Mitogenomics, phylogeny and morphology reveal two new entomopathogenic species of Ophiocordyceps (Ophiocordycipitaceae, Hypocreales) from south-western China.

Ophiocordyceps encompasses over 300 species, demonstrating a wide range of morphological features, hosts and habitats within its species diversity. In this study, two novel species in Ophiocordyceps were revealed parasitising Hepialidae larva buried in soil. Ophiocordycepsalbastroma was morphologically characterised by white stromata, solitary and cylindrical conidiogenous cells and smooth ovoid or ellipsoidal conidia. Ophiocordycepsnigristroma was characterised by woody and dark brown stromata, monophialidic, swollen base and lageniform conidiogenous cells and smooth fusiform or oval conidia. The two new species formed a separate clade, respectively, based on the phylogenetic analyses of a combined dataset including nrSSU, nrLSU, rpb1, rpb2, and tef-1α, as well as a dataset of mitochondrial 14 protein coding genes (PCGs). They were all closely grouped with O.sinensis. The mitochondrial genomes of them were first reported. Their mitogenomes were all typical of circular molecules, with positive AT and GC skew, similar GC content, similar genetic composition, similar codon usage and conservative gene positions. However, the length of the mitogenomes varied. Changes in the length of the genes were the leading cause of changes in the length of mitochondrial genome of Ophiocordyceps. The discovery and identification of new Ophiocordyceps species and analysis their mitochondrial genomes may serve as foundations for phylogeny and diversity research within the genus Ophiocordyceps.

Compositional and functional diversities of core microbial communities in wild and artificial Ophiocordyceps sinensis / Diversidades compositivas y funcionales de comunidades microbianas centrales en Ophiocordyceps sinensis silvestre y artificial

Ophiocordyceps sinensis is an entomogenous fungus, and its wildlife resource is very insufficient, as it is widely traded as a natural health product. The artificial culture of O. sinensis is a remarkably effective progress in addressing the problem. Adding microorganisms may improve the process of artificial culture. To analyse the composition and function of the microbial community, high-throughput sequencing was used to explore the microbial community inhabiting wild and artificial O. sinensis and surrounding soil. Significant differences in the microbial communities across groups were revealed by the PCoA analysis. There were 51 fungal and 598 bacterial operational taxonomic units only being assigned to the fruiting bodies of wild O. sinensis (Wf) by the Venn diagram. From the LEfSe analysis, 39 fungal taxa and 75 bacterial taxa were enriched in Wf. Enzymes that were highly abundant in the core fungi were involved in physiological metabolic processes. Metabolic pathways were dominated in the core bacteria, followed by environmental information processing. The core microorganisms, with the marked differences between Wf and the other three groups, were essential for wild O. sinensis. Functional analysis verified their involvement in the growth, development, and infection of O. sinensis. These core microorganisms may be a valuable resource for the artificial culture of O. sinensis.(AU)

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.

Compositional and functional diversities of core microbial communities in wild and artificial Ophiocordyceps sinensis.

Ophiocordyceps sinensis is an entomogenous fungus, and its wildlife resource is very insufficient, as it is widely traded as a natural health product. The artificial culture of O. sinensis is a remarkably effective progress in addressing the problem. Adding microorganisms may improve the process of artificial culture. To analyse the composition and function of the microbial community, high-throughput sequencing was used to explore the microbial community inhabiting wild and artificial O. sinensis and surrounding soil. Significant differences in the microbial communities across groups were revealed by the PCoA analysis. There were 51 fungal and 598 bacterial operational taxonomic units only being assigned to the fruiting bodies of wild O. sinensis (Wf) by the Venn diagram. From the LEfSe analysis, 39 fungal taxa and 75 bacterial taxa were enriched in Wf. Enzymes that were highly abundant in the core fungi were involved in physiological metabolic processes. Metabolic pathways were dominated in the core bacteria, followed by environmental information processing. The core microorganisms, with the marked differences between Wf and the other three groups, were essential for wild O. sinensis. Functional analysis verified their involvement in the growth, development, and infection of O. sinensis. These core microorganisms may be a valuable resource for the artificial culture of O. sinensis.

