Abstract
Background Ophiocordyceps sinensis (syn. Cordyceps sinensis), which is a parasite of caterpillars and is endemic to alpine regions on the Tibetan Plateau, is one of the most valuable medicinal fungi in the world. “Natural O. sinensis specimens” harbor various other fungi. Several of these other fungi that have been isolated from natural O. sinensis specimens have similar chemical components and/or pharmaceutical effects as O. sinensis. Nevertheless, the mycobiota of natural O. sinensis specimens has not been investigated in detail.Methodology/Principal FindingsBased on the technique of PCR-single-strand conformation polymorphism (PCR-SSCP), the mycobiota of three different sections (stromata, sclerotia, and mycelial cortices) from natural O. sinensis specimens were investigated using both culture-dependent and -independent methods. For the culture-dependent method, 572 fungal strains were isolated, and 92 putative operational taxonomic units (OTUs) were identified from 226 sequenced strains with the threshold of 97%. For the culture-independent method, 490 fungal clones were identified from about 3000 clones of ITS fragments from the whole-community DNA; based on PCR-SSCP analyses, 266 of these clones were selected to be sequenced, and 118 putative OTUs were detected. The overwhelming majority of isolates/clones and OTUs were detected from mycelial cortices; only a few were detected from stromata and sclerotia. The most common OTUs detected with both methods belonged to Ascomycota; however, only 13 OTUs were detected simultaneously by both methods. Potential novel lineages were detected by each of the two methods.Conclusions/SignificanceA great number of fungal species present in the mycobiota of naturally-occurring O. sinensis specimens were detected, and many of them may represent undescribed lineages. That only a few of the same OTUs were detected by both methods indicated that different methods should be used. This study increased our understanding about the fungal community structure of this valuable medicinal herb.
Highlights
As a large, diverse, and economically important group of organisms, fungi impact humankind in a myriad of ways and are vital components of most ecosystems and communities
A large fraction of microorganisms in nature have so far not been cultivated in the laboratory [5], and culture-dependent as well as culture-independent methods should be used together to obtain a holistic view of fungal community structures [6]
We investigated the fungal community structures of naturally-occurring O. sinensis
Summary
Diverse, and economically important group of organisms, fungi impact humankind in a myriad of ways and are vital components of most ecosystems and communities. In culture-dependent studies, fungal strains are isolated and identified based on morphological characteristics and/or certain DNA sequences. In culture-independent studies, molecular techniques are used to construct libraries of certain DNA fragments, and clones are subsequently sequenced [6]. The ITS regions are valuable for studies of fungal communities [9] and DNA barcoding [10]. ‘‘Natural O. sinensis specimens’’ harbor various other fungi. Cordyceps sinensis), which is a parasite of caterpillars and is endemic to alpine regions on the Tibetan Plateau, is one of the most valuable medicinal fungi in the world. Several of these other fungi that have been isolated from natural O. sinensis specimens have similar chemical components and/or pharmaceutical effects as O. sinensis. The mycobiota of natural O. sinensis specimens has not been investigated in detail
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