Morphological and Phylogenetic Analyses Reveal Three New Species of Didymella (Didymellaceae, Pleosporales) from Jiangxi, China.

Didymella contains numerous plant pathogenic and saprobic species associated with a wide range of hosts. Over the course of our mycological surveys of plant pathogens from terrestrial plants in Jiangxi Province, China, eight strains isolated from diseased leaves of four host genera represented three new species of Didymella, D. bischofiae sp. nov., D. clerodendri sp. nov., and D. pittospori sp. nov. Phylogenetic analyses of combined ITS, LSU, RPB2, and TUB2 sequence data, using maximum-likelihood (ML) and Bayesian inference (BI), revealed their taxonomic placement within Didymella. Both morphological examinations and molecular phylogenetic analyses supported D. bischofiae, D. clerodendri, and D. pittospori as three new taxa within Didymella. Illustrations and descriptions of these three taxa were provided, along with comparisons with closely related taxa in the genus.

Three New Species of Microdochium (Microdochiaceae, Xylariales) on Bambusaceae sp. and Saprophytic Leaves from Hainan and Yunnan, China.

Species of the genus Microdochium (Microdochiaceae, Xylariales) have been reported from the whole world and separated from multiple plant hosts. The primary aim of the present study is to describe and illustrate three new species isolated from the leaf spot of Bambusaceae sp. and saprophytic leaves in Hainan and Yunnan provinces, China. The proposed three species, viz., Microdochium bambusae, M. nannuoshanense and M. phyllosaprophyticum, are based on multi-locus phylogenies from a combined dataset of ITS rDNA, LSU, RPB2 and TUB2 in conjunction with morphological characteristics. Descriptions and illustrations of three new species in the genus are provided.

Multi-locus phylogenetic analyses reveal eight novel species of Distoseptispora from southern China.

IMPORTANCE: Distoseptispora as a single genus in Distoseptisporaceae was introduced by morphological and phylogenetic analyses. Members of this genus occur mainly as asexual morphs, forming effuse, hairy colonies on decaying wood, plant stems, bamboo culms, and fallen leaves and shafts in terrestrial and freshwater habitats. In the present study, saprobic hyphomycetes from plant debris were investigated, and eight new Distoseptispora species were introduced based on morphology and phylogenetic analyses of LSU, ITS, TEF1, and RPB2 sequence data. This study provides important data on the species diversity, ecological environment, and geographical area of Distoseptispora, greatly updates the classification of Distoseptispora, and improves our understanding of the taxonomy of Distoseptispora.

First Report of Muskmelon Fruit Rot Caused by Fusarium sulawesiense in China.

Muskmelon (Cucumis melo L.) is a widely cultivated and economically important fruit crop worldwide. In June 2022, fruit rot symptoms were observed on ripening muskmelons (cv. Boyang) in Shouguang City (36.81°N 118.90°E) of China. To determine the causal agent, we surveyed 200 muskmelon plants in about 1000 m2 of planting area and collected diseased muskmelons. Approximately 20% of muskmelon fruits had symptoms, and yield loss averaged 20%. Water-soaked lesions were observed on the surface and the fruit rotted from inside. Lesions were covered with white mycelium. Rotted fruit were surface-disinfested with 1% NaOCl for 1 min, 75% ethanol for 30 s, and washed three times with sterile water. Pieces (1 cm3) were cut from the disinfested fruit, placed on potato dextrose agar (PDA), and incubated at 25°C for 1 week. Ten isolates with similar morphology were obtained and isolates SG66 and SG68 were selected for further characterization. Colonies maintained on PDA in the dark had an average radial growth rate of 10-12 mm/d at 25°C. Surface was white, velvety to felty mycelium. Reverse was white to pale wheat. Diffusible pigments were absent. On carnation leaf agar, sporodochia appeared as slimy dots, macroconidia were 3- to 5-septate, 20-35 × 3-5 µm, falcate, with a pronounced dorsiventral curvature, with blunt to papillate apical cell, and barely to distinctly notched basal cell. Microconidia and chlamydospores were not observed. These morphological characteristics were consistent with descriptions of Fusarium sp. DNA was extracted from isolates SG66 and SG68 using a CTAB method. Nucleotide sequences of the internal transcribed spacers (ITS) (White et al. 1990), calmodulin (CAM), RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1-α gene (TEF1) (Xia et al. 2019) were amplified using generic primers, the products sequenced, and sequences deposited in GenBank (ITS: OP251362, OP251363; CAM: OP266024, OP266025; RPB2: OP266028, OP266029; TEF1: OP266026, OP266027). Isolates SG66 and SG68 clustered with Fusarium sulawesiense (85% bootstrap) (Maryani et al. 2019). The Fusarioid-ID database pairwise alignment of ITS (526 bp), CAM (534 bp), RPB2 (861 bp), and TEF1 (636 bp) sequences from isolate SG66 showed 99.6% (98.9% coverage), 100% (100% coverage), 100% (100% coverage) and 100% (98.4% coverage) similarity with the corresponding sequences (GQ505730, LS479422, LS479855 and GQ505641), respectively, of the reference strains of F. sulawesiense (InaCC F940 and NRRL 34059). To perform a pathogenicity test, 10 µl of conidial suspensions (1 × 106 conidia/ml) were injected into ten muskmelon fruit using a syringe, and ten control fruit were inoculated with 10 µl of sterile distilled water. The test was repeated three times. After 7 days at 25°C, the pulp of all inoculated muskmelons began to rot, and the lesion expanded from the inside to the fruit surface at the injection site and became covered with white mycelia. No symptoms developed on the control fruit. The fungus was successfully re-isolated from infected tissues and confirmed as F. sulawesiense by morphological and phylogenetic analyses. F. sulawesiense has previously been reported on yellow melon (Canary) in Brazil (Lima et al. 2021) and on a range of hosts, including Luffa aegyptiaca, in China (Wang et al. 2019). To our knowledge, this is the first report of muskmelon fruit rot caused by F. sulawesiense in China.

