Taxonomic analysis of Amphisphaeriales in China: discovery of multiple novel taxa

Qi-Yun Liu; Zhao-Xue Zhang; Ya-Ling Wang; Meng-Yuan Zhang; Zi-Xu Dong; Yu-Xin Shang; Du-Hua Li; Jin-Yang Yuan; Xian-Hao Li; Yang Jiang; Wen-Wen Liu; Xing-Sheng Wang; Ji-Wen Xia; Cong-Cong Ai; Shi Wang; Xiao-Yong Liu; Zhuang Li; Xiu-Guo Zhang; Qi-Yun Liu; Zhao-Xue Zhang; Ya-Ling Wang; Meng-Yuan Zhang; Zi-Xu Dong; Yu-Xin Shang; Du-Hua Li; Jin-Yang Yuan; Xian-Hao Li; Yang Jiang; Wen-Wen Liu; Xing-Sheng Wang; Ji-Wen Xia; Cong-Cong Ai; Shi Wang; Xiao-Yong Liu; Zhuang Li; Xiu-Guo Zhang

doi:10.48130/mycosphere-0026-0010

Amphisphaeriales primarily contains saprobes, pathogens, and endophytes. This order of fungi is difficult to identify because of its taxonomic and morphological complexities. However, it has significant ecological and economic value and has recently become a major focus in fungal research. A total of 164 strains, isolated from decaying plant materials, diseased leaves, and healthy leaves, were used to explore the taxonomic diversity of Amphisphaeriales in China based on analyses of its morphology and multi-locus phylogenies (internal transcribed spacer [ITS], large subunit of ribosomal DNA [LSU], translation elongation factor 1-alpha [tef-1α], and beta tubulin [tub2]). The results of these analyses showed that all these collected strains belonged to 71 species, including Apiosporaceae (Apiospora and Nigrospora), Beltraniaceae (Beltraniella), and Sporocadaceae (Neopestalotiopsis, Pestalotiopsis, and Pseudopestalotiopsis). These analyses reveal three major findings: 38 new species, 33 known species (with A. locuta-pollinis (syn. A. fuxianhuensis) included among the known species. Through this study, we confirm a primary association of Apiospora with Poaceae in tropical and subtropical climates and documents multi-species co-infections. Our findings significantly contribute to the understanding of the placement of new taxa in Apiosporaceae, Beltraniaceae, and Sporocadaceae. Moreover, the results of this study contribute to the future taxonomic revisions and systematic studies of Amphisphaeriales. Keywords: Amphisphaeriales, new species, Apiosporaceae, Beltraniaceae, Sporocadaceae, Phylogeny, Taxonomy

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Introduction

The order Amphisphaeriales was introduced by Eriksson and Hawksworth in 1986 and was taxonomically classified within the subclass Xylariomycetidae, class Sordariomycetes, and subphylum Pezizomycotina^[1]. Amphisphaeriales and Xylariales had historically been treated as closely related orders or even synonyms^[2,3], and were later formally considered synonymous^[4,5]. Amphisphaeriales was resurrected based on morphological and phylogenetic data^[6]. Previous studies have provided key evidence supporting the recognition of Amphisphaeriales and Xylariales as distinct orders, with divergence time serving as a critical criterion^[7−11]. The sexual morph of Amphisphaeriales is defined as follows: ascomata are predominantly solitary, globose to subglobose, coriaceous, and erumpent, occurring with or without stromatic tissues; asci are cylindrical or clavate with a J+ or J− apical ring; ascospores are ellipsoid, apiosporous, fusiform, or oval in shape, unicellular to trans-septate, variously colored, and possess either smooth or ornamented walls with or without appendages or mucilaginous sheaths^[6,11,12]. The asexual morph of Amphisphaeriales is characterized by acervular or pycnidial conidiomata with holoblastic, annellidic, ampulliform or lageniform conidiogenous cells; conidia range from aseptate to transeuseptate, come in various colors, and often bear appendages^[6,12]. Some species of Amphisphaeriales are mainly saprobes, pathogens or endophytes on plants^[6]. Some Arthrinium species, such as Arthrinium phaeospermum, were reported as human pathogens causing cutaneous mycosis^[13]. Members of Amphisphaeriales exhibit diverse bioactivities with practical applications; for example, Arthrinium arundinis has been utilized as an antifungal agent in pharmaceuticals^[14]. Similarly, Seiridium species produce seiridins because of their phytotoxicity and antimicrobial potential^[15]. Currently, Amphisphaeriales comprises 16 families and two incertae sedis genera^[16]. In this study, we investigate three families (Apiosporaceae, Beltraniaceae, and Sporocadaceae) by refining their taxonomic boundaries and clarifying phylogenetic relationships. This study addresses the gaps in knowledge about this fungal group in China and establishes a robust classification framework, demonstrating significant academic value.

