Research Article
Print
Research Article
Two new species of Parastagonospora and a new species of Phaeoseptoriella (Phaeosphaeriaceae, Pleosporales) from grasslands in Yunnan Province, China
expand article infoYing Gao‡§|, Tingfang Zhong§, Prapassorn Damrongkool Eungwanichayapant‡, Ruvishika S. Jayawardena‡, Kevin D. Hyde#‡, Turki Kh. Faraj#, Dhanushka N. Wanasinghe#§|, Heng Gui§|#
‡ Mae Fah Luang University, Chiang Rai, Thailand
§ Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
| Center for Mountain Futures, Kunming Institute of Botany, Honghe, China
¶ University of Chinese Academy of Sciences, Beijing, China
# King Saud University, Riyadh, Saudi Arabia

Corresponding author: Dhanushka N. Wanasinghe ( dnadeeshan@gmail.com )

Corresponding author: Heng Gui ( guiheng@mail.kib.ac.cn )

Academic editor: Samantha C. Karunarathna
© 2024 Ying Gao, Tingfang Zhong, Prapassorn Damrongkool Eungwanichayapant, Ruvishika S. Jayawardena, Kevin D. Hyde, Turki Kh. Faraj, Dhanushka N. Wanasinghe, Heng Gui.
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.
Citation: Gao Y, Zhong T, Eungwanichayapant PD, Jayawardena RS, Hyde KD, Faraj TKh, Wanasinghe DN, Gui H (2024) Two new species of Parastagonospora and a new species of Phaeoseptoriella (Phaeosphaeriaceae, Pleosporales) from grasslands in Yunnan Province, China. MycoKeys 109: 239-263. https://doi.org/10.3897/mycokeys.109.134136
Open Access

Abstract

During our investigation of microfungi on grasslands in Yunnan Province, China, three new fungal taxa associated with grasses were collected. Morphological observations and phylogenetic analyses of the combined SSU, LSU, ITS, tef1-α, and rpb2 loci based on maximum likelihood and Bayesian inference were used to reveal the taxonomic placement of these fungal taxa. This study introduces Parastagonospora yunnanensis, Para. zhaotongensis, Phaeoseptoriella poaceicola. Parastagonospora yunnanensis is characterized by ampulliform or globose to subglobose conidiogenous cells, with conidia that are cylindrical to subcylindrical, 0–1-septate, rounded at the apex and slightly truncate at the base. Parastagonospora zhaotongensis features similar globose to subglobose conidiogenous cells but with 0–3-septate, cylindrical to subcylindrical conidia. Phaeoseptoriella poaceicola is distinguished by its globose to subglobose conidiogenous cells and phragmosporous conidia that are initially hyaline, turn pale yellowish at maturity, and are 7-septate, cylindrical to subcylindrical, either straight or slightly curved. These discoveries underscore the significance of exploring and accurately identifying fungal taxa within Ascomycota, highlighting the species richness and potential for new species discoveries in grass-based habitats. The findings from this study expand our understanding of the taxonomy and phylogeny of grassland-associated Ascomycota, providing a foundation for further ecological and taxonomic studies of these fungi within their natural environments.

Key words

Ascomycota, coelomycetes, phragmosporous conidia, Poaceae, taxonomy, 3 new species

Introduction

Phaeosphaeriaceae was introduced by Barr (1979) with Phaeosphaeria as the type genus and belongs in Dothideomycetes (Wijayawardene et al. 2022). The family is one of the most species-rich families in Dothideomycetes and it includes species that inhabit a wide range of ecosystems, including marine, terrestrial, freshwater, and mangroves (Phookamsak et al. 2014, 2017; Bakhshi et al. 2019; Jones et al. 2019; Luo et al. 2019; Tennakoon et al. 2019). The taxa of Phaeosphaeriaceae are typically endophytic, saprobic, and pathogenic on a wide range of hosts (Barr 1992; Ariyawansa et al. 2013; Phookamsak et al. 2014, 2017, 2019; Hyde et al. 2017; Wanasinghe et al. 2018; Maharachchikumbura et al. 2019; Dissanayake et al. 2022).

Parastagonospora was introduced by Quaedvlieg et al. (2013) in Phaeosphaeriaceae (Pleosporales, Dothideomycetes), with Parastagonospora nodorum as the type species. There are 27 Parastagonospora species listed in Index Fungorum (2024). Parastagonospora species are generally identified through their asexual forms. However, only seven species viz. Para. arcana, Para. elymi, Para. forlicesenica, Para. fusiformis, Para. jasniorum, Para. poaceicola and Para. zildae have been documented in their sexual forms (Li et al. 2016; Thambugala et al. 2017; Goonasekara et al. 2019; Croll et al. 2021).

Species of Parastagonospora have been reported from Australia, China, Denmark, Germany, Italy, Iran, the Netherlands, New Zealand, Russia, Turkey, the UK, and the USA as pathogens or saprobes of grasses (Table 1). Parastagonospora species are directly and indirectly responsible for significant annual crop losses worldwide. Cunfer (2000) reported that Parastagonospora avenae was a significant pathogen in oats and caused leaf spots in barley and rye. Parastagonospora nodorum was reported as a critical pathogen in many countries where wheat and barley are cultivated (Cunfer 2000; Quaedvlieg et al. 2013; Goonasekara et al. 2019; Croll et al. 2021). The list of Parastagonospora species reported worldwide is provided in Table 1.

Table 1.
Download as
CSV
XLSX

List of Parastagonospora species reported worldwide. NA: data not available.

Species name Host Country Life-Mode References
Para. allouniseptata Dactylis glomerata Italy Saprobic Li et al. (2015)
Para. Arcana Triticum aestivum Iran Pathogenic Croll et al. (2021)
Para. Avenae Lolium multiflorum, Avena sativa Australia, Germany Pathogenic Quaedvlieg et al. (2013); Croll et al. (2021)
Para. bromicola Bromus inermis USA NA Croll et al. (2021)
Para. cumpignensis Dactylis glomerata Italy Saprobic Li et al. (2016)
Para. Caricis Phalaris arundinacea, Carex acutiformis Cyperaceae sp. Netherlands, USA NA Quaedvlieg et al. (2013); Fulcher et al. (2018)
Para. dactylidicola Dactylis glomerata Italy Saprobic Brahmanage et al. (2020)
Para. dactylidigena Dactylis glomerata Iran NA Croll et al. (2021)
Para. dactylidis Dactylis sp. Italy Saprobic Li et al. (2015)
Para. Elymi Elymus repens Russia Saprobic Goonasekara et al. (2019)
Para. forlicesenica Dactylis glomerata Italy Saprobic Thambugala et al. (2017)
Para. fusiformis Dactylis glomerata Italy Saprobic Thambugala et al. (2017)
Para. golestanensis Agropyron tauri Iran NA Croll et al. (2021)
Para. Italica Dactylis sp. Italy Saprobic Li et al. (2015)
Para. Jasniorum Triticum aestivum Iran Pathogenic Croll et al. (2021)
Para. macrouniseptata Dactylis glomerata Italy Saprobic Goonasekara et al. (2019)
Para. Minima Dactylis sp. Italy Saprobic Li et al. (2015)
Para. nodorum Lolium perenne, Triticum aestivum, Leymus chinensis Africa, Australia, China, Denmark, North Iran, Turkey, UK, USA Pathogenic Quaedvlieg et al. (2013); Zhang and Nan (2018); Croll et al. (2021)
Para. novozelandica Poaceae sp. New Zealand NA Marin-Felix et al. (2019)
Para.phragmitis Phragmites sp. Australia NA Marin-Felix et al. (2019)
Para.poaceicola Dactylis glomerata Italy Saprobic Thambugala et al. (2017)
Para. Poae Poa sp. Netherlands NA Quaedvlieg et al. (2013)
Para. poagena Poa sp. Netherlands NA Crous et al. (2014)
Para. pseudonodorum Triticum aestivum Iran Pathogenic Croll et al. (2021)
Para. Stipae Stipa pulchra USA NA Croll et al. (2021)
Para. uniseptata Daucus sp. Italy Saprobic Li et al. (2015)
Para.yunnanensis Lolium perenne China Saprobic In this study
Para. zildae Triticum aestivum Iran Pathogenic Croll et al. (2021)
Para. zhaotongensis Dactylis glomerata China Saprobic In this study

Phaeoseptoriella was introduced by Crous et al. (2019), with Phaeoseptoriella zeae as the type species, on the leaves of Zea mays (Poaceae) from South Africa. The asexual morphs of Phaeoseptoriella species are characterized by globose, solitary, brown conidiomata with a central ostiole, pale brown, ampulliform to doliiform conidiogenous cells that are smooth with percurrent proliferation at the apex, and pale brown, solitary, fusoid-ellipsoid, straight to slightly curved conidia that are finely roughened with a subobtuse apex, truncate base and are septate (Crous et al. 2019). Tan and Shivas (2023) reported two new species of Phaeoseptoriella, Ph. emmelinepankhurstiae and Ph. vidagoldsteiniae, based on phylogenetic analysis, however, the morphological description was provided. Phaeoseptoriella emmelinepankhurstiae and Ph. vidagoldsteiniae were collected from leaves of Sporobolus natalensis (Poaceae) in Australia (Tan and Shivas 2023). Phaeoseptoriella edithcowaniae was collected from a leaf spot of Heteropogon triticeus (Poaceae) in Australia (Tan and Shivas 2024).

Grasslands comprise a biome subjected to alternating droughts, where grass and grass-like species dominate (Risser 1988). In the grassland biome, several living organisms, such as insects, herbivorous mammals and fungi (saprobic, pathogenic, and symbiotic), play essential roles in maintaining biodiversity and biomass (Karunarathna et al. 2021). A checklist of Ascomycetes on grasses, which lists 3,165 fungal species, was provided by Karunarathna et al. (2022). Studies of fungi on grasses include those of Thambugala et al. (2017), Goonasekara et al. (2018), and Brahmanage et al. (2020). Some fungi from grass have also been reported in Yunnan province, China. Dactylella crassa was introduced by Miao et al. (1999) from Oryza sp. Hypogymnia congesta was reported by McCune et al. (2003) from the Poaceae hosts. Yuan et al. (2010) introduced a new species, Harpophora oryzae collected from Oryza granulate. Xia et al. (2013) introduced a novel species Heteroconium bannaense from Phragmites sp. Yunnanensis phragmitis was reported by Karunarathna et al. (2017). Based on morphology and phylogeny, Gao et al (2022) introduced Microdochium graminearum, M. shilinens, and M. bolleyi to the genus Microdochium. They were collected from grasses in Yunnan, China. In recent studies, Li et al. (2024) introduced Anthostomella yunnanensis, Astrocystis heterocyclae and Collodiscula baoshanensis, while Dissanayake et al. (2024) introduced Apiospora guangdongensis, A. locuta-pollinis, A. menglaensis, A. pseudoparenchymatica, Collodiscula yunnanensis, Digitodochium ailaoshanense and D. yunnanense from bamboo species in Yunnan.