Morphological and Phylogenetic Analyses Reveal Three New Species of Pestalotiopsis (Sporocadaceae, Amphisphaeriales) from Hainan, China.

Species of Pestalotiopsis were mainly introduced as endophytes, plant pathogens or saprobes from various hosts. In this study, ten strains were isolated from Ficus macrocarpa, Phoebe zhennan and Spatholobus suberectus in China. Based on multilocus phylogenies from the internal transcribed spacer (ITS), the partial translation elongation factor 1-alpha gene (tef1α) and the partial beta-tubulin gene (tub2), in conjunction with morphological characteristics, we describe three new species, viz., Pestalotiopsis ficicola sp. nov., P. phoebes sp. nov. and P. spatholobi sp. nov.

Morphological and phylogenetic analyses reveal two new species and a new record of Apiospora (Amphisphaeriales, Apiosporaceae) in China.

The genus Apiospora includes endophytes, pathogens and saprobes, with a wide host range and geographic distribution. In this paper, six Apiospora strains isolated from diseased and healthy tissues of bamboo leaves from Hainan and Shandong provinces in China were classified using a multi-locus phylogeny based on a combined dataset of ITS, LSU, tef1 and tub2, in conjunction with morphological characters, host association and ecological distribution. Two new species, Apiosporadongyingensis and A.hainanensis, and a new record of A.pseudosinensis in China, are described based on their distinct phylogenetic relationships and morphological analyses. Illustrations and descriptions of the three taxa are provided, along with comparisons with closely related taxa in the genus.

Morphological and Phylogenetic Analyses Reveal Three New Species of Distoseptispora (Distoseptisporaceae, Distoseptisporales) from Yunnan, China.

Three new species of Distoseptispora, viz. D. mengsongensis, D. nabanheensis, and D. sinensis, are described and illustrated from specimens collected on dead branches of unidentified plants in Yunnan Province, China. Phylogenetic analyses of LSU, ITS, and TEF1 sequence data, using maximum-likelihood (ML) and Bayesian inference (BI), reveal the taxonomic placement of D. mengsongensis, D. nabanheensis, and D. sinensis within Distoseptispora. Both morphological observations and molecular phylogenetic analyses supported D. mengsongensis, D. nabanheensis, and D. sinensis as three new taxa. To extend our knowledge of the diversity of Distoseptispora-like taxa, a list of recognized species of Distoseptispora with major morphological features, habitat, host, and locality is also provided.

Taxonomy, Phylogeny, Divergence Time Estimation, and Biogeography of the Family Pseudoplagiostomataceae (Ascomycota, Diaporthales).

