layer always fertile, monophialidic, rarely polyphialidic, collarette on conidiogenous cells and aseptate, hyaline, falcate conidia without setulae. Since then, more and more species have been discovered and classified as or transferred to Dictyochaeta and its molecular and morphological data have been expanded. According to literature and herbarium records, the Dictyochaeta-like fungi are globally distributed in the Holarctic Region and the Tropics and grow on decaying plant, such as bark, wood, bamboo culms, palm fronds, fallen leaves and petioles in freshwater and terrestrial environments. They also occur as plant pathogens or endophytes in living plants. (Agnihothrudu 1968, Lunghini et al. 1971, Shearer and Crane 1971, Sutton and Hodges 1975, Hewings and Crane 1981, Holubova-Jechova 1984, Kuthubutheen 1987, Kuthubutheen and Nawawi 1990, Kuthubutheen and Nawawi 1991a, Kuthubutheen and Nawawi 1991b, Reblova et al. 1999, Kirschner and Chen 2002, Crous et al. 2014, Crous et al. 2015, Maharachchikumbura et al. 2016). There are always different opinions on the classification of Dictyochaeta, which is considered as a synonym of several genera, such as Codinaea Maire., Menispora Pers. and Menisporopsis S. Hughes. Previously, Dictyochaeta was usuallly regarded as a synonym of Codinaea (Maire 1937), because they share greatly similar features on phialidic conidiogenous cells and setae, but differ mainly in the conidia without setulae (Cai et al. 2006). Crous et al. (2018) thought that priority should be given to the older name, Dictyochaeta. Reblova (2000) recommended that species with setulae should be classified into Codinaea and those without setulae into Dictyochaeta. Réblova et al. (2021a) re- evaluated the concept of Dictyochaeta and revised species delimitation, based on six loci A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae)... 3 (ITS, LSU, SSU, RPB2, TEF1-a, TUB2) along with comparative morphological and cultivation studies. In their study, some species of Dictyochaeta, such as D. siamensis, D. simplex etc. clustered within the clade Codinaea with a high support. As for the demarcation between Dictyochaeta and Codinaea, Reblova et al. (2021a) supported using conidial appendages as a Classification criterion to distinguish Dictyochaeta from Codinaea (Réeblova and Winka 2000). Based on revised species, morphological characteristics of conidia (shape, septation, absence or presence of setulae), collarettes (Shape) and setae (presence or absence) and extension of the conidiogenous cell proved to be important at the generic level. To date, Dictyochaeta-like fungi, together with Codinaea-like fungi, were divided into five lineages in the phylogenetic analyses (Reéeblova et al. 2021b). Dual DNA barcoding and ancestral reconstruction of ecological and geographic distribution facilitated re-assessment of Dictyochaeta-like fungi. Reblova et al. (2021b) introduced five genera (Codinaeella, Nimesporella, Stilbochaeta, Tainosphaeriella and Xyladelphia) to accommodate Codinaea-like fungi and retained the taxonomic status of Dictyochaeta sensu stricto. Materials and methods Sample collection and specimen examination Submerged wood samples were collected in a stream from Jishui County, Ji’an City, Jiangxi Province, China on 9 April 2018. The samples were taken to the laboratory in ziplock bags and placed in plastic boxes. The microscopic analysis was performed by a stereomicroscope to observe the fungal fruiting body on a natural substrate. Micro- examination and photomicrographs were taken under a compound microscope (Nikon Ni). The specimens were deposited in the Herbarium of Fungi, Jiangxi Agricultural University (HFJAU), Nanchang, China. DNA extraction, PCR amplification and sequencing Cultures were grown at room temperature on potato-dextrose agar (PDA). Mycelia were directly scraped off from