Two new species of Parastagonospora and a new species of Phaeoseptoriella (Phaeosphaeriaceae, Pleosporales) from grasslands in Yunnan Province, China

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 Parastagonosporayunnanensis, Para.zhaotongensis, Phaeoseptoriellapoaceicola. Parastagonosporayunnanensis 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. Parastagonosporazhaotongensis features similar globose to subglobose conidiogenous cells but with 0–3-septate, cylindrical to subcylindrical conidia. Phaeoseptoriellapoaceicola 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.

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 Parastagonosporanodorum 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 Parastagonosporaavenae was a significant pathogen in oats and caused leaf spots in barley and rye. Parastagonosporanodorum 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.

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

Species name	Host	Country	Life-Mode	References
Para.allouniseptata	Dactylisglomerata	Italy	Saprobic	Li et al. (2015)
Para.Arcana	Triticumaestivum	Iran	Pathogenic	Croll et al. (2021)
Para.Avenae	Loliummultiflorum, Avenasativa	Australia, Germany	Pathogenic	Quaedvlieg et al. (2013); Croll et al. (2021)
Para.bromicola	Bromusinermis	USA	NA	Croll et al. (2021)
Para.cumpignensis	Dactylisglomerata	Italy	Saprobic	Li et al. (2016)
Para.Caricis	Phalarisarundinacea, CarexacutiformisCyperaceae sp.	Netherlands, USA	NA	Quaedvlieg et al. (2013); Fulcher et al. (2018)
Para.dactylidicola	Dactylisglomerata	Italy	Saprobic	Brahmanage et al. (2020)
Para.dactylidigena	Dactylisglomerata	Iran	NA	Croll et al. (2021)
Para.dactylidis	Dactylis sp.	Italy	Saprobic	Li et al. (2015)
Para.Elymi	Elymusrepens	Russia	Saprobic	Goonasekara et al. (2019)
Para.forlicesenica	Dactylisglomerata	Italy	Saprobic	Thambugala et al. (2017)
Para.fusiformis	Dactylisglomerata	Italy	Saprobic	Thambugala et al. (2017)
Para. golestanensis	Agropyrontauri	Iran	NA	Croll et al. (2021)
Para.Italica	Dactylis sp.	Italy	Saprobic	Li et al. (2015)
Para.Jasniorum	Triticumaestivum	Iran	Pathogenic	Croll et al. (2021)
Para.macrouniseptata	Dactylisglomerata	Italy	Saprobic	Goonasekara et al. (2019)
Para.Minima	Dactylis sp.	Italy	Saprobic	Li et al. (2015)
Para.nodorum	Loliumperenne, Triticumaestivum, Leymuschinensis	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	Dactylisglomerata	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	Triticumaestivum	Iran	Pathogenic	Croll et al. (2021)
Para.Stipae	Stipapulchra	USA	NA	Croll et al. (2021)
Para.uniseptata	Daucus sp.	Italy	Saprobic	Li et al. (2015)
Para.yunnanensis	Loliumperenne	China	Saprobic	In this study
Para.zildae	Triticumaestivum	Iran	Pathogenic	Croll et al. (2021)
Para.zhaotongensis	Dactylisglomerata	China	Saprobic	In this study

Phaeoseptoriella was introduced by Crous et al. (2019), with Phaeoseptoriellazeae as the type species, on the leaves of Zeamays (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. Phaeoseptoriellaemmelinepankhurstiae and Ph.vidagoldsteiniae were collected from leaves of Sporobolusnatalensis (Poaceae) in Australia (Tan and Shivas 2023). Phaeoseptoriellaedithcowaniae was collected from a leaf spot of Heteropogontriticeus (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. Dactylellacrassa was introduced by Miao et al. (1999) from Oryza sp. Hypogymniacongesta was reported by McCune et al. (2003) from the Poaceae hosts. Yuan et al. (2010) introduced a new species, Harpophoraoryzae collected from Oryzagranulate. Xia et al. (2013) introduced a novel species Heteroconiumbannaense from Phragmites sp. Yunnanensisphragmitis was reported by Karunarathna et al. (2017). Based on morphology and phylogeny, Gao et al (2022) introduced Microdochiumgraminearum, 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 Anthostomellayunnanensis, Astrocystisheterocyclae and Collodisculabaoshanensis, while Dissanayake et al. (2024) introduced Apiosporaguangdongensis, A.locuta-pollinis, A.menglaensis, A.pseudoparenchymatica, Collodisculayunnanensis, Digitodochiumailaoshanense 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, Parastagonosporayunnanensis and Para.zhaotongensis, in Parastagonospora and a novel species, Phaeoseptoriellapoaceicola, to the Phaeoseptoriella. The specimens from which these species were collected on Loliumperenne and Dactylisglomerata 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.