In Yunnan, China, we are continuously surveying the grassland-associated microfungi. Many fungal species may be nearing extinction because they cannot adapt quickly enough to the rapid ecological changes (Wanasinghe et al. 2022; Yasanthika et al. 2023). To mitigate this loss and understand their ecological significance, extensive fungal sampling across various grasslands in different geographic regions is urgently required. Our recent efforts have already yielded several strains of unidentified species isolated from different grass-based hosts (Gao et al. 2022, 2024; Dissanayake et al. 2024), suggesting that there are potentially many new fungal species yet to be discovered in these habitats. Based on morphological illustrations and multi-gene phylogenetic analyses employing ML, and BI, this study introduces three novel species within the Phaeosphaeriaceae. We describe two new species, Parastagonospora yunnanensis and Para. zhaotongensis, in Parastagonospora and a novel species, Phaeoseptoriella poaceicola, to the Phaeoseptoriella. The specimens from which these species were collected on Lolium perenne and Dactylis glomerata were from the grassland areas of Qujing and Zhaotong in Yunnan Province, China.

Materials and methods

Sample collection, isolation, and morphological observations

Fresh fungal materials were collected from grasslands in Zhaotong and Qujing City, Yunnan Province, China, during the autumn from August to October 2022. The local environment in Zhaotong is characterized by Poaceae as the predominant plant species and features typical plateau vegetation. This area is influenced by a three-dimensional monsoon climate and reaches a maximum elevation of approximately 4000 m (Pei 2022). In contrast, Qujing is characterized by a typical subtropical plateau monsoon climate, with an annual mean temperature of 14.5 °C and average annual precipitation around 1000 mm (Deng et al. 2016). Specimens were stored in plastic Ziplock bags and returned to the mycology laboratory at the Kunming Institute of Botany. Samples were examined using an Olympus SZ-61 dissecting microscope. Fungal fruiting structures were manually sectioned and mounted in water on a slide to observe their microscopic features. Micro-morphological characteristics were examined using a Nikon ECLIPSE Ni-U complex microscope with differential interference contrast (DIC) and phase contrast (PC) illumination. Photos of microscopic structures were captured using a Nikon DS-Ri2 camera. Photo plates and measurements were processed using Adobe Photoshop CS6 Extended version 13.0.1 (Adobe Systems, CA, USA). Single spore isolation of conidia was conducted, and germinated spores were processed by following the methods described in Senanayake et al. (2020). Pure cultures were incubated at 27 °C for two weeks. The living cultures were deposited in the duplicates, which were maintained in the China General Microbiological Culture Collection Center (CGMCC). Herbarium specimens were deposited in the herbarium of the Kunming Institute of Botany Academia Sinica (HKAS). The new taxa have been registered and can be referenced using their Index Fungorum and Faces of Fungi (FoF) numbers as reported by Jayasiri et al. (2015) and in the Index Fungorum (2024). These new taxa are also documented on the Greater Mekong Subregion website (https://gmsmicrofungi.org), as detailed by Chaiwan et al. (2021). Taxonomic novelties were introduced based on a polyphasic approach that integrates morphological, molecular and ecological data, aligning with contemporary taxonomic standards (Chethana et al. 2021; Jayawardena et al. 2021; Maharachchikumbura et al. 2021).

DNA extraction, PCR amplification, and sequencing

The extraction of genomic DNA was performed using these fresh mycelia following the methods of Wanasinghe et al. (2016) and Hyde et al. (2023), using the Biospin Fungus Genomic DNA Extraction Kit (BioFlux, Hangzhou, P.R. China) and following manufacturer guidelines. The DNA for the polymerase chain reaction (PCR) was stored at 4 °C for regular use and at -20 °C for long-term usage. Polymerase chain reaction (PCR) was carried out for five genetic markers. The primers and amplification conditions used are listed in Table 2. The total volume of PCR mixtures for amplification was 25 μL containing 8.5 μL ddH2O, 12.5 μL 2 × F8FastLong PCR MasterMix (Beijing Aidlab Biotechnologies Co. Ltd), 2 μL of DNA template, and 1 μL of each forward and reverse primers (stock of 10 pM). The amplified PCR fragments were sent to the Qingke Company, Kunming City, Yunnan Province, China, and Shanghai Sangon Biological Engineering Technology and Service Co., Ltd., China, for sequencing. Sequences were deposited in GenBank.

Table 2.
Download as
CSV
XLSX

Details of genetic markers with PCR primers and thermal cycling program for PCR amplification.

Genetic Marker Primers PCR thermal cycle protocols References
The 18S small subunit rDNA (SSU) NS1 aAnnealing at 55 °C for 15 sc White et al. (1990)
NS4
The 28S large subunit rDNA (LSU) LR0R Rehner and Samuels (1994)
LR5 Vilgalys and Hester (1990)
The internal transcribed spacers (ITS) ITS5 White et al. (1990)
ITS4
The translation elongation factor 1-alpha (tef1-α) EF1-983F aAnnealing at 55 °C for 30 sc Rehner and Buckley (2005)
EF1-2218R
The partial RNA polymerase second largest subunit (rpb2) fRPB2-5F bAnnealing at 57 °C for 50 sc Liu et al. (1999)
fRPB2-7cR

Notes: a initial denaturation at 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 10 s, elongation at 72 °C for 20 s; b initial denaturation at 95 °C for 3 min, followed by 35 cycles at 95 °C for 45 s, elongation at 72 °C for 1.5 min; c final extension at 72 °C for 10 min.

Phylogenetic analyses

Sequences obtained from different primers targeting the relevant genes were compared with other sequences sourced from GenBank. A BLAST search identified sequences with high similarity, indicating the closest matches within the Phaeosphaeriaceae taxa and referencing previously published data (Quaedvlieg et al. 2013; Li et al. 2015; Thambugala et al. 2017; Goonasekara et al. 2019; Marin-Felix et al. 2019). The sequences of SSU, LSU, ITS, tef1-α, and rpb2 were downloaded from GenBank (Table 3). Some of the sequences from the study by Croll et al. (2021), which introduced Parastagonospora bromicola, Para. dactylidigena, Para. golestanensis, Para. jasniorum, Para. pseudonodorum to Parastagonospora, were not available in the GenBank. Therefore, we obtained these sequences directly from the first author. The sequences in this study were assembled and manually refined using BioEdit 7.0.9.0 (Hall 1999). The multiple alignments, which included both consensus sequences and reference sequences, were initially generated using MAFFT v. 7 (Kuraku et al. 2013; Katoh et al. 2019). The multiple alignments, which included both consensus sequences and reference sequences, were initially generated using MAFFT v.7. online platform (Katoh et al. 2019) and trimmed with TrimAl v. 1.3 (Capella-Gutiérrez et al. 2009) via the web server Phylemon2 (http://phylemon.bioinfo.cipf.es/utilities.html; accessed on July 10, 2024). and multi-gene alignments were made by the SequenceMatrix program (1.7.8) (Vaidya et al. 2011). Phylogenetic reconstructions of individual and combined datasets were performed using maximum likelihood (ML) and Bayesian inference (BI) analyses on the CIPRES Science Gateway portal (https://www.phylo.org/) (Miller et al. 2012).

Table 3.
Download as
CSV
XLSX

GenBank accession numbers of the strains used for phylogenetic analysis in this study. The new sequences are indicated in bold. Ex-type strains are indicated with the superscript “T”. “NA” is unavailable.

