Investigation and analysis of microbiological communities in natural Ophiocordyceps sinensis

Abstract

Ophiocordyceps sinensis is a fungus that parasitizes caterpillars, and more than 30 species of filamentous fungi have been isolated from its fruiting body. However, its microbiological diversity remains unclear. Based on the clone library and quantitative PCR techniques, the bacterial flora and mycobiota of 3 different samples (larva, stromata/sclerotia, and surface soil) from natural O. sinensis specimens were investigated using primer sets that targeted the 16S rRNA gene and internal transcribed spacer region of ribosomal DNA. The results showed that the abundance of bacterial and fungal communities in the soil attached to the surface of O. sinensis was (6.4 ± 1.4) × 10(6) and (6.0 ± 0.3) × 10(7) copies/g dry matter, respectively, which was the highest compared with that in the larva and stromal samples. The main groups of bacteria in the O. sinensis samples were Proteobacteria and Actinobacteria, while Ascomycota was the most dominant fungal group in the 3 samples. At the genus level, Geomyces, Phoma, and Trichocladium were the dominant genera in the larval sample, while Geomyces and Cladosporium were the dominant genera in the stromal sample. In conclusion, a great number of bacterial and fungal species were present in naturally occurring O. sinensis specimens, and there was a high diversity of bacterial and fungal communities. These findings contribute to the understanding of the bacterial and fungal community structure of this valuable medicinal fungus and lay the foundation for the future discovery of new medicinal microorganism resources.

Pyrosequencing analysis revealed complex endogenetic microorganism community from natural DongChong XiaCao and its microhabitat

“…So there was a little piece of information on endogenetic microorganism from natural DCX in the database. The bacterial population of DCXC was only one report available [30], and the effects of bacteria on the growth process and medicinal composition of natural DCXC are unclear. Based on the results analysis of this study, the main groups of bacteria in the different samples were Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria and Firmicutes.…”

“…The samples were kept in aseptic valve bags and transported in an ice box to laboratory of Plant Biotechnology R&D Center of Shanghai Jiao Tong University, Shanghai, China. To get rid of the microorganisms inhabiting the surface of natural DCXC, the samples were treated with different disinfectants, including 75 % ( v/v ) ethanol, 2.5 % sodium hypochlorite, 0.01 % mercuric chloride individually, and then washed with sterilized water three times [30]. Subsequently, all the natural DCXC samples collected from three sampling locations were homogenized and divided into stroma, sclerotia and its microhabitat samples, including external mycelial cortices (membrane covering the larvae) and the soil adhering to the surface of the membrane covering DCXC, to investigate the microorganism community (Fig.…”

“…According to traditional Chinese medicine theory, the geoherbalism formation of TCMM is based on many factors, among which the microenvironment of rhizosphere soil plays an important role in the formation of secondary metabolism in medicinal plants. Therefore, based on the diversity of microbial communities by the clone library approach [30], the natural DCXC couple with its microhabitat soil was divided into four parts: stroma, larva, external mycelial cortices and soil, to in each sample using high throughput pyrosequencing platform (Roche/454 GS-FLX Titanium System) and quantitative real-time PCR (qPCR) techniques. The aim of the study was to detect the microbial diversity and community structure of natural DCXC and its microhabitats.…”

Identification and Bioactive Potential of Endogenetic Fungi Isolated from Medicinal Caterpillar Fungus Ophiocordyceps sinensis from Tibetan Plateau

“…For the research of Chinese Cordyceps, more and more investigators focused on their scientific interest in endogenetic fungi of Chinese Cordyceps and the fungal diversity (Zhang et al, 2010;Xia et al, 2015;Xia et al, 2016a;Xia et al, 2016b). Although endogenetic fungi of Chinese Cordyceps had been investigated and studied extensively, there was a little minor fault existing in the research process.…”

Suitability and Advantages of Abundance-Weighted Algorithms for the Holistic Comparison of Sample Systems

“…Scientists have reported the molecular heterogeneity of C. sinensis-associated fungi of the genera Hirsutella, Paecilomyces and Tolypocladium as well as Geomyces pannorum, Cladosporium macrocarpum, Phaeosphaeria pontiformis and Neosetophoma samarorum (in total, more than 90 species spanning at least 37 genera) [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21]. SNP mass spec genotyping and amplicon sequencing with or without molecular cloning have identified the mutant ITS sequences of at least 17 genotypes of O. sinensis fungi with multiple, scattered point mutations or DNA segment substitution hereditary variations from the natural C. sinensis insect-fungi complex [1][2][3][4][10][11][12][13]19,[22][23][24][25][26][27][28].…”