Multi-locus molecular phylogenetic analysis reveals two new species of Amphichorda (Bionectriaceae, Hypocreales).

Amphichorda has been previously accepted as a member of the Cordycipitaceae and currently it is considered a member of the Bionectriaceae. The substrates of Amphichorda were complex and varied, being mainly animal faeces. This study reports two new species of Amphichorda from Yunnan Province in south-western China. Based on the five-gene (nrSSU, nrLSU, tef-1α, rpb1 and rpb2) sequence and ITS data phylogenetic analysis, two new species, namely A.excrementa and A.kunmingensis, are proposed and a detailed description of the new species is provided. Amphichordaexcrementa and A.kunmingensis were isolated from animal faeces in the park. The morphological characteristics of two novel species and seven known species in Amphichorda are also compared.

Molecular Phylogeny and Morphology Reveal Cryptic Species in the Cordyceps militaris Complex from Vietnam.

The Cordyceps militaris complex, which is a special group in the genus Cordyceps, is rich in species diversity and is widely distributed in nature. Throughout the investigations of arthropod-pathogenic fungi in the national reserves and in Vietnam parks, collections of C. militaris attacking lepidopteran pupae or larvae were located in the soil and on the leaf litter. The phylogenetic analyses of the combined nrSSU, nrLSU, TEF, RPB1, and RPB2 sequence data indicated that the fungal materials collected in Vietnam belonged to C. militaris and two hidden species in the C. militaris complex. The phylogenetic analyses and morphological comparisons presented here strongly supported the descriptions of C. polystromata and C. sapaensis as new taxa as well as C. militaris as a known species. The morphological characteristics of 11 species in the C. militaris complex, which included two novel species and nine known taxa, were also compared.

Six new species of zombie-ant fungi from Yunnan in China.

Some Ophiocordyceps species infecting ants are able to manipulate the host behavior. The hosts are manipulated in order to move to location that are advantageous for fungal spore transmission. Ophiocordyceps species that are able to manipulate the ant's behavior are called "zombie-ant fungi". They are widespread within tropical forests worldwide, with relatively few reports from subtropical monsoon evergreen broad-leaf forest. Zombie-ant fungi have been described and reported in different countries worldwide. However, there were a few reports from China. This study proposed six new species of zombie-ant fungi from China based on multi-gene (SSU, LSU, TEF, RPB1 and RPB2) phylogenetic analyses and morphological characteristics. Six novel species of Ophiocordyceps from China were identified as the Ophiocordyceps unilateralis core clade, forming a separate lineage with other species. Six novel species of Ophiocordyceps with hirsutella-like asexual morphs exclusively infecting ants were presented herein, namely, Ophiocordyceps acroasca, Ophiocordyceps bifertilis, Ophiocordyceps subtiliphialida, Ophiocordyceps basiasca, Ophiocordyceps nuozhaduensis and Ophiocordyceps contiispora. Descriptions and illustrations for six taxon were provided. Five of these species were collected from the subtropical monsoon evergreen broad-leaf forest, and one was collected from the rainforest and subtropical monsoon evergreen broad-leaf forest. This work proposes that the same host of Camponotus can be infected by multiple ant pathogenic fungi, while multiple ants of Polyrhachis can be infected by the same pathogenic fungi at the same time. This study contributes towards a better understanding of the evolutionary relationship between hosts and fungi, and provides novel insights into the morphology, distribution, parasitism, and ecology of Ophiocordyceps unilateralis sensu lato. We have provided a method for obtaining living cultures of Ophiocordyceps unilateralis complex species and their asexual morphs based on the living cultures, which is of significant value for further studies of Ophiocordyceps unilateralis complex species in the future.

Phylogeny and Systematics of the Genus Tolypocladium (Ophiocordycipitaceae, Hypocreales).

The taxonomy and phylogeny of the genus Tolypocladium are herein revised based on the most comprehensive dataset to date. Two species-level phylogenies of Tolypocladium were constructed: a single-gene phylogeny (ITS) of 35 accepted species and a multigene phylogeny (nrSSU, nrLSU, tef-1α, rpb1, and rpb2) of 27 accepted species. Three new species, Tolypocladium pseudoalbum sp. nov., Tolypocladium subparadoxum sp. nov., and Tolypocladium yunnanense sp. nov., are described in the present study. The genetic divergences of four markers (ITS, tef-1α, rpb1 and rpb2) among Tolypocladium species are also reported. The results indicated that species of Tolypocladium were best delimited by rpb1 sequence data, followed by the sequence data for the rpb2, tef-1α, and ITS provided regions. Finally, a key to the 48 accepted species of Tolypocladium worldwide is provided.

Genomic Comparative Analysis of Cordyceps pseudotenuipes with Other Species from Cordyceps.

