乳酸杆菌分离与分子鉴定（.docVIP

乳酸杆菌的分离与分子鉴定( 刘芸1，刘波1*，朱育菁1，马丽娜1，刘丹莹1，张衡宇2，刘贯锋2 （1.福建省农业科学院农业生物资源所，福建 福州 350003；2.福建农林大学，福建 福州 350002） 摘要：从市售酸奶样品中分离，筛选出2株乳酸杆菌CF7和CF9，它们能较好地利用豆类基质发酵形成乳酸饮料。本文利用形态学观察、生理生化方法对菌株进行了初步的分类和鉴定。菌株CF7，CF9均为杆状，无芽孢，革兰氏阳性，过氧化氢酶阴性。菌株CF7的生理生化特征与植物乳杆菌(Lactobacillus plantarum)一致，菌株CF9的生理生化特征与干酪乳杆菌(Lactobacillus casei)基本一致。并进一步通过菌株的16S rDNA基因扩增、克隆测序分析。测序结果在GenBank中进行同源性比较，并下载部分具有代表性属或种的16S rDNA序列，利用软件DNAMAN进行分子系统发育树分析，其鉴定结果与生理生化鉴定结果一致，表明菌株CF7为植物乳杆菌，菌株CF9为干酪乳杆菌。 关键词: 乳酸杆菌；16S rDNA；分子鉴定；植物乳杆菌；干酪乳杆菌 中图分类号: S879.1 文献标识码: A　　 Isolation and identification of Lactic acid bacteria strains LIU Yun1，LIU Bo1, ZHU Yu-jing1, XIAO Rong-feng1, MA Li-na1, ZHANG Heng-Yu2, LIU Guang-feng2 Agricultural Bio-Resources Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China; 2. Fujian Agricultural and Forestry University, Fuzhou 350002, China) Abstract: Two lactic acid bacteria strains CF7 and CF9 were isolated from a jogurt sample. They can be used to prepare bean lactobacillus beverage. In present research, they were observed under scanning and transmission electronic microscope and identified according to their physiological and bio-chemical properties. Their 16 S rDNA were subjected to amplification by PCR, clonning and sequencing. The results revealed that both of CF7 and CF9 were rod-shaded, Gram-negative and catalase-negative without spore. The biochemical characteristics of CF7 and CF9 were similar to those of Lactobacillus plantarum and Lactobacillus casei, respectively. The clonned 16 S rDNA were 1528 bp for CF7 (GenBank access #: HQ318714) and 1531 bp for CF9 (GenBank access #: HQ318715). The tested DNA sequences were compared with those in GenBank. The phylogenetic tree of 16 S rRNA, which was carried out with software of DNAMAN, indicated that CF7 and CF9 were Lactobacillus plantarum and Lactobacillus casei, respectively. Morphological charateristics, physiological and bio-chemical properties supported the results from the an