16S rRNA基因序列方法分析传统发酵菜中乳酸菌多样性

以发酵甘蓝卤水中微生物的总DNA为模板,用扩增革兰氏阳性菌16S rRNA基因的方法研究中国发酵甘蓝菜卤中乳酸菌的多样性。获得了42种含有1.5kb的16S rRNA基因插入核苷酸片段的克隆菌。利用克隆菌落核苷酸的部分序列和NCBI数据库中搜索到的相似序列进行对比后作出鉴定,结果得出4种独特的菌落,鉴定为Lactobacillus acidifarinae、植物乳杆菌(Lactobacillus Plantarum)、融合乳杆菌(Weissella confusa)和足球菌属(Pediococcus sp.)。此外,还发现两种非核糖体序列。用这种分子方法发现了常规分离培养技术未鉴定的乳酸菌,如Lactobacillus acidifarinae。

Determination of Lactic Acid Bacterial Diversity in the Traditional Fermented Vegetable by 16S rRNA Sequence Analysis

In order to study the lactic acid bacteria diversity of fermented vegetable,a total DNA of microbe in fermented cabbage brine as a template and PCR amplification of 16S rRNA for Gram-positive and related bacterial groups were used. Forty-two clones were obtained and sequenced to 1.5 kb nucleotides. These partial sequences were identified by submission to the non-redundant nucleotide database of NCBI. The identified sequences were aligned with reference sequences of the closest related organisms. This analysis indicated that only 4 sequences were unique clones and these were identified as Lactobacillus acidifarinae,Lactobacillus Plantarum,Weissella confusa,and Pediococcus sp. Two non-ribosomal sequences were also detected. Use of this molecular method permitted detection of lactic acid bacteria different from those previously isolated and identified by culture techniques.