Morpho-Molecular Characterization Reveals a New Genus, Three Novel Species and Two New Host Records in Xylariomycetidae.

Xylariomycetidae comprises extremely diverse taxa that are widespread on decaying wood worldwide. An investigation of the diversity of microfungi on oil tree plantations in Sichuan Province was conducted during 2020-2021. Twelve saprobic taxa representing five species were identified as members of Amphisphaeriales and Xylariales through morphological comparisons. Phylogenetic analyses of combined ITS, LSU, rpb2, tub2 and tef1 sequence data indicated a distinct clade formed by three strains within Xylariomycetidae, unrelated to any currently recognized families. Thus, a novel anthostomella-like genus, Bicellulospora, is proposed and treated as Xylariales genera incertae sedis. Bicellulospora is characterized by dark brown to black, immersed, subglobose ascomata with a clypeus, cylindrical asci, and hyaline to yellowish brown, inequilaterally ellipsoidal ascospores with a large upper cell and a dwarf lower cell. Two new species of Amphisphaeria, namely A. oleae and A. verniciae, are introduced based on multi-gene phylogenetic analyses (ITS, LSU, rpb2 and tub2) coupled with morphological characteristics. Amphisphaeria micheliae and Endocalyx ptychospermatis are reported as new host records.

Morpho-Phylogenetic Evidence Reveals Novel Species and New Records of Botryosphaeriaceae in China and Thailand.

Species in the Botryosphaeriaceae are common plant pathogens, endophytes, and saprobes found on a variety of mainly woody hosts. Botryosphaeriaceae is a high-profile fungal family whose genera have been subjected to continuous revisions in recent years. Surveys conducted during 2019 and 2020 on several decaying woody hosts (from dead arial twigs, branches, stems, bark, and seed pods) in China and Thailand revealed a high diversity of Botryosphaeriaceae fungi. Identification of 16 Botryosphaeriaceae isolates was carried out based on both morphological characteristics and phylogenetic analyses of combined ITS, LSU, tef1-α, and tub2 sequence data. Four novel species (Dothiorella ovata, Do. rosacearum, Do. septata, and Lasiodiplodia delonicis) and seven previously known species (Botryosphaeria fujianensis, Diplodia mutila, Di. seriata, L. crassispora, L. mahajangana, Macrophomina euphorbiicola and Sphaeropsis eucalypticola) were identified while new hosts and geographical records were reported. This study indicates that the fungal family Botryosphaeriaceae seems to be common and widespread on a broad range of hosts in China and Thailand.

Botryosphaerialean fungi associated with woody oil plants cultivated in Sichuan Province, China.

Woody oil plants are important economic trees which are widely cultivated and distributed throughout China. Surveys conducted during 2020 and 2021 on several woody oil plantations from five regions of Sichuan Province, China, revealed a high diversity of Botryosphaerialean fungi. The identification of 50 botryosphaeriaceous isolates was carried out based on both morphology and multi-gene phylogenetic analysis of internal transcribed spacer region (ITS), translation elongation factor 1-alpha gene (tef1) and ß-tubulin gene (tub2). This allowed the identification of twelve previously known Botryosphaeriales species: Aplosporellaprunicola, A.ginkgonis, Barriopsistectonae, Botryosphaeriadothidea, Bo.fabicerciana, Diplodiamutila, Di.seriata, Dothiorellasarmentorum, Neofusicoccumparvum, Sardiniellaguizhouensis, Sphaeropsiscitrigena, and Sp.guizhouensis, and four novel species belonging to the genera Diplodia and Dothiorella, viz. Di.acerigena, Di.pistaciicola, Do.camelliae and Do.zanthoxyli. The dominant species isolated across the surveyed regions were Botryosphaeriadothidea, Sardiniellaguizhouensis and Diplodiamutila, representing 20%, 14% and 12% of the total isolates, respectively. In addition, most isolates were obtained from Pistaciachinensis (14 isolates), followed by Camelliaoleifera (10 isolates). The present study enhances the understanding of Botryosphaeriales species diversity on woody oil plants in Sichuan Province, China.