Taxon Strain numbers GenBank accession numbers
SSU LSU ITS tef1-α rpb2
Dematiopleospora mariae MFLU 16-0121 MT226689 MT214576 MT310621 MT394635 NA
Dematiopleospora mariae MFLUCC 13-0612T KJ749652 KJ749653 KX274244 KJ749655 NA
Dematiopleospora salsolae MFLUCC 17-0828T NG_063679 NG_059184 NR_157514 MG829201 MG829254
Neosphaerellopsis thailandica CPC 21659T NA NG_067289 NR_137954 NA NA
Nodulosphaeria aconiti MFLUCC 13-0728T KU708840 KU708844 NR_154236 KU708852 KU708856
Nodulosphaeria guttulatum MFLUCC 15-0069 KY501115 KY496726 KY496746 KY514394 KY514405
Nodulosphaeria scabiosae MFLUCC 14-1111T NG_063602 KU708846 NR_154237 KU708854 KU708857
Paraloratospora marina MFLUCC 19-0691T OQ130107 OQ130110 OQ130046 OQ357219 OQ162221
Paraloratospora sichuanensis KUNCC 23-14218T OR206405 OR206415 OR206396 OR195712 OR195721
Paraloratospora sichuanensis HKAS 129218 OR206406 OR206416 OR206397 OR195713 OR195722
Parastagonospora allouniseptata MFLUCC 13-0386T NA KU058721 KU058711 MG520914 NA
Parastagonospora avenae CBS 289.69 NA KF251678 KF251174 NA KF252182
Parastagonospora avenae CBS 290.69 NA KF251679 KF251175 NA KF252183
Parastagonospora caricis CBS 135671T NA KF251680 KF251176 NA KF252184
Parastagonospora dactylidicola MFLU 20-0387T NA MT370430 MT370412 NA NA
Parastagonospora dactylidis MFLUCC 13-0375T NA KU058722 KU058712 NA NA
Parastagonospora dactylidis MFLUCC 13-0376 MG520986 KU058723 KU058713 MG520916 NA
Parastagonospora dactylidis MFLUCC 13-0573 KU842390 KU842389 KU842388 NA NA
Parastagonospora elymi KUMCC 16-0125T NA MN002870 MN002867 NA NA
Parastagonospora forlicesenica MFLUCC 13-0557T NA KY769661 KY769660 NA NA
Parastagonospora fusiformis MFLUCC 13-0215T NG_068367 NG_068235 NR_165848 NA KX863711
Parastagonospora italica MFLUCC 13-0377T MG520985 KU058724 KU058714 MG520915 NA
Parastagonospora macrouniseptata KUMCC 16-0111T NA MN002868 MN002869 NA MN019669
Parastagonospora nodorum CBS 110109 EU754076 KF251681 KF251177 NA KF252185
Parastagonospora novozelandica CPC 29613T NA MK540028 MK539957 NA MK540088
Parastagonospora phragmitis CPC 32075T NA NG_066451 NR_164454 NA MK540089
Parastagonospora poaceicola MFLUCC 15-0471T NG_068368 NG_068537 NA NA KX880499
Parastagonospora poae CBS 135091 NA KF251683 KF251179 NA KF252187
Parastagonospora poae CBS 135089T NA KF251682 KF251178 NA KF252186
Parastagonospora poagena CBS 136776T NA KJ869174 KJ869116 NA NA
Parastagonospora stipae pn1617 NA NA MW263184 NA NA
Parastagonospora uniseptata MFLUCC 13-0387T MG520987 KU058725 KU058715 MG520917 NA
Parastagonospora yunnanensis CGMCC 3.24527T PQ046289 PQ046315 PQ046302 PQ058300 PQ058313
Parastagonospora yunnanensis CGMCC 3.24528 PQ046290 PQ046316 PQ046303 PQ058301 PQ058314
Parastagonospora yunnanensis CGMCC 3.24529 PQ046291 PQ046317 PQ046304 PQ058302 PQ058315
Parastagonospora yunnanensis CGMCC 3.24530 PQ046292 PQ046318 PQ046305 PQ058303 PQ058316
Parastagonospora yunnanensis CGMCC 3.24511 PQ046285 PQ046311 PQ046298 PQ058296 PQ058309
Parastagonospora yunnanensis CGMCC 3.24512 PQ046286 PQ046312 PQ046299 PQ058297 PQ058310
Parastagonospora zhaotongensis CGMCC 3.24519T PQ046287 PQ046313 PQ046300 PQ058298 PQ058311
Parastagonospora zhaotongensis CGMCC 3.24520 PQ046288 PQ046314 PQ046301 PQ058299 PQ058312
Phaeoseptoriella edithcowaniae BRIP 75864aT NA PP708933 PP707905 NA NA
Phaeoseptoriella emmelinepankhurstiae BRIP65639aT NA NA OR673891 NA NA
Phaeoseptoriella poaceicola CGMCC 3.24561T PQ046283 PQ046309 PQ046296 PQ058294 PQ058307
Phaeoseptoriella poaceicola CGMCC 3.24562 PQ046284 PQ046310 PQ046297 PQ058295 PQ058308
Phaeoseptoriella poaceicola CGMCC 3.25058 PQ046293 PQ046319 PQ046306 PQ058304 PQ058317
Phaeoseptoriella poaceicola CGMCC 3.25059 PQ046294 PQ046320 PQ046307 PQ058305 PQ058318
Phaeoseptoriella poaceicola CGMCC 3.25060 PQ046295 PQ046321 PQ046308 PQ058306 PQ058319
Phaeoseptoriella vidagoldsteiniae BRIP65641aT NA NA OR673892 NA NA
Phaeoseptoriella zeae CBS 144614T NA NG_067869 NR_163371 NA MK442674
Phaeosphaeria chengduensis KUNCC 23-13571T OR206401 OR206411 OR206392 OR195708 OR195717
Phaeosphaeria chiangraina MFLUCC 13-0231T KM434289 NG_069237 NR_155643 KM434298 KM434307
Phaeosphaeria sichuanensis KUNCC 23-13569T OR206399 OR206409 OR206390 OR195706 OR195715
Phaeosphaeria thysanolaenicola MFLUCC 10-0563T KM434286 NG_069236 NR_155642 KM434295 KM434303
Quixadomyces hongheensis KUMCC 20-0215T NG_074964 MW264194 NR_172441 MW256816 MW269529
Quixadomyces hongheensis HKAS 112346 MW541833 MW541822 MW541826 MW556134 MW556136
Sclerostagonospora lathyri MFLUCC 14-0958T NG_063692 NG_069566 NR_158956 MG829235 NA
Sclerostagonospora rosicola MFLUCC 15-0129T NG_063693 MG829068 MG828957 MG829237 NA
Septoriella arundinicola MFLU 16-0225T NG_062199 MG829056 MG828946 MG829228 MG829261
Septoriella asparagicola MFLUCC 16-0379T NG_067708 NG_070081 NR_165908 MK443385 MK443387
Septoriella neodactylidis MFLUCC 14-0966T NG_061288 NG_069554 NR_157511 MG829199 MG829253
Wojnowiciella clematidis MFLUCC 17-2159T MT226695 MT214582 NR_170812 MT394641 MT394698
Wojnowiciella kunmingensis KUMCC 18-0159T NG_067701 NG_070079 NR_164446 MK359071 MK359078

Maximum likelihood trees were inferred using RAxML-HPC2 on XSEDE v. 8.2.12 (Stamatakis 2014) and used the GTR+GAMMA model of nucleotide evolution with 1000 bootstrap replicates. Bayesian inference analysis was conducted using MrBayes on XSEDE (3.2.7a) (Ronquist et al. 2012). The alignments containing SSU, LSU, ITS, tef1-α, and rpb2 were converted to NEXUS format (.nxs) using CLUSTAL X (2.0) and PAUP v. 4.0b10 (Thompson et al. 1997; Swofford 2002). The evolutionary models for BI analysis were selected independently for each locus using MrModeltest v. 2.3 (Nylander et al. 2008) under the Akaike information criterion as follows: GTR+I+G substitution model was chosen for ITS, LSU, tef1-α, and rpb2, HKY substitution model was selected for SSU. Markov Chain Monte Carlo sampling (MCMC) was used to determine posterior probabilities (PP) (Zhaxybayeva and Gogarten 2002). Six simultaneous Markov chains were run for five million generations, and trees were sampled every 200th generation. The first 25% of trees were considered burn-in and discarded. The two runs were considered convergent when the standard deviation of split frequencies dropped below 0.01. The Fig. Tree version 1.4.0 program (Rambaut and Drummond 2012) was used to visualize the phylogenetic trees and reorganized them in Microsoft PowerPoint before being saved in PDF format and, finally, converted to TIFF format using Adobe Photoshop CS6 Extended version 13.0.1 (Adobe Systems, CA, USA).

Results

Phylogenetic analysis

The combined sequence data of SSU, LSU, ITS, tef1-α, and rpb2, comprised 82 strains including the outgroup (Fig. 1). A total of 4,292 characters, including gaps, were obtained in the phylogenetic analysis viz. SSU = 1–965 bp, LSU = 966–1,756 bp, ITS = 1,757–2,378 bp, tef1-α = 2,379–3,160 bp, rpb2 = 3,161–4,292 bp. The RAxML analysis of the combined dataset yielded a best-scoring tree with a final ML optimization likelihood value of -25260.798404. The matrix had 1,382 distinct alignment patterns, with 31.47% undetermined characters or gaps. The estimated base frequencies were as follows: A = 0.245359, C = 0.244397, G = 0.264796, T = 0.245448; substitution rates: AC = 1.259544, AG = 3.943431, AT = 1.705789, CG = 0.844343, CT = 7.018866, GT = 1.000000, proportion of invariable sites I = 0.644641; and gamma distribution shape parameter α = 0.619642.

Figure 1. 

Phylogenetic tree obtained from combined SSU, LSU, ITS, tef1-α, and rpb2 sequence data. The tree is rooted with Quixadomyces hongheensis (HKAS 112346 and KUMCC 20-0215). Bootstrap support values for ML equal to or greater than 60% and the Bayesian posterior probabilities equal to or higher than 0.95 PP are indicated above the nodes as ML/PP. Ex-type strains are indicated in bold, and the new isolates are highlighted in blue.

The Bayesian analysis proceeded for 783,000 generations, achieving an average standard deviation for split frequencies below 0.01 (0.009957). This analysis produced a total of 7,831 trees. After discarding the first 25% as burn-in, 5,874 trees were sampled for further consideration. The alignment included 1,383 unique site patterns. Both BI and ML trees were consistent with each other; the ML tree is presented in Fig. 1. Where relevant, the phylogenetic findings shown in Fig. 1 are further discussed in the descriptive notes that follow.

Except for Parastagonospora allouniseptata (MFLUCC 13-0386), Para. macrouniseptata (KUMCC 16-0111) and Para. novozelandica (CPC 29613), all other Parastagonospora strains nested within a monophyletic clade supported by 64% ML and 1.00 BYPP. Within this clade, the new strains CGMCC 3.24511, CGMCC 3.24512, CGMCC 3.24527, CGMCC 3.24528, CGMCC 3.24529, and CGMCC 3.24530 formed a distinct monophyletic clade, achieving 100% ML and 1.00 BYPP bootstrap support (Clade A, Fig. 1). Additionally, CGMCC 3.24519 and CGMCC 3.24520 grouped together with 100% ML and 1.00 BYPP bootstrap support (Clade B, Fig. 1), positioned as sister to Parastagonospora poae (CBS 135089, CBS 135091) and Para. uniseptata (MFLUCC 13-0387). However, this sister relationship was not statistically supported (Fig. 1).

Phaeoseptoriella edithcowaniae (BRIP 75864a), Ph. emmelinepankhurstiae (BRIP65639a), Ph. vidagoldsteiniae (BRIP65641a), Ph. zeae (CBS 144614) grouped with our new isolates, CGMCC 3.24561, CGMCC 3.24562, CGMCC 3.25058, CGMCC 3.25059 and CGMCC 3.25060. All of these new isolates clustered in a distinct monophyletic clade, achieving 100% ML and 1.00 BYPP bootstrap support (Clade C, Fig. 1).

Taxonomy

Parastagonospora yunnanensis Y. Gao, H. Gui & K.D. Hyde, sp. nov.

Index Fungorum: IF902468
Facesoffungi Number: FoF16258
Fig. 2

Etymology

The specific epithet “yunnanensis” refers to Yunnan Province, where the holotype was collected.

Holotype

HKAS 128771.

