Molecular phylogeny and morphology of Sporodiniella sinensis sp. nov. (Syzygitaceae, Mucorales), an invertebrate-associated species from Yunnan, China.

During investigations of invertebrate-associated fungi in Yunnan Province of China, a new species, Sporodiniella sinensis sp. nov., was collected. Morphologically, S. sinensis is similar to Sporodiniella umbellata; however, it is distinguished from S. umbellata by its greater number of sporangiophore branches, longer sporangiophores, larger sporangiospores, and columellae. The novel species exhibits similarities of 91.62 % for internal transcribed spacer (ITS), 98.66-99.10 % for ribosomal small subunit (nrSSU), and 96.36-98.22 % for ribosomal large subunit (nrLSU) sequences, respectively, compared to S. umbellata. Furthermore, phylogenetic analyses based on combined sequences of ITS, nrLSU and nrSSU show that it forms a separate clade in Sporodiniella, and clusters closely with S. umbellata with high statistical support. The phylogenetic and morphological evidence support S. sinensis as a distinct species. Here, it is formally described and illustrated, and compared with other relatives.

Molecular phylogeny and morphology reveal two new entomopathogenic species of Ophiocordyceps (Ophiocordycipitaceae, Hypocreales) parasitic on termites from China.

Two new termite-pathogenic species, Ophiocordycepsglobiperitheciata and O.longistipes, are described from Yunnan Province, China. Six-locus (ITS, nrSSU, nrLSU, tef-1α, rpb1 and rpb2) phylogenetic analyses in combination with morphological observations were employed to characterize these two species. Phylogenetically, O.globiperitheciata is most closely related to Hirsutellacryptosclerotium and O.communis, whereas O.longistipes shares a sister relationship with O.fusiformis. However, O.globiperitheciata differs from H.cryptosclerotium by parasitizing Blattodea and producing clavate, unbifurcated stromata. Ophiocordycepsglobiperitheciata is distinguished from O.communis by multiple stromata, shorter asci and ascospores. Ophiocordycepslongistipes differs from O.fusiformis in producing larger stromata, perithecia, asci and ascospores, as well as smaller citriform or oval conidia. Morphological descriptions of the two new species and a dichotomous key to the 19 termite-pathogenic Ophiocordyceps species are presented.

Phylogeny and species delimitations in the economically, medically, and ecologically important genus Samsoniella (Cordycipitaceae, Hypocreales).

Samsoniella is a ubiquitous genus of cosmopolitan arthropod-pathogenic fungi in the family Cordycipitaceae. The fungi have economic, medicinal, and ecological importance. Prior taxonomic studies of these fungi relied predominantly on phylogenetic inferences from five loci, namely, the nuclear ribosomal small and large subunits (nr SSU and nr LSU), the 3' portion of translation elongation factor 1 alpha (3P_TEF), and RNA polymerase II subunits 1 and 2 (RPB1 and RPB2). Despite many new species being described, not all of the recognized species inside this group formed well-supported clades. Thus, the search for new markers appropriate for molecular phylogenetic analysis of Samsoniella remains a challenging problem. In our study, we selected the internal transcribed spacer regions of the rDNA (ITS rDNA) and seven gene regions, namely, 3P_TEF, the 5' portion of translation elongation factor 1 alpha (5P_TEF), RPB1, RPB2, γ-actin (ACT), ß-tubulin (TUB), and a gene encoding a minichromosome maintenance protein (MCM7), as candidate markers for species identification. Genetic divergence comparisons showed that the ITS, RPB2, ACT, and TUB sequences provided little valuable information with which to separate Samsoniella spp. In contrast, sequence data for 3P_TEF, 5P_TEF, RPB1, and MCM7 provided good resolution of Samsoniella species. The phylogenetic tree inferred from combined data (5P_TEF + 3P_TEF + RPB1 + MCM7) showed well-supported clades for Samsoniella and allowed for the delimitation of 26 species in this genus. The other two species (S.formicae and S.lepidopterorum) were not evaluated, as they had abundant missing data.