Taxonomic Novelties and New Records of Amanita Subgenus Amanitina from Thailand.

The Amanita subgenus Amanitina contains six sections, and the species diversity of this subgenus has still not been explored in Thailand. Twenty samples collected in 2019 and 2020, which had the morphological characteristics of the Amanita subgen. Amanitina, were observed in this study. Both the microscopical characteristics and multi-gene phylogenetic analyses of the ITS, nrLSU, RPB2, TEF1-α, and TUB gene regions revealed that the 20 samples represented nine species and dispersed into four sections. Remarkably, three taxa were different from any other currently known species. Here, we describe them as new to science, namely A. albifragilis, A. claristriata, and A. fulvisquamea. Moreover, we also recognized six interesting taxa, including four records that were new to Thailand, viz. A. cacaina, A. citrinoannulata, A. griseofarinosa, and A. neoovoidea, as well as two previously recorded species, A. caojizong and A. oberwinkleriana. Moreover, we provide the first RPB2 and TEF1-α gene sequences for A. cacaina. Detailed descriptions, illustrations as line drawings, and comparisons with related taxa are provided.

Additions to Bambusicolous Fungi of Savoryellaceae from Southwest China.

Asexual and sexual morphs of saprobic bambusicolous fungi were collected from freshwater and terrestrial habitats in Sichuan Province, China. Taxonomic identification of these fungi was carried out on the basis of morphological comparison, culture characteristics, and molecular phylogeny. Multi-gene phylogeny based on combined SSU, ITS, LSU, rpb2, and tef1α sequence data was performed to determine their phylogenetic placement, and the result showed that these fungi belong to Savoryellaceae. Morphologically, four asexual morphs are similar to Canalisporium and Dematiosporium, while a sexual morph well-fits to Savoryella. Three new species, Canalisporium sichuanense, Dematiosporium bambusicola, and Savoryella bambusicola are identified and described. Two new records, C. dehongense and D. aquaticum, were recovered from the bamboo hosts in terrestrial and freshwater habitats, respectively. In addition, the nomenclatural confusion of C. dehongense and C. thailandense is discussed.

Re-Evaluating Botryosphaeriales: Ancestral State Reconstructions of Selected Characters and Evolution of Nutritional Modes.

Botryosphaeriales (Dothideomycetes, Ascomycota) occur in a wide range of habitats as endophytes, saprobes, and pathogens. The order Botryosphaeriales has not been subjected to evaluation since 2019 by Phillips and co-authors using phylogenetic and evolutionary analyses. Subsequently, many studies introduced novel taxa into the order and revised several families separately. In addition, no ancestral character studies have been conducted for this order. Therefore, in this study, we re-evaluated the character evolution and taxonomic placements of Botryosphaeriales species based on ancestral character evolution, divergence time estimation, and phylogenetic relationships, including all the novel taxa that have been introduced so far. Maximum likelihood, maximum parsimony, and Bayesian inference analyses were conducted on a combined LSU and ITS sequence alignment. Ancestral state reconstruction was carried out for conidial colour, septation, and nutritional mode. Divergence times estimates revealed that Botryosphaeriales originated around 109 Mya in the early epoch of the Cretaceous period. All six families in Botryosphaeriales evolved in the late epoch of the Cretaceous period (66-100 Mya), during which Angiosperms also appeared, rapidly diversified and became dominant on land. Families of Botryosphaeriales diversified during the Paleogene and Neogene periods in the Cenozoic era. The order comprises the families Aplosporellaceae, Botryosphaeriaceae, Melanopsaceae, Phyllostictaceae, Planistromellaceae and Saccharataceae. Furthermore, current study assessed two hypotheses; the first one being "All Botryosphaeriales species originated as endophytes and then switched into saprobes when their hosts died or into pathogens when their hosts were under stress"; the second hypothesis states that "There is a link between the conidial colour and nutritional mode in botryosphaerialean taxa". Ancestral state reconstruction and nutritional mode analyses revealed a pathogenic/saprobic nutritional mode as the ancestral character. However, we could not provide strong evidence for the first hypothesis mainly due to the significantly low number of studies reporting the endophytic botryosphaerialean taxa. Results also showed that hyaline and aseptate conidia were ancestral characters in Botryosphaeriales and supported the relationship between conidial pigmentation and the pathogenicity of Botryosphaeriales species.