plates and transferred into centrifugal tube after fragmentation. DNA was extracted with the CTAB method following Doyle and Doyle (1987). Approximately 500 mg of mycelium was mixed with ca. 0.2 g of white quartz sand and ground with preheated (ca. 65°C) 2 x CTAB buffer [2% (w/v) CTAB; 100 mM Tris-HCl; 1.4 M NaCl; 20 mM EDTA, pH 8.0]. DNA was extracted by chloroform:isoamyl alcohol (24:1) and precipitated by isopropanol at -20°C. The DNA precipitation was purified by 70% ethanol to remove remaining impurities. Approximately 50-100 ul TE buffer or deionised water were added and stored at -20°C. Dried DNA was dissolved in deionised water at 37°C and stored at -20°C. DNA amplification was performed by polymerase chain reaction (PCR). LSU, TUB2, EF1-a and ITS regions were amplified using primers LROR and LR6d (Vilgalys and Hester 1990, Rehner and Samuels 1995), EF1-983F and EF1-2218R (van den Brink et al. 2012), ITS1 and ITS4 (White et al. 1990) and T1 & Bt2b (Glass and Donaldson 1995, O'Donnell and 4 Yan X et al Cigelnik 1997) with 25 ul of the final volume including 9.5 wl ddH 20, 12.5 pl 2 xTaq PCR MasterMix (Qingke, Changsha, China), 1 yl of DNA template and 1 ul of each primer (10 uM). The PCR reaction was under the following conditions: 94°C for 4 min, then 35 cycles of 94°C for 60 s, 53°C (ITS, LSU, TEF1-a), 55°C (RPB2) for 60 s and 72°C for 80 s, followed by a final extension step of 72°C for 8 min (Wu et al. 2014). PCR products were checked on 2% agarose electrophoresis gels stained with GelRed. DNA sequencing was performed using the primers mentioned above by Tsingke, Changsha, China. Phylogenetic analyses The novel sequences and reference sequences collected from GenBank were aligned with MAFFT v.7.036 (http://mafft.cbrc.jp/alignment/server, Katoh et al. (2019)). The multilocus sequences were concatenated by PhyloSuite v.1.2.2 (Zhang et al. 2020). The concatenated aligned datasets were analysed separately using Maximum Likelihood (ML) and Bayesian Inference (Bl). The best-fit models of evolution for the two loci tested were estimated by PhyloSuite v.1.2.2 (Zhang et al. 2020). The ML analyses were conducted with RAXML v.7.2.6 (Stamatakis and Alachiotis 2010) using a GTRGAMMA substitution model with 1000 bootstrap replicates. The robustness of the analyses was evaluated by bootstrap support (MLBS). Markov Chain Monte Carlo (MCMC) methods in MrBayes was used to estimate the posterior probabilities (PP) (Zhaxybayeva and Gogarten 2002). Trees were sampled every 100 generations. The MCMC sampling was set as four chains (three hot chains and one cold chain) running 2,000,000 generations simultaneously, resulting in 20001 total trees. The first 25% of trees were discarded as burn-in trees and the remaining trees were used to calculate posterior probabilities. Posterior probabilities values of the BI analyses (BPP) over 0.95 were regarded to be important. Sequences generated in this study were displayed in GenBank (Table 1). Table 1. Strains used in this study and their GenBank numbers. Note: Type strains are in bold. The underlined species indicated the new taxa in this study. The sequences of new species are indicated as underlined and unavailable sequences in GenBank are indicated by hyphen "-". Species Strain number GenBank accession numbers ITS LSU Achrochaeta talbotii ICMP 15161 MT454480 MT454495 Adautomilanezia caesalpiniae CCLAMIC KX821777 KU170671 Arcuatospora novae-zelandiae CBS 109474 MW984569 MW984552 Arcuatospora novae-zelandiae CBS 109476 MW984570 MW984553 Arcuatospora seorsa CBS 147509 MW984571 MW984554 Arcuatospora seorsa CBS 147510 MW984572 MW984555 Brunneodinemasporium brasiliense CBS 112007 JQ889272 JQ889288 A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) ... Species Cacumisporium capitulatum Calvolachnella guaviyunis Catenularia cubensis Chaetosphaeria catenulat Chaetosphaeria