传统分离培养结合PCR-DGGE技术分析传统乳制品中的乳酸菌

Analysis of lactic acid bacteria in traditional dairy products by culture-independent and denaturing gradient gel electrophoresis(DGGE) methods

采用传统培养分离方法结合聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对传统乳制品中乳酸菌种群结构进行研究。结果表明:用传统分离与分子鉴定方法得到5种乳酸菌,其中,乳酸片球菌(Pediococcus acidilactici)为优势菌群,对通过PCR-DGGE方法得到的9条16S rDNA条带序列进行了比对,结果表明瑞士乳杆菌(Lactobacillus helveticus)的丰度最高,是酸奶样品中主要的优势菌群,乳酸片球菌(Pediococcus acidilactici)为次优势菌群。传统分离法与PCR-DGGE技术结合能够更有效、更全面地分析传统乳制品中微生物的群落结构及优势菌群。

In order to analyze lactic acid bacteria community structure in traditional dairy products and explore the predominant microbial species, both culture-dependent and culture-independent methods and denaturing gradient gel electrophoresis （DGGE） based on the sequence of 16S rDNA V6-V8 region gene were used to examine the total lactic acid bacteria counts. The results showed that 5 kinds of lactic acid bacteria were identified from traditional dairy products by the Culture-independent method, wherein the dominant bacteria belonged to Pediococcus acidilactici. By 16S rDNA V6-V8 region gene DGGE method, the most dominant bacteria was Lactobacillus helveticus, the second is Pediococcus acidilactici. This suggested that Independent culture combined with PCR-DGGE could be applied to analyze bacterial community structure and predominant bacteria in traditional dairy products effectively, thus provide a theoretical reference for the development of culture starter and that will give us more information of microorganism diversity.