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.

Phylogeny and systematics of the genus Clonostachys.

Introduction: Clonostachys, a genus with rich morphological and ecological diversity in Bionectriaceae, has a wide distribution among diverse habitats. Methods and Results: In the present study, a phylogenetic framework is reconstructed for the family Bionectriaceae focusing on Clonostachys through increased taxon-sampling using the nrLSU sequence. Through surveying Clonostachys in China, Vietnam, and Thailand over the past 3 years, seven Clonostachys spp. were found and identified. Two new species, C. chuyangsinensis and C. kunmingensis, are described and illustrated based on morphological characteristics and molecular data. The phylogenetic positions of the seven species were evaluated based on four genomic loci (ITS, nrLSU, TUB2, and TEF1). Discussion: Moreover, the genetic divergence comparisons of Clonostachys species for three markers (ITS, TUB2, and TEF1) are also provided. The results indicated that the TEF1 sequence data provided the best resolution for distinguishing species of Clonostachys, followed by sequence data for the TUB2 and ITS regions.

Phylogeny and morphology of Ophiocordyceps puluongensis sp. nov. (Ophiocordycipitaceae, Hypocreales), a new fungal pathogen on termites from Vietnam.

Termites are serious pests in agriculture and forestry, causing significant economic losses to property and the construction industry. However, only a few entomopathogenic fungi attack termites that are dominant members of most terrestrial biomes. This study contributes to the taxonomic knowledge of entomopathogenic fungi with the description of a new pathogen on termites collected from the Pu Luong Nature Reserve in Vietnam. The new termite pathogen, Ophiocordyceps puluongensis, is introduced on the basis of morphological and multigene phylogenetic evidence. Based on the combined dataset of five genes including the nuclear ribosomal small and large subunits (nrSSU and nrLSU), the elongation factor 1α (tef-1α), and the largest and the second largest subunits of RNA polymerase II (rpb1 and rpb2), phylogenetic analyses were performed by Maximum Likelihood and Bayesian inference methods to determine the phylogenetic position of O. puluongensis. Three samples of O. puluongensis are clustered in the Hirsutella thompsonii subclade of Hirsutella lineages in Ophiocordyceps, and clustered together with O. asiatica to form a separate clade from other Ophiocordyceps species. Morphologically, O. puluongensis differs from O. asiatica by its smaller and shorter perithecia, asci and ascospores, pink to reddish-orange stipes of stromata, as well as smaller fusiform or citriform conidia. The distinctiveness of this termite pathogen is strongly supported by both molecular phylogeny and morphology. The entomopathogenic fungus O. puluongensis could have the potential to be used as bioinsecticides to control termites.

https://invertebratefungi.org/: an expert-curated web-based platform for the identification and classification of invertebrate-associated fungi and fungus-like organisms.

Fungi are the major decomposers in terrestrial and aquatic ecosystems, playing essential roles in biogeochemical cycles and food webs. The Fungi kingdom encompasses a diverse array of taxa that often form intimate relationships with other organisms, including plants, insects, algae, cyanobacteria and even other fungi. Fungal parasites of insects are known as entomopathogenic fungi and are the causative agents of serious disease and/or mortality of their hosts. Entomopathogens produce distinct metabolic compounds with roles in pathogenicity, virulence and host-parasite interactions. Thus, the potential of discovering new bioactive compounds useful in biocontrol and pharmaceutical industries is high. Given the significance of entomopathogenic fungi, the rapid research advances and the increased interest, it has become necessary to organize all available and incoming data. The website https://invertebratefungi.org/ has been developed to serve this purpose by gathering and updating entomopathogenic genera/species information. Notes of entomopathogenic genera will be provided with emphasis on their taxonomic status. Information on other invertebrates, such as rotifers, will also be included. Descriptions, photographic plates, information on distribution and host (where applicable) along with molecular data and other interesting details will also be provided. The website is easily and freely accessible to users. Instructions concerning the platform architecture and functionality of the website are introduced herein. The platform is currently being expanded and will be continuously updated as part of the effort to enrich knowledge on this group of fungi. Database URL: https://invertebratefungi.org/.