The family Apiosporaceae was established to accommodate fungal genera that was characterized by apiosporous hyaline ascospores and a basauxic, Arthrinium-like mode of conidiogenesis^[17]. Saccardo established the genus Apiospora, designating A. montagnei as its type species^[18]. The genus Arthrinium was established with Arthrinium caricicola as its type species by Schmidt and Kunze^[19] and validated by Fries^[20]. Based on the principles advocated by Hawksworth^[21], Crous and Groenewald^[22] synonymized the sexual genus Apiospora with Arthrinium and prioritized the use of the latter. Five genera were established in Apiosporaceae: Appendicospora, Arthrinium, Dictyoarthrinium, Endocalyx, and Nigrospora^[10]. Samarakoon and Konta have subsequently segregated the genera Dictyoarthrinium, Endocalyx, and Appendicospora from the Apiosporaceae based on a series of phylogenetic analyses^[11,23,24]. Pintos and Alvarado^[25] proposed the taxonomic separation of Apiospora and Arthrinium by integrating genetic fragments and morphological and ecological data. Thus, the family Apiosporaceae currently accommodates three genera: Apiospora, Arthrinium, and Nigrospora. Notably, the Apiospora contains various species that are difficult to distinguish because of their highly similar morphological characteristics. However, Apiospora and Arthrinium can be distinguished based on three primary characteristics: 1) conidial morphology: Apiospora appears nearly spherical and lenticular with a pale equatorial slit, whereas Arthrinium exhibits an angular, curved, boat-shaped, fusiform, and polygonal morphology^[22,25−28]; 2) host preference: Apiospora predominantly colonizes Poaceae and other plants, while Arthrinium occurs mainly on Cyperaceae or Juncaceae^[28,29]; and 3) biogeography: Apiospora is common in tropical and subtropical regions, but Arthrinium is frequently reported from temperate, cold, or alpine habitats^[25,28,29].

The fungal tribe Beltranieae was established by a single genus Beltrania^[30]. Nannizzi^[31] later introduced the Beltraniaceae to accommodate Beltrania and its similar genera, thereby treating the earlier tribe as a synonym. Five genera, including Beltrania, Beltraniella, Beltraniopsis, Parapleurotheciopsis, and Pseudobeltrania, were emended to Beltraniaceae by Crous et al.^[32]. Then, Rajeshkumar et al. expanded the family Beltraniaceae^[33] by adding two other genera: Hemibeltrania and Porobeltraniella. Subsequently, four other Beltrania-like genera were proposed: Pseudosubramaniomyces was established by its type species, Pseudosubramaniomyces fusisaprophyticus^[34]; Subsessila was established by its type species Subsessila turbinate, collected from an unidentified decaying leaf in Thailand^[35]; Anabeltraniomyces and Parabeltrania were established with Anabeltraniomyces maximus as the type species from Clusia sp. and with Parabeltrania persianiae as the type species from indeterminate dead leaves, respectively, in Costa Rica in 2024^[36]. Liu et al.^[37] developed a dichotomous key to facilitate the identification of 11 genera within Beltraniaceae.