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 Parastagonosporabromicola, 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.

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
Dematiopleosporamariae	MFLU 16-0121	MT226689	MT214576	MT310621	MT394635	NA
Dematiopleosporamariae	MFLUCC 13-0612T	KJ749652	KJ749653	KX274244	KJ749655	NA
Dematiopleosporasalsolae	MFLUCC 17-0828T	NG_063679	NG_059184	NR_157514	MG829201	MG829254
Neosphaerellopsisthailandica	CPC 21659T	NA	NG_067289	NR_137954	NA	NA
Nodulosphaeriaaconiti	MFLUCC 13-0728T	KU708840	KU708844	NR_154236	KU708852	KU708856
Nodulosphaeriaguttulatum	MFLUCC 15-0069	KY501115	KY496726	KY496746	KY514394	KY514405
Nodulosphaeriascabiosae	MFLUCC 14-1111T	NG_063602	KU708846	NR_154237	KU708854	KU708857
Paraloratosporamarina	MFLUCC 19-0691T	OQ130107	OQ130110	OQ130046	OQ357219	OQ162221
Paraloratosporasichuanensis	KUNCC 23-14218T	OR206405	OR206415	OR206396	OR195712	OR195721
Paraloratosporasichuanensis	HKAS 129218	OR206406	OR206416	OR206397	OR195713	OR195722
Parastagonosporaallouniseptata	MFLUCC 13-0386T	NA	KU058721	KU058711	MG520914	NA
Parastagonosporaavenae	CBS 289.69	NA	KF251678	KF251174	NA	KF252182
Parastagonosporaavenae	CBS 290.69	NA	KF251679	KF251175	NA	KF252183
Parastagonosporacaricis	CBS 135671T	NA	KF251680	KF251176	NA	KF252184
Parastagonosporadactylidicola	MFLU 20-0387T	NA	MT370430	MT370412	NA	NA
Parastagonosporadactylidis	MFLUCC 13-0375T	NA	KU058722	KU058712	NA	NA
Parastagonosporadactylidis	MFLUCC 13-0376	MG520986	KU058723	KU058713	MG520916	NA
Parastagonosporadactylidis	MFLUCC 13-0573	KU842390	KU842389	KU842388	NA	NA
Parastagonosporaelymi	KUMCC 16-0125T	NA	MN002870	MN002867	NA	NA
Parastagonosporaforlicesenica	MFLUCC 13-0557T	NA	KY769661	KY769660	NA	NA
Parastagonosporafusiformis	MFLUCC 13-0215T	NG_068367	NG_068235	NR_165848	NA	KX863711
Parastagonosporaitalica	MFLUCC 13-0377T	MG520985	KU058724	KU058714	MG520915	NA
Parastagonosporamacrouniseptata	KUMCC 16-0111T	NA	MN002868	MN002869	NA	MN019669
Parastagonosporanodorum	CBS 110109	EU754076	KF251681	KF251177	NA	KF252185
Parastagonosporanovozelandica	CPC 29613T	NA	MK540028	MK539957	NA	MK540088
Parastagonosporaphragmitis	CPC 32075T	NA	NG_066451	NR_164454	NA	MK540089
Parastagonosporapoaceicola	MFLUCC 15-0471T	NG_068368	NG_068537	NA	NA	KX880499
Parastagonosporapoae	CBS 135091	NA	KF251683	KF251179	NA	KF252187
Parastagonosporapoae	CBS 135089T	NA	KF251682	KF251178	NA	KF252186
Parastagonosporapoagena	CBS 136776T	NA	KJ869174	KJ869116	NA	NA
Parastagonosporastipae	pn1617	NA	NA	MW263184	NA	NA
Parastagonosporauniseptata	MFLUCC 13-0387T	MG520987	KU058725	KU058715	MG520917	NA
Parastagonosporayunnanensis	CGMCC 3.24527T	PQ046289	PQ046315	PQ046302	PQ058300	PQ058313
Parastagonosporayunnanensis	CGMCC 3.24528	PQ046290	PQ046316	PQ046303	PQ058301	PQ058314