curvispora Chaetosphaeria dilabens Chaetosphaeria hebetiseta Chaetosphaeria inaequalis Chaetosphaeria innumera Chaetosphaeria mangrovei Chaetosphaeria myriocarpa Chaetosphaeria pygmaea Chaetosphaeria submersa Chloridium lignicola Codinaea acaciae Codinaea lambertiae Codinaea paniculata Codinaea pini Codinaea yunnanensis Codinaeopsis gonytrichodes Conicomyces pseudotransvaalensis Cryptophiale hamulata Cryptophiale udagawae Cryptophialoidea fasciculata Dendrophoma cytisporoides Dictyochaeta aquatica Dictyochaeta assamica Dictyochaeta brevis Dictyochaeta callimorpha Dictyochaeta callimorpha Dictyochaeta callimorpha Dictyochaeta cangshanensis Dictyochaeta curvispora Dictyochaeta detriticola Dictyochaeta detriticola Strain number FMR 11339 CBS 134695 S.M.H. 3258 $891 CBS 113644 CBS 712.88 CBS 102340 MR 1450 MenisporaR. 1175 MCD 069 CBS 264.76 MenisporaR. 1365 MFLUCC 181342 CBS 143.54 CBS 139907 CBS 143419 CBS 145098 CBS 138866 MFLU:18-1611 CBS 593.93 GS20 MFLU 17-1975 MFLU:18-1497 MFLU 18-1499 CBS 223.95 MFLU 152691 CBS 242.66 MFLU 190216 ICMP 15155 ICMP 15170 ICMP 15130 MFLU:181614 CBS 114070 ICMP 14948 EXPO560F GenBank accession numbers ITS HF677176 KJ834524 MW987826 AF 178557 AF 178549 AF 178564 AF 178551 MG813821 AF 178552 AF 178545 MK828634 AF 178544 KR476732 MG386052 MT118230 KP004465 MK828623 AF 178556 LC001710 MH758198 MH758195 JQ889273 MH476572 MH858788 MN104614 MT454484 MT454485 MT454483 MK828632 MH862954 MT454486 DQ914666 LSU KJ834525 MK835838 GU180636 AF 178557 AF 178549 AF 178564 AF 178551 MG813820 AF 178552 AF 178545 MK835835 AF 178544 MG386105 M1T118201 KP004493 MK835823 AF 178556 LC001708 MG386756 MH758211 MH758208 JQ889289 MH476569 MH870426 MN104625 MT454499 MT454500 MT454498 MK835832 MT454501 Species Dictyochaeta ellipsoidea Dictyochaeta ellipsoidea Dictyochaeta fuegiana Dictyochaeta fuegiana Dictyochaeta jiangxiensis Dictyochaeta lignicola Dictyochaeta mimusopis Dictyochaeta montana Dictyochaeta pandanicola Dictyochaeta querna Dictyochaeta querna Dictyochaeta septata Dictyochaeta siamensis Dictyochaeta siamensis Dictyochaeta simplex Dictyochaeta simplex Dictyochaeta sp. Dictyochaeta stratosa Dictyochaeta stratosa Dictyochaeta submersa Dictyochaeta terminalis Dinemasporium americanum Dinemasporium decipiens Dinemasporium morbidum Dinemasporium morbidum Dinemasporium nelloi Dinemasporium polygonum Dinemasporium pseudoindicum Ellisembia aurea Ellisembia brachypus Endoxyla operculata Ericiosphaeria spinosa Eucalyptostroma eucalypti Exserticlava vasiformis Flectospora laminata Yan X et al Strain number MFLU:181612 $304 ICMP 15153 FMR_13126 JAUCC 2824 MFLU:181613 CBS 143435 CBS 145342 KUMCC 160153 CBS 146103 CBS 145503 CBS 143386 MFLUCC 160371 MFLUCC 150614 CBS 966.69 MFLU 190202 CBS 138684 CBS 138739 FMR 11228 MFLU:182321 GZCC 180085 CBS 127127 CBS 592.73 CBS 129.66 CBS 995.97 MFLUCC 130482 CBS516.95 CBS 127402 CBS 144403 HKUCC10555 UAMH 11085 S.M.H. 2754 CBS 142074 TAMA450 CBS 112964 GenBank accession numbers ITS MK828628 MK828627 MT454487 KY853440 MN619652 MK828630 MH107888 MT454488 MH388338 MT454490 MT454489 MH107889 MH388339 KX609955 AF178559 MN104609 MT454493 MT454491 MT454492 MK828631 MN104613 JQ889274 JQ889275 JQ889280 JQ889281 KP711358 JQ889276 JQ889277 MH836375 MW984575 KY173408 MW984576 LSU MK835828 MK835827 EF063574 KY853500 MN607224 MK835830 MH107935 MT454502 MH376710 MT454504 MT454503 MH107936 MH376711 KX609952 AF178559 MN104620 MT454507 MT454505 MT454506 MK835831 MN104624 JQ889290 JQ889291 JQ889296 JQ889297 KP711363 JQ889292 JQ889293 MH836376 DQ408563 JX460992 AF 466079 KY173500 AB753846 MW984558 A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) ... Species Infundibulomyces cupulata Infundibulomyces oblongisporus Kionochaeta castaneae Kionochaeta microspora Kionochaeta ramifera Lecythothecium duriligni Melanochaeta aotearoae Melanochaeta hemipsila Melanochaeta