Cordycepin as a Promising Inhibitor of SARS-CoV-2 RNA Dependent RNA Polymerase (RdRp).

BACKGROUND: SARS-CoV-2, which emerged in Wuhan, China, is a new global threat that has killed millions of people and continues to do so. This pandemic has not only threatened human life but has also triggered economic downturns across the world. Researchers have made significant strides in discovering molecular insights into SARSCoV- 2 pathogenesis and developing vaccines, but there is still no successful cure for SARS-CoV-2 infected patients. OBJECTIVE: The present study has proposed a drug-repositioning pipeline for the design and discovery of an effective fungal-derived bioactive metabolite as a drug candidate against SARS-CoV-2. METHODS: Fungal derivative "Cordycepin" was selected for this study to investigate the inhibitory properties against RNA-dependent RNA polymerase (RdRp) (PDB ID: 6M71) of SARS-CoV-2. The pharmacological profile, intermolecular interactions, binding energy, and stability of the compound were determined utilizing cheminformatic approaches. Subsequently, molecular dynamic simulation was performed to better understand the binding mechanism of cordycepin to RdRp. RESULTS: The pharmacological data and retrieved molecular dynamics simulations trajectories suggest excellent drug-likeliness and greater structural stability of cordycepin, while the catalytic residues (Asp760, Asp761), as well as other active site residues (Trp617, Asp618, Tyr619, Trp800, Glu811) of RdRp, showed better stability during the overall simulation span. CONCLUSION: Promising results of pharmacological investigation along with molecular simulations revealed that cordycepin exhibited strong inhibitory potential against SARSCoV- 2 polymerase enzyme (RdRp). Hence, cordycepin should be highly recommended to test in a laboratory to confirm its inhibitory potential against the SARS-CoV-2 polymerase enzyme (RdRp).

The complete mitochondrial genome of Ophiocordyceps gracilis and its comparison with related species.

In this study, the complete mitochondrial genome of O. gracilis was sequenced and assembled before being compared with related species. As the second largest mitogenome reported in the family Ophiocordycipitaceae, the mitogenome of O. gracilis (voucher OG201301) is a circular DNA molecule of 134,288 bp that contains numerous introns and longer intergenomic regions. UCA was detected as anticodon in tRNA-Sec of O. gracilis, while comparative mitogenome analysis of nine Ophiocordycipitaceae fungi indicated that the order and contents of PCGs and rRNA genes were considerably conserved and could descend from a common ancestor in Ophiocordycipitaceae. In addition, the expansion of mitochondrial organization, introns, gene length, and order of O. gracilis were determined to be similar to those of O. sinensis, which indicated common mechanisms underlying adaptive evolution in O. gracilis and O. sinensis. Based on the mitochondrial gene dataset (15 PCGs and 2 RNA genes), a close genetic relationship between O. gracilis and O. sinensis was revealed through phylogenetic analysis. This study is the first to investigate the molecular evolution, phylogenetic pattern, and genetic structure characteristics of mitogenome in O. gracilis. Based on the obtained results, the mitogenome of O. gracilis can increase understanding of the genetic diversity and evolution of cordycipitoid fungi.

Morphological and Molecular Phylogenetic Data of the Chinese Medicinal Fungus Cordyceps liangshanensis Reveal Its New Systematic Position in the Family Ophiocordycipitaceae.

A cordycipitoid fungus infecting Hepialidae sp. in Nepal was supposed to be identical to Cordyceps liangshanensis, originally described from southwestern China, and thus, transferred to the genus Metacordyceps or Papiliomyces in previous studies. However, our multi-gene (nrSSU-nrLSU-tef-1α-rpb1-rpb2) phylogenetic and morphological studies based on the type specimen and additional collections of C. liangshanensis revealed that the fungus belongs to the genus Ophiocordyceps (Ophiocordycipitaceae). Therefore, a new combination O. liangshanensis was made, and a detailed description of this species was provided.