Description

Saprobic on decaying stem of Lolium perenne (Poaceae). Sexual morph: Undetermined. Asexual morph: Conidiomata 35–45 µm high, 120–140 μm diam. (x– = 40.6 × 132.7 μm, n = 10), solitary, flattened, subglobose to irregular oval, brown to dark brown spots, immersed in the epidermis of the host, ostiolate. Conidiomata wall 4–13 µm wide (x– = 9 μm, n = 25), composed of brown cells of textura angularis, with an inner layer comprising hyaline cells. Conidiogenous cells (3.2–)3.5–4.7(–5.3) × (3.4–)4–5.3(–6.1) μm (x– = 4 ± 0.57 × 4.77 ± 0.59 μm, n = 30), hyaline, ampulliform or globose to subglobose, smooth-walled. Conidia (16.2–)18–20(–20.4) × (3–)3.2–3.7(–4) μm (x– = 19 ± 1.1 × 3.4 ± 0.24 μm, n = 35), hyaline, 0–1-septate, cylindrical to subcylindrical, rounded at apex, slightly truncate at base, guttulate, smooth-walled.

Culture characteristics

Conidia germinated on PDA within 24 hours, and a germ tube was initially produced from the ends of the conidia. Colonies on PDA reaching 20 mm in 3 weeks at room temperature (25–27 °C), regular, floccose, white from the above and light grey from the centre and below, smooth with a filamentous edge.

Figure 2. 

Asexual morph of Parastagonospora yunnanensis (HKAS 128771, holotype) on a dead stalk of Lolium perenne a, b conidiomata on the host c, d vertical section of conidiomata e conidioma wall f–h conidiogenous cells arise from the wall and develop conidia i–l conidia m germinating conidium n cultures on PDA from above o cultures on PDA from the reverse. Scale bars: 30 μm (c, d); 10 μm (e, h–m); 5 μm (f, g).

Material examined

China • Yunnan Province, Zhaotong City, (26°56'39"N, 103°8'53"E), on a decaying stem of Lolium perenne (Poaceae), 25 August 2022, Ying Gao, QG69A (HKAS 128771, holotype), ex-type (CGMCC 3.24527) • ibid. QG69B (HKAS 128772, paratype), ex-paratype (CGMCC 3.24528) • ibid. QG71A (HKAS 128773), culture (CGMCC 3.24529) • ibid. QG71B (HKAS 128774), living culture (CGMCC 3.24530); ibid. • Qujing City, (26°21'31"N, 103°14'13"E), on a decaying stem of Lolium perenne (Poaceae), 27 August 2022, Ying Gao, QG19A (HKAS 128799), living culture (CGMCC 3.24511) • ibid. QG19B (HKAS 128800), living culture (CGMCC 3.24512).

Notes

Parastagonospora yunnanensis is introduced as a new species based on its distinct morphology and phylogenetic analysis of combined SSU, LSU, ITS, tef1-α, and rpb2 datasets. We have collected six isolates of this fungus from both the Qujing and Zhaotong regions. Parastagonospora yunnanensis is phylogenetically related to Parastagonospora elymi (KUMCC 16-0125). Parastagonospora elymi was introduced as a saprobic fungus from Elymus repens in Russia, the asexual morph of Parastagonospora elymi has not been determined (Goonasekara et al. 2019). In addition, the ITS pairwise nucleotide comparison of these species showed 18/497 bp differences (3.62%, with 2 gaps), the comparison of base pairs in LSU showed 0.36% differences (3/834 bp, without gaps), SSU, tef1-α, and rpb2 of Parastagonospora elymi were not provided.

Parastagonospora zhaotongensis Y. Gao, H. Gui & K.D. Hyde, sp. nov.

Index Fungorum: IF902469
Facesoffungi Number: FoF16259
Fig. 3

Etymology

The specific epithet “zhaotongensis” refers to Zhaotong City, where the holotype was collected.

Holotype

HKAS 132983.

Description

Saprobic on decaying stem of Dactylis glomerata (Poaceae). Sexual morph: Undetermined. Asexual morph: Conidiomata 70–85 μm high × 80–110 μm diam (x– = 76 × 94 μm, n = 10) 80–110 μm diam × 70–85 μm high (x– = 94 × 76 μm, n = 10), flattened, solitary, immersed in the epidermis of the host, globose to subglobose, brown to dark brown spots. Conidiomatal wall 5–13 µm wide (x– = 8 μm, n = 30), thin wall, 2–3 layered, composed of pale brown cells of textura angularis, with inner layer comprising hyaline cells. Conidiogenous cells (3.5–)4.5–6(–6.5) × (3–)4–5.5(–6) μm (x– = 5.5 ± 0.74 × 5 ± 0.77 μm, n = 20), hyaline, globose to subglobose, smooth-walled. Conidia (22–)25–30(–32)× (2.7–)3–3.4(–3.7) μm (x– = 28 ± 2.45 × 3.3 ± 0.21 μm, n = 35), hyaline, 0–3-septate, cylindrical to subcylindrical, smooth-walled, rounded at apex, slightly truncate at base, guttulate.

Figure 3. 

Asexual morph of Parastagonospora zhaotongensis (HKAS 132983, holotype) on a dead stalk of Dactylis glomerata a, b conidiomata on the host c vertical section of conidioma d conidioma wall e conidiogenous cells arise from the wall and develop conidia f conidia g germinating conidium h cultures on PDA from above and reverse. Scale bars: 30 μm (c); 15 μm (d); 10 μm (e–g).

Culture characteristics

Colonies on PDA, reaching 20–25 mm diam., after three weeks at 25–27 °C, with circular, floccose, white from the above and in reverse pale yellow.

Material examined

China • Yunnan Province, Qujing City (26°37'38"N, 103°15'29"E), on decaying stem of Dactylis glomerata (Poaceae), 27 August 2022, Ying Gao, QG44A (HKAS 132983, holotype), ex-type (CGMCC 3.24519) • ibid. QG44B (HKAS 132984, paratype), ex-paratype (CGMCC 3.24520).

Note

Based on multi-locus phylogenetic analyses, our strains of Parastagonospora zhaotongensis (CGMCC 3.24519 and CGMCC 3.24520) are closely related to Para. uniseptata (MFLUCC 13-0387) and Para. poae. Parastagonospora uniseptata was reported on Daucus sp. from Italy by Li et al. (2015). Pairwise nucleotide comparison indicates that our strains differ from Parastagonospora uniseptata in 17/573 bp of ITS (2.97%, with 4 gaps), 2/834 bp of LSU (0.24%, without gaps) and 20/906 bp of tef1-α (2.21%, without gaps). The rpb2 sequence of Parastagonospora uniseptata was not available for comparisons. Morphologically, Parastagonospora zhaotongensis is distinguished by its conidiogenous cells (globose to subglobose vs. ampulliform to broadly conical, phialidic), and conidia (22–32 μm long, 0–3-septate vs. 14–18 μm long, 1-septate) (Table 4). The pairwise nucleotide comparison showed that our strains (CGMCC 3.24519 and CGMCC 3.24520) differ from Parastagonospora poae (CBS 135089) in 15/562 bp of ITS (2.67%, with 4 gaps), 2/828 bp of LSU (0.24%, without gaps), and 10/250 bp of rpb2 (4.00%, without gaps). SSU and tef1-α data for Parastagonospora poae were not provided. Parastagonospora zhaotongensis differs from Para. poae in conidiomata (80–110 μm in diam., brown to dark brown spots vs. up to 250 μm in diam. black), conidiogenous cells (3.5–6.5 μm long, globose to subglobose vs. 6–10 μm long, ampulliform to subcylindrical) and conidia (2.7–3.7 μm wide, 0–3-septate vs. 2–2.5 μm wide, 1-septate) (Table 4).

Table 4.
Download as
CSV
XLSX

Synopsis of asexual morphological characters of Parastagonospora species.

Name of Taxon Conidiomata size (μm) Conidiogenous cells Conidia Reference
Shape Size (μm) Shape Size (μm) septa
Para. allouniseptata 60–90 × 70–90 Ampulliform, phialidic 3–5 × 3–5.5 Subcylindrical, subobtuse apex, truncate base 16–22 × 2.5–3.5 1 Li et al. (2015)
Para. avenae 60–90 Ampulliform 7–10 × 3–5 Subcylindrical, truncate base with obtuse apex 4–6 × 2 0 Croll et al. (2021)
Para. bromicola 150–200 Ampulliform to subcylindrical 4–6 × 4–5 Subcylindrical, subobtuse apex, truncate base 12–18 × 2–3 1(–3) Croll et al. (2021)
Para. caricis Up to 250 Ampulliform, phialidic 8–15 × 4–6 Subcylindrical, subobtuse apex, truncate base, scolecosporous 50–75 × 5–6 7–15 Quaedvlieg et al. (2013)
Para. dactylidicola 100–110 × 85–115 Ampulliform to subcylindrical, broadly cylindrical or conical, phialidic – Hyaline or subhyaline, ellipsoid to oblong, or subcylindrical, with obtuse or subobtuse apex 7.5–10 × 2.5–3.5 1 Brahmanage et al. (2020)
Para. dactylidigena 250–350 Ampulliform to subcylindrical 5–7 × 4–5 Subcylindrical, subobtuse apex, truncate base 25–42 × 4–5 3(–6) Croll et al. (2021)
Para. dactylidis 50–100 × 100–150 Ampulliform, phialidic 2–6 × 3–8 Fusiform, curved, rounded at both ends 25–40 × 4–5.5 3 Li et al. (2015)
Para. golestanensis 200–350 Ampulliform 5–10 × 4–5 Subcylindrical, subobtuse apex, truncate base 22–35 × 2.5–3 (1–)3 Croll et al. (2021)
Para. italica 65–80 × 40–150 Broadly cylindrical, phialidic – Cylindric-fusiform, with narrow and obtuse apex, truncate base 25–32 × 3–4 3-euseptate Li et al. (2015)
Para. jasniorum 250–300 Ampulliform, phialidic, aggregated 5–6 × 4–5 Subcylindrical, apical cell with slight taper to subobtuse apex 22–35 × 2.5–3 (1–)3(–5) Croll et al. (2021)
Para. macrouniseptata 120–160 × 150–190 Ampulliform to lageniform, phialidic, discrete 4.2 × 3 Cylindrical to subcylindrical, rounded at apex, truncate base 14–20 × 1–2.5 1 Goonasekara et al. (2019)
Para. minima 40–70 × 50–100 Ampulliform, phialidic 3–6.5 × 3–7 Subcylindrical, slightly curved, wider at the basal half, narrow, and rounded at both ends 20–28 × 3.5–4.5 3-euseptate Li et al. (2015)
Para. nodorum 10–15 Globose to ampulliform 5–7 × 4–6 Subcylindrical, subobtuse apex, truncate base 11–28 × 2.5–4 1–3 Croll et al. (2021)
Para. novozelandica 180–200 Ampulliform to subcylindrical 6–8 × 2.5–5 Subobtuse apex, truncate base, subcylindrical 9–16 × 2–3 1 Marin-Felix et al. (2019)
Para.phragmitis 250–300 Ampulliform to doliiform 7–10 × 8–9 Hyaline to pale olivaceous, subcylindrical-fusoid 18–27 × 3–4 3 Marin-Felix et al. (2019)
Para. poae up to 250 Ampulliform to subcylindrical, phialidic, aggregated 6–10 × 3–5 Truncate base, cylindrical, thin-walled, with obtuse apex 20–32 × 2–2.5 1 Quaedvlieg et al. (2013)
Para. poagena up to 350 Ampulliform to subcylindrical 4–6 × 3–6 Subcylindrical, truncate base, sigmoid 30–60 × 3–4 3–9 Crous et al. (2014)
Para. pseudonodorum 200–350 Ampulliform to subcylindrical 4–9 × 4–6 Cylindrical, subobtuse apex 27–36 × 2.5–4 3 Croll et al. (2021)
Para. stipae 150–180 Ampulliform to subcylindrical 5–6 × 3–4 Subcylindrical, subobtuse apex 8–18 × 2.5–3 1 Croll et al. (2021)
Para. uniseptata 60–100 × 70–100 Ampulliform to broadly conical, phialidic 3–6 × 3–6.5 Subcylindrical, truncate base with obtuse apex 14–18 × 2–3 1 Li et al. (2015)
Para.yunnanensis 120–140 × 35–45 Ampulliform or globose to subglobose 3.2–5.3 × 3.4–6.1 Cylindrical to subcylindrical 16–20 × 3–4 0–1 In this study
Para. zhaotongensis 80–110 × 70–85 Globose to subglobose 3.5–6.5 × 3–6 Subcylindrical, rounded at apex, truncate base 22–32 × 3–4 0–3 In this study