taitensis Melanochaeta taitensis Melanopsammella gonytrichii Melanopsammella vermicularioides Menispora caesia Menispora caesia Menispora ciliata Menispora ciliata Menispora tortuosa Menisporopsis anisospora Menisporopsis breviseta Menisporopsis dushanensis Menisporopsis pirozynskii Menisporopsis theobromae Menisporopsis theobromae Menisporopsis theobromae Multiguttulispora dimorpha Multiguttulispora triseptata Multiguttulispora triseptata Nawawia filiformis Neopseudolachnella magnispora Neopseudolachnella uniseptata Paliphora intermedia Paragaeumannomyces garethjonesii Paragaeumannomyces longisporus Paragaeumannomyces raciborskii Paragaeumannomyces rubicundus Strain number BCC 11929 BCC 13400 MFLU 19-0204 MFLU 19-0206 MUCL 39164 CBS 101317 SMH 3551 SMH 2125 GKM156N GKM150N SMH 3785 FC 404 M.R. 1120 CBS 144659 ICMP 18253 CBS 122131 DAOM 231154 CBS 109475 MFLU 19-0212 GZCC 180084 MUCL 47217 MFLUCC 150055 MUCL 41079 MUCL 40984 CBS 140002 CBS 487.92 IMI 353690 MFLUCC 17-2394 MAFF 244359 MAFF 244360 CBS 896.97 MFLUCC 15-1012 ILLS00121385 S.M.H. 3119 S.M.H. 3221 GenBank accession numbers ITS EF 113976 EF 113977 MN104610 MN104607 MW144421 AF 178543 MW984578 EU488736 KT225527 MH862827 MN104612 MN104615 MW984579 KX609957 MW984580 MW984581 MW984582 MW984583 MW984584 MH758196 AB934066 AB934067 MH862682 KY212751 M1T118237 AY906953 M1118242 LSU EFI 3979 EF 113980 MN104621 MN104618 MW144404 AF261071 AF 466082 AY346292 EU583220 EU583219 AF 466085 AF 466087 MW984560 GU180637 AY544682 MH874421 MN104623 MN104626 MW984561 KX609954 MW984562 MW984563 MW984564 MW984565 MW984566 MH758209 AB934042 AB934043 MH874289 KY212759 MT118211 AY436402 MT118224 7 Species Phaeostalagmus cyclosporus Phialogeniculata guadalcanalensis Phialosporostilbe scutiformis Phialoturbella aseptata Phialoturbella calva Phialoturbella lunata Polynema podocarpi Pseudodinemasporium fabiforme Pseudolachnea fraxini Pseudolachnea hispidula Pseudolachnella asymmetrica Pseudolachnella botulispora Pyrigemmula aurantiaca Pyrigemmula aurantiaca Rattania setulifera Sporidesmium minigelatinosa Sporidesmium parvum Sporoschisma longicatenatum Sporoschisma mirabile Striatosphaeria castanea Striatosphaeria codinaeophora Striatosphaeria codinaeophora Tainosphaeria cecropiae Tainosphaeria crassiparies Tainosphaeria jonesii Tainosphaeria jonesii Tainosphaeria siamensis Thozetella fabacearum Thozetella nivea Thozetella tocklaiensis Tracylla aristata Tracylla eucalypti Umbrinosphaeria caesariata Zanclospora iberica Zanclospora novae-zelandiae Yan X et al Strain number CBS 663.70 CBS:346.76 MFLUCC 17-0227 MFLU 19-0208 ICMP 23826 MFLUCC 18-0642 CPC:32761 MAFF 244361 CBS 113701 MFLU:19-2863 MAFF 244366 MAFF 244367 CBS 126743 CBS 126744 GUFCC 15501 NN 47497 HKUCC 10836 MFLUCC 160180 FMR 11247 CBS 145352 M.R. 1230 S.M.H. 1524 CBS 101687 S.M.H. 1934 GZCC 160065 GZCC 16-0053 MFLUCC 150607 MFLU 16-1021 EU825201 CBS 378.58 CPC 25500 CPC:31806 CBS 102664 CBS 130426 ICMP 15781 GenBank accession numbers ITS MH859892 MH860986 MH758194 MN104611 MW984585 MK828624 MH327797 AB934068 JQ889287 MT185550 AB934073 AB934074 HM241692 HM241693 GU191794 KX505871 HF677174 MT118244 AF 178546 M1T118245 MW984586 MW984587 KY026060 MN121305 KX609956 KY212754 EU825201 MH857817 KX306770 MH327810 KY853480 MW144429 LSU MH871680 MH872756 MH758207 MN104622 MW984567 MK835824 MH327833 AB934044 JQ889301 M1T183515 AB934049 AB934050 HM241692 HM241693 HM171322 DQ408567 DQ408558 KX358077 HF677183 M1T118229 AF 178546 AF 466088 MW984568 AF466089 KY026057 KY026056 KX609953 KY212762 EU825200 MH869349 KX306795 MH327846 AF261069 KY853544 MW144411 A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae)... 