PCR-DGGE技术对中华开菲尔微生物菌群的分析

Analysis of Microbial Community Structures in Kefir Grains By PCR-DGGE Techniques

为了解中华开菲尔微生物菌群的结构特征,本论文运用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对开菲尔菌株发酵过程中微生物菌群的结构变化进行了实验分析,结果表明:细菌菌群DGGE图谱上出现有三种不同迁移位置的斑带,而酵母菌群DGGE图谱上只有一条斑带;经过DNA序列的对比分析可知:细菌菌群分别为肠膜明串珠菌(Leuconostoc mesenteroides)、马乳样乳杆菌(Lactobacillus kefiranofaciens)和开菲尔乳杆菌(Lactobacillus kefir),它们的序列同源性都达到100%;酵母菌群为德尔布有孢圆酵母(Torulaspora delbrueckii),其序列同源性为99%。本论文首次报道了德尔布有孢圆酵母在开菲尔菌落中的存在。

The dynamic variation of microbial flora in kefir liquor during its fermentation was studied by PCR-DGGE technique.The analysis result showed that 3 different visible bands in bacteria community and only 1 band in yeast community were obtained in their electrophoresis pattern.Anaylsis of 16SrDNA sequence blast showed that these sequences of all 3 different visible bands had a high homology with the sequence of 16S rDNA of lactic acid bacteria（LAB）.They belonged to Leuconostoc mesenteroides,Lactobacillus kefiranofacien and Lactobacillus kefir.The sequence of the band in yeast community had a high homology with the 16S rDNA of yeast Torulaspora delbrueckii.Yeast Torulaspora delbrueckii was,for the first time,found in kefir grains.