Phaeoseptoriella poaceicola Y. Gao, H. Gui & K.D. Hyde, sp. nov.

Index Fungorum: IF902470
Facesoffungi Number: FoF16260
Fig. 4

Etymology

in reference to the holotype occurring on grasses (Poaceae)

Holotype

HKAS 128741.

Description

Saprobic on decaying stem of Dactylis glomerata (Poaceae). Sexual morph: Undetermined. Asexual morph: Conidiomata 60–75 μm high × 90–100 μm diam. (x– = 70 × 97 μm, n = 15), flattened, solitary, immersed in the epidermis of the host, globose to subglobose, brown to black spots. Conidiomatal wall 5.5–13 µm wide (x– = 9.5 μm, n = 25), thin wall, 1–4 layered, composed of pale brown cells of textura angularis, with inner layer comprising hyaline cells. Conidiogenous cells (4.1–)4–7.7(–9.1) × (4–)4.3–5.7(5.5) μm (x– = 5.9 ± 1.82 × 5 ± 0.70 μm, n = 25), hyaline, globose to subglobose, smooth-walled. Conidia (30–)33–39(–41) × (4.3–)5–6(–6.7) μm (x– = 36 ± 3.23 × 5.5 ± 0.56 μm, n = 35), phragmosporous, initially hyaline, becoming pale yellowish at maturity, 7-septate, cylindrical to subcylindrical, straight or slightly curved, smooth-walled, rounded at apex, slightly truncate at base, guttulate.

Figure 4. 

Asexual morph of Phaeoseptoriella poaceicola (HKAS 128741, holotype) on a dead stalk of Dactylis glomerata a, b conidiomata on the host c, d vertical section of conidiomata e conidioma wall i conidiogenous cell arise from the wall and develop conidium f–h, j conidia k germinating conidium l cultures on PDA from above m cultures on PDA from the reverse. Scale bars: 30 μm (c); 20 μm (d, e, k); 10 μm (f–h, j); 5 μm (i).

Culture characteristics

Colonies on PDA, reaching 10–20 mm diam., after three weeks at 25–27 °C, with irregular, floccose, raised, white from the above and in reverse yellow.

Material examined

China • Yunnan Province, Zhaotong City (27°38'37"N, 103°37'5"E), on decaying stem of Dactylis glomerata (Poaceae), 25 September 2022, Ying Gao, LG7A (HKAS 128741, holotype), ex-type (CGMCC 3.24561) • ibid. LG7B (HKAS 128742, paratype), ex-paratype (CGMCC 3.24562). ibid. • China, Yunnan Province, Zhaotong City, (27°26'34"N,103°19'16"E), on decaying stems of Anaphalis tenuisissima, 20 August 2021, Ying Gao, living cultures: ZY356B (CGMCC 3.25058), ZY359A (CGMCC 3.25059), ZY359B (CGMCC 3.25060).

Note

Phaeoseptoriella poaceicola is introduced as a new species based on morphology and phylogenetic analysis of combined SSU, LSU, ITS, tef1-α, and rpb2 datasets. Our strains of Phaeoseptoriella poaceicola (CGMCC 3.24561, CGMCC 3.24561, CGMCC 3.25058, CGMCC 3.25059, and CGMCC 3.25060) distinct clade (100% ML, 1.00 PP, Clade C, Fig. 1). The ITS pairwise nucleotide comparison of our isolate Phaeoseptoriella poaceicola (CGMCC 3.24561) with Phaeoseptoriella edithcowaniae (PP707905), Ph. emmelinepankhurstiae (OR673891), Ph. vidagoldsteiniae (OR673892), Ph. zeae (NR_163371) showed 123/601 bp, 83/537 bp, 81/537 bp, 88/534 bp, differences (20.47%, with 37 gaps; 15.46%, with 27 gaps; 15.08%, with 25 gaps; 16.48%, with 28 gaps), respectively. Our isolate differs from Ph. edithcowaniae (PP708933) and Ph. zeae (NG_067869) in 13/836 bp and 4/836 (1.56% and 0.48% without gaps) in the LSU regions, respectively. It differs from Ph. zeae (MK442674) in 130/878 bp (14.81%, without gaps) in the rpb2 regions, SSU and tef1-α of other Parastagonospora species were not provided. Phaeoseptoriella only included four species, however, Ph. emmelinepankhurstiae, Ph. edithcowaniae, and Ph. eidagoldsteiniae have not provided with morphological analyses (Tan and Shivas 2023, 2024). Phaeoseptoriella poaceicola differs from Ph. zeae in comparatively smaller conidiomata (90–100 μm diam. vs. 200–250 μm diam.), conidiogenous cells (hyaline, globose to subglobose vs. pale brown, ampulliform to doliiform), bigger conidia (30–41 μm long, 4–7 μm wide, cylindrical to subcylindrical, 7-septate, vs. 14–23 μm long, 3–4 μm wide, fusoid-ellipsoid, 3-septate) (Crous et al. 2019). Based on the guidelines for a polyphasic approach recommended for species boundary delimitation (Chethana et al. 2021; Maharachchikumbura et al. 2021), we introduce Phaeoseptoriella poaceicola as a novel taxon.

Discussion

This study refines the taxonomic classification of microfungi in grasslands across Yunnan Province, southwestern China, by identifying and characterizing three new fungal species viz. Parastagonospora yunnanensis, Para. zhaotongensis, and Phaeoseptoriella poaceicola. Our taxonomic approach incorporates a multi-locus sequence analysis utilizing five gene loci (SSU, LSU, ITS, tef1-α, and rpb2) crucial for discerning species boundaries in genera where morphological characteristics are either overlapping or inadequate for clear species differentiation (Wanasinghe and Maharachchikumbura 2023, Dissanayake et al. 2024; Wanasinghe et al. 2024). Notably, our results underscore the effectiveness of ITS and rpb2 loci in distinguishing species within the genera Parastagonospora and Phaeoseptoriella, supporting prior research on their importance for precise species identification (Quaedvlieg et al. 2013; Crous et al. 2019). However, we found that sequences from LSU and SSU alone often do not provide adequate differentiation. There is a notable lack of comprehensive data regarding the tef1-α region among the species studied, suggesting that its phylogenetic utility requires further investigation.

Morphologically, Parastagonospora species are primarily identified from their asexual states in natural settings, with sexual morphs either rarely observed or under-documented. The identified sexual morphs resemble didymella-like and phaeosphaeria-like structures, characterized by immersed ascomata with slightly papillate ostioles, bitunicate asci, and fusoid, septate ascospores that range from subhyaline to pale brown (Quaedvlieg et al. 2013). In this study, we have isolated eight collections of Parastagonospora, all reported from their asexual morphs. We have comprehensively summarized the asexual characteristics of all known Parastagonospora species in Table 4. The most common characteristics of these asexual morphs are globose to subglobose, brown or black, and semi- or fully immersed conidiomata, ampulliform, subcylindrical, lageniform, or doliiform conidiogenous cells proliferating percurrently at the apex and cylindrical or subcylindrical, subhyaline or hyaline, granular to multi-guttulate septate conidia (Quaedvlieg et al. 2013; Li et al. 2016; Thambugala et al. 2017; Goonasekara et al. 2019; Brahmanage et al. 2020; Croll et al. 2021). Our two new species fit well within the morphological features representing the genus.

Lolium perenne is an important pasture and forage plant used in many pasture seed mixes (Wei et al. 2023) and has been reported to have the potential for phytoremediation of contaminated soils (Yarahmadi et al. 2017; Madni et al. 2021). Parastagonospora nodorum was reported on Lolium perenne in Denmark by Quaedvlieg et al. (2013). In this study, Parastagonospora yunnanensis, is reported on the host plant Lolium perenne in China. Dactylis glomerata (Poaceae) is considered an economically important grass in grasslands (Nösberger and Opitz von Boberfeld 1986). At present, eight species from the genus Parastagonospora have been reported on Dactylis glomerata in Italy (Li et al. 2015, 2016; Thambugala et al. 2017; Goonasekara et al. 2019; Brahmanage et al. 2020; Croll et al. 2021). This study also reports Parastagonospora zhaotongensis, on the host plant Dactylis glomerata in China. Our findings, coupled with the host information for this fungal species presented in Table 1, suggest that there may be widespread interactions between Parastagonospora and various grass species across diverse geographic regions. The discovery of Phaeoseptoriella poaceicola on Dactylis glomerata marks the first report of any Phaeoseptoriella species on this host, suggesting a previously unrecognized host-fungus interaction.