9 Species Strain number GenBank accession numbers ITS LSU Zanclospora xylophila ICMP 22737 MW144437 MW144417 Zignoélla pulviscula MUCL 15710 — AF466090 Zignoélla pulviscula SMH 3289 — AF466091 Taxon treatment Dictyochaeta jiangxiensis J.E. Huang, X.Y. Yan, H.Y. Song & D.M. Hu, sp. nov. ° MycoBank 846403 Material Holotype: a. scientificName: Dictyochaeta jiangxiensis; acceptedNameUsage: Dictyochaeta jiangxiensis J.E. Huang, X.Y. Yan & D.M. Hu; kingdom: Fungi; phylum: Ascomycota; class: Sordariomycetes; order: Chaetosphaeriales; family: Chaetosphaeriaceae; taxonRank: species; verbatimTaxonRank: species; genus: Dictyochaeta; specificEpithet: jiangxiensis; scientificNameAuthorship: J.E. Huang, X.Y. Yan & D.M. Hu; continent: Asia; country: China; stateProvince: Jiangxi Province; county: Jishui county; locality: Dingjiang; verbatimLatitude: 27.127397 N; verbatimLongitude: 115.276527 E; identifiedBy: J.E Huang, X.Y. Yan; type: PhysicalObject; language: en; rightsHolder: Dian-Ming Hu; institution|ID: HFJAU 3175; collection!|D: HJ0108-1; institutionCode: the Herbarium of Fungi, Jiangxi Agricultural University (HFJAU); collectionCode: Fungi; ownertnstitutionCode: the Herbarium of Fungi, Jiangxi Agricultural University (HFJAU); basisOfRecord: PreservedSpecimen; occurrence!D: B23E3165-B465-54BD-ADAE- FB318AEB9F70 Description Saprobic on decaying submerged wood. Sexual morph Undetermined. Asexual morph Hyphomycetous. Colonies effuse, aggregate, spreading very widely, glistening white to transparent spots and short dark brown hairs. Mycelium composed of partly immersed and partly superficial, brown to dark brown, septate. Setae of the upper layer sterile, brown to black, usually associated with the conidiophores and together these can form small clusters originating from a knot of superficial hyphae, 200—420 =x 4.6— 7.2 ym, cylindrical, straight or slightly flexuous, septate, smooth, thick-walled, base swollen 9-12 ym wide, tapering to terminal. Conidiophores of the lower layer always fertile, mononematous, macronematous, erect or flexuous, unbranched, 26-60 x 3.5—5 um (av. = 48.3 x 4.0 um, n = 20), 3-8-septate, smooth, thin-walled, base brown 4.8—7.5 um, apex pale brown with single phialide. Conidiogenous cells monophialidic (15—) 24— 34 x 3.6—-5 um (av. = 27.6 x 4 um, n = 20), subcylindrical, light brown, narrowing below the collarette. Collarettes light brown, funnel-shaped, 2.2—4.9 ym at the opening, 0.8— 1.1 um at deep. Conidia accumulating at the heads white, 23-32 x 2.5-3.2 um (av. = 10 Yan X et al 26.1 x 2.9 um, n = 30), hyaline, unicellular, thin-walled, smooth, abundant guttulate, falcate to subclavate, rarely straight, narrowly rounded at the both ends, with 6-11 um long hair-like appendages at both ends, smooth (Fig. 2). Figure 1. EESI Phylogenetic tree based on combined ITS, LSU sequences of most taxa of the Chaetosphaeriaceae. Species name given in bold red is a new taxon in this study; species name given in bold indicates a type strain, respectively. Asterisk (*) indicates branches with MLBS = 100% and PP value = 1.0. The ML bootstrap support values and Bayesian posterior probabilities are given above the branches (MLBS/BPP). The tree is rooted to Tracylla aristata CPC 25500 and Tracylla eucalypti CPC:31806. Culture characteristics: Conidia germinating on PDA within 12 h. Colonies growing on PDA, reaching 20-30 mm diam. after 3 weeks at 28°C, circular, white to pale grey mycelium with hyaline margin, centre lightly raised, pale brown to dark brown in reverse, with smooth margin. Material examined: CHINA, Jiangxi Province, Jian, Dingjiang, on submerged wood in a stream, 9 April 2018, J.-E. Huang (HFJAU 3175, Holotype); ex-type living culture (JAUCC 2824). Etymology yilangxiensis' refering to the host location, Jiangxi Province, where the holotype was collected. A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) ... 