Multiverruca sinensis gen. nov., sp. nov., a thermotolerant fungus isolated from soil in China.

During a survey of thermotolerant fungi in China, three isolates were obtained from soil samples. Phylogenetic analysis of a combined internal transcribed spacer and large subunit dataset showed that these isolates belong to the same species, which form a well-separated lineage distinct from the other genera in Latoruaceae. Morphologically, the isolates are characterized by having globose and smooth conidiogenous cells, verruculose mycelium and cymbiform conidia. Combining the phylogenetic analyses and morphological characteristics, Multiverruca gen. nov. is proposed and introduced to accommodate a single new species, Multiverruca sinensis sp. nov. Detailed descriptions, illustrations and notes are provided for the new genus and species.

Keratinophyton chongqingense sp. nov. and Keratinophyton sichuanense sp. nov., from soil in China.

Keratinophyton is a genus of well-known keratinophilic fungi found in various terrestrial habitats. During a survey of keratinolytic fungi in China, a total of 12 isolates of Keratinophyton species, representing eight taxa, were obtained from the soil. Two of these isolates were described as new species based on their morphological characteristics and molecular analyses of the internal transcribed spacer region and the rRNA gene of the nuclear large subunit. Descriptions and illustrations of these two novel species, which are named Keratinophyton chongqingense sp. nov. and Keratinophyton sichuanense sp. nov., are provided herein.

New taxonomic framework for Arthrodermataceae: a comprehensive analysis based on their phylogenetic reconstruction, divergence time estimation, phylogenetic split network, and phylogeography.

The Arthrodermataceae, or dermatophytes, are a major family in the Onygenales and important from a public health safety perspective. Here, based on sequenced and downloaded from GenBank sequences, the evolutionary relationships of Arthrodermataceae were comprehensively studied via phylogenetic reconstruction, divergence time estimation, phylogenetic split network, and phylogeography analysis. These results showed the clades Ctenomyces, Epidermophyton, Guarromyces, Lophophyton, Microsporum, Paraphyton, and Trichophyton were all monophyletic groups, whereas Arthroderma and Nannizzia were polyphyletic. Among them, Arthroderma includes at least four different clades, Arthroderma I, III and IV are new clades in Arthrodermataceae. Nannizzia contains at least two different clades, Nannizzia I and Nannizzia II, but Nannizzia II was a new clade in Arthrodermataceae. The unclassified group, distributed in Japan and India, was incorrectly identified; it should be a new clade in Arthrodermataceae. The phylogenetic split network based on the ITS sequences provided strong support for the true relationships among the lineages in the reconstructed phylogenetic tree. A haplotype phylogenetic network based on the ITS sequences was used to visualize species evolution and geographic lineages relationships in all genera except Trichophyton. The new framework provided here for the phylogeny and taxonomy of Arthrodermataceae will facilitate the rapid identification of species in the family, which should useful for evaluating the results of preventive measures and interventions, as well as for conducting epidemiological studies.

Effects of Medicinal Plants on Fungal Community Structure and Function in Hospital Grassplot Soil.

Hospital grassplot soil is an important repository of pathogenic fungi exposed to the hospital environment, and the diffusion of these fungi-containing soil particles in the air increases the risk of nosocomial fungal infections. In this study, from the perspective of soil microbes-plant holobiont, four medicinal plants Mirabilis jalapa, Artemisia argyi, Viola philippica, and Plantago depressa were used as materials, based on ITS high-throughput amplicon sequencing and simulated pot experiments to explore the effect of medicinal plants on the fungal community in hospital grassplot soil, in order to provide a new exploration for hospital grassplot soil remediation. The results showed that the fungal community ecological guilds in primary test soil was mainly pathogen, and the abundance of animal pathogen with potential threats to human reached 61.36%. After planting medicinal plants, the composition and function of soil fungal community changed significantly. Although this change varied with plant species and growth stages, all samples collected in the pot experiment showed that the pathogen abundance decreased and the saprotroph abundance increased. In addition, 45 of the 46 core fungal genera defined in all potted samples were present in primary test soil, and many of them were human potential pathogens. These findings imply that the idea of enhancing soil quality in hospital grassplot soil by planting specific plants is feasible. However, the initial fungal community of the hospital grassplot soil has a certain stability, and it is difficult to completely eliminate the threat of pathogenic fungi by planting medicinal plants.

