Zuo-Yi Liu's research while affiliated with Zhongkai University of Agriculture and Engineering and other places

This study documents the morphology and phylogeny of ascomycetes collected from karst landscapes of Guizhou Province, China. Based on morphological characteristics in conjunction with DNA sequence data, 70 species are identified and distributed in two classes (Dothideomycetes and Sordariomycetes), 16 orders, 41 families and 60 genera. One order Planisphaeriales, four families Leptosphaerioidaceae, Neoleptosporellaceae, Planisphaeriaceae and Profundisphaeriaceae, ten genera Conicosphaeria, Karstiomyces, Leptosphaerioides, Neoceratosphaeria, Neodiaporthe, Neodictyospora, Planisphaeria, Profundisphaeria, Stellatus and Truncatascus, and 34 species (Amphisphaeria karsti, Anteaglonium hydei, Atractospora terrestris, Conicosphaeria vaginatispora, Corylicola hydei, Diaporthe cylindriformispora, Dictyosporium karsti, Hysterobrevium karsti, Karstiomyces guizhouensis, Leptosphaerioides guizhouensis, Lophiotrema karsti, Murispora hydei, Muyocopron karsti, Neoaquastroma guizhouense, Neoceratosphaeria karsti, Neodiaporthe reniformispora, Neodictyospora karsti, Neoheleiosa guizhouensis, Neoleptosporella fusiformispora, Neoophiobolus filiformisporum, Ophioceras guizhouensis, Ophiosphaerella karsti, Paraeutypella longiasca, Paraeutypella karsti, Patellaria guizhouensis, Planisphaeria karsti, Planisphaeria reniformispora, Poaceascoma herbaceum, Profundisphaeria fusiformispora, Pseudocoleophoma guizhouensis, Pseudopolyplosphaeria guizhouensis, Stellatus guizhouensis, Sulcatispora karsti and Truncatascus microsporus) are introduced as new to science. Moreover, 13 new geographical records for China are also reported, which are Acrocalymma medicaginis, Annulohypoxylon thailandicum, Astrosphaeriella bambusae, Diaporthe novem, Hypoxylon rubiginosum, Ophiosphaerella agrostidis, Ophiosphaerella chiangraiensis, Patellaria atrata, Polyplosphaeria fusca, Psiloglonium macrosporum, Sarimanas shirakamiense, Thyridaria broussonetiae and Tremateia chromolaenae. Additionally, the family Eriomycetaceae was resurrected as a non-lichenized family and accommodated within Monoblastiales. Detailed descriptions and illustrations of all these taxa are provided.

A survey of the taxonomy and diversity of freshwater fungi from Karst Plateau wetlands in Guizhou Province, China, three fresh collections were collected from a freshwater river. They have cheiroid, digitate, dictyosporous conidia with one or two hyaline appendages. Phylogenetic analysis of the combined ITS, LSU and tef1-α sequence data support their placement with genera Dictyocheirospora and Dictyosporium (Dictyosporiaceae, Pleosporales). Two collections represent the same species, Dictyocheirospora rotunda. The other collection is introduced as a new species Dictyosporium duliujiangense. An updated phylogenetic backbone tree, descriptions and illustrations of the two taxa are provided for Dictyosporiaceae.