Acknowledgments

We thank Yunnan Department of Sciences and Technology of China (Grant No: 202302AE090023, 202303AP140001). We would also like to thank the support from the Youth Innovation Promotion Association of CAS, China (Grant No.: 2022396), and the Yunnan Revitalization Talent Support Program “Young Talent” Project.Dhanushka Wanasinghe, Kevin Hyde and Turki Faraj gratefully acknowledge the financial support provided by the Distinguished Scientist Fellowship Program (DSFP) at King Saud University in Riyadh, Saudi Arabia. We thank the Chinese Academy of Sciences for providing molecular laboratory facilities.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

No funding was reported.

Author contributions

Conceptualization: DNW, YG. Data curation: YG. Formal analysis: DNW, YG, RSJ. Funding acquisition: HG, TKF. Investigation: YG. Methodology: YG. Project administration: KDH, HG. Supervision: PDE, RSJ, HG, KDH, DNW. Writing - original draft: YG. Writing - review and editing: HG, PDE, RSJ, DNW, KDH, TKF.

Author ORCIDs

Ying Gao https://orcid.org/0000-0001-8671-1978

Tingfang Zhong https://orcid.org/0009-0000-2767-1347

Prapassorn Damrongkool Eungwanichayapant https://orcid.org/0000-0001-8005-4137

Ruvishika S. Jayawardena https://orcid.org/0000-0001-7702-4885

Kevin D. Hyde https://orcid.org/0000-0002-2191-0762

Turki Kh. Faraj https://orcid.org/0000-0002-6012-8474

Dhanushka N. Wanasinghe https://orcid.org/0000-0003-1759-3933

Heng Gui https://orcid.org/0000-0002-0946-1589

Data availability

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

References

  • Ariyawansa HA, Kang J-C, Alias SA, Chukeatirote E, Hyde KD (2013) Towards a natural classification of Dothideomycetes: The genera Dermatodothella, Dothideopsella, Grandigallia, Hysteropeltella and Gloeodiscus (Dothideomycetes incertae sedis). Phytotaxa 147(2): 35–47. https://doi.org/10.11646/phytotaxa.147.2.1
  • Bakhshi M, Arzanlou M, Groenewald JZ, Quaedvlieg W, Crous PW (2019) Parastagonosporella fallopiae gen. et sp. nov. (Phaeosphaeriaceae) on Fallopia convolvulus from Iran. Mycological Progress 18(1–2): 203–214. https://doi.org/10.1007/s11557-018-1428-z
  • Barr ME (1992) Additions to and notes on the Phaeosphaeriaceae (Pleosporales, Loculoascomycetes). Mycotaxon 43: 371–400.
  • Brahmanage RS, Dayarathne MC, Wanasinghe DN, Thambugala KM, Jeewon R, Chethana KWT, Samarakoon MC, Tennakoon DS, De Silva NI, Camporesi E, Raza M, Yan JY, Hyde KD (2020) Taxonomic novelties of saprobic Pleosporales from selected dicotyledons and grasses. Mycosphere 11(1): 2481–2541. https://doi.org/10.5943/mycosphere/11/1/15
  • Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T (2009) trimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15): 1972–1973. https://doi.org/10.1093/bioinformatics/btp348
  • Chaiwan N, Gomdola D, Wang S, Monkai J, Tibpromma S, Doilom M, Wanasinghe DN, Mortimer PE, Lumyong S, Hyde KD (2021) An online database providing updated information of microfungi in the Greater Mekong Subregion. Mycosphere 12(1): 1513–1526. https://doi.org/10.5943/mycosphere/12/1/19
  • Chethana KT, Manawasinghe IS, Hurdeal VG, Bhunjun CS, Appadoo MA, Gentekaki E, Raspé O, Promputtha I, Hyde KD (2021) What are fungal species and how to delineate them? Fungal Diversity 109(1): 1–25. https://doi.org/10.1007/s13225-021-00483-9
  • Croll D, Crous PW, Pereira D, Mordecai EA, McDonald BA, Brunner PC (2021) Genome-scale phylogenies reveal relationships among Parastagonospora species infecting domesticated and wild grasses. Persoonia 46: 116–128. https://doi.org/10.3767/persoonia.2021.46.04
  • Crous PW, Shivas RG, Quaedvlieg W, van der Bank M, Zhang Y, Summerell BA, Guarro J, Wingfield MJ, Wood AR, Alfenas AC, Braun U, Cano-Lira JF, García D, Marin-Felix Y, Alvarado P, Andrade JP, Armengol J, Assefa A, den Breeÿen A, Camele I, Cheewangkoon R, De Souza JT, Duong TA, Esteve-Raventós F, Fournier J, Frisullo S, García-Jiménez J, Gardiennet A, Gené J, Hernández-Restrepo M, Hirooka Y, Hospenthal DR, King A, Lechat C, Lombard L, Mang SM, Marbach PAS, Marincowitz S, Marin-Felix Y, Montaño-Mata NJ, Moreno G, Perez CA, Pérez Sierra AM, Robertson JL, Roux J, Rubio E, Schumacher RK, Stchigel AM, Sutton DA, Tan YP, Thompson EH, Vanderlinde E, Walker AK, Walker DM, Wickes BL, Wong PTW, Groenewald JZ (2014) Fungal Planet Description Sheets: 214–280. Persoonia 32(1): 184–306. https://doi.org/10.3767/003158514X682395
  • Crous PW, Schumacher RK, Akulov A, Thangavel R, Hernández-Restrepo M, Carnegie AJ, Cheewangkoon R, Wingfield MJ, Summerell BA, Quaedvlieg W, Coutinho TA, Roux J, Wood AR, Giraldo A, Groenewald JZ (2019) New and Interesting Fungi. 2. Fungal Systematics and Evolution 3(1): 57–134. https://doi.org/10.3114/fuse.2019.03.06
  • Deng M, Chen J, Liu G, Wang H (2016) Risk assessment of drought based on IEAPP‐IDM in Qujing, Yunnan Province, China. Advances in Meteorology 7082467(1): 1–10. https://doi.org/10.1155/2016/7082467
  • Dissanayake LS, Marasinghe DS, Thambugala KM, Kang JC (2022) Neostagonosporella bambusicola sp. nov. (Phaeosphaeriaceae, Pleosporales) from bamboo in China. Phytotaxa 573(2): 262–274. https://doi.org/10.11646/phytotaxa.573.2.6
  • Dissanayake LS, Samarakoon MC, Maharachchikumbura SSN, Hyde KD, Tang X, Li QR, Mortimer PE, Faraj TK, Xu JC, Kang JC, Wanasinghe DN (2024) Exploring the taxonomy and phylogeny of Sordariomycetes taxa emphasizing Xylariomycetidae in southwestern China. Mycosphere : Journal of Fungal Biology 15(1): 1675–1793.
  • Fulcher MR, Winans JB, Bergstrom GC (2018) First report of tawny blotch caused by Parastagonospora caricis on Phalaris arundinacea in New York. Plant Disease 102(8): 1659–1659. https://doi.org/10.1094/PDIS-09-17-1348-PDN
  • Gao Y, Ren GC, Wanasinghe DN, Xu J-C, de Farias ARG, Gui H (2022) Two new species and a new record of Microdochium from grasses in Yunnan Province, South-West China. Journal of Fungi 8(12): 1297. https://doi.org/10.3390/jof8121297
  • Gao Y, Thiyagaraja V, Eungwanichayapant PD, de Farias ARG, Xu JC, Gui H, Wanasinghe DN (2024) Two new Stictidaceae species from grasslands in Yunnan province, China. New Zealand Journal of Botany 62(2–3): 1–15. https://doi.org/10.1080/0028825X.2024.2314562
  • Goonasekara ID, Jayawardene RS, Saichana N, Hyde KD (2018) Checklist of microfungi on grasses in Thailand (excluding bambusicolous fungi). Asian Journal of Mycology 1(1): 88–105. https://doi.org/10.5943/ajom/1/1/7
  • Goonasekara ID, Camporesi E, Bulgakov TS, Phookamsak R, Jayawardena RS, Saichana N, McKenzie EHC (2019) Two novel species of Parastagonospora (Phaeosphaeriaceae, Pleosporales) on grasses from Italy and Russia. Asian Journal of Mycology 2(1): 170–182. https://doi.org/10.5943/ajom/2/1/8
  • Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.
  • Hyde KD, Norphanphoun C, Abreu VP, Bazzicalupo A, Chethana KWT, Clericuzio M, Dayarathne MC, Dissanayake AJ, Ekanayaka AH, He MQ, Hongsanan S, Huang SK, Jayasiri SC, Jayawardena RS, Karunarathna A, Konta S, Kušan I, Lee H, Li JF, Lin CG, Liu NG, Lu YZ, Luo ZL, Manawasinghe IS, Mapook A, Perera RH, Phookamsak R, Phukhamsakda C, Siedlecki I, Soares AM, Tennakoon DS, Tian Q, Tibpromma S, Wanasinghe DN, Xiao YP, Yang J, Zeng XY, Abdel-Aziz FA, Li WJ, Senanayake IC, Shang QJ, Daranagama DA, de Silva NI, Thambugala KM (2017) Fungal diversity notes 603–708: Taxonomic and phylogenetic notes on genera and species. Fungal Diversity 87(1): 1–235. https://doi.org/10.1007/s13225-017-0391-3
  • Hyde KD, Norphanphoun C, Ma J, Yang HD, Zhang JY, Du TY, Gao Y, Gomes de Farias AR, He SC, He YK, Li CJY, Li JY, Liu XF, Lu L, Su HL, Tang X, Tian XG, Wang SY, Wei DP, Xu RF, Xu RJ, Yang YY, Zhang F, Zhang Q, Bahkali AH, Boonmee S, Chethana KWT, Jayawardena RS, Lu YZ, Karunarathna SC, Tibpromma S, Wang Y, Zhao Q (2023) Mycosphere notes 387–412 – novel species of fungal taxa from around the world. Mycosphere 14(1): 663–744. https://doi.org/10.5943/mycosphere/14/1/8