The whole genome of Cordyceps pseudotenuipes was sequenced, annotated, and compared with three related species to characterize the genome. The antibiotics and Secondary Metabolites Analysis Shell (antiSMASH) and local BLAST analysis were used to explore the secondary metabolites (SMs) and biosynthesis gene clusters (BGCs) of the genus Cordyceps. The genome-wide basic characteristics of C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris revealed unequal genome size, with C. cicadae as the largest (34.11 Mb), followed by C. militaris (32.27 Mb). However, the total gene lengths of C. pseudotenuipes and C. tenuipes were similar (30.1 Mb and 30.06 Mb). The GC contents of C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris genomes differed slightly (51.40% to 54.11%). AntiSMASH and local BLAST analysis showed that C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris had 31, 28, 31, and 29 putative SM BGCs, respectively. The SM BGCs contained different quantities of polyketide synthetase (PKS), nonribosomal peptide synthetase (NRPS), terpene, hybrid PKS + NRPS, and hybrid NRPS + Other. Moreover, C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris had BGCs for the synthesis of dimethylcoprogen. C. pseudotenuipes, C. tenuipes, and C. cicadae had BGCs for the synthesis of leucinostatin A/B, neosartorin, dimethylcoprogen, wortmanamide A/B, and beauvericin. In addition, the SM BGCs unique to C. pseudotenuipes were clavaric acid, communesin, and deoxynivalenol. Synteny analysis indicated that the scaffolds where the SM BGC was located were divided into more than 70 collinear blocks, and there might be rearrangements. Altogether, these findings improved our understanding of the molecular biology of the genus Cordyceps and will facilitate the discovery of new biologically active SMs from the genus Cordyceps using heterologous expression and gene knockdown methods.

Morphological and Phylogenetic Characterization Reveals Five New Species of Samsoniella (Cordycipitaceae, Hypocreales).

Samsoniella is a very important fungal resource, with some species in the genus having great medical, economic and ecological value. This study reports five new species of Samsoniella from Yunnan Province and Guizhou Province in Southwestern China and Dole Province in Vietnam, providing morphological descriptions, illustrations, phylogenetic placements, associated hosts and comparisons with allied taxa. Based on morphological observations and phylogenetic analyses of combined nrSSU, nrLSU, tef-1α, rpb1 and rpb2 sequence data, it was determined that these five new species were located in the clade of Samsoniella and different from other species of Samsoniella. The five novel species had morphologies similar to those of other species in the genus, with bright orange cylindrical to clavate stromata (gregarious). The fertile part lateral sides usually had a longitudinal ditch without producing perithecia, and superficial perithecia. The phialides had a swollen basal portion, tapering abruptly into a narrow neck and oval or fusiform one-celled conidia, often in chains. The morphological characteristics of 23 species in Samsoniella, including five novel species and 18 known taxa, were also compared in the present study.

Morphology and Phylogeny Reveal Five Novel Species in the Genus Cordyceps (Cordycipitaceae, Hypocreales) From Yunnan, China.

The current study was aimed to introduce five new species of Cordyceps from Yunnan, with morphological descriptions, illustrations, color photographs, phylogenetic placement, associated host, and a comparison with allied taxa. The five new species were morphologically distinct from all other Cordyceps sensu lato species, and it was also suggested that they should differ from other species in the genus Cordyceps based on combined multigene analyses. Employing DNA nucleotide sequences of the nrLSU, nrSSU, tef-1α, rpb1, and rpb2, the five new species were recognized in the clade of Cordyceps by using molecular phylogenetic analyses, including five well-supported subclades: three new species, Cordyceps bullispora, Cordyceps longiphialis, and Cordyceps nabanheensis, were found in the subclade of C. pruinosa, and two new species, Cordyceps pseudotenuipes and Cordyceps simaoensis, were located in the subclade of C. tenuipes. The five novel species shared similar morphologies to other species in the genus Cordyceps, with fleshy and brightly pigmented stromata; perithecia superficial to completely immersed, ordinal in arrangement; and hyaline asci, with thickened cylindrical ascus apex. The morphological characteristics of 66 species in Cordyceps sensu stricto, namely, 5 novel species and 61 known taxa, were also compared.

Morphology, phylogeny, mitogenomics and metagenomics reveal a new entomopathogenic fungus Ophiocordycepsnujiangensis (Hypocreales, Ophiocordycipitaceae) from Southwestern China.

Ophiocordyceps contains the largest number of Cordyceps sensu lato, various species of which are of great medicinal value. In this study, a new entomopathogenic fungus, Ophiocordycepsnujiangensis, from Yunnan in southwestern China, was described using morphological, phylogenetic, and mitogenomic evidence, and its fungal community composition was identified. It was morphologically characterized by a solitary, woody, and dark brown stromata, smooth-walled and septate hyphae, solitary and gradually tapering conidiogenous cells with plenty of warty protrusions, and oval or fusiform conidia (6.4-11.2 × 3.7-6.4 µm) with mucinous sheath. The phylogenetic location of O.nujiangensis was determined based on the Bayesian inference (BI) and the maximum likelihood (ML) analyses by concatenating nrSSU, nrLSU, tef-1a, rpb1, and rpb2 datasets, and ten mitochondrial protein-coding genes (PCGs) datasets (atp6, atp9, cob, cox2, nad1, nad2, nad3, nad4, nad4L, and nad5). Phylogenetic analyses revealed that O.nujiangensis belonged to the Hirsutellasinensis subclade within the Hirsutella clade of Ophiocordyceps. And O.nujiangensis was phylogenetically clustered with O.karstii, O.liangshanensis, and O.sinensis. Simultaneously, five fungal phyla and 151 fungal genera were recognized in the analysis of the fungal community of O.nujiangensis. The fungal community composition differed from that of O.sinensis, and differences in the microbial community composition of closely related species might be appropriate as further evidence for taxonomy.