Isolation, identification and fingerprinting of lactobacillus bacteria in dairy and fermented products of Lorestan province and determination of genetic relationships between them using RAPD-PCR marker
Subject Areas : MicrobiologyS. Daraie 1 , B. Doosty 2 , K. Samiei 3
1 - MSc Student of the Department of Biology, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran.
2 - Associate Professor of the Department of Biology, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran.
3 - Assistant Professor of the Department of Agronomy and Plant Breeding, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran.
Keywords: RAPD-PCR, phylogenetic, Lactic Acid, Lactobacillus,
Abstract :
Introduction: Lactic acid bacteria (LAB) are the main and natural components of the microflora of the digestive system. Identification of lactic acid bacteria and their use in the food industry is very important. Material and Methods: In order to isolate natural lactic acid bacteria, samples of local yogurt and traditional tarkhineh were collected from different regions of Lorestan province, with respect to suitable geographical distances and different weather conditions. After separating gram-positive and catalase-negative bacteria, 20 lactic acid bacteria were selected and then using specific primers, the 16SrDNA gene was amplified and after alignment and comparison with the NCBI database, the type of bacteria was determined. In order to estimate the genetic diversity between the studied bacteria using the RAPD marker, 4 primers were used. Finally, a phylogenetic tree was made between the studied bacteria