Sporocadaceae was originally established by Corda^[38] to accommodate the genera Sporocadus, Pestalozzia (currently Pestalotia), and Prosthemium. For a considerably long period, Sporocadaceae was synonymized with Amphisphaeriaceae^[39]. Sporocadaceae was resurrected by Jaklitsch et al.^[4]. Liu et al.^[40] established a relatively robust classification system for Sporocadaceae based on morphology and multi-gene phylogenetics. Pestalotiopsis, as an important taxon in Sporocadaceae, was established to accommodate species with 4-septate conidia^[41]. However, these pestalotioid taxa pose a challenge for accurate identification due to their high degree of morphological similarity, particularly in the absence of molecular data^[12]. Phylogenetic analysis of large subunit of ribosomal DNA (LSU) alignment, together with literature and morphological appraisal, defined the segregation of Neopestalotiopsis and Pseudopestalotiopsis from Pestalotiopsis^[42]. Liu et al.^[43] reported that the internal transcribed spacer (ITS) sequence lengths of Neopestalotiopsis was 480–484 bp, that of Pestalotiopsis was 489–495 bp, and that of Pseudopestalotiopsis was 536–540 bp. Razaghi et al. further revealed that whole-genome sizes of Neopestalotiopsis (49.73–53.78 Mbp) and Pseudopestalotiopsis (50.41–51.91 Mbp) are generally larger, but that of Pestalotiopsis is the smallest (< 50 Mbp)^[44].

In this study, we collected decaying plant materials, diseased leaves, and healthy leaves from eight provinces in China from 2022 to 2025. Through morphological comparisons and multi-gene phylogenetic analysis, a total of 71 species in the Amphisphaeriales were identified, comprising 38 new species and 33 known species, with A. locuta-pollinis (syn. A. fuxianhuensis) included among the known species.These findings significantly contribute to our understanding of taxa diversity in the Amphisphaeriales and help in refining its taxonomic framework.

Materials and methods

Sample collection, strain isolation, and morphological study

Between 2022 and 2025, plant specimens were collected from eight provinces across China. The distribution and predominant habitat types of samples were as follows: Fujian (1,230 specimens, natural forest and farmland), Guangdong (372 specimens, natural forest and farmland), Guangxi (500 specimens, natural forest and farmland), Guizhou (203 specimens, natural forest), Hainan (3,090 specimens, natural forest, farmland, riparian wetland, bamboo forest, and tea plantation), Shandong (345 specimens, natural forest, and farmland), Sichuan (183 specimens, alpine meadow, and bamboo forest), and Yunnan (304 specimens, natural forest, and alpine meadow). The time, geography, and host plant information were recorded for all these samples. Then, they were all preserved for further analysis. Pure colonies were isolated using the tissue method. The strains were deposited in 15% sterile glycerol at 4 °C. Typical specimens were preserved in the Herbarium of the Department of Plant Pathology, Shandong Agricultural University, Shandong, China (HSAUP). Living cultures were deposited in the Shandong Agricultural University Culture Collection (SAUCC) and China General Microbiological Culture Collection Center (CGMCC). After 7 or 14 days of growth on PDA, we recorded the morphological information of the isolates colony and photographed the plate surfaces (obverse and reverse) with a Canon G7X digital camera (Canon, Tokyo, Japan). Morphological features were described using two media: water agar (WA) with or without pine needles and potato dextrose agar (PDA) without pine needles. Fungal microstructures were observed using a stereomicroscope (Olympus SZ61, Olympus Corporation, Tokyo, Japan) and microscope (Olympus BX53, Olympus Corporation, Tokyo, Japan) with differential interference contrast (DIC), and subsequently captured using attached digital cameras (BioHD-A20c, FluoCa Scientific, Shanghai, China). We randomly measured the microstructures using Digimizer software v5.6.0 (www.digimizer.com) and calculated the average size (av.), with "n" indicating the total number of measurements.

The abbreviations of genera are as follows: A. = Apiospora, B. = Beltraniella, Neo. = Neopestalotiopsis, Nig. = Nigrospora, P. = Pestalotiopsis, and Ps. = Pseudopestalotiopsis. Newly introduced fungal species were registered with the Fungal Names database (https://nmdc.cn/fungalnames/)^[45].