Morpho-Phylogenetic Evidence Reveals New Species of Fuscosporellaceae and Savoryellaceae from Freshwater Habitats in Guizhou Province, China.

During a survey of freshwater fungi in Guizhou Province, China, six hyphomycetous collections were founded on decaying wood from freshwater habitats. These taxa were characterized and identified based on morphology, phylogeny, and culture characteristics. Phylogenetic analysis of combined LSU, SSU, ITS, RPB2 and TEF1α sequence data indicated that our six isolates formed three distinct lineages and were distributed within Fuscosporellaceae and Savoryellaceae. They can be organized as three new species: Fuscosporella guizhouensis, Mucisporaaquatica and Neoascotaiwaniaguizhouensis. Fuscosporella guizhouensis and Neoascotaiwania guizhouensis have sporodochial conidiomata, micronematous conidiophores and dark brown conidia. The former possesses irregularly ellipsoidal conidia with apical appendages, while the latter has fusiform to obovoid conidia. Mucispora aquatica is characterized by macronematous conidiophores, elongating percurrently and dark brown, narrowly obovoid conidia. The detailed, illustrated descriptions and notes for each new taxon are provided, and the species of Fuscosporella is reported for the first time in China.

Identification and Pathogenicity of Pestalotiod Fungi Associated with Woody Oil Plants in Sichuan Province, China.

Pestalotiod fungi are associated with a wide variety of plants worldwide and occur as endophytes, pathogens, and saprobes. The present study provides an updated phylogeny for genera Neopestalotiopsis, Pestalotiopsis, and Seiridium using fresh collections from woody oil plants (Camellia oleifera, Olea europaea, Paeonia suffruticosa, Sapium sebiferum, and Vernicia fordii) in Sichuan Province, China. We coupled morphology and combined sequence data analyses of ITS, tub2, and tef1-α for Neopestalotiopsis and Pestalotiopsis, with ITS, LSU, tub2, tef1-α, and rpb2 for Seiridium. Three novel species of Neopestalotiopsis (N. mianyangensis, N. paeonia-suffruticosa, N. terricola) and three of Seiridium (S. guangyuanum, S. vernicola, S. oleae), were found. Three other species, Pestalotiopsis kenyana, Seiridium ceratosporum, and S. rosarum were identified and reported as new records. All isolated species are fully described and illustrated. Additionally, the sexual morph of Pestalotiopsis kenyana is described for the first time. Pathogenicity tests revealed that Neopestalotiopsis mianyangensis, N. paeonia-suffruticosa, N. terricola, Pestalotiopsis kenyana, Seiridium guangyuanum, S. vernicola, and S. oleae are pathogenic on detached olive leaves.

Morpho-Phylogenetic Evidence Reveals Novel Pleosporalean Taxa from Sichuan Province, China.

Pleosporales is the largest and most morphologically diverse order in Dothideomycetes, including a large proportion of saprobic fungi. During the investigation of microfungi from decaying wood in Sichuan Province, several novel fungal taxa of asexual and sexual morphs were collected, identified, and well-described. Phylogenetic analyses based on SSU, ITS, LSU, RPB2 and TEF1α gene sequences suggested that these new taxa were related to Pleosporales and distributed in five families, viz. Amorosiaceae, Bambusicolaceae, Lophiostomataceae, Occultibambusaceae and Tetraplosphaeriaceae. The morphological comparison and molecular phylogeny evidence justify the establishment of six new taxa, namely Bambusicola guttulata sp. nov., Flabellascoma sichuanense sp. nov., Neoangustimassarina sichuanensis gen. et sp. nov., Occultibambusa sichuanensis sp. nov. and Pseudotetraploa bambusicola sp. nov. Among them, Neoangustimassarina was introduced as the second sexual morph genus in Amorosiaceae; Bambusicola guttulata, O. sichuanensis and P. bambusicola were isolated from bamboos, which contributed to the diversity of bambusicolous fungi. The detailed, illustrated descriptions and notes for each new taxon are provided, as well as a brief note for each family. The potential richness of fungal diversity in Sichuan Province is also discussed.