Freshwater fungi comprises a highly diverse group of organisms occurring in freshwater habitats throughout the world. During a survey of freshwater fungi on submerged wood in streams and lakes, a wide range of sexual and asexual species were collected mainly from karst regions in China and Thailand. Phylogenetic inferences using partial gene regions of LSU, ITS, SSU, TEF1α, and RPB2 sequences revealed that most of these fungi belonged to Dothideomycetes and Sordariomycetes and a few were related to Eurotiomycetes. Based on the morphology and multi-gene phylogeny, we introduce four new genera, viz. Aquabispora, Neocirrenalia, Ocellisimilis and Uvarisporella, and 47 new species, viz. Acrodictys chishuiensis, A. effusa, A. pyriformis, Actinocladium aquaticum, Annulatascus tratensis, Aquabispora setosa, Aqualignicola setosa, Aquimassariosphaeria vermiformis, Ceratosphaeria flava, Chaetosphaeria polygonalis, Conlarium muriforme, Digitodesmium chishuiense, Ellisembia aquirostrata, Fuscosporella atrobrunnea, Halobyssothecium aquifusiforme, H. caohaiense, Hongkongmyces aquisetosus, Kirschsteiniothelia dushanensis, Monilochaetes alsophilae, Mycoenterolobium macrosporum, Myrmecridium splendidum, Neohelicascus griseoflavus, Neohelicomyces denticulatus, Neohelicosporium fluviatile, Neokalmusia aquibrunnea, Neomassariosphaeria aquimucosa, Neomyrmecridium naviculare, Neospadicoides biseptata, Ocellisimilis clavata, Ophioceras thailandense, Paragaeumannomyces aquaticus, Phialoturbella aquilunata, Pleurohelicosporium hyalinum, Pseudodactylaria denticulata, P. longidenticulata, P. uniseptata, Pseudohalonectria aurantiaca, Rhamphoriopsis aquimicrospora, Setoseptoria bambusae, Shrungabeeja fluviatilis, Sporidesmium tratense, S. versicolor, Sporoschisma atroviride, Stanjehughesia aquatica, Thysanorea amniculi, Uvarisporella aquatica and Xylolentia aseptata, with an illustrated account, discussion of their taxonomic placement and comparison with morphological similar taxa. Seven new combinations are introduced, viz. Aquabispora grandispora (≡ Boerlagiomyces grandisporus), A. websteri (≡ Boerlagiomyces websteri), Ceratosphaeria suthepensis (≡ Pseudohalonectria suthepensis), Gamsomyces aquaticus (≡ Pseudobactrodesmium aquaticum), G. malabaricus (≡ Gangliostilbe malabarica), Neocirrenalia nigrospora (≡ Cirrenalia nigrospora), and Rhamphoriopsis glauca (≡ Chloridium glaucum). Ten new geographical records are reported in China and Thailand and nine species are first reported from freshwater habitats. Reference specimens are provided for Diplocladiella scalaroides and Neocirrenalia nigrospora (≡ Cirrenalia nigrospora). Systematic placement of the previously introduced genera Actinocladium, Aqualignicola, and Diplocladiella is first elucidated based on the reference specimens and new collections. Species recollected from China and Thailand are also described and illustrated. The overall trees of freshwater Dothideomycetes and Sordariomycetes collected in this study are provided respectively and genera or family/order trees are constructed for selected taxa.

Photinia × fraseri is a well-known evergreeen ornamental tree. Owing to its flower-like red leaves and its ability to tolerate stressful environments, P. fraseri is widely cultured as a fast-growing hedge in southern China. From July to September in 2021, a disease with symptoms similar to leaf spot was extensively observed on P. fraseri in Daozhen county (28° 51 ’N, 107° 57 ’E), Zunyi, Guizhou province, China. About 500 plants were surveyed and the incidence of leaf spot on P. fraseri leaves was 35% to 70%, significantly reducing the ornamental and economic value. The symptomatic leaves displayed irregular, watery dark brown lesions with black conidiomata in gray centers, and 10 symptomatic leaves were collected from 10 trees. After surface sterilization (0.5 min in 75% ethanol and 2 min in 3% NaOCl, washed three times with sterilized distilled water) (Fang 2007), small pieces of symptomatic leaf tissue (0.2 × 0.2 cm) were plated on potato dextrose agar (PDA) and incubated at 25°C for about 7 days. Three single-spore isolates, GZAAS 21-0327, GZAAS 21-0328 and GZAAS 21-0329, were obtained, which were identical in morphology and molecular analysis. Therefore, the representative isolate GZAAS 21-0328 was used for further study. The pathogenicity of GZAAS 21-0328 was tested through a pot assay. Ten healthy plants were scratched with a sterilized needle on the leaves. Plants were inoculated by spraying a spore suspension (106 spores mL−1) of GZAAS 21-0328 onto leaves until runoff, and the control leaves sprayed with sterile water. The plants were maintained at 28°C with high relative humidity (95%) in a growth chamber. The pathogenicity test was carried out three times (Fang 2007). The symptoms developed on all inoculated leaves but not on the control leaves. The lesions were first visible 72 h after inoculation, and typical lesions similar to those observed on field plants appeared after 15 days. The same fungus was reisolated and identified based on the morphological characterization and molecular analyses (ITS, TUB and TEF) from the infected leaves but not from the noninoculated leaves. Results of pathogenicity experiments of isolated fungi fulfilled Koch’s postulates. Fungal colonies on PDA were villiform, creamy-white and sparse aerial mycelium on the surface with black, gregarious conidiomata. The conidia were fusoid, ellipsoid, straight to slightly curved, 4-septate, septa darker than the rest of the cell, and 23.0 (21.0 to 27.0) × 6.0 (5.0 to 7.0) µm (n=50). The morphological features were consistent with the descriptions of Neopestalotiopsis asiatica Maharachch. & K.D. Hyde (Maharachchikumbura et al. 2012; Maharachchikumbura et al. 2014; Farr et al. 2022). The pathogen was confirmed to be N. asiatica by amplification and sequencing of the internal transcribed spacer region (ITS), the partial β-tubulin (TUB) and partial translation elongation factor 1-alpha (TEF) genes using primers ITS4/ITS5, T1/Bt-2b and EF1-728F/EF-2, respectively (Maharachchikumbura et al. 2014). The sequences of PCR products were deposited in GenBank with accession numbers OK563071 (ITS), OK584020 (TUB) and OK663023 (TEF). BLAST searches of the obtained sequences revealed 100% (482/482 nucleotides), 99.05% (419/421 nucleotides), and 99.33% (891/897 nucleotides) homology with those of N. asiatica in GenBank (JX398983, JX399018 and JX399049, respectively). Phylogenetic analysis (MEGA 6.0) using the maximum likelihood method placed the isolate GZAAS 21-0328 in a well-supported cluster with N. asiatica. The pathogen was thus identified as N. asiatica based on the morphological characterization and molecular analyses. To our knowledge, this is the first report of leaf spot on P. fraseri caused by N. asiatica in China. This study provides valuable information for the identification and control of the leaf spot on Photinia × fraseri.