DNA extraction, PCR amplification, and sequencing

Genomic DNA was extracted using the modified CTAB method^[46] or magnetic fungi DNA extraction kit (DC112-C7, VAMNE Magnetic Bacteria/Fungi DNA Extraction Kit [Prepackaged], Vazyme Biotech Co.,Ltd). The respective primers and different conditions for the PCR are detailed in Table 1. The PCR amplification reactions were performed in a 25 μL mixture containing 12.5 μL of 2 × Hieff Canace Plus PCR Master Mix (AG12202, Accurate Biotechnology, ChangSha, China), 1 μL each of forward and reverse primers (TsingKe, Qingdao, China), 1 μL of template genomic DNA, and nuclease-free water to adjust the final volume. The amplified products were separated by electrophoresis on a 1% agarose gel (Cat. Nos. RM02852, ABclonal Biotechnology Co., Ltd., Wuhan, China) stained with GelStain (GS101; TransGen Biotech, China) and then visualized under UV light. Target bands were excised from the gel, purified using a commercial gel extraction kit (DM1200, Beijing Solarbio Science & Technology Co., Ltd.), and subsequently sent for sequencing to Youkang Company Limited (Zhejiang, China).

Table 1. Molecular markers, and all PCR primers and programs used in this study.

Loci	PCR primers	Sequence (5′→3′)	PCR cycles	Ref.
ITS	ITS5	GGA AGT AAA AGT CGT AAC AAG G	95 °C 5 min; (95 °C 30 s, 55 °C 30 s, 72 °C 1 min) × 35 cycles	[47]
ITS	ITS4	TCC TCC GCT TAT TGA TAT GC		[47]
LSU	LR0R	GTA CCC GCT GAA CTT AAG C	95 °C 5 min; (95 °C: 30 s, 52 °C: 30 s, 72 °C: 1 min) × 35 cycles	[48,49]
LSU	LR5	TCC TGA GGG AAA CTT CG		[48,49]
tef1-α	EF1-728F	CAT CGA GAA GTT CGA GAA GG	95 °C 5 min; (95 °C 30 s, 55 °C 60 s, 72 °C 1 min) × 30 cycles	[50,51]
tef1-α	EF2	GGA RGT ACC AGT SAT CAT GTT		[50,51]
tub2	Bt-2a	GGT AAC CAA ATC GGT GCT GCT TTC	95 °C 5 min; (95 °C: 30 s, 53 °C: 30 s, 72 °C: 1 min) × 35 cycles	[52]
tub2	Bt-2b	ACC CTC AGT GTA GTG ACC CTT GGC		[52]

Pairwise homoplasy index (PHI) test

PHI tests were conducted on concatenated multilocus datasets using SplitsTree software, with Φw values <0.05, indicating significant recombination^[53,54]. Phylogenetic relationships were visualized through split graphs generated via Log-Det transformation and split decomposition methods.

Sequence alignments and phylogenetic analysis
Species identity was confirmed for all assembled sequences through BLAST searches against the GenBank database^[55]. All sequences were aligned using MAFFT v.7 online service (https://mafft.cbrc.jp/alignment/server/) and then adjusted by MEGA v7.0^[56]. We deposited the nucleotide sequences in the NCBI’s GenBank nucleotide database (www.ncbi.nlm.nih.gov). The newly-generated sequences (Supplementary Information 2) were aligned with related sequences (Supplementary Information 3–8). The phylogenetic trees were generated using maximum likelihood (RAxML-HPC2 on XSEDE v.8.2.12) and Bayesian inference (MrBayes v.3.2.7a, Linux)^[57]. In the phylogenetic trees, all tested isolates are highlighted in red. The phylogenetic tree visualizations were generated using ITOL (Interactive Tree of Life: https://itol.embl.de)^[58] and final layout adjustments made in Adobe Illustrator CS6 (Adobe Systems Inc., USA).

