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冯广达,邓名荣,郭俊,王东东,朱红惠,张曦,梁浩亮,朱昌雄.利用PCR-DGGE溯源典型农村塘坝饮用水中的污染[J].环境科学学报,2011,31(3):468-475

Microbial source tracking of pollution in typical rural pond-drinking water using PCR-DGGE

张曦
1. 广东省微生物研究所, 广东省菌种保藏与应用重点实验室, 广东省微生物应用新技术公共实验室, 广州 510070; 2. 中国农业科学院农业环境与可持续发展研究所, 北京 100081

摘要：以华南地区典型农村塘坝饮用水为研究对象,采用基于大肠杆菌的两种特异性基因(β-D-葡萄糖苷酶编码基因uidA和膜外周磷通道蛋白编码基因phoE)的群体差异分析(PCR-DGGE),对饮用水中的污染情况进行了溯源研究.结果表明,采用富集培养的混合菌体DNA作为模板扩增大肠杆菌特异性基因更加实用.uidA和phoE的引物均具有较强的特异性.对Genebank中已有的两种特异性基因序列的分析结果表明,phoE的平均变异程度约为uidA的2倍,基于phoE的PCR-DGGE分析技术能够更好地反映样品之间的联系,适合在微生物溯源中应用.研究还表明,塘坝饮用水及其周围环境污染呈现面源特征,养殖废弃物是水体污染的主要来源.

Abstract：Taking the rural pond-drinking water in south China as the object,microbial source tracking of water pollution was done using PCR-DGGEto analyze the differences inEscherichia colicommunities based on two specific genes (the beta-D-glucuronidase coding gene uidAand the outer membrane phosphoporin protein coding gene phoE). According to our results,it is more practical to take all the cultured microbes DNAas templates in PCR. The primers of the gene uidAand phoEare of high specificity. Analysis of the two specific gene sequences in GenBank showed that the average degree of variation of phoEwas about twice that of uidA.PCR-DGGEbased on phoEcould exactly reflect the difference and connection among the samples and be more adapted to microbial source tracking than uidA. The pond-drinking water and its surrounding water were polluted with non-point pollution and breeding waste might be the main source of pollution.

Key words：microbial source tracking Escherichia coli PCR-DGGE beta-D-glucuronidase coding gene (uidA) outer membrane phosphoporin protein coding gene (phoE)