Structure and Function of Bacterial Microbiota in Eucommia ulmoides Bark.

The study aimed to explore the bacterial community composition and the functions of core microbiota in Eucommia ulmoides bark. The bark samples of E. ulmoides were collected from Wangcang Sichuan Province, Cili Hunan Province, and Zunyi Guizhou Province, in China, respectively. Through the high-throughput sequencing methods and techniques, the community composition, core microbiota, and function of the bacteria were studied. The bacterial community of E. ulmoides bark consisted of 9 phyla, 11 classes, 22 orders, 28 families, 31 genera, and 37 OTUs. At the genus level, the dominant genus was the unclassified bacteria of Cyanobacteria, with a relative abundance of 97.01%. The bacterial communities of E. ulmoides bark from different areas have their unique units except for the common microbiota. The core microbiota of bacteria included an unclassified genus of Cyanobacteria, an unclassified genus of Mitochondria, Pseudomonas, Sphingobium, Rhizobium, Novosphingobium, Enterobacter, Rhodococcus, Curtobacterium, and Ralstonia. FAPROTAX function prediction suggested that the core microbiota has a substantial potential for photoautotrophy, phototrophy, aerobic chemoheterotrophy, chemoheterotrophy. Ten taxa composed the core microbiota, and the majority of them were related to the pharmacologically active ingredients of E. ulmoides bark. The research provides a scientific basis for the biological marker of genuineness and microbial technology for improving the content of medicinal ingredients of E. ulmoides.

[Community composition and ecological functional structural analysis of endophytic fungi in bark of Eucommia ulmoides in different areas].

Based on high-throughput sequencing and metagenomic technology,the community composition and ecological functions of endophytic fungi in the bark of Eucommia ulmoides from three producing areas,Fengxiang town in Zunyi county of Guizhou province,Lingyang town of Cili county of Hunan province and Mumen town of Wangcang county of Sichuan province,were analyzed. A total of110 865 effective sequences of endophytic fungi were obtained in the study. The corresponding fungal group of OTUs after clustering belonged to 3 phyla( Ascomycota,Basidiomycota,Zygomycota),10 classes,25 orders,41 families,57 genera and 74 species. Among them,the dominant genera of Sichuan Wangcang Bark of E. ulmoides( EWP) was an unclassified genus in the Nectriaceae of the Crimsonaceae,with a relative abundance of 54. 79%; The dominant species of Hunan Cili Bark of E. ulmoides( ECP) was the unclassified genus of Ascomycota,with a relative abundance of 39. 97% and the dominant species of Guizhou Zunyi bark( EZP) was Lophiostoma,and its relative abundance was 47. 07%. The analysis of α diversity indicated that the shannon diversity index of endophytic fungi from different places was as follows: ECP: 1. 340 2>EZP: 1. 380 4 > EWP: 1. 168 3. The simpson diversity index was: EWP( 0. 427 3) >EZP( 0. 332 5) > ECP( 0. 313 6). FUNGuild software platform analysis displayed that endophytic fungi of E. ulmoides bark from three producing areas contained the following 14 functional groups: plant pathogen,animal pathogen and endophyte et al.,the number of functional groups in the 3 samples of E. ulmoides reached up 8 genera and exceeded one half of the total number. Correlation analysis of Canonical correspondence analysis( CCA) between endophytic fungal community diversity and four active compounds of E. ulmoides were analyzed,the results showed that the contents of pinoresinol diglucoside and chlorogenic acid,to a certain extend,had a positive correlation with an unclassified genus of Davidiellaceae,Mortierella,Chaetomium and Pestalotiopsis from the endophytic fungi in EWP sample.

Nitric oxide mediates hypocrellin accumulation induced by fungal elicitor in submerged cultures of Shiraia bambusicola.