Supplementary Information 1 Details of known species.
Supplementary Information 2 Names, strains and corresponding GenBank accession numbers of all the taxa are listed in this study.
Supplementary Information 3 GenBank accession numbers used for the Apiospora phylogenetic reconstruction.
Supplementary Information 4 GenBank accession numbers used for the Nigrospora phylogenetic reconstruction.
Supplementary Information 5 GenBank accession numbers used for the Beltraniella phylogenetic reconstruction.
Supplementary Information 6 GenBank accession numbers used for the Neopestalotiopsis phylogenetic reconstruction.
Supplementary Information 7 GenBank accession numbers used for the Pestalotiopsis phylogenetic reconstruction.
Supplementary Information 8 GenBank accession numbers used for the Pseudopestalotiopsis phylogenetic reconstruction.
Supplementary Information 9 Dichotomous key for Amphisphaeriales.
Supplementary Information 10 Single-locus phylograms of the genus Apiospora based on alignments of the ITS, LSU, tef1-α, and tub2 regions, respectively.
Supplementary Information 11 Asexual morphological characters in selected Apiospora species.
Supplementary Information 12 Phylogram of the genus Neopestalotiopsis based on a concatenated ITS, tef1-α, and tub2 sequence alignment, with Pestalotiopsis colombiensis and P. diversiseta serving as outgroups.
Supplementary Information 13 Single-locus phylograms of the genus Pestalotiopsis based on alignments of the ITS, tef1-α, and tub2 regions, respectively.
Supplementary Information 14 The phi test for Pestalotiopsis yuxiensis (p = 0.5).
Supplementary Information 15 Apiospora isolates obtained from hosts in China.
Supplementary Information 16 Record of microbial co-infection from individual plant leaves.