Parastagonosporayunnanensis	CGMCC 3.24529	PQ046291	PQ046317	PQ046304	PQ058302	PQ058315
Parastagonosporayunnanensis	CGMCC 3.24530	PQ046292	PQ046318	PQ046305	PQ058303	PQ058316
Parastagonosporayunnanensis	CGMCC 3.24511	PQ046285	PQ046311	PQ046298	PQ058296	PQ058309
Parastagonosporayunnanensis	CGMCC 3.24512	PQ046286	PQ046312	PQ046299	PQ058297	PQ058310
Parastagonosporazhaotongensis	CGMCC 3.24519T	PQ046287	PQ046313	PQ046300	PQ058298	PQ058311
Parastagonosporazhaotongensis	CGMCC 3.24520	PQ046288	PQ046314	PQ046301	PQ058299	PQ058312
Phaeoseptoriellaedithcowaniae	BRIP 75864aT	NA	PP708933	PP707905	NA	NA
Phaeoseptoriellaemmelinepankhurstiae	BRIP65639aT	NA	NA	OR673891	NA	NA
Phaeoseptoriellapoaceicola	CGMCC 3.24561T	PQ046283	PQ046309	PQ046296	PQ058294	PQ058307
Phaeoseptoriellapoaceicola	CGMCC 3.24562	PQ046284	PQ046310	PQ046297	PQ058295	PQ058308
Phaeoseptoriellapoaceicola	CGMCC 3.25058	PQ046293	PQ046319	PQ046306	PQ058304	PQ058317
Phaeoseptoriellapoaceicola	CGMCC 3.25059	PQ046294	PQ046320	PQ046307	PQ058305	PQ058318
Phaeoseptoriellapoaceicola	CGMCC 3.25060	PQ046295	PQ046321	PQ046308	PQ058306	PQ058319
Phaeoseptoriellavidagoldsteiniae	BRIP65641aT	NA	NA	OR673892	NA	NA
Phaeoseptoriellazeae	CBS 144614T	NA	NG_067869	NR_163371	NA	MK442674
Phaeosphaeriachengduensis	KUNCC 23-13571T	OR206401	OR206411	OR206392	OR195708	OR195717
Phaeosphaeriachiangraina	MFLUCC 13-0231T	KM434289	NG_069237	NR_155643	KM434298	KM434307
Phaeosphaeriasichuanensis	KUNCC 23-13569T	OR206399	OR206409	OR206390	OR195706	OR195715
Phaeosphaeriathysanolaenicola	MFLUCC 10-0563T	KM434286	NG_069236	NR_155642	KM434295	KM434303
Quixadomyceshongheensis	KUMCC 20-0215T	NG_074964	MW264194	NR_172441	MW256816	MW269529
Quixadomyceshongheensis	HKAS 112346	MW541833	MW541822	MW541826	MW556134	MW556136
Sclerostagonosporalathyri	MFLUCC 14-0958T	NG_063692	NG_069566	NR_158956	MG829235	NA
Sclerostagonosporarosicola	MFLUCC 15-0129T	NG_063693	MG829068	MG828957	MG829237	NA
Septoriellaarundinicola	MFLU 16-0225T	NG_062199	MG829056	MG828946	MG829228	MG829261
Septoriellaasparagicola	MFLUCC 16-0379T	NG_067708	NG_070081	NR_165908	MK443385	MK443387
Septoriellaneodactylidis	MFLUCC 14-0966T	NG_061288	NG_069554	NR_157511	MG829199	MG829253
Wojnowiciellaclematidis	MFLUCC 17-2159T	MT226695	MT214582	NR_170812	MT394641	MT394698
Wojnowiciellakunmingensis	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 200^th 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 Quixadomyceshongheensis (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 Parastagonosporaallouniseptata (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 Parastagonosporapoae (CBS 135089, CBS 135091) and Para.uniseptata (MFLUCC 13-0387). However, this sister relationship was not statistically supported (Fig. 1).