Complex microbial communities inhabiting natural Cordyceps militaris and the habitat soil and their predicted functions.

Cordyceps militaris is a traditional Chinese medicinal food that is challenging to quality maintaining while mass cultivation. Many studies have found that abundant microbes inhabit Ophiocordyceps sinensis and perform important functions for their host. In this study, our objective was to reveal the microbial communities that inhabit C. militaris and analyze their potential functions. High-throughput sequencing of 16S rRNA and ITS genes was used to compare the diversity and composition of the bacterial and fungal communities associated with naturally occurring C. militaris collected from Yunnan Province, southwestern China. The diversity and richness of the microbial communities and the number of function genes of the bacteria were significantly higher in the habitat soil than in the fruiting body. The sclerotia and stromata samples shared the same microbiota and functions. The main bacterial phyla were Proteobacteria, Acidobacteria, Bacteroidetes and Actinobacteria, and Ascomycota was the main fungal phylum. The growth-promoting bacteria Herbaspirillum and the plant probiotic Phyllobacterium, which may enhance C. militaris quality and facilitate its cultivation, were detected in the fruiting body samples. Genes related to metabolism were more abundant in the soil bacteria, while membrane transport genes were more abundant in the endophytic bacteria of C. militaris. Our study is the first to reveal the unexpectedly high diversity of the microbial communities and the bacterial functions inhabiting the natural C. militaris using high-throughput sequencing, and our results provide insights into mining the functions of microorganisms in the development and quality of C. militaris.

Multigene Phylogeny and Morphology of Ophiocordyceps alboperitheciata sp. nov., A New Entomopathogenic Fungus Attacking Lepidopteran Larva from Yunnan, China.

A new fungus, Ophiocordyceps alboperitheciata, parasitic on the larva of Noctuidae (Lepidoptera) was identified from a survey of entomopathogenic fungi in Kunming Wild Duck Forest Park, Yunnan Province, China. It can be primarily distinguished from relatives by its longer fertile parts, sterile tips, superficial perithecia, narrower asci, and smaller septa of ascospores. As revealed from phylogenetic analyses inferred from nrSSU, nrLSU, tef-1α, rpb1, and rpb2 sequence data, O. alboperitheciata belongs to the Hirsutella citriformis clade in the genus Ophiocordyceps of Ophiocordycipitaceae, and forms a separated clade from other related species. The uniqueness of the taxon is significantly evidenced by both molecular phylogeny and morphology. Furthermore, the interspecific relationships in the H. citriformis clade are discussed.

Prospects for Discovering the Secondary Metabolites of Cordyceps Sensu Lato by the Integrated Strategy.

BACKGROUND: Some species of Cordyceps sensu lato are famous Chinese herbs with significant biological activities, often used as edible food and traditional medicine in China. Cordyceps represents the largest entomopathogenic group of fungi, including 40 genera and 1339 species in three families and incertae sedis of Hypocreales. OBJECTIVE: Most of the Cordyceps-derivatives have been approved clinically for the treatment of various diseases such as diabetes, cancers, inflammation, cardiovascular, renal and neurological disorders and are used worldwide as supplements and herbal drugs, but there is still need for highly efficient Cordyceps-derived drugs for fatal diseases with approval of the U.S. Food and Drug Administration. METHODS: Computer-aided drug design concepts could improve the discovery of putative Cordyceps- derived medicine within less time and low budget. The integration of computer-aided drug design methods with experimental validation has contributed to the successful discovery of novel drugs. RESULTS: This review focused on modern taxonomy, active metabolites, and modern drug design techniques that could accelerate conventional drug design and discovery of Cordyceps s. l. Successful application of computer-aided drug design methods in Cordyceps research has been discussed. CONCLUSION: It has been concluded that computer-aided drug design techniques could influence the multiple target-focused drug design, because each metabolite of Cordyceps has shown significant activities for the various diseases with very few or no side effects.