During an investigation of fungi on bamboos in Guizhou Province of China, a new ascomycete inhabiting decaying bamboo is introduced based on morpho-molecular evidence. Morphologically, our new species is characterized by hemispherical ascomata with slit-like ostiole, trabeculate pseudoparaphyses, obclavate to cylindrical asci, and fusiform, hyaline, 1-septate ascospores, surrounded by a distinctive thin sheath, which fit well with Aigialaceae. The phylogenetic analyses based on a combined LSU, ITS, SSU, and TEF1-α sequence data showed that the new collections are affiliated with the genus Fissuroma and can be recognized as a new species Fissuroma bambusicola. The comprehensive description and notes of F. bambusicola are provided.

An asexual fungus, collected from diseased leaves of Paphiopedilum sp. from Guizhou Province, China, and based on the phylogenetic analyses and morphological characters, it was identified as a new species in Curvicladiella . The genus Curvicladiella are recorded for the first time for China. The morphology of Curvicladiella paphiopedili sp. nov. is characterised by penicillate conidiophores with a stipe, dull, tapering towards the apex, the curved stipe extension and cylindrical conidia. In the phylogenetic analyses of combined cmdA, his3, ITS, LSU, tef1 and tub2 sequence data, this taxon was clustered as sister to Curvicladiella cignea within Nectriaceae.

Paphiopedilum is known as “slipper orchids”, which has a high ornamental value and can be used as household bonsai and garden plants. Paphiopedilum is also one of the most beautiful plants in the world due to their exotic and unique flowers. An asexual fungus was collected from diseased leaves of Paphiopedilum sp. from Guizhou Province, China, is described and illustrated on the basis of morphological characters and molecular evidence. The morphologies of Curvicladiella paphiopedili sp. nov. were characterized by penicillate conidiophores with a stipe, dull, tapering towards the apex and the curved stipe extension, cylindrical conidia. In the phylogenetic analyses of combined LSU, cmdA, his3, ITS, tef1 and tub2 sequence data, this taxon was clustered as sister to Curvicladiella cignea within Nectriaceae.

Distoseptispora is one of the sporidesmium-like taxa with great variation in asexual morphology and delineation of species. Phylogenetic analyses of four gene regions LSU, ITS, TEF1α, and RPB2 revealed the placement of several sporidesmium-like species in Distoseptispora (Distoseptisporaceae, Distoseptisporales, Sordariomycetes), collected on submerged decaying twigs from streams in China and Thailand. Based on morphological examination and molecular DNA data, six new species, Distoseptispora amniculi, D. atroviridis, D. effusa, D. fusiformis, D. hyalina, and D. verrucosa, are proposed. Among them, D. hyalina is the first sexual morph confirmed in the genus. A new geographical record is reported for D. lignicola in China. Conidial length proved to be of less taxonomic significance for some Distoseptispora species, whereas the type of conidial septa is informative at species level.