  • Jayasiri SC, Hyde KD, Ariyawansa HA, Bhat J, Buyck B, Cai L, Dai YC, Abd-Elsalam KA, Ertz D, Hidayat I, Jeewon R, Jones EBG, Bahkali AH, Karunarathna SC, Liu JK, Luangsa-ard JJ, Lumbsch HT, Maharachchikumbura SSN, McKenzie EHC, Moncalvo JM, Ghobad-Nejhad M, Nilsson H, Pang K, Pereira OL, Phillips AJL, Raspé O, Rollins AW, Romero AI, Etayo J, Selçuk F, Stephenson SL, Suetrong S, Taylor JE, Tsui CKM, Vizzini A, Abdel-Wahab MA, Wen TC, Boonmee S, Dai DQ, Daranagama DA, Dissanayake AJ, Ekanayaka AH, Fryar SC, Hongsanan S, Jayawardena RS, Li WJ, Perera RH, Phookamsak R, De Silva NI, Thambugala KM, Tian Q, Wijayawardene NN, Zhao RL, Zhao Q, Kang JC, Promputtha I (2015) The Faces of Fungi database: Fungal names linked with morphology, phylogeny and human impacts. Fungal Diversity 74(1): 3–18. https://doi.org/10.1007/s13225-015-0351-8
  • Jayawardena RS, Hyde KD, de Farias AR, Bhunjun CS, Ferdinandez HS, Manamgoda DS, Udayanga D, Herath IS, Thambugala KM, Manawasinghe IS, Gajanayake AJ, Samarakoon BC, Bundhun D, Gomdola D, Huanraluek H, Sun Y-R, Tang X, Promputtha I, Thines M (2021) What is a species in fungal plant pathogens? Fungal Diversity 109(1): 239–266. https://doi.org/10.1007/s13225-021-00484-8
  • Jones EBG, Pang KL, Abdel-Wahab MA, Scholz B, Hyde KD, Boekhout T, Ebel R, Rateb ME, Henderson L, Sakayaroj J, Suetrong S, Dayarathne MC, Kumar V, Raghukumar S, Sridhar KR, Bahkali AH, Gleason FH, Norphanphoun C (2019) An online resource for marine fungi. Fungal Diversity 96(1): 347–433. https://doi.org/10.1007/s13225-019-00426-5
  • Karunarathna A, Papizadeh M, Senanayake IC, Jeewon R, Phookamsak R, Goonasekara ID, Wanasinghe DN, Wijayawardene NN, Amoozegar MA, Shahzadeh Fazeli SA, Camporesi E, Hyde KD, Weerahewa HLD, Lumyong S, McKenzie EHC (2017) Novel fungal species of Phaeosphaeriaceae with an asexual/sexual morph connection. Mycosphere 8(10): 1818–1834. https://doi.org/10.5943/mycosphere/8/10/8
  • Karunarathna A, Tibpromma S, Jayawardena RS, Nanayakkara C, Asad S, Xu J-C, Hyde KD, Karunarathna SC, Stephenson SL, Lumyong S, Kumla J (2021) Fungal pathogens in grasslands. Frontiers in Cellular and Infection Microbiology 11: 695087. https://doi.org/10.3389/fcimb.2021.695087
  • Karunarathna A, Withee P, Pakdeeniti P, Haituk S, Tanakaew N, Senwanna C, Działak P, Karunarathna SC, Tibpromma S, Promthep T, Monkhung S, Cheewangkoon R (2022) Worldwide checklist on grass Fungi: What do we know so far in Ascomycota. Warasan Khana Witthayasat Maha Witthayalai Chiang Mai 49(3): 742–984. https://doi.org/10.12982/CMJS.2022.058
  • Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: Multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20(4): 1160–1166. https://doi.org/10.1093/bib/bbx108
  • Kuraku S, Zmasek CM, Nishimura O, Katoh K (2013) ALeaves facilitates on-demand exploration of metazoan gene family trees on MAFFT sequence alignment server with enhanced interactivity. Nucleic Acids Research 41(W1): 22–28. https://doi.org/10.1093/nar/gkt389
  • Li WJ, Bhat DJ, Camporesi E, Tian Q, Wijayawardene NN, Dai DQ, Phookamsak R, Chomnunti P, Bahkali AH, Hyde KD (2015) New asexual morph taxa in Phaeosphaeriaceae. Mycosphere 6(6): 681–708. https://doi.org/10.5943/mycosphere/6/6/5
  • Li GJ, Hyde KD, Zhao RL, Hongsanan S, Abdel-Aziz FA, Abdel-Wahab MA, Alvarado P, Alves-Silva G, Ammirati JF, Ariyawansa HA, Baghela A, Bahkali AH, Beug M, Bhat DJ, Bojantchev D, Boonpratuang T, Bulgakov TS, Camporesi E, Boro MC, Ceska O, Chakraborty D, Chen JJ, Chethana KWT, Chomnunti P, Consiglio G, Cui BK, Dai DQ, Dai YC, Daranagama DA, Das K, Dayarathne MC, De Crop E, De Oliveira RJV, de Souza CAF, de Souza JI, Dentinger BTM, Dissanayake AJ, Doilom M, Drechsler-Santos ER, Ghobad-Nejhad M, Gilmore SP, Góes-Neto A, Gorczak M, Haitjema CH, Hapuarachchi KK, Hashimoto A, He MQ, Henske JK, Hirayama K, Iribarren MJ, Jayasiri SC, Jayawardena RS, Jeon SJ, Jerônimo GH, Jesus AL, Jones EBG, Kang JC, Karunarathna SC, Kirk PM, Konta S, Kuhnert E, Langer E, Lee HS, Lee HB, Li WJ, Li XH, Liimatainen K, Lima DX, Lin CG, Liu JK, Liu XZ, Liu ZY, Luangsa-ard JJ, Lücking R, Lumbsch HT, Lumyong S, Leaño EM, Marano AV, Matsumura M, McKenzie EHC, Mongkolsamrit S, Mortimer PE, Nguyen TTT, Niskanen T, Norphanphoun C, O’Malley MA, Parnmen S, Pawłowska J, Perera RH, Phookamsak R, Phukhamsakda C, Pires-Zottarelli CLA, Raspé O, Reck MA, Rocha SCO, de Santiago ALCMA, Senanayake IC, Setti L, Shang QJ, Singh SK, Sir EB, Solomon KV, Song J, Srikitikulchai P, Stadler M, Suetrong S, Takahashi H, Takahashi T, Tanaka K, Tang LP, Thambugala KM, Thanakitpipattana D, Theodorou MK, Thongbai B, Thummarukcharoen T, Tian Q, Tibpromma S, Verbeken A, Vizzini A, Vlasák J, Voigt K, Wanasinghe DN, Wang Y, Weerakoon G, Wen HA, Wen TC, Wijayawardene NN, Wongkanoun S, Wrzosek M, Xiao YP, Xu JC, Yan JY, Yang J, Da Yang S, Hu Y, Zhang JF, Zhao J, Zhou LW, Peršoh D, Phillips AJL, Maharachchikumbura SSN (2016) Fungal diversity notes 253–366: Taxonomic and phylogenetic contributions to fungal taxa. Fungal Diversity 78(1): 1–237. https://doi.org/10.1007/s13225-016-0366-9
  • Li QR, Habib K, Long SH, Wu YP, Zhang X, Hu HM, Wu QZ, Liu LL, Zhou SX, Shen XC, Kang JC (2024) Unveiling fungal diversity in China: New species and records within the Xylariaceae family. Mycosphere : Journal of Fungal Biology 15(1): 275–364. https://doi.org/10.5943/mycosphere/15/1/2
  • Luo ZL, Hyde KD, Liu JK, Maharachchikumbura SSN, Jeewon R, Bao DF, Bhat DJ, Lin CG, Li WL, Yang J, Liu NG, Lu YZ, Jayawardena RS, Li JF, Su HY (2019) Freshwater Sordariomycetes. Fungal Diversity 99(1): 451–660. https://doi.org/10.1007/s13225-019-00438-1
  • Madni MA, Rasool SJ, Naeem M, Najeeb HU, Maqbool AI, Tariq A, Ijaz-ul-Haq S, Ali S, Rizwan M (2021) Phytoremediation of Anthracene and Pyrene Contaminated Soil by Lolium Perenne. Plant Science 10(11): 4294–4302. https://doi.org/10.21746/aps.2021.10.11.1
  • Maharachchikumbura SSN, Ariyawansa HA, Wanasinghe DN, Dayarathne MC, Al-Saady NA, Al-Sadi AM (2019) Phylogenetic classification and generic delineation of Hydeomyces desertipleosporoides gen. et sp. nov., (Phaeosphaeriaceae) from Jebel Akhdar Mountain in Oman. Phytotaxa 391(1): 28–38. https://doi.org/10.11646/phytotaxa.391.1.2
  • Maharachchikumbura SS, Chen Y, Ariyawansa HA, Hyde KD, Haelewaters D, Perera RH, Samarakoon MC, Wanasinghe DN, Bustamante DE, Liu JK, Lawrence DP, Cheewangkoon R, Stadler M (2021) Integrative approaches for species delimitation in Ascomycota. Fungal Diversity 109(1): 155–179. https://doi.org/10.1007/s13225-021-00486-6
  • Marin-Felix Y, Hernández-Restrepo M, Iturrieta-González I, García D, Gené J, Groenewald JZ, Cai L, Chen Q, Quaedvlieg W, Schumacher RK, Taylor PWJ, Ambers C, Bonthond G, Edwards J, Krueger-Hadfield SA, Luangsa-ard JJ, Morton L, Moslemi A, Sandoval-Denis M, Tan YP, Thangavel R, Vaghefi N, Cheewangkoon R, Crous PW (2019) Genera of phytopathogenic fungi: GOPHY 3. Studies in Mycology 94(1): 1–124. https://doi.org/10.1016/j.simyco.2019.05.001
  • Miao ZQ, Lei LP, Liu XZ (1999) Dactylella crassa, a new species of nematode-trapping fungi. Mycosystema 18(4): 354–356.
  • Miller MA, Pfeiffer W, Schwartz T (2012) The CIPRES science gateway: Enabling high-impact science for phylogenetics researchers with limited resources. Proceedings of the 1st Conference of the Extreme Science and Engineering Discovery Environment: Bridging from the Extreme to the Campus and Beyond 1–8. https://doi.org/10.1145/2335755.2335836
  • Nösberger J, Opitz von Boberfeld W (1986) Gründfutter-produktion. Verlag Paul Parey, Hamburg.
  • Nylander JAA, Wilgenbusch JC, Warren DL, Swofford DL (2008) AWTY: A system for graphical exploration of MCMC convergence in Bayesian phylogenetics. Bioinformatics 24(4): 581–583. https://doi.org/10.1093/bioinformatics/btm388