11 iaaret nee TAM . , Pas. is ¥ 9 she . } ~Saeggai gas ChENt.0 2.” = eqn Fea * Figure 2. EE Dictyochaeta jiangxiensis (HFJAU 3175, holotype). a Colonies on submerged wood; b, c Setae and conidiophores; d, e Conidiophores and phialides with a developing conidia; f, g Apex and base of setae; h, i Conidia; j, k Colony on PDA from above and below. Scale bars: a = 200 um, b, c= 50 um, d—h = 5 um. Notes Dictyochaeta jiangxiensis is a distinct species in the genus as supported by molecular phylogenetic analysis and it clusters with D. brevis, but the latter has smaller conidia (7.5—11.4 um long, 2.0—2.9 ym wide; Lin et al. (2019)). We found that there was 8% nucleotide difference of ITS sequences and about 2% nucleotide difference between 12 Yan X et al the LSU sequences of Dictyochaeta jiangxiensis sp. nov. JAUCC2824 and Dictyochaeta brevis MFLU 19-0216. Morphologically, D. jiangxiensis matches Dictyochaeta well, especially the setae surrounded by several conidiophores and conidia with setulae at both ends. D. jiangxiensis is similar to D. fuegiana (Chaetosphaenia fuegiana), D. occidentalis, C. siamensis (Dictyochaeta siamensis) and C. lignicola (Dictyochaeta lignicola) in having multi-septae and a single phialide at the apex, subcylindrical conidiogenous cells with funnel-shaped collarette and guttulate conidia with hair-like appendages. However, D. fuegiana (Chaet. fuegiana) has smaller conidia (15—23 x 2—2.5 um) without hair-like appendages at both ends (Spegazzini 1923). C. lignicola also has smaller conidia (13-15 ym long, 4.5—5.5 um wide) and has no setae (Luo et al. 2019). D. occidentalis has wider conidia (24—32 x 3—4 ym; Whitton et al. (2000)) with degenerated appendages. C. siamensis has mono- or polyphialidic conidiogenous cells and samller conidia (8-17 x 2—5 um; Tibpromma et al. (2018)). Analysis Phylogenetic analyses Based on ITS and LSU, a multi-locus phylogenetic tree was established to demonstrate the relationships between the new species and related taxa in Chaetosphaeriaceae (Fig. 1). The alignment has 1767 characters (including alignment gaps), with 715 characters for ITS and 1052 characters for LSU. The ML analysis result showed coincident topology with BI. Fig. 1 shows the ML tree based on the combined dataset, along with the fully supported bootstrap values and Bayesian posterior probabilities. All phylogenetic trees were similar in topologies. The new species Dictyochaeta jiangxiensis, together with Dictyochaeta brevis MFLU 19-0216, formed a well-supported clade (BPP = 1.00; MLBS = 100%), but formed a separate branch and there were obvious differences between them. By comparing the ITS and LSU sequences of Dictyochaeta jiangxiensis sp. nov. JAUCC2824 and Dictyochaeta brevis MFLU 19-0216 respectively in NCBI, we found that there was 8% nucleotide difference of ITS sequences and about 2% nucleotide difference between the LSU sequences of Dictyochaeta jiangxiensis sp. nov JAUCC2824 and Dictyochaeta brevis MFLU 19-0216. Discussion Freshwater fungi refer to the fungi that rely on the freshwater habitats for the whole life cycle or part of the life cycle. Phylogenetic studies on freshwater ascomycetes have shown that some species cluster with terrestrial ascomycete lineages, while others cluster with exclusive aquatic lineages (Raja et al. 2018). However, Shearer (1993) defined freshwater fungi as “fungi that must rely on the freshwater environment to complete their life cycle”. The concept of aquatic fungi in a broad sense was adopted in this’ study. Chaetosphaeriaceae is a huge and diverse group with overwhelmingly phialidic fungi and A new species of Dictyochaeta (Sordariomycetes, Chaetosphaeriales, Chaetosphaeriaceae) ... 