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Phylum: Ascomycota
Subphylum: Pezizomycotina
Class: Sordariomycetes O.E. Erikss. & Winka
Order: Amphisphaeriales D. Hawksw. & O.E. Erikss.
Family: Apiosporaceae K.D. Hyde, J. Fröhl., Joanne E. Taylor & M.E. Barr
Genus: Apiospora Sacc.
1.	Apiospora adinandrae X.Y. Liu, Z.X. Zhang and X.G. Zhang, Journal of Fungi 10 (1, no. 74): 6 (2024) (new host record)
2.	Apiospora alpiniae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
3.	Apiospora arecacearum L.S. Dissan., K.D. Hyde & J.C. Kang, Mycosphere 15 (1): 1675–1793 (2024) (new host and geographical records)
4.	Apiospora babylonica H. Sheng, Z.X. Zhang & X.G. Zhang, Microorganisms 12: 1372 (2024) (new host and geographical records)
5.	Apiospora baihualingensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
6.	Apiospora bambusicaulis K.M. Yu & Y.L. Jiang, MycoKeys 112: 233–252 (2025) (new geographical record)
7.	Apiospora baotingensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
8.	Apiospora yinggelingensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
9.	Apiospora campsidis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
10.	Apiospora changjiangensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
11.	Apiospora chiangraiense X.G. Tian & Tibpromma S, Life 11: 1071 (2021) (new host and geographical records)
12.	Apiospora clavata Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
13.	Apiospora danzhouensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
14.	Apiospora dematiacea C.F. Liao & Doilom, Journal of Fungi 9 (no. 1087): 10 (2023) (new host and geographical records)
15.	Apiospora diaoluoshanensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
16.	Apiospora dryopteris Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
17.	Apiospora elongata Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
18.	Apiospora guangdongensis J.Y. Zhang & Y.Z. Lu, Journal of Fungi 9 (no. 1096): 12 (2023) (new geographical record)
19.	Apiospora guangzhouensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
20.	Apiospora hainanensis R.Y. Liu, J.W. Xia & X.G. Zhang, MycoKeys 95: 37 (2023)
21.	Apiospora hydei (Crous) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 207 (2021)
22.	Apiospora hysterina (Sacc.) Pintos & Alvarado, Fungal Systematics and Evolution 7:206P. (2021)
23.	Apiospora intestini (Kajale, Sonawane & Rohit Sharma) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 206 (2021) (new host and geographical records)
24.	Apiospora pseudosasae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
25.	Apiospora jiangxiensis (Mei Wang & L. Cai) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 206 (2021) (new geographical record)
26.	Apiospora lageniformis S.L. Kwon & J.J. Kim, Mycobiology 50 (5): 302–316 (2022) (new host and geographical records)
27.	Apiospora armeniaca H. Sheng, Z.X. Zhang & X.G. Zhang, Microorganisms 2024, 12, 1372 (new host and geographical records)
28.	Apiospora lingshuiensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
29.	Apiospora locuta-pollinis (F. Liu & L. Cai) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 207 (2021) (new host record)
30.	Apiospora longistroma (D.Q. Dai & K.D. Hyde) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 207 (2021) (new geographical record)
31.	Apiospora multiplex Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
32.	Apiospora pingtangensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
33.	Apiospora pseudoparenchymatica (M. Wang & L. Cai) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 207 (2021) (new geographical record)
34.	Apiospora cyclobalanopsidis (Yao Feng & Jian K. Liu) X.G. Tian & Tibpromma, in Tian, Karunarathna, Mapook, Promputtha, Xu, Bao & Tibpromma, Life 11 (no. 1071): 17 (2021) (new host and geographical records)
35.	Apiospora qiongzhongensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
36.	Apiospora renhuaensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
37.	Apiospora rutila Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
38.	Apiospora shaoguanensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
39.	Apiospora taianensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
40.	Apiospora teapae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
41.	Apiospora ventricosa Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
42.	Apiospora wuyishanensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
43.	Apiospora xiangxiensis X.Y. Chang & M.J. Chen, MycoKeys 116: 214 (2025) (new host and geographical records)
44.	Apiospora zingiberis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
Genus: Nigrospora Zimm.
45.	Nigrospora bambusae Mei Wang & L. Cai, Persoonia 39: 127 (2017) (new geographical record)
46.	Nigrospora brasiliensis A.C.Q. Brito, C. Conforto, A.R. Machado, Persoonia 42: 439 (2019) (new host and geographical records)
47.	Nigrospora buddlejae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
48.	Nigrospora camelliae-sinensis Mei Wang & L. Cai, Persoonia 39: 127 (2017) (new host record)
49.	Nigrospora chinensis Mei Wang & L. Cai, Persoonia 39: 129 (2017) (new host record)
50.	Nigrospora helwingiae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
51.	Nigrospora lacticolonia Mei Wang & L. Cai, Persoonia 39: 06 (2017) (new host record)
52.	Nigrospora officinalis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
53.	Nigrospora oryzae (Berk. & Broome) Petch (1924) (new host record)
54.	Nigrospora penniseti Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
55.	Nigrospora pseudosasae Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
56.	Nigrospora pyriformis Mei Wang & L. Cai, Persoonia 39: 126 (2017) (new host record)
57.	Nigrospora rubi Mei Wang & L. Cai, Persoonia 39: 127 (2017) (new host and geographical records)
58.	Nigrospora sacchari-officinarum M. Raza & L. Cai, Fungal Diversity 99: 95 (2019) (new host record)
59.	Nigrospora vesicularifera M. Raza & L. Cai, Fungal Diversity 99: 96 (2019) (new host record)
Family: Beltraniaceae Nann.
Genus: Beltraniella Subram.
60.	Beltraniella obovata D.H. Li, J.W. Xia & X.G. Zhang, sp. nov.
Family: Sporocadaceae Corda
Genus: Neopestalotiopsis Maharachch., K.D. Hyde & Crous
61.	Neopestalotiopsis bambooensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
62.	Neopestalotiopsis mianyangensis W.L. Li & Jian K. Liu, Journal of Fungi 8: 10 (2022) (new host record)
Genus: Pestalotiopsis Steyaert
63.	Pestalotiopsis castanopsis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
64.	Pestalotiopsis dracaenae Yong Wang bis, Y. Song, K. Geng & K.D. Hyde, Fungal Diversity 75: 164 (2015) (new host record)
65.	Pestalotiopsis hydei Huanraluek & Jayaward, Phytotaxa 479 (1): 023–043 (2021) (new host and geographical records)
66.	Pestalotiopsis nanpingensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
67.	Pestalotiopsis rhizomaticola Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
68.	Pestalotiopsis rostrata Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
69.	Pestalotiopsis smilacicola Y.R. Sun & Yong Wang bis, Microbiology Spectrum 11(1): e03987-22, 15 (2023) (new host and geographical records)
70.	Pestalotiopsis yuxiensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.
Genus: Pseudopestalotiopsis Maharachch., K.D. Hyde & Crous
71.	Pseudopestalotiopsis xishuangbannaensis Q.Y. Liu, Z. Li & X.G. Zhang, sp. nov.

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Taxonomic analysis of Amphisphaeriales in China: discovery of multiple novel taxa

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