  • Pei Y (2022) Analysis of temperature variation characteristics in Zhaotong City in recent 50 years. Journal of Agricultural Catastrophology 12: 3.
  • Phookamsak R, Liu JK, McKenzie EHC, Manamgoda DS, Ariyawansa HA, Tambugala KM, Dai DQ, Camporesi E, Chukeatirote E, Wijayawardene NN, Bahkali AH, Mortimer PE, Xu JC, Hyde KD (2014) Revision of Phaeosphaeriaceae. Fungal Diversity 68(1): 159–238. https://doi.org/10.1007/s13225-014-0308-3
  • Phookamsak R, Wanasinghe DN, Hongsanan S, Phukhamsakda C, Huang SK, Tennakoon DS, Norphanphoun C, Camporesi E, Bulgakov TS, Promputtha I, Mortimer PE, Xu J-C, Hyde KD (2017) Towards a natural classification of Ophiobolus and ophiobolus-like taxa; introducing three novel genera Ophiobolopsis, Paraophiobolus and Pseudoophiobolus in Phaeosphaeriaceae (Pleosporales). Fungal Diversity 87(1): 299–339. https://doi.org/10.1007/s13225-017-0393-1
  • Phookamsak R, Hyde KD, Jeewon R, Bhat DJ, Jones EBJ, Maharachchikumbura SSN, Raspé O, Karunarathna SC, Wanasinghe DN, Hongsanan S, Doilom M, Tennakoon DS, Machado AR, Firmino AL, Ghosh A, Karunarathna A, Mešić A, Dutta AK, Thongbai B, Devadatha B, Norphanphoun C, Senwanna C, Wei D, Pem D, Ackah FK, Wang GN, Jiang HB, Madrid H, Lee HB, Goonasekara ID, Manawasinghe IS, Kušan I, Cano J, Gené J, Li J, Das K, Acharya K, Raj KNA, Latha KPD, Chethana KWT, He MQ, Dueñas M, Jadan M, Martín MP, Samarakoon MC, Dayarathne MC, Raza M, Park MS, Telleria MT, Chaiwan N, Matočec N, de Silva NI, Pereira OL, Singh PN, Manimohan P, Uniyal P, Shang QJ, Bhatt RP, Perera RH, Alvarenga RLM, Nogal-Prata S, Singh SK, Vadthanarat S, Oh SY, Huang SK, Rana S, Konta S, Paloi S, Jayasiri SC, Jeon SJ, Mehmood T, Gibertoni TB, Nguyen TTT, Singh U, Thiyagaraja V, Sarma VV, Dong W, Yu XD, Lu YZ, Lim YW, Chen Y, Tkalčec Z, Zhang ZF, Luo ZL, Daranagama DA, Thambugala KM, Tibpromma S, Camporesi E, Bulgakov T, Dissanayake AJ, Senanayake IC, Dai DQ, Tang LZ, Khan S, Zhang H, Promputtha I, Cai L, Chomnunti P, Zhao RL, Lumyong S, Boonmee S, Wen TC, Mortimer PE, Xu J-C (2019) Fungal diversity notes 929–1036: Taxonomic and phylogenetic contributions on genera and species of fungal taxa. Fungal Diversity 95(1): 1–273. https://doi.org/10.1007/s13225-019-00421-w
  • Quaedvlieg W, Verkley GJM, Shin HD, Barreto RW, Alfenas AC, Swart WJ, Groenewald JZ, Crous PW (2013) Sizing up Septoria. Studies in Mycology 75: 307–390. https://doi.org/10.3114/sim0017
  • Rambaut A, Drummond AJ (2012) FigTree: Tree Figure Drawing Tool. Institute of Evolutionary Biology, University of Edinburgh, Edinburgh.
  • Rehner SA, Buckley E (2005) A Beauveria phylogeny inferred from nuclear ITS and EF1-αsequences: Evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97(1): 84–98. https://doi.org/10.3852/mycologia.97.1.84
  • Risser PG (1988) Diversity in and Among Grasslands. In: Wilson E, Peter F (Eds) Biodiversity, (Washington, DC: National Academics Press), 176–180.
  • 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(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • 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 11(1): 2678–2754. https://doi.org/10.5943/mycosphere/11/1/20
  • Tennakoon DS, Jeewon R, Gentekaki E, Kuo CH, Hyde KD (2019) Multigene phylogeny and morphotaxonomy of Phaeosphaeria ampeli sp. nov. from Ficus ampelas and a new record of P. musae from Roystonea regia. Phytotaxa 406(2): 111–128. https://doi.org/10.11646/phytotaxa.406.2.3
  • Thambugala KM, Wanasinghe DN, Phillips AJL, Camporesi E, Bulgakov TS, Phukhamsakda C, Ariyawansa HA, Goonasekara ID, Phookamsak R, Dissanayake A, Tennakoon DS, Tibpromma S, Chen YY, Liu ZY, Hyde KD (2017) Mycosphere notes 1–50: Grass (Poaceae) inhabiting Dothideomycetes. Mycosphere 8(4): 697–796. https://doi.org/10.5943/mycosphere/8/4/13
  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24(24): 4876–4882. https://doi.org/10.1093/nar/25.24.4876
  • Vaidya G, Lohman DJ, Meier R (2011) SequenceMatrix: Concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27(2): 171–180. https://doi.org/10.1111/j.1096-0031.2010.00329.x
  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172(8): 4238–4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990
  • Wanasinghe DN, Maharachchikumbura SSN (2023) Exploring the diversity and systematics of Phaeosphaeriaceae: Taxonomic novelties from ecologically diverse habitats and their phylogenetic resolution. Journal of Fungi 9(8): 853. https://doi.org/10.3390/jof9080853
  • Wanasinghe DN, Jones EBG, Dissanayake AJ, Hyde KD (2016) Saprobic Dothideomycetes in Thailand: Vaginatispora appendiculata sp. nov. (Lophiostomataceae) introduced based on morphological and molecular data. Studies in Fungi 1(1): 56–68. https://doi.org/10.5943/sif/1/1/5
  • Wanasinghe DN, Phukhamsakda C, Hyde KD, Jeewon R, Lee HB, Jones EBG, Tibpromma S, Tennakoon DS, Dissanayake AJ, Jayasiri SC, Gafforov Y, Camporesi E, Bulgakov TS, Ekanayake AH, Perera RH, Samarakoon MC, Goonasekara ID, Mapook A, Li WJ, Senanayake IC, Li J, Norphanphoun C, Doilom M, Bahkali AH, Xu J-C, Mortimer PE, Tibell L, Savic ST, Karunarathna SC (2018) Fungal diversity notes 709–839: Taxonomic and phylogenetic contributions to fungal taxa with an emphasis on fungi on Rosaceae. Fungal Diversity 89(1): 1–236. https://doi.org/10.1007/s13225-018-0395-7
  • Wanasinghe DN, Nimalrathna TS, Qin Xian L, Faraj TK, Xu J, Mortimer PE (2024) Taxonomic novelties and global biogeography of Montagnula (Ascomycota, Didymosphaeriaceae). MycoKeys 101: 191–232. https://doi.org/10.3897/mycokeys.101.113259
  • Wei H, Wang J, Wang Q, He W, Liao S, Huang J, Hu W, Tang M, Chen H (2023) Role of melatonin in enhancing arbuscular mycorrhizal symbiosis and mitigating cold stress in perennial ryegrass (Lolium perenne L.). Frontiers in Microbiology 14: 1123632. https://doi.org/10.3389/fmicb.2023.1123632
  • Wijayawardene NN, Hyde KD, Dai DQ, Sánchez-García M, Goto BT, Saxena RK, Erdoğdu M, Selçuk F, Rajeshkumar KC, Aptroot A, Błaszkowski J, Boonyuen N, da Silva GA, de Souza FA, Dong W, Ertz D, Haelewaters D, Jones EBG, Karunarathna SC, Kirk PM, Kukwa M, Kumla J, Leontyev DV, Lumbsch HT, Maharachchikumbura SSN, Marguno F, Martínez-Rodríguez P, Mešić A, Monteiro JS, Oehl F, Pawłowska J, Pem D, Pfliegler WP, Phillips AJL, Pošta A, He MQ, Li JX, Raza M, Sruthi OP, Suetrong S, Suwannarach N, Tedersoo L, Thiyagaraja V, Tibpromma S, Tkalčec Z, Tokarev YS, Wanasinghe DN, Wijesundara DSA, Wimalaseana SDMK, Madrid H, Zhang GQ, Gao Y, Sánchez-Castro I, Tang LZ, Stadler M, Yurkov A, Thines M (2022) Outline of Fungi and fungus-like taxa – 2021. Mycosphere 13(1): 53–453. https://doi.org/10.5943/mycosphere/13/1/2
  • Xia JW, Ren SC, Ma LG, Zhang XG (2013) Heteroconium bannaense sp. nov. and a new record of the genus from China. Mycotaxon 121(1): 413–417. https://doi.org/10.5248/121.413
  • Yarahmadi Z, Baharlouei J, Shokoohi R, Alikhani MY, Shirmohammadi-Khorram N (2017) The efficiency of Lolium perenne for phytoremediation of anthracene in polluted soils in the presence of Bacillus aerophilus. Petroleum Science and Technology 35(7): 647–652. https://doi.org/10.1080/10916466.2016.1252771
  • Yasanthika WAE, Gomes de Farias AR, Wanasinghe DN, Chethana KWT, Zare R, Cai L, Maharachchikumbura SSN, Tennakoon DS, Perera RH, Luangharn T, Chomnunti P (2023) a web-based platform for soil inhabiting Ascomycota species. Studies in Fungi 8(1): 16. https://doi.org/10.48130/SIF-2023-0016
  • Yuan ZL, Lin FC, Zhang CL, Kubicek CP (2010) A new species of Harpophora (Magnaporthaceae) recovered from healthy wild rice (Oryza granulata) roots, representing a novel member of a beneficial dark septate endophyte. FEMS Microbiology Letters 307(1): 94–101. https://doi.org/10.1111/j.1574-6968.2010.01963.x
  • Zhaxybayeva O, Gogarten JP (2002) Bootstrap, Bayesian probability and maximum likelihood mapping: Exploring new tools for comparative genome analyses. BMC Genomics 3(1): 1–4. https://doi.org/10.1186/1471-2164-3-4
login to comment