Species of Pseudoplagiostomataceae were mainly introduced as endophytes, plant pathogens, or saprobes from various hosts. Based on multi-locus phylogenies from the internal transcribed spacers (ITS), the large subunit of nuclear ribosomal RNA gene (LSU), partial DNA-directed RNA polymerase II subunit two gene (rpb2), the partial translation elongation factor 1-alpha gene (tef1α), and the partial beta-tubulin gene (tub2), in conjunction with morphological characteristics, we describe three new species, viz. Pseudoplagiostoma alsophilae sp. nov., P. bambusae sp. nov., and P. machili sp. nov. Molecular clock analyses on the divergence times of Pseudoplagiostomataceae indicated that the conjoint ancestor of Pseudoplagiostomataceae and Apoharknessiaceae occurred in the Cretaceous period. and had a mean stem age of 104.1 Mya (95% HPD of 86.0-129.0 Mya, 1.0 PP), and most species emerged in the Paleogene and Neogene period. Historical biogeography was reconstructed for Pseudoplagiostomataceae by the RASP software with a S-DEC model, and suggested that Asia, specifically Southeast Asia, was probably the ancestral area.

Molecular Phylogeny and Morphology Reveal Four Novel Species of Corynespora and Kirschsteiniothelia (Dothideomycetes, Ascomycota) from China: A Checklist for Corynespora Reported Worldwide.

Plant debris are habitats favoring survival and multiplication of various microbial species. During continuing mycological surveys of saprobic microfungi from plant debris in Yunnan Province, China, several Corynespora-like and Dendryphiopsis-like isolates were collected from dead branches of unidentified perennial dicotyledonous plants. Four barcodes, i.e., ITS, LSU, SSU and tef1-α, were amplified and sequenced. Morphological studies and multigene phylogenetic analyses by maximum likelihood and Bayesian inference revealed three new Corynespora species (C. mengsongensis sp. nov., C. nabanheensis sp. nov. and C. yunnanensis sp. nov.) and a new Kirschsteiniothelia species (K. nabanheensis sp. nov.) within Dothideomycetes, Ascomycota. A list of identified and accepted species of Corynespora with major morphological features, host information and locality was compiled. This work improves the knowledge of species diversity of Corynespora and Kirschsteiniothelia in Yunnan Province, China.

Morphological and Phylogenetic Analyses Reveal Four New Species of Acrodictys (Acrodictyaceae) in China.

During our ongoing survey of dematiaceous hyphomycetes associated with dead branches in tropical forests, eight Acrodictys isolates were collected from Hainan, China. Morphology from the cultures and phylogeny based on partial small subunit (SSU), entire internal transcribed spacer regions with intervening 5.8S (ITS), partial large subunit (LSU) of rRNA gene, partial beta-tubulin (tub2), and partial RNA polymerase II second largest subunit (rpb2) genes were employed to identify these isolates. As a result, four new species, namely Acrodictys bawanglingensis sp. nov., A. diaoluoshanensis sp. nov., A. ellisii sp. nov., and A. pigmentosa sp. nov., are introduced. Illustrations and descriptions of the four taxa are provided, along with comparisons with closely related taxa in the genus. For facilitating relative studies, an updated key to all accepted species of this genus is also compiled.

Three New Species of Microdochium (Sordariomycetes, Amphisphaeriales) on Miscanthus sinensis and Phragmites australis from Hainan, China.

Species in Microdochium, potential agents of biocontrol, have often been reported as plant pathogens, occasionally as endophytes and fungicolous fungi. Combining multiple molecular markers (ITS rDNA, LSU rDNA, TUB2 and RPB2) with morphological characteristics, this study proposes three new species in the genus Microdochium represented by seven strains from the plant hosts Miscanthus sinensis and Phragmites australis in Hainan Island, China. These three species, Microdochium miscanthi sp. Nov., M. sinense sp. Nov. and M. hainanense sp. Nov., are described with MycoBank number, etymology, typification, morphological features and illustrations, as well as placement on molecular phylogenetic trees. Their affinity with morphologically allied and molecularly closely related species are also analyzed. For facilitating identification, an updated key to the species of Microdochium is provided herein.

Morphological and phylogenetic analyses reveal two new species of Sporocadaceae from Hainan, China.

