PCR-DGGE技术分析生物膜工艺微生物群落多样性  

Application of PCR-DGGE to analysis the microbial community diversity in biological biofilm reactor

作　　者：易绿云[1] 李红阳[1] 宋汕珂[1] 浦跃武[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《广东农业科学》2013年第11期153-156,共4页Guangdong Agricultural Sciences

摘　　要：为了揭示生物膜法工艺处理模拟生活废水过程总细菌群落结构的多样性,分别取系统启动初期和运行后期瓷板滑面、瓷板粗面、白胶板和黑实验台板上的附着生物膜,通过细胞裂解直接提取基因组DNA,以细菌通用引物进行16S rRNA基因V3区域PCR扩增,将PCR产物进行变性梯度凝胶电泳,获得微生物群落的DNA特征指纹图谱。结果表明,不同区段微生物群落间相似度最高达78.3%,最低达41.4%,不同生物膜样品既存在共同的微生物种属也存在特异微生物种属。但系统启动初期运行后期的生物膜群落结构较稳定,演替不明显。另外,对该生物膜群落的部分优势细菌进行了克隆测序和系统发育树分析,通过鉴定获得5条细菌16S rDNA序列,它们分别与梭状芽孢杆菌属、梭菌属和单胞菌属的同源性在97%以上,这些优势微生物在生物膜工艺去除有机物的过程中起重要作用。In ＂order to reveal microbial community structure diversity of bioflim reactor processing simulation domestic wastewater, the start-up initial stage and operation later stage biofilm from porcelain plate sliding surface, porcelain plate coarse surface, white rubber sheet and black experiment countertops were taken and the genomic DNA of microbial communities were extracted by the method of cell lysis. PCR-amplified 16S rRNA which using the universal primers and denaturing gradient gel electrophoresis （DGGE） were applied to obtain the DNA fingerprint profile of the microbial communities. The results showed that similarity between microbial communities of different blocking material were up to 78.3%, the lowest of 41.4%, the profile of DGGE showed that different biofilm samples had both different microbial species and common species. However, the biofilm community structure of both start-up initial stage and operation later stage are maintaining stability and the succession was obvious. In addition, part of the biofilm microbial community of dominant bacteria was analyzed through claning sequencing and phylogenetic analysis. Five bacterial 16S rDNA sequences were identified and with a homology over than97% when compared with the bacteria of Clostridia bacterium sp., Clostridium sp. and Brevundimonas sp.. These dominant microorganisms played important roles in the removal of organic pollutants in the bioflim system.

关 键 词：微生物多样性 斜板 生物膜法 变性梯度凝胶电泳 16S RRNA 

分 类 号：Q789[生物学—分子生物学]