Multiple responses of Shiraia bambusicola, including nitric oxide (NO) generation, hypocrellins production and salicylic acid (SA) biosynthesis, were induced by a fungal elicitor prepared from the mycelium of Aspergillum niger. Both the NO donator, sodium nitroprusside, and SA enhanced hypocrellin production without the fungal elicitor. However, the NO scavenger, 2,4-carboxyphenyl-4,4, 5,5- tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and the SA biosynthesis inhibitor, cinnamic acid (CA), inhibited hypocrellin accumulation in the presence of the elicitor. cPTIO also inhibited SA production induced by the A. niger elicitor. CA failed to inhibit NO production but it significantly inhibited hypocrellin accumulation. Aspergillum niger elicitor induced an NO burst, SA accumulation, and hypocrellin production in S. bambusicola. Therefore, the fungal elicitor was involved in the signaling pathway, which is a mechanism different from that of higher plants.

Polysaccharides from the Medicinal Mushroom Cordyceps taii Show Antioxidant and Immunoenhancing Activities in a D-Galactose-Induced Aging Mouse Model.

Cordyceps taii, an edible medicinal mushroom native to south China, is recognized as an unparalleled resource of healthy foods and drug discovery. In the present study, the antioxidant pharmacological properties of C. taii were systematically investigated. In vitro assays revealed the scavenging activities of the aqueous extract and polysaccharides of C. taii against various free radicals, that is, 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and superoxide anion radical. The EC(50) values for superoxide anion-free radical ranged from 2.04 mg/mL to 2.49 mg/mL, which was at least 2.6-fold stronger than that of antioxidant thiourea. The polysaccharides also significantly enhanced the antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase) and markedly decreased the malondialdehyde production of lipid peroxidation in a D-galactose-induced aging mouse model. Interestingly, the immune function of the administration group was significantly boosted compared with the D-galactose-induced aging model group. Therefore, the C. taii polysaccharides possessed potent antioxidant activity closely associated with immune function enhancement and free radical scavenging. These findings suggest that the polysaccharides are a promising source of natural antioxidants and antiaging drugs. Consequently, a preliminary chemical investigation was performed using gas chromatography-mass spectroscopy and revealed that the polysaccharides studied were mainly composed of glucose, mannose, and galactose. Fourier-transform infrared spectra also showed characteristic polysaccharide absorption bands.

Antibacterial activity of hypocrellin A against Staphylococcus aureus.

The antibacterial activity and acting mechanism of hypocrellin A (HA) were conducted regarding in vitro activity of HA on Staphylococcus aureus GZ86 by analyzing the growth, permeability, and morphology of the bacterial cells following treatment with HA. The experimental results indicated 1.5 mg/l HA could completely inhibit the growth of 107 CFU/ml S. aureus cells in liquid beef extract-peptone medium under a halogen-tungsten lamp for 120 min. Meanwhile, HA resulted in the leakage of reducing sugars and proteins and induced the respiratory chain dehydrogenases into inactive state, suggesting that HA were able to destroy the permeability of the bacterial membranes. When the cells of S. aureus were exposed to 2.5 mg/l HA under a halogen-tungsten lamp for 120 min, many pits and gaps were observed in bacterial cells by scanning electron microscopy, and the cell wall was fragmentary, indicating the bacterial cells were damaged severely. The experiments strongly confirmed the contribution of multiform reactive oxygen species (ROS) to bactericidal effect. In conclusion, the combined results suggested that ROS may damage the structure of bacterial cell wall and depress the activity of some membranous enzymes, which cause S. aureus bacteria to die eventually.

Multigene phylogeny, phylogenetic network, and morphological characterizations reveal four new arthropod-associated Simplicillium species and their evolutional relationship.