Reassessment of Dyfrolomyces and Four New Species of Melomastia from Olive (Olea europaea) in Sichuan Province, China.

Pleurotremataceae species are saprobes on decaying wood in terrestrial, mangrove, and freshwater habitats. The generic boundary of the family has traditionally been based on morphology. All genera of Pleurotremataceae have a high degree of morphological overlap, of which the generic circumscription of Melomastia and Dyfrolomyces has not been well resolved. Thus, the delimitation of genera has always been challenging. Melomastia traditionally differs from Dyfrolomyces in having 2-septate, oblong, with obtuse-ends ascospores. These main characteristics have been used to distinguish Melomastia from Dyfrolomyces for a long time. However, the above characteristics sometimes overlap among Dyfrolomyces and Melomastia species. Based on the morphology and multigene phylogeny with newly obtained data, we synonymized Dyfrolomyces under Melomastia following up-to-date results. Four novel species (i.e., Melomastia fusispora, M. oleae, M. sichuanensis and M. winteri) collected from the dead branches of Olea europaea L. in Chengdu Olive Base, Sichuan Province in China are introduced based on detailed morphological characterization and phylogenetic analyses of sequences based on nuclear ribosomal (LSU and SSU) and protein-coding gene (tef1-α). The 11 new combinations proposed are Melomastia aquatica (=Dyfrolomyces aquaticus), M. chromolaenae (=D. chromolaenae), M. distoseptata (=D. distoseptatus), M. mangrovei (=D. mangrovei), M. marinospora (=D. marinosporus), M. neothailandica (=D. neothailandicus), M. phetchaburiensis (=D. phetchaburiensis), M. sinensis (=D. sinensis), M. thailandica (=D. thailandica), M. thamplaensis (=D. thamplaensis) and M. tiomanensis (=D. tiomanensis).

Occurrence and Morpho-Molecular Identification of Botryosphaeriales Species from Guizhou Province, China.

Botryosphaeriales is an important order of diverse fungal pathogens, saprobes, and endophytes distributed worldwide. Recent studies of Botryosphaeriales in China have discovered a broad range of species, some of which have not been formerly described. In this study, 60 saprobic isolates were obtained from decaying woody hosts in southwestern China. The isolates were compared with other species using morphological characteristics, and available DNA sequence data was used to infer phylogenetic analyses based on the internal transcribed spacer (ITS), large subunit rRNA gene (LSU), and translation elongation factor 1-α (tef) loci. Three novel species were illustrated and described as Botryobambusa guizhouensis, Sardiniella elliptica, and Sphaeropsis guizhouensis, which belong to rarely identified genera within Botryosphaeriaceae. Botryobambusa guizhouensis is the second species identified from the respective monotypic genus. The previously known species were identified as Aplosporella hesperidica, Barriopsis tectonae, Botryosphaeria dothidea, Diplodia mutila, Di. neojuniperi, Di. pseudoseriata, Di. sapinea, Di. seriata, Dothiorella sarmentorum, Do. yunnana, Lasiodiplodia pseudotheobromae, Neofusicoccum parvum, Sardiniella celtidis, Sa. guizhouensis, and Sphaeropsis citrigena. The results of this study indicate that numerous species of Botryosphaeriales are yet to be revealed in southwestern China.

https://botryosphaeriales.org/, an online platform for up-to-date classification and account of taxa of Botryosphaeriales.