Species of Sporocadaceae have often been reported as plant pathogens, endophytes or saprophytes and are commonly isolated from a wide range of plant hosts. The isolated fungi were studied through a complete examination, based on multilocus phylogenies from combined datasets of ITS/tub2/tef1, in conjunction with morphological characteristics. Nine strains were isolated from Ficusmicrocarpa, Ilexchinensis and Schimasuperba in China which represented four species, viz., Monochaetiaschimae sp. nov., Neopestalotiopsishaikouensis sp. nov., Neopestalotiopsispiceana and Pestalotiopsislicualicola. Neopestalotiopsispiceana was a new country record for China and first host record from Ficusmacrocarpa. Pestalotiopsislicualicola was first report from Ilexchinensis in China.

First Report of Muskmelon Fruit Rot Caused by Fusarium nanum in China.

Muskmelon (Cucumis melo L.) is one of the most widely cultivated and economically important fruit crops in the world. However, many pathogens can cause decay of muskmelon fruit, including Fusarium asiaticum, F. equiseti, F. incarnatum and F. lateritium (Hao et al. 2021; Wang et al. 2019). Fusarium spp. are the most important pathogens affecting muskmelon fruit yield and quality (Wang et al. 2011). In August 2020, fruit rot symptoms were observed on ripening muskmelons (cv. Tianbao) in several fields in Jiyang District, Jinan City of Shandong Province, China. The incidences of infected muskmelon ranged from 15% to 30% and caused an average 20% yield loss. Symptoms appeared as pale brown, water-soaked lesions that were irregular in shape, with the lesion sizes ranging from a small spot (1 to 2 cm) to decay of the entire fruit. The core and surface of infected fruit were colonized and covered with white mycelia. Two infected muskmelons were collected from two fields, 7 km apart. Tissues removed from inside the infected fruit were surface disinfected with 75% ethanol for 30 s, and cultured on potato dextrose agar (PDA) at 25°C in the dark for 5 days. Four purified cultures were obtained using the single spore method. On carnation leaf agar (CLA), macroconidia were 1 to 5 septate, falcate, with a pronounced dorsiventral curvature with blunt to papillate apical cell, and barely to distinctly notched basal cell, measuring 12 to 35 × 3.5 to 6 µm. Microconidia and chlamydospores were not observed. These morphological characteristics were consistent with the description of Fusarium sp. Because these isolates had similar morphology, two representative isolates (XP9 and XP10) were selected for multilocus phylogenetic analyses. DNA was extracted from the representative isolates using a CTAB method. Nucleotide sequences of the internal transcribed spacers (ITS) (White et al. 1990), calmodulin (CAM), RNA polymerase II second largest subunit (RPB2), translation elongation factor 1-α gene (TEF1) (Xia et al. 2019) were amplified using specific primers, sequenced, and deposited in GenBank (ITS: MW391507 and MW391508, CAM: MW392787 and MW392788, RPB2: MW392795 and MW392796, TEF1: MW392791 and MW392792). The Fusarium MLST database pairwise alignment of ITS (546 bp), CAM (628 bp), RPB2 (902 bp) and TEF1 (718 bp) sequences from isolate XP9 showed 99.63%, 99.33%, 100.00% and 99.71% similarity with the corresponding sequences (GQ505685, GQ505508, GQ505774 and GQ505596) of the reference strain of F. nanum (NRRL 22244), respectively. The overlap of ITS, CAM, RPB2 and TEF1 sequences from XP9 and NRRL 22244 were 100.00%, 95.06%, 97.45% and 94.99%, respectively. Alignments of a combined dataset of ITS, CAM, RPB2 and TEF1 were made using MAFFT v. 7, and phylogenetic analyses were conducted in MEGA v. 7.0 using the maximum likelihood method. The muskmelon isolates (XP9 and XP10) clustered together with the F. nanum reference strain CGMCC3.19498 and NRRL 22244 (100% bootstrap) (Wang et al., 2019). To perform a pathogenicity test, 10 µl of conidial suspensions (1 × 106 conidia/ml) were injected into each muskmelon fruit using a syringe, and the control fruit was inoculated with 10 µl of sterile distilled water. There were ten replicated fruits for each treatment. The test was repeated three times. After 7 days at 25°C, the interior of the inoculated muskmelons begun to rot, and the rot lesion expanded from the core towards the surface of the fruit, then white mycelia were produced on the surface. Ten isolations were re-isolated from the infected tissues and identified by morphological and phylogenetic analyses and confirmed to fulfill Koch's postulates. No symptoms were observed on the control muskmelons. To our knowledge, this is the first report of muskmelon fruit rot caused by F. nanum in China. Considering the economic value of the muskmelon crop, correct identification can help farmers select appropriate field management measures for control of this disease.