Simplicillium species are widely distributed and commonly found on various substrates. A minority of species are associated with arthropods. A spider-associated species Simplicillium araneae, and three insect-associated species, Simplicillium coleopterorum, Simplicillium guizhouense, and Simplicillium larvatum, are proposed as novel species based on a multi-locus phylogenetic analysis and morphological characteristics. These Simplicillium species completely fit the nutritional model of Hypocreales fungi and could be used as a model to study their evolutionary relationship. A phylogenetic network analysis based on ITS sequences suggests that a host jump was common among Simplicillium species, and S. araneae may have originally come from an insect host and then jumped to a spider host. However, the evolutionary relationship of S. coleopterorum, S. guizhouense, and S. larvatum was not clear in the phylogenetic network and more sequencing information should be added to the network. In addition, strain CBS 101267 was identified as Simplicillium subtropicum.

Assembly, Core Microbiota, and Function of the Rhizosphere Soil and Bark Microbiota in Eucommia ulmoides.

Medicinal plants are inhabited by diverse microbes in every compartment, and which play an essential role in host growth and development, nutrient absorption, synthesis of secondary metabolites, and resistance to biological and abiotic stress. However, the ecological processes that manage microbiota assembly and the phenotypic and metabolic characteristics of the core microbiota of Eucommia ulmoides remain poorly explored. Here, we systematically evaluated the effects of genotypes, compartment niches, and environmental conditions (climate, soil nutrition, and secondary metabolites) on the assembly of rhizosphere soil and bark associated bacterial communities. In addition, phenotypic and metabolic characteristics of E. ulmoides core microbiota, and their relationship with dominant taxa, rare taxa, and pharmacologically active compounds were deciphered. Results suggested that microbiota assembly along the two compartments were predominantly shaped by the environment (especially pH, relative humidity, and geniposide acid) and not by host genotype or compartment niche. There were 690 shared genera in the rhizosphere soil and bark, and the bark microbiota was mainly derived from rhizosphere soil. Core microbiota of E. ulmoides was a highly interactive "hub" microbes connecting dominant and rare taxa, and its phenotypic characteristics had a selective effect on compartment niches. Metabolic functions of the core microbiota included ammonia oxidation, nitrogen fixation, and polyhydroxybutyrate storage, which are closely related to plant growth or metabolism. Moreover, some core taxa were also significantly correlated with three active compounds. These findings provide an important scientific basis for sustainable agricultural management based on the precise regulation of the rhizosphere soil and bark microbiota of E. ulmoides.

Species Diversity of Cordyceps-Like Fungi in the Tiankeng Karst Region of China.

Tiankeng acts as a refugium for biodiversity amid a changing global climate, and a previous study has shown that some ancient (Alsophila spinulosa) and unique plants (cool-adapted plants) are present in Tiankeng. However, there are few reports on Ascomycota from the Tiankeng karst region. In this research, the species diversity of Cordyceps-like fungi in Monkey-Ear Tiankeng was investigated. Seven species in the genera Akanthomyces, Beauveria, Cordyceps, and Samsoniella were identified based on internal transcribed spacer sequences and morphological characteristics. Eight new species in the genera Akanthomyces, Cordyceps, and Samsoniella were established and described according to a multilocus phylogenetic analysis and morphological characteristics. Our results revealed that Cordyceps-like fungi were abundant in Monkey-Ear Tiankeng, providing new insights into the diversity of Ascomycota in this special eco-environment. IMPORTANCE Karst Tiankeng has a special eco-environment and acts as a refugium for biodiversity. However, there are few reports on Ascomycota from the Tiankeng karst region. In this research, seven known species and eight new species in the genera Akanthomyces, Beauveria, Cordyceps, and Samsoniella were reported. The results showed that Cordyceps-like fungi are abundant in Monkey-Ear Tiankeng. Interestingly, the month of the sampling was November, which is not an active period of growth and reproduction for Cordyceps-like fungi. These results revealed that unconventional time sampling should not be ignored, especially for a special eco-environment, and provided new insights into the diversity of Ascomycota in this special eco-environment.

Phylogenetic, ecological and morphological characteristics reveal two new spider-associated genera in Clavicipitaceae.

