Key Laboratory of Systematic Mycology and Lichenology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
PLoS One. 2010 Dec 15;5(12):e15570. doi: 10.1371/journal.pone.0015570.
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 FINDINGS: Based 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/SIGNIFICANCE: A 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.
冬虫夏草(又名中华虫草菌)是一种寄生于高山地区鳞翅目幼虫的寄生虫,是世界上最有价值的药用真菌之一。“天然冬虫夏草标本”中还存在其他各种真菌。从天然冬虫夏草标本中分离出的一些其他真菌具有与冬虫夏草相似的化学成分和/或药用功效。然而,天然冬虫夏草标本的真菌群尚未得到详细研究。
方法/主要发现:基于 PCR-单链构象多态性(PCR-SSCP)技术,采用培养依赖和非依赖的方法,对天然冬虫夏草标本的三个不同部位(子实体、菌核和菌丝皮层)的真菌群进行了研究。对于培养依赖的方法,从 226 株测序菌株中分离出 572 株真菌菌株,鉴定出 92 个假定的分类操作单元(OTUs),阈值为 97%。对于非培养依赖的方法,从整个群落 DNA 的 ITS 片段中约 3000 个克隆中鉴定出 490 个真菌克隆;基于 PCR-SSCP 分析,选择其中的 266 个克隆进行测序,检测到 118 个假定的 OTUs。绝大多数分离株/克隆和 OTUs 是从菌丝皮层中检测到的,只有少数是从子实体和菌核中检测到的。两种方法检测到的最常见的 OTUs 属于子囊菌门;然而,只有 13 个 OTUs同时被两种方法检测到。两种方法都检测到了潜在的新谱系。
结论/意义:从天然冬虫夏草标本的真菌群中检测到了大量的真菌物种,其中许多可能代表未描述的谱系。只有少数相同的 OTUs 被两种方法同时检测到,这表明应该使用不同的方法。本研究增加了我们对这种有价值的药用植物真菌群落结构的理解。