Investigating the Diversity of Lactic Acid Bacteria in Tibetan Traditional Fermented Dairy Products by PCR-DGGE

doi:10.7506/spkx1002-6630-201401033

Abstract

Abstract:

Nineteen samples of Tibetan fermented dairy products were collected from eight pasturing areas and a polymerase
chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was established to investigate the biodiversity of
lactic acid bacteria in Tibetan fermented dairy products. After total DNA was extracted from dairy samples by a commercial
kit (Tiangen DP326), the V3 region of the 16S rRNA gene was amplified by nested PCR and touchdown PCR, and the PCRamplified
products were subjected to DGGE electrophoresis. All bands were excised from the gel and sequenced, and the
species of strains were identified and a phylogenetic tree was constructed. NTsys 2.10e software was used to analyze the
similarity of band profiles. Results showed that the microbial flora of lactic acid bacteria in Tibetan fermented dairy products
was composed of Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus crispatus,
Lactobacillus delbrueckii, Lactobacillus buchneri, Lactococcus raffinolactis, Leuconostoc mesenteroides, Lactobacillus
plantarum, Pediococcus pentosaceus, Lactococcus lactis and Streptococcus thermophilus. Therefore the conclusion is drawn
that Lactobacillus delbrueckii is the dominant species in most of the samples, and it simultaneously distributes in all eight
pasturing areas. This study has successfully established a PCR-DGGE method to investigate the biodiversity of lactic acid
bacteria in Tibetan fermented dairy products.

Key words: lactic acid bacteria, biodiversity, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), phylogenetic tree, dominant species

CLC Number:

TS252.54

JIANG Hou-yang1, CHEN Zhi-lan2, ZHAO Guo-hua1,3, YANG Ji-xia1,3,*. Investigating the Diversity of Lactic Acid Bacteria in Tibetan Traditional Fermented Dairy Products by PCR-DGGE[J]. FOOD SCIENCE, 2014, 35(1): 167-173.

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