Clavicipitaceous fungi are pathogenic to scale insects, white flies and other insect orders. However, a few species are spider-associated. Two new genera from China, Neoaraneomyces and Pseudometarhizium, are described based on phylogenetic, ecological and morphological characteristics. Two spider-associated species, Neoaraneomycesaraneicola, Pseudometarhiziumaraneogenum, and an insect-associated species Pseudometarhiziumlepidopterorum are included. The morphological characteristics of paecilomyces-like conidiogenous structures, present in many insect/spiders associated species make species-level identifications difficult. A phylogenetic analysis of the combined dataset (ITS, LSU, RPB2 and TEF), placed the two new genera in Clavicipitaceae. The new spider-associated species may be the result of convergent evolution to adapt to the ecological environment and may have undergone host jumping or altered their nutritional preferences.

Morphological characteristics and phylogenetic evidence reveal two new species of Acremonium (Hypocreales, Sordariomycetes).

Using chicken feathers as bait, Acremoniumglobosisporum sp. nov. and Acremoniumcurvum sp. nov. were collected from the soil of Yuncheng East Garden Wildlife Zoo and Zhengzhou Zoo in China. They were identified by combining the morphological characteristics and the two-locus DNA sequence (LSU and ITS) analyses. In the phylogenetic tree, both new species clustered into separate subclades, respectively. They were different from their allied species in their morphology. The description, illustrations, and phylogenetic tree of the two new species were provided.

Two new species of Hirsutella (Ophiocordycipitaceae, Sordariomycetes) that are parasitic on lepidopteran insects from China.

Hirsutella are globally distributed entomopathogenic fungi that offer important economic applications in biological control and biomedicine. Hirsutella was suppressed in favour of Ophiocordyceps affected by the ending of dual nomenclature for pleomorphic fungi in 2011. Currently, Hirsutella has been resurrected as a genus under Ophiocordycipitaceae. In this study, we introduce two new species of Hirsutella, based on morphological and phylogenetic analyses. Hirsutellaflava and H.kuankuoshuiensis are pathogenic on different species of larval Lepidoptera in China. Hirsutellaflava primarily differs from related species by its awl-shaped base; long and narrow neck, 24-40.8 × 2.2-2.5 µm; long and narrow cymbiform or fusoid conidia, 6.5-10 × 2.1-4.3 µm. Hirsutellakuankuoshuiensis has two types of phialides and distinctive 9.9-12.6 × 2.7-4.5 µm, clavate or botuliform conidia. The distinctions amongst the new species and phylogenetic relationships with other Hirsutella species are discussed.

Cryptic Diversity of Isaria-like Species in Guizhou, China.

Many Isaria-like species have recently been moved into more appropriate genera. However, more robust molecular phylogenetic analyses are still required for Isaria-like fungi to ensure accurate taxonomic identification. We analyzed these Isaria-like strains using multi-gene phylogenetics. Cryptic diversity was discovered in several Isaria farinosa strains, and two new species, Samsoniella pseudogunnii and S. pupicola, are proposed. Our results reveal that more attention needs to be paid to cryptic intraspecific diversity across different isolates and genotypes of the Isaria-like species, some of which will need to be transferred to Samsoniella. Interestingly, S. hepiali, with a very broad host distribution, has been widely used as a medicinal and edible cordycipitoid fungus.

Taxonomic and phylogenetic characterizations reveal four new species of Simplicillium (Cordycipitaceae, Hypocreales) from Guizhou, China.

Simplicillium species are commonly found from soil, seawater, rock surface, decayed wood, air and as symbiotic, endophytic, entomopathogenic and mycoparasitic fungi. Minority insect-associated species was reported. Simplicillium coccinellidae, S. hymenopterorum, S. neolepidopterorum and S. scarabaeoidea were introduced as the newly insect-associated species. The phylogenetic analyses of two combined datasets (LSU + RPB1 + TEF and SSU + ITS + LSU) revealed that S. coccinellidae and S. hymenopterorum were both nested in an independent clade. S. neolepidopterorum and S. scarabaeoidea have a close relationship with S. formicidae and S. lepidopterorum, respectively. S. neolepidopterorum can be easily distinguished from S. formicidae by ellipsoidal to cylindrical, solitary conidia which occasionally gather in short imbricate chains. S. scarabaeoidea could be easily distinguished from S. lepodopterorum by having longer phialides and larger conidia. Based on the morphological and phylogenetic conclusion, we determine the four newly generated isolates as new species of Simplicillium and a new combination is proposed in the genus Leptobacillium.

