Abstract
BackgroundThe fungal communities inhabiting natural Ophiocordyceps sinensis play critical ecological roles in alpine meadow ecosystem, contribute to infect host insect, influence the occurrence of O. sinensis, and are repertoire of potential novel metabolites discovery. However, a comprehensive understanding of fungal communities of O. sinensis remain elusive. Therefore, the present study aimed to unravel fungal communities of natural O. sinensis using combination of high-throughput sequencing and culture-dependent approaches.ResultsA total of 280,519 high-quality sequences, belonging to 5 fungal phyla, 15 classes, 41 orders, 79 families, 112 genera, and 352 putative operational taxonomic units (OTUs) were obtained from natural O. sinensis using high-throughput sequencing. Among of which, 43 genera were identified in external mycelial cortices, Ophiocordyceps, Sebacinia and Archaeorhizomyces were predominant genera with the abundance of 95.86, 1.14, 0.85%, respectively. A total of 66 genera were identified from soil microhabitat, Inocybe, Archaeorhizomyces, unclassified Thelephoraceae, Tomentella, Thelephora, Sebacina, unclassified Ascomycota and unclassified fungi were predominant genera with an average abundance of 53.32, 8.69, 8.12, 8.12, 7.21, 4.6, 3.08 and 3.05%, respectively. The fungal communities in external mycelial cortices were significantly distinct from soil microhabitat. Meanwhile, seven types of culture media were used to isolate culturable fungi at 16 °C, resulted in 77 fungal strains identified by rDNA ITS sequence analysis, belonging to 33 genera, including Ophiocordyceps, Trichoderma, Cytospora, Truncatella, Dactylonectria, Isaria, Cephalosporium, Fusarium, Cosmospora and Paecilomyces, etc.. Among all culturable fungi, Mortierella and Trichoderma were predominant genera.ConclusionsThe significantly differences and overlap in fungal community structure between two approaches highlight that the integration of high-throughput sequencing and culture-dependent approaches would generate more information. Our result reveal a comprehensive understanding of fungal community structure of natural O. sinensis, provide new insight into O. sinensis associated fungi, and support that microbiota of natural O. sinensis is an untapped source for novel bioactive metabolites discovery.
Highlights
The fungal communities inhabiting natural Ophiocordyceps sinensis play critical ecological roles in alpine meadow ecosystem, contribute to infect host insect, influence the occurrence of O. sinensis, and are repertoire of potential novel metabolites discovery
Quality analysis of sequencing data A total of 280,519 high quality fungal sequences were generated from six samples using Illumina Miseq sequencing
Richness and diversity of fungal community in natural O. sinensis In all six samples derived from external mycelial cortices and soil microhabitat, the coverage values close to 100%, and the rarefaction curves were asymptotic, indicating that overwhelming majority of the fungal species were covered (Fig. 1)
Summary
The fungal communities inhabiting natural Ophiocordyceps sinensis play critical ecological roles in alpine meadow ecosystem, contribute to infect host insect, influence the occurrence of O. sinensis, and are repertoire of potential novel metabolites discovery. A comprehensive understanding of fungal communities of O. sinensis remain elusive. The present study aimed to unravel fungal communities of natural O. sinensis using combination of high-throughput sequencing and culture-dependent approaches. The caterpillar fungus Ophiocordyceps sinensis parasitizes the larvae of Thitarodes that belongs to family Hepialidae (Lepidoptera) form the fungus-caterpillar complex (Chinese Cordyceps) [1]. O. sinensis harbour a variety of bioactive metabolites, e.g., cordycepin, cordycepic acid, polysaccharides, ergosterol, mannitol and macrolides, etc. It may be considered as key factors in the coevolution of O. sinensis and its host, coevolution of symbiotic and associated microbiota. Illuminating mysterious drive factor in the occurrence and maturation of O. sinensis, enhancing the yield and discovery of its alternative substitutes, have already become hot topics
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