Impact of Traditional Plants and their Secondary Metabolites in the Discovery of COVID-19 Treatment.

BACKGROUND: Coronavirus Disease-2019 belongs to the family of viruses which cause serious pneumonia along with fever, breathing issues and infection of lungs, and was first reported in China and later spread worldwide. OBJECTIVE: Several studies and clinical trials have been conducted to identify potential drugs and vaccines for Coronavirus Disease-2019. The present study listed natural secondary metabolites identified from plant sources with antiviral properties and could be a safer and tolerable treatment for Coronavirus Disease-2019. METHODS: A comprehensive search on the reported studies was conducted using different search engines such as Google Scholar, SciFinder, Sciencedirect, Medline PubMed, and Scopus for the collection of research articles based on plant-derived secondary metabolites, herbal extracts, and traditional medicine for coronavirus infections. RESULTS: Status of COVID-19 worldwide and information of important molecular targets involved in COVID- 19 are described, and through literature search, it is highlighted that numerous plant species and their extracts possess antiviral properties and are studied with respect to coronavirus treatments. Chemical information, plant source, test system type with a mechanism of action for each secondary metabolite are also mentioned in this review paper. CONCLUSION: The present review has listed plants that have presented antiviral potential in the previous coronavirus pandemics and their secondary metabolites, which could be significant for the development of novel and a safer drug which could prevent and cure coronavirus infection worldwide.

Diversity and composition of microbiota during fermentation of traditional Nuodeng ham.

The microbial community is one of the most important factors in shaping the characteristics of fermented food. Nuodeng ham, traditionally produced and subjected to 1-4 years of fermentation, is a dry fermented food product with cultural and economic significance to locals in southwestern China. In this study, we aimed to characterize the microbiota and physicochemical profiles of Nuodeng ham across different stages of fermentation. Ham samples from each of the four years were analyzed by sequencing bacterial 16S rRNA gene and fungal internal transcribed spacer sequence, in order to characterize the diversity and composition of their microflora. A total of 2,679,483 bacterial and 2,983,234 fungal sequences of high quality were obtained and assigned to 514 and 57 genera, respectively. Among these microbes, Staphylococcus and Candida were the most abundant genera observed in the ham samples, though samples from different years showed differences in their microbial abundance. Results of physicochemical properties (pH, water, amino acid, NaCl, nitrate and nitrite contents, and the composition of volatile compounds) revealed differences among the ham samples in the composition of volatile compounds, especially in the third year samples, in which no nitrite was detected. These results suggest that the structure and diversity of microbial communities significantly differed across different stages of fermentation. Moreover, the third year hams exhibits a unique and balanced microbial community, which might contribute to the special flavor in the green and safe food products. Thus, our study lends insights into the production of high quality Nuodeng ham.

Cordycipitoid Fungi Powders Promote Mycelial Growth and Bioactive-Metabolite Production in Liquid Cultures of the Stout Camphor Medicinal Mushroom Taiwanofungus camphoratus (Agaricomycetes).