Morphological and phylogenetic analyses reveal a new genus and two new species of Tubakiaceae from China.

Species of Tubakiaceae have often been reported as plant pathogens or endophytes, commonly isolated from a wide range of plant hosts. The isolated fungi were studied through a complete examination, based on multilocus phylogenies from combined datasets of ITS/LSU/rpb2 and ITS/tef1/tub2, in conjunction with morphological characteristics. Five strains isolated from Lithocarpusfohaiensis and Quercuspalustris in China represented a new genus of Tubakiaceae, Obovoideisporodochium and three species, viz. Obovoideisporodochiumlithocarpi sp. nov., Tubakialushanensis sp. nov. and T.dryinoides.

Morphological and phylogenetic analyses reveal three new species of Diaporthe from Yunnan, China.

Species of Diaporthe have often been reported as plant pathogens, endophytes or saprobes, commonly isolated from a wide range of plant hosts. Sixteen strains isolated from species of ten host genera in Yunnan Province, China, represented three new species of Diaporthe, D. chrysalidocarpi, D. machili and D. pometiae as well as five known species D. arecae, D. hongkongensis, D. middletonii, D. osmanthi and D. pandanicola. Morphological comparisons with known species and DNA-based phylogenies based on the analysis of a multigene (ITS, TUB, TEF, CAL and HIS) dataset support the establishment of the new species. This study reveals that a high species diversity of Diaporthe with wide host ranges occur in tropical rainforest in Yunnan Province, China.

Morphological and molecular identification of Diaporthe species in south-western China, with description of eight new species.

Diaporthe species have often been reported as plant pathogens, endophytes and saprophytes, commonly isolated from a wide range of infected plant hosts. In the present study, twenty strains obtained from leaf spots of twelve host plants in Yunnan Province of China were isolated. Based on a combination of morphology, culture characteristics and multilocus sequence analysis of the rDNA internal transcribed spacer region (ITS), translation elongation factor 1-α (TEF), ß-tubulin (TUB), calmodulin (CAL), and histone (HIS) genes, these strains were identified as eight new species: Diaporthe camelliae-sinensis, D. grandiflori, D. heliconiae, D. heterostemmatis, D. litchii, D. lutescens, D. melastomatis, D. pungensis and two previously described species, D. subclavata and D. tectonendophytica. This study showed high species diversity of Diaporthe in tropical rain forests and its hosts in south-western China.

Morphological and molecular phylogenetic analyses reveal three species of Colletotrichum in Shandong province, China.

Colletotrichum has numerous host range and distribution. Its species are important plant pathogens, endophytes and saprobes. Colletotrichum can cause regular or irregular depressions and necrotic lesions in the epidermal tissues of plants. During this research Colletotrichum specimens were collected from Mengyin County, Shandong Province, China. A multi-locus phylogenetic analysis of ITS, GAPDH, CHS-1, ACT, TUB2, CAL and GS sequence data combined with morphology, revealed a new species and two known species, viz. C.mengyinense sp. nov., C.gloeosporioides and C.pandanicola, belonging to the C.gloeosporioides species complex. The new species is described and illustrated in this paper and compared with taxa in the C.gloeosporioides species complex.

Two new species of Microdochium from Indocalamus longiauritus in south-western China.

Microdochium species have often been reported as plant pathogens and saprophytes and are commonly isolated from some diseased plant hosts. The primary aim of the present study was to describe and illustrate two new Microdochium species isolated from the leaf spot of Indocalamus longiauritus in Yunnan Province, China, namely Microdochium yunnanense and M. indocalami, spp. nov., based on their morphology and multilocus phylogenetic analyses of the combined ITS, LSU, TUB2, and RPB2. DNA sequence data indicate that six strains represent three independent groups from related and similar species in Microdochium. Microdochium indocalami sp. nov. clustered with M. fisheri, M. lycopodinum, M. rhopalostylidis, and M. phragmitis. Microdochium yunnanense sp. nov. grouped with M. bolleyi. In addition, the strain SAUCC1017 is recorded as an unidentified species in Microdochium. Descriptions and illustrations of the new species in the genus and Microdochium sp. indet. are provided.