13 some members of Chaetosphaeriaceae possess known teleomorphs. The family has a world-wide distribution. They are predominantly isolated from soil and plant debris, some are endophytic and have been isolated from herbaceous plants (Hughes and Kendrick 1968, Reblova 2004, Fernandez and Huhndorf 2005, Huhndorf and Fernandez 2005, Crous et al. 2012, Yang et al. 2018, Lin et al. 2019, Luo et al. 2019). In this study, the new Dictyochaeta species in the family Chaetosphaeriaceae was isolated from a freshwater environment. Most known species in Dictyochaeta were reported from rotting parts of plants, such as decaying leaf, bark or stem and submerged wood, partly from soil. Previously, Dictyochaeta and Codinaea were hard to demarcate, not only because of their morphological and ecological similarities, but their closely-related phylogenetic relationship. The difference between Dictyochaeta and Codinaea lies in the presence or absence of setae. The taxonomy of these fungi has relied mainly on morphological criteria. However, it is hard to treat setae as a criterion for identification as setae are always irregular amongst these similar taxa as mentioned above. Codinaea was introduced to accommodate a single species, C. aristata. Since then, the type species of Codinaea has become a taxonomic bottleneck. This species has not been recorded in any literature since its initial description. The holotype material and molecular data could not be traced. That is why the phylogenetic statuses of the Dictyochaeta-like fungi are still ambiguous. Hughes and Kendrick (1968) made an attempt at using the name Dictyochaeta instead of Codinaea on account of the principle of priority and suggested to adopt the name Codinaea as the type material for D. fuegiana of Dictyochaeta. Since Gamundi et al. (1977) redescribed D. fuegiana from fresh material, Dictyochaeta became a precedently used name. Crous et al. (2018) accepted this treatment. Simultaneously, the name Codinaea was suggested to be treated as the type material for D. fuegiana of Dictyochaeta by Hughes and Kendrick (1968), but this view has great limitations. Shortage of abundant original descriptions and loss of the type material of Codinaea were the factors (Liu et al. 2016) which indicated that the molecular phylogeny of Dictyochaeta has not been solved due to the small number of sequences in GenBank. The species, thus, need recollecting, epitypifying and sequencing to establish which morphological characters are of taxonomic significance and generic boundaries. Recently, Dictyochaeta has still not been classified as monophyletic even though most Dictyochaeta-like and Codinaea-like species were re-assessed and recognised as five genera: Codinaeella, Nimesporella, Stilbochaeta, Tainosphaeriella and Xyladelphia (Reblova et al. 2021b). Reblova et al. (2021b) indicated Codinaea is a highly polyphyletic taxon unrelated to Dictyochaeta and that its original delimitation, based on a single morphotype of C. arstata, is too narrow and unsustainable and they emphasise the importance of combination of microscopic morphological characters developed in culture and under a natural substrate for identification. In our analysis, the phylogenetic position of some taxa, such as Zignoélla and Menispora, are unclear in the tree with low BS and PP values. Kionochaeta formed a sister clade with Dictyochaeta and they were clustered together, but with low BS and PP values. Quite a number of species have not been adopted to establish the phylogenetic tree on account of the absence of molecular data and type materials. The molecular database and materials of Chaetosphaeriaceae species needs to be supplemented and improved. 14 Yan X et al Acknowledgements Funds for research were provided by the National Natural Science Foundation of China (NSFC 32070023, NSFC 32060014), the Key Projects of Youth Fund of Jiangxi Science and Technology Department of China (20192ACBL21017) and the Natural Science Foundation of Education Department of Jiangxi Province of China (GJJ190168). References ° Agnihothrudu V (1968) Notes on fungi from North-east India. XVII. Menisporella assamica gen. et sp. nov. 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