Preferences for core microbiome composition and function by different definition methods: Evidence for the core microbiome of Eucommia ulmoides bark.

The core microbiome, as a unique group of microorganisms, is an emerging research hotspot that provides a new opportunity to improve growth and production of a host. However, the subjectivity associated with the concept of "core microbiome" means there is currently no uniform definition method for the core microbiome. In this study, the strengths and limitations of four commonly used definition methods for the core microbiome were explored from composition to function based on the 16S rRNA gene dataset of Eucommia ulmoides bark from 25 different biogeographical regions in China. There were differences in the composition of the core microbiomes defined by the different methods. The four definition methods of phylogeny, membership, composition, and network connection contained 274, 10, 5, and 5 core OTUs (operational taxonomic units), respectively. In contrast, the core microbiomes defined by different methods displayed similarities in function. In addition, different definition methods showed varying preferences for abundant taxa, intermediate taxa, and rare taxa. Some core taxa defined by the definition method of phylogeny were significantly associated with pharmacologically active ingredients of E. ulmoides bark. The findings of this study suggest that although the core microbiomes defined by different methods have preferences in composition and function, the term refers to a group of microbes that are particularly notable and important for host-associated microbiomes. Therefore, we propose: (I) The definition method of the core microbiome should be selected according to the ecological problems faced; (II) A combination of multiple methods may comprehensively reveal the core microbiome at different levels of the host, and may also facilitate understanding of the ecological and evolutionary processes that govern host-microbe interactions.

Multi-gene phylogenetic evidence indicates that Pleurodesmospora belongs in Cordycipitaceae (Hypocreales, Hypocreomycetidae) and Pleurodesmospora lepidopterorum sp. nov. on pupa from China.

A new species, Pleurodesmospora lepidopterorum, isolated from a pupa, is introduced. Morphological comparisons and phylogenetic analyses based on multigene datasets (ITS+RPB1+RPB2+TEF) support the establishment of the new species. Pleurodesmospora lepidopterorum is distinguished from P. coccorum by its longer conidiogenous pegs located in the terminal or lateral conidiophores, and smaller subglobose or ellipsoidal conidia. A combined dataset of RPB1, RPB2, and TEF confirmed the taxonomic placement of Pleurodesmospora in Cordycipitaceae for the first time.

Bacterial Community Analysis and Potential Functions of Core Taxa in Different Parts of the Fungus Cantharellus cibarius.

Cantharellus cibarius is a widely distributed, popular, edible fungus with high nutritional and economic value. However, significant challenges persist in the microbial ecology and artificial cultivation of C. cibarius. Based on the 16S rRNA sequencing data, this study analyzed bacterial community structures and diversity of fruit bodies and rhizomorph parts of C. cibarius and mycosphere samples (collected in the Wudang District, Guiyang, Guizhou Province, China). It explored the composition and function of the core bacterial taxa. The analyzed results showed that the rhizomorph bacterial community structure was similar to mycosphere, but differed from the fruit bodies. Members of the Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium complex had the highest abundance in the fruit bodies. However, they were either absent or low in abundance in the rhizomorphs and mycosphere. At the same time, members of the Burkholderia-Caballeronia-Paraburkholderia complex were abundant in the fruit bodies and rhizomorphs parts of C. cibarius, as well as mycosphere. Through functional annotation of core bacterial taxa, we found that there was an apparent trend of potential functional differentiation of related bacterial communities in the fruit body and rhizomorph: potential functional groups of core bacterial taxa in the fruit bodies centered on nitrogen fixation, nitrogen metabolism, and degradation of aromatic compounds, while those in rhizomorphs focused on aerobic chemoheterotrophy, chemoheterotrophy, defense against soil pathogens, decomposition of complex organic compounds, and uptake of insoluble inorganic compounds. The analysis of functional groups of bacteria with different structures is of great significance to understand that bacteria promote the growth and development of C. cibarius.