Comparison of Widely Targeted Metabolomics and Untargeted Metabolomics of Wild Ophiocordyceps sinensis.

The authors of this paper conducted a comparative metabolomic analysis of Ophiocordyceps sinensis (OS), providing the metabolic profiles of the stroma (OSBSz) and sclerotia (OSBSh) of OS by widely targeted metabolomics and untargeted metabolomics. The results showed that 778 and 1449 metabolites were identified by the widely targeted metabolomics and untargeted metabolomics approaches, respectively. The metabolites in OSBSz and OSBSh are significantly differentiated; 71 and 96 differentially expressed metabolites were identified by the widely targeted metabolomics and untargeted metabolomics approaches, respectively. This suggests that these 71 metabolites (riboflavine, tripdiolide, bromocriptine, lumichrome, tetrahymanol, citrostadienol, etc.) and 96 metabolites (sancycline, vignatic acid B, pirbuterol, rubrophen, epalrestat, etc.) are potential biomarkers. 4-Hydroxybenzaldehyde, arginine, and lumichrome were common differentially expressed metabolites. Using the widely targeted metabolomics approach, the key pathways identified that are involved in creating the differentiation between OSBSz and OSBSh may be nicotinate and nicotinamide metabolism, thiamine metabolism, riboflavin metabolism, glycine, serine, and threonine metabolism, and arginine biosynthesis. The differentially expressed metabolites identified using the untargeted metabolomics approach were mainly involved in arginine biosynthesis, terpenoid backbone biosynthesis, porphyrin and chlorophyll metabolism, and cysteine and methionine metabolism. The purpose of this research was to provide support for the assessment of the differences between the stroma and sclerotia, to furnish a material basis for the evaluation of the physical effects of OS, and to provide a reference for the selection of detection methods for the metabolomics of OS.

Species Diversity and Virulence Potential of the Beauveria bassiana Complex and Beauveria scarabaeidicola Complex.

Beauveria is a very important fungal resource. Some Beauveria species have great economic and ecological value. Through surveying Beauveria in China and Thailand over the past 4 years, 15 Beauveria spp. were collected and identified. Three new species-namely, B. polyrhachicola, B. songmingensis, and B. subscarabaeidicola-were described and illustrated based on morphological characteristics and molecular data. The phylogenetic positions of the 15 species were evaluated according to phylogenetic inferences based on six loci (nrSSU, nrLSU, TEF, RPB1, RPB2, and Bloc). Nine species of Beauveria in our study were isolated from adult scarab beetles (Coleoptera: Scarabaeidae). The pathogenicity of the isolates from the B. bassiana complex and B. scarabaeidicola complex was determined with three bioassays using B. mori and T. molitor larvae, in addition to Protaetia brevitarsis adults. The results indicated that the B. bassiana complex isolates had great potential in the biocontrol of the three insects; by contrast, the B. scarabaeidicola complex isolates showed obvious host specificity and low virulence.

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.

Multigene phylogeny and morphology reveal Ophiocordycepshydrangea sp. nov. and Ophiocordycepsbidoupensis sp. nov. (Ophiocordycipitaceae).

Ophiocordyceps species have a wide range of insect hosts, from solitary beetle larva to social insects. However, among the species of Ophiocordyceps, only a few attack cicada nymphs. These species are mainly clustered in the Ophiocordycepssobolifera clade in Ophiocordyceps. A new entomopathogenic fungus parasitic on cicada nymphs, and another fungus parasitic on the larva of Coleoptera, are described in this study. The two new species viz. Ophiocordycepshydrangea and Ophiocordycepsbidoupensis were introduced based on morphology and multigene phylogenetic evidence. The phylogenetic framework of Ophiocordyceps was reconstructed using a multigene (nrSSU, nr LSU, tef-1α, rpb1, and rpb2) dataset. The phylogenetic analyses results showed that O.hydrangea and O.bidoupensis were statistically well-supported in the O.sobolifera clade, forming two separate subclades from other species of Ophiocordyceps. The distinctiveness of these two new species was strongly supported by both molecular phylogeny and morphology.