Taiwanofungus camphoratus, a medicinal mushroom indigenous to Taiwan, has attracted the attention of pharmaceutical companies due to its remarkable properties and considerable commercial value. Since it grows slowly and most of its components are chemically unstable, its production and distribution have been problematic. In the present study, cordycipitoid fungi powders of seven species were used to cultivate T. camphoratus, and optimal conditions for biomass production were determined by response surface methodology. The initial liquid medium was enriched with cordycipitoid fungi powders, inoculated with a spore suspension of T. camphoratus, and then incubated on a rotary shaker (120 r/min at 27°C) for 14 days. The effects of cordycipitoid fungi powders on T. camphoratus production were investigated by examining the biomass production of T. camphoratus. The key factors influencing biomass production, as identified by a two-level Plackett-Burman design with eight variables, were (1) powder content of cordycipitoid fungi, (2) glucose content, and (3) bottling volume. Box-Behnken design and response surface analysis were applied to further investigate the mutual interactions among these factors, and to obtain optimal values leading to maximal biomass yields. Levels of triterpenoids, polysaccharides, mannitol, adenosine, and ergosterol were determined as estimates of the medicinal value of T. camphoratus cultured with or without cordycipitoid fungi powders. The results showed that Paecilomyces hepiali, Metacordyceps neogunnii, and Beauveria bassiana promoted mycelial growth of T. camphoratus, with P. hepiali showing the most prominent effect. The optimal conditions promoting maximal biomass production were found to be as follows: 6.93 g/L P. hepiali powder content; 26.48 g/L glucose content; 180.55 mL of bottling volume (in a 500-mL Erlenmayer flask). Under these conditions, the biomass production was increased by 38.32%, from 13.10 to 18.12 g/L. The polysaccharide, mannitol, adenosine and ergosterol contents, but not the triterpenoid contents of T. camphoratus cultured with P. hepiali powder, were noticeably higher than when cultured with no powder (control condition), and were higher than those of the P. hepiali powder itself. These results indicate the feasibility of large-scale fermentation of T. camphoratus to produce valuable substances that may be used in pharmaceutical products.

Morphology, molecular characterization, and virulence of Beauveria pseudobassiana isolated from different hosts.

Beauveria pseudobassiana has great potential for use in the management of various insect pests. In the present study, we aimed to explore the the virulence of B. pseudobassiana isolated from a diversity of hosts to Bombyx mori and Tenebrio molitor larvae. To this end, 15B. pseudobassiana isolates from 10 different geographical locations were identified based on morphological characteristics and molecular data. The phylogenetic positions of the isolates were evaluated according to morphological features and phylogenetic inferences based on six loci (nrSSU, nrLSU, TEF, RPB1, RPB2 and Bloc). In addition to growth in soil, the B. pseudobassiana isolates in our study were isolated from a wide host range that extended to 5 orders, 11 families, and 14 species. Moreover, anamorphically typified B. pseudobassiana was grown for the first time from teleomorph stromata. Pathogenicity of the B. pseudobassiana isolates from the different hosts was determined with two bioassays using B. mori and T. molitor larvae. The results indicated that mortality of B. mori caused by the lepidopteran isolates was significantly higher than that of isolates from other hosts, and virulence of the coleopteran isolates to T. molitor was significantly higher than that of isolates from other hosts. The host specificity of B. pseudobassiana should be studied in more detail before future consideration of isolates for use in biological control of pests.

Distribution of nucleosides in populations of Cordyceps cicadae.

A rapid HPLC method had been developed and used for the simultaneous determination of 10 nucleosides (uracil, uridine, 2'-deoxyuridine, inosine, guanosine, thymidine, adenine, adenosine, 2'-deoxyadenosine and cordycepin) in 10 populations of Cordyceps cicadae, in order to compare four populations of Ophicordyceps sinensis and one population of Cordyceps militaris. Statistical analysis system (SAS) 8.1 was used to analyze the nucleoside data. The pattern of nucleoside distribution was analyzed in the sampled populations of C. cicadae, O. sinensis and C. militaris, using descriptive statistical analysis, nested analysis and Q cluster analysis. The total amount of the 10 nucleosides in coremium was 1,463.89-5,678.21 µg/g in 10 populations of C. cicadae, 1,369.80-3,941.64 µg/g in sclerotium. The average contents of the 10 analytes were 4,392.37 µg/g and 3,016.06 µg/g in coremium and sclerotium, respectively. The coefficient of variation (CV) of nucleosides ranged from 8.36% to 112.36% in coremium of C. cicadae, and from 10.77% to 155.87% in sclerotium of C. cicadae. The CV of the nucleosides was wide within C. cicadae populations. The nested variation analysis by the nine nucleosides' distribution indicated that about 42.29% of the nucleoside variability in coremium was attributable to the differentiation among populations, and the remaining 57.71% resided in the populations. It was also shown that about 28.94% of the variation in sclerotium was expressed between populations, while most of the variation (71.06%) corresponded to the populations.