Chinese rose (Rosa chinensis Jacq.) is cultivated for edible flowers in southwestern China (Zhang et al. 2014). In March 2020, a leaf spot disease was observed on about 3-5% leaves of Chinese rose cultivar ‘Mohong’ in Guizhou Botanical Garden (26°37' 45'' N, 106°43' 10'' E), Guiyang, Guizhou province, China. The symptomatic plants displayed circular, dark brown lesions with black conidiomata in grey centers on leaves, and leaf samples were collected. After surface sterilization (0.5 min in 75% ethanol and 2 min in 3% NaOCl, washed 3 times with sterilized distilled water) (Fang 2007), small pieces of symptomatic leaf tissue (0.3 × 0.3 cm) were plated on potato dextrose agar (PDA) and incubated at 28oC for about 7 days. Two single-spore isolates, GZUMH01 and GZUMH02, were obtained, which were identical in morphology and molecular analysis. Therefore, the representative isolate GZUMH01 was used for further study. The pathogenicity of GZUMH01 was tested through a pot assay. Ten healthy plants were scratched with a sterilized needle on the leaves. Plants were inoculated by spraying a spore suspension (106 spores ml-1) onto leaves until runoff, and the control leaves sprayed with sterile water. The plants were maintained at 25°C with high relative humidity (90 to 95%) in a growth chamber. The pathogenicity test was carried out three times using the method described in Fang (2007). The symptoms developed on all inoculated leaves but not on the control leaves. The lesions were first visible 48 h after inoculation, and typical lesions similar to those observed on field plants after 7 days. The same fungus was re-isolated from the infected leaves but not from the non-inoculated leaves, fulfilling Koch’s postulates. Fungal colonies on PDA were villiform and greyish. The conidia were abundant, oval-ellipsoid, aseptate, 15.8 (13.7 to 18.8) × 5.7 (4.3 to 6.8) µm. The fungal colonies, hyphae, and conidia were consistent with the descriptions of Colletotrichum boninense Moriwaki, Toy. Sato & Tsukib. (Damm et al. 2012; Moriwaki et al. 2003). The pathogen was confirmed to be C. boninense by amplification and sequencing of the internal transcribed spacer region (ITS), the glyceraldehyde-3-phosphate dehydrogenase (GADPH), actin (ACT), and chitin synthase 1 (CHS-1) genes using primers ITS1/ITS4, GDF1/GDR1, ACT512F/ACT783R, and CHS-79F/CHS-345R, respectively (Damm et al. 2012; Moriwaki et al. 2003). The sequences of the PCR products were deposited in GenBank with accession numbers MT845879 (ITS), MT861006 (GADPH), MT861007 (ACT), and MT861008 (CHS-1). BLAST searches of the obtained sequences of the ITS, GADPH, ACT, and CHS-1 genes revealed 100% (554/554 nucleotides), 100% (245/245 nucleotides), 97.43% (265/272 nucleotides), and 99.64% (279/280 nucleotides) homology with those of C. boninense in GenBank (JQ005160, JQ005247, JQ005508, and JQ005334, respectively). Phylogenetic analysis (MEGA 6.0) using the maximum likelihood method placed the isolate GZUMH01 in a well-supported cluster with C. boninense. The pathogen was thus identified as C. boninense based on its morphological and molecular characteristics. To our knowledge, this is the first report of the anthracnose disease on R. chinensis caused by C. boninense in the world.

Arthrinium has a widespread distribution occurring in various substrates (e.g., air, soil debris, plants, lichens, marine algae and even human tissues). It is characterized by the basauxic conidiogenesis in the asexual morph, with apiospores in the sexual morph. In this study, seventeen isolates of Arthrinium were collected in China. Based on their morphology and phylogenetic characterization, four new species (A. biseriale, A. cyclobalanopsidis, A. gelatinosum, and A. septatum) are described and seven known species (A. arundinis, A. garethjonesii, A. guizhouense, A. hydei, A. neosubglobosa, A. phyllostachium and A. psedoparenchymaticum) are identified, of which the sexual morph of three species (A. guizhouense, A. phyllostachium and A. psedoparenchymaticum) and asexual morph of A. garethjonesii are reported for the first time. The detailed descriptions, illustrations and comparisons with related taxa of these new collections are provided. Phylogenetic analyses of combined ITS, LSU, TUB2, and TEF sequence data support their placements in the genus Arthrinium and justify the new species establishments and identifications of known species.