Phaeoseptoriellaedithcowaniae (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.

249C7016-9A4F-5429-8E7E-98B6C065A69C

Index Fungorum: IF902468

Facesoffungi Number: FoF16258

Fig. 2

Figure 2.

Asexual morph of Parastagonosporayunnanensis (HKAS 128771, holotype) on a dead stalk of Loliumperennea, 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).

Etymology.

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

Holotype.

HKAS 128771.

Description.

Saprobic on decaying stem of Loliumperenne (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.

Material examined.

China • Yunnan Province, Zhaotong City, (26°56'39"N, 103°8'53"E), on a decaying stem of Loliumperenne (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 Loliumperenne (Poaceae), 27 August 2022, Ying Gao, QG19A (HKAS 128799), living culture (CGMCC 3.24511) • ibid. QG19B (HKAS 128800), living culture (CGMCC 3.24512).

Notes.

Parastagonosporayunnanensis 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. Parastagonosporayunnanensis is phylogenetically related to Parastagonosporaelymi (KUMCC 16-0125). Parastagonosporaelymi was introduced as a saprobic fungus from Elymusrepens in Russia, the asexual morph of Parastagonosporaelymi 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 Parastagonosporaelymi were not provided.

. Parastagonospora zhaotongensis

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

C80654EA-E062-5F6A-874C-F9742BCB1737

Index Fungorum: IF902469

Facesoffungi Number: FoF16259

Fig. 3

Figure 3.

Asexual morph of Parastagonosporazhaotongensis (HKAS 132983, holotype) on a dead stalk of Dactylisglomerataa, 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).

Etymology.

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

Holotype.

HKAS 132983.

Description.

Saprobic on decaying stem of Dactylisglomerata (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.

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 Dactylisglomerata (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 Parastagonosporazhaotongensis (CGMCC 3.24519 and CGMCC 3.24520) are closely related to Para.uniseptata (MFLUCC 13-0387) and Para.poae. Parastagonosporauniseptata was reported on Daucus sp. from Italy by Li et al. (2015). Pairwise nucleotide comparison indicates that our strains differ from Parastagonosporauniseptata 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 Parastagonosporauniseptata was not available for comparisons. Morphologically, Parastagonosporazhaotongensis 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 Parastagonosporapoae (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 Parastagonosporapoae were not provided. Parastagonosporazhaotongensis 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.

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.

247AD12B-F2A1-5475-8FDA-35DFA8C9CD47

Index Fungorum: IF902470

Facesoffungi Number: FoF16260

Fig. 4

Figure 4.

Asexual morph of Phaeoseptoriellapoaceicola (HKAS 128741, holotype) on a dead stalk of Dactylisglomerataa, 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).

Etymology.

in reference to the holotype occurring on grasses (Poaceae)

Holotype.

HKAS 128741.

Description.

Saprobic on decaying stem of Dactylisglomerata (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.

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 Dactylisglomerata (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 Anaphalistenuisissima, 20 August 2021, Ying Gao, living cultures: ZY356B (CGMCC 3.25058), ZY359A (CGMCC 3.25059), ZY359B (CGMCC 3.25060).

Note.

Phaeoseptoriellapoaceicola 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 Phaeoseptoriellapoaceicola (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 Phaeoseptoriellapoaceicola (CGMCC 3.24561) with Phaeoseptoriellaedithcowaniae (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). Phaeoseptoriellapoaceicola 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 Phaeoseptoriellapoaceicola 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. Parastagonosporayunnanensis, Para.zhaotongensis, and Phaeoseptoriellapoaceicola. 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.

Loliumperenne 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). Parastagonosporanodorum was reported on Loliumperenne in Denmark by Quaedvlieg et al. (2013). In this study, Parastagonosporayunnanensis, is reported on the host plant Loliumperenne in China. Dactylisglomerata (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 Dactylisglomerata 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 Parastagonosporazhaotongensis, on the host plant Dactylisglomerata 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 Phaeoseptoriellapoaceicola on Dactylisglomerata marks the first report of any Phaeoseptoriella species on this host, suggesting a previously unrecognized host-fungus interaction.

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

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.