云南江城和黑井盐矿沉积物未培养放线菌多样性比较

Comparison of Actinobacterial Diversity in Jiangcheng and Heijing Saline Mines in Yunnan by Using Culture-independent Approach

摘要:

类群特异性引物的应用使得研究者可以对感兴趣的微生物类群进行针对性研究。围绕云南江城和黑井两个地区的3个盐矿样点沉积物中放线菌的多样性和群落组成, 我们通过放线菌特异性引物对总DNA进行16S rRNA基因扩增,经过克隆文库构建, 利用酶切并选择其中不同带型的133个克隆的16S rRNA基因插入片段进行测序。系统发育分析和统计学结果表明, 两地放线菌16S rRNA基因克隆广泛分布于整个放线菌门, 同时发现部分序列可能属于放线菌的新类群。分析结果还预示, 江城和黑井两地盐矿虽处云南不同地域含盐区, 但两地未培养放线菌物种多样性和系统发育关系均较为相似。

关键词:盐矿, 极端环境, 放线菌多样性, 免培养技术

Abstract:

Actinobacterial diversity in samples of saline mines was investigated by constructing three actinobacterium-specific 16S rRNA gene clone libraries from Jiangcheng salt mine and Heijing brine pits. The primers for the Class Actinobacteria were used to construct the 16S rRNA gene clone libraries. At least 469 purified clones were digested with Hae III for RFLP analysis, in order to separate the clones into groups according to their restriction patterns. Among of them, 133 sequences were selected and determined for their partial 16S rRNA gene sequences. The result of phylogenetic and statistic analyses indicated that actinobac- terial OTUs were closely related to most members of the Class Actinobacteria, and some OTUs may represent some unknown groups. Heijing and Jiangcheng located in different saline places of Yunnan, China. However, the communities, species diversity of Actinobacteria and Phylogenetic relationship from the two saline mines were similar.

Key words:Saline mine, Extreme environments, Actinobacterial diversity, Culture-independent approach