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基于16SrRNA基因克隆文库技术分析广西富钟水牛瘤胃产甲烷菌组成及多样性

DOI: 10.3969/j.issn.1006-267x.2014.12.012

Keywords: 富钟水牛,瘤胃,产甲烷菌,多样性

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Abstract:

本研究旨在利用16SrRNA基因克隆库技术分析广西富钟水牛瘤胃产甲烷菌组成及多样性.选取3头体况基本一致的健康雌性富钟水牛作为试验动物,采用机械破壁法提取瘤胃内容物总DNA,采用产甲烷菌引物Met86F/Met1340R扩增16SrRNA基因,构建16SrRNA基因克隆文库.结果表明,本试验共获得93个非嵌合体16SrRNA序列,按照97%的相似性划分为39个分类操作单元(OTU).其中,60个序列(15个OTU)与已培养细菌16SrRNA序列相似性≥97%,占总序列的64.5%;32个序列(23个OTU)与已培养菌16SrRNA序列相似性处于90%~(<97%);仅有1个序列与Methanomassiliicoccusluminyensis相似性<90%.系统发育树分析表明,98.9%的序列均属于甲烷杆菌目(Methanobacteriales),部分序列与Methanobacteriales中任何已知相似序列都相隔较远,它们可能代表Methanobacteriales中新的属或种.由上述结果可见,富钟水牛瘤胃产甲烷菌以Methanobacteriales为优势菌群,其中有许多未知的产甲烷菌需进一步分离培养并对其功能进行分析.

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