Fungi are eukaryotes that inhabit various ecosystems worldwide and have a decomposing effect that other organisms cannot replace. Fungi are divided into two main groups depending on how their sexual spores are formed, viz. Ascomycota and Basidiomycota. The members of Botryosphaeriales (Dothideomycetes, Ascomycota) are ubiquitous. They are pathogenic on a wide range of hosts, causing diverse diseases including dieback, canker, leaf spots and root rots and are also reported as saprobes and endophytes worldwide. As an important fungal group, of which most are plant pathogens, it is necessary to organize data and information on Botryosphaeriales so that scientific literature can be used effectively. For this purpose, a new website, https://botryosphaeriales.org is established to gather all published data together with updates on the present taxonomy of Botryosphaeriales. The website consists of an easy-to-operate searching system and provides an up-to-date classification together with accounts of Botryosphaeriales taxa, including colour illustrations, descriptions, notes and numbers of species in each genus, as well as their classification. Thus, readers will be able to obtain information on botryosphaerialean taxa through this platform. Database URL: https://botryosphaeriales.org/.

Additions to the Genus Arthrinium (Apiosporaceae) From Bamboos in China.

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.

Unravelling Diaporthe Species Associated with Woody Hosts from Karst Formations (Guizhou) in China.

Though several Diaporthe species have been reported in China, little is known about the species associated with nature reserves in Guizhou province. During a survey of fungi in six nature reserves in Guizhou province of China, thirty-one Diaporthe isolates were collected from different woody hosts. Based on morphology, culture characteristics and molecular phylogenetic analysis, these isolates were characterized and identified. Phylogenetic analysis of internal transcribed spacer region (ITS), combined with translation elongation factor 1-alpha (tef), ß-tubulin (tub), calmodulin (cal) and histone H3 (his) gene regions identified five known Diaporthe species and seven distinct lineages representing novel Diaporthe species. The details of five known species: Diaporthe cercidis, D. cinnamomi, D. conica, D. nobilis and D. sackstonii are given and the seven new species D. constrictospora, D. ellipsospora, D. guttulata, D. irregularis, D. lenispora, D. minima, and D. minusculata are introduced with detailed descriptions and illustrations. This study revealed a high diversity of previously undescribed Diaporthe species associated with woody hosts in various nature reserves of Guizhou province, indicating that there is a potential of Diaporthe species remains to be discovered in this unique landform (Karst formations) in China. Interestingly, the five known Diaporthe species have been reported as pathogens of various hosts, and this could indicate that those newly introduced species in this study could be potentially pathogenic pending further studies to confirm.

Molecular Phylogeny and Morphology of Amphisphaeria (= Lepteutypa) (Amphisphaeriaceae).

Amphisphaeriaceous taxa (fungi) are saprobes on decaying wood in terrestrial, mangrove, and freshwater habitats. The generic boundaries of the family have traditionally been based on morphology, and the delimitation of genera has always been challenging. Amphisphaeria species have clypeate ascomata and 1-septate ascospores and a coelomycetous asexual morph. Lepteutypa is different from Amphisphaeria in having eutypoid stromata and more than 1-septate ascospores. These main characters have been used for segregation of Lepteutypa from Amphisphaeria for a long time. However, the above characters are overlapping among Amphisphaeria and Lepteutypa species. Therefore, here we synonymized Lepteutypa under Amphisphaeria based on holomorphic morphology and multigene phylogeny. Further, our cluster analysis reveals the relationship between seven morphological traits among Amphisphaeria/Lepteutypa species and suggests those morphologies are not specific to either genus. Three new species (i.e., Amphisphaeria camelliae, A. curvaticonidia, and A. micheliae) are introduced based on morphology and LSU-ITS-RPB2-TUB2 phylogenies. Furthermore, the monotypic genus Trochilispora, which had been accepted in Amphisphaeriaceae, is revisited and synonymized under Hymenopleella and placed in Sporocadaceae.

Morpho-phylogenetic evidence reveals new species in Rhytismataceae (Rhytismatales, Leotiomycetes, Ascomycota) from Guizhou Province, China.

