应用16S rDNA克隆文库解析人工快速渗滤系统细菌种群多样性

Analysis of Bacterial Community Composition by 16S rDNA Clone Library Sampling from Constructed Rapid Infiltration System(CRI)

摘要:

本文通过构建16S rDNA克隆文库开展了人工快速渗滤(CRI)系统表层(10 cm深)细菌种群多样性研究, 结果表明, 在CRI系统表层填料中细菌多样性十分丰富, 存在7个主要类群, 其中不可培养的酸杆菌纲(uncultured Acidobacteria)和其它不可培养细菌(uncultured bacteria)在整个克隆文库中比例最大, 比例为53.72%, 其次是浮霉菌纲(uncultured planctomycete)和β-变形菌纲(β-proteobacterium), 分别占文库比例的13.89%和8.33%; 克隆文库中反硝化菌的比例高于亚硝化单胞菌属细菌(0.925%), 没有出现Nitrospira属的克隆子。本文通过16S rDNA克隆文库揭示了在生物膜上存在的优势菌为不可培养菌, 而假单胞菌(Pseudomonas aeruginosa)和紫色色杆菌(Chromobacterium sp.)等在平板分离培养中出现的频率较高, 两种方法之间的结果存在差异。本研究采用16S rDNA克隆文库方法揭示的结果, 将为CRI系统生物降解的进一步提高提供依据。

关键词:人工快速渗滤系统(CRI), 16S rDNA克隆文库, 细菌种群多样性

Abstract:

In order to analyze the bacterial population diversity in surface layer(10 cm depth) of CRI, a bacterial 16S rDNA gene clone library was constructed. The results indicated that the bacterial community composition in surface Layer of CRI was high diversity and could be divided into 7 groups. The uncultured Acidobacteria and uncultured bacteria were the largest fraction(53.72% of total clones), followed by uncultured planctomycete (13.89%) and β-proteobacterium (8.33%); In clone library, the proportion of denitrifying bacteria was larger than that of nitrosobacteria and Nitrospira-like bacteria wasn’t found. The high percentage of uncultured bacterial clones predominated in clone library, but by plating culture method the majority colonies were only Pseudomonas aeroginosa and Chromobacterium sp., Which were the minority presented in clone library. The results demonstrated that either the bacterial community composition diversity or the abundance of phytogenetic groups analyzed by 16S rDNA clone library were much higher than those by plating culture method. The bacterial population diversity shown by 16S rDNA gene library is certain meaningful to reveal the activity profile of biodegradation in CRI.

Key words:Constructed Rapid Infiltration system (CRI), 16S rDNA clone library, Bacterial community composition