基于MiSeq测序分析新疆泥火山土壤细菌群落多样性

【目的】以新疆乌苏泥火山土壤为研究对象,了解泥火山细菌群落结构及其时空动态变化。【方法】选择泥火山4种不同生境土壤在4、7、11月份采样,应用Illumina Mi Seq测序技术测定泥火山土壤细菌的16S r RNA基因V3–V4变异区序列,分析乌苏泥火山不同生境土壤细菌群落组成。【结果】泥火山土壤细菌在97%的相似水平下共得到OTU个数为29 005,在细菌门水平上共有38种细菌类群,Proteobacteria、Actinobacteria、Bacteroidetes为优势菌群,在属水平上共有72种细菌类群,其中含量最高的是未分类细菌;多样性分析表明生境D的丰度指数和多样性指数最高,将泥火山细菌群落多样性与理化因子结合分析,发现其多样性随着土壤养分的增加而基本降低,说明物种多样性指数与理化因子之间呈负相关关系;OTU水平的分析表明生境A的群落组成在时空动态上没有显著差异,其样品群落组成较为相似,而生境C的物种组成差异较大。【结论】相比较于传统方法,Mi Seq测序能够更全面解析环境样品中微生物多样性,揭示了乌苏泥火山群蕴含着丰富的微生物资源,这将为深入研究泥火山生态系统奠定基础,为合理利用和开发泥火山微生物资源提供指导。

Analysis the diversity of soil bacterial community from mud volcano in Xinjiang using MiSeq sequencing

Objective] In order to study soil bacterial community diversity and space-time analysis, we chose Usu mud volcano in Xinjiang province as the research object. Methods] we selected four different soil habitats to take samples in April, July, November, 2014. The Illumina MiSeq sequencing with 16S rRNA V3-V4 variable region was adopted to analyze the bacterial community structures of Usu mud volcano in Xinjiang province. Results] a total number of 29 005 operational taxonomic units (OTUs) were obtained from mud volcano soil under the similarity level of 97%. For phyla level, we have got 38 groups of bacterial that the Proteobacteria, Actinobacteria and Bacteroidetes were the dominant groups. For genus level, we have got 72 groups, but the most abundant genera were unclassified. Diversity index showed that the type D has a higher abundance effect and diversity index. Compared with the bacteria community diversity and physicochemical property, we found that bacteria diversity was decreased with the increasing of soil fertility, which shows negative correlations between bacteria diversity and physicochemical property. The results of PCA and heat map indicated that the community composition of type A has no significant variation with space-time dynamic changes, but type C is opposite. Conclusion] Compared with traditional method, Illumina MiSeq sequencing can provide a more comprehensive understanding of microbial diversity in the environmental samples and these results reveal clues that it is possible to contain abundant microbial resources, and it lays the foundation of studying ecosystem of mud volcano and provides guidance for exploitation and utilization of microbial resources more reasonably in Usu mud volcano.