Karst formations represent a unique eco-environment. Research in the microfungi inhabiting this area is limited. During an ongoing survey of ascomycetous microfungi from karst terrains in Guizhou Province, China, we discovered four new species, which are introduced here as Hypoderma paralinderae, Terriera karsti, T. meitanensis and T. sigmoideospora placed in Rhytismataceae, based on phylogenetic analyses and morphological characters. Molecular analyses, based on concatenated LSU-ITS-mtSSU sequence data, were used to infer phylogenetic affinities. Detail descriptions and comprehensive illustrations of these new taxa are provided and relationships with the allied species are discussed, based on comparative morphology and molecular data.

Phylogeny and morphology of Lasiodiplodia species associated with Magnolia forest plants.

Two new species of Lasiodiplodia (Lasiodiplodia endophytica and Lasiodiplodia magnoliae) are described and illustrated from Magnolia forests in Yunnan, China. Endophytic and saprobic Lasiodiplodia pseudotheobromae and endophytic L. thailandica are new records from this host. The internal transcribed spacers (ITS), part of the translation elongation factor-1α (tef1) and partial ß-tubulin (tub2) sequence data were analyzed to investigate the phylogenetic relationships of the new species with other Lasiodiplodia species. Lasiodiplodia magnoliae is phylogenetically sister to L. mahajangana and L. pandanicola but morphologically distinct from L. mahajangana in having larger conidia. Lasiodiplodia endophytica is most closely related to L. iraniensis and L. thailandica and the three species can be distinguished from one another by 2 base pair differences in ITS and three or four base pair differences in tef1. The new collections suggest that Magnolia forest plants are good hosts for Lasiodiplodia species with endophytic and saprobic life-styles.

The genus Castanediella.

Two new species, Castanediellabrevis and C.monoseptata, are described, illustrated and compared with other Castanediella taxa. Evidence for the new species is provided by morphological comparison and sequence data analyses. Castanediellabrevis can be distinguished from other Castanediella species by the short hyaline conidiophores and fusiform, aseptate hyaline conidia, while C.monoseptata differs from other Castanediella species by its unbranched conidiophores and fusiform, curved, 0-1-sepatate, hyaline conidia. Phylogenetic analysis of combined ITS and LSU sequence data was carried out to determine the phylogenetic placement of the species. A synopsis of hitherto described Castanediella species is provided. In addition, Castanediella is also compared with morphologically similar-looking genera such as Idriella, Idriellopsis, Microdochium, Neoidriella, Paraidriella and Selenodriella.

Striatiguttulaceae, a new pleosporalean family to accommodate Longicorpus and Striatiguttula gen. nov. from palms.

Palms represent the most morphological diverse monocotyledonous plants and support a vast array of fungi. Recent examinations of palmicolous fungi in Thailand led to the discovery of a group of morphologically similar and interesting taxa. A polyphasic approach based on morphology, multi-gene phylogenetic analyses and divergence time estimates supports the establishment of a novel pleosporalean family Striatiguttulaceae, which diversified approximately 39 (20-63) MYA (crown age) and 60 (35-91) MYA (stem age). Striatiguttulaceae is characterized by stromata or ascomata with a short to long neck, trabeculate pseudoparaphyses and fusiform to ellipsoidal, 1-3-septate ascospores, with longitudinal striations and paler end cells, surrounded by a mucilaginous sheath. Multi-gene phylogenetic analysis showed that taxa of Striatiguttulaceae form a well-supported and distinct monophyletic clade in Pleosporales, and related to Ligninsphaeriaceae and Pseudoastrosphaeriellaceae. However, these families can be morphologically demarcated by the slit-like ascomata and extremely large ascospores in Ligninsphaeriaceae and the rather narrow fusiform ascospores in Pseudoastrosphaeriellaceae. Eight strains of Striatiguttulaceae formed two monophyletic sub-clades, which can be recognized as Longicorpus gen. nov. and Striatiguttula gen. nov. Morphologically, the genus Longicorpus can be differentiated from Striatiguttula by its elongated immersed ascomata and fusiform ascospores with relatively larger middle cells and paler end cells. Two new species Striatiguttulanypae and S.phoenicis, and one new combination, Longicorpusstriataspora are introduced with morphological details, and phylogenetic relationships are discussed based on DNA sequence data.