采用高通量测序和基于培养的方法揭示冬虫夏草自然栖息地中的真菌群落组成。
Uncovering fungal community composition in natural habitat of Ophiocordyceps sinensis using high-throughput sequencing and culture-dependent approaches.
机构信息
School of Life Sciences, Guizhou Normal University, Huaxi University Town, Gui'an New District, Guiyang, 550025, China.
出版信息
BMC Microbiol. 2020 Nov 2;20(1):331. doi: 10.1186/s12866-020-01994-2.
BACKGROUND
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. 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.
RESULTS
A 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.
CONCLUSIONS
The 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.
背景
栖息于天然冬虫夏草中的真菌群落对高山草甸生态系统起着至关重要的生态作用，它们有助于感染宿主昆虫，影响冬虫夏草的发生，并蕴藏着潜在新型代谢物发现的宝库。然而，我们对冬虫夏草真菌群落的全面了解仍然难以捉摸。因此，本研究旨在结合高通量测序和培养依赖方法来揭示天然冬虫夏草中的真菌群落。
结果
通过高通量测序从天然冬虫夏草中获得了 280519 条高质量序列，它们属于 5 个真菌门、15 个纲、41 个目、79 个科、112 个属和 352 个假定的操作分类单位（OTU）。其中，在外部菌丝皮层中鉴定出 43 个属，优势属为 Ophiocordyceps、Sebacinia 和 Archaeorhizomyces，丰度分别为 95.86%、1.14%和 0.85%。从土壤微生境中共鉴定出 66 个属，优势属为 Inocybe、Archaeorhizomyces、未分类的 Thelephoraceae、Tomentella、Thelephora、Sebacina、未分类的子囊菌门和未分类的真菌，平均丰度分别为 53.32%、8.69%、8.12%、8.12%、7.21%、4.6%、3.08%和 3.05%。外部菌丝皮层中的真菌群落与土壤微生境有显著差异。同时，在 16°C 下使用七种培养基分离可培养真菌，通过 rDNA ITS 序列分析得到 77 株真菌菌株，属于 33 个属，包括 Ophiocordyceps、Trichoderma、Cytospora、Truncatella、Dactylonectria、Isaria、Cephalosporium、 Fusarium、Cosmospora 和 Paecilomyces 等。在所有可培养真菌中，Mortierella 和 Trichoderma 是优势属。
结论
两种方法得到的真菌群落结构的显著差异和重叠表明，高通量测序和培养依赖方法的结合将产生更多的信息。我们的研究结果全面了解了天然冬虫夏草的真菌群落结构，为冬虫夏草相关真菌提供了新的见解，并支持天然冬虫夏草的微生物群是新型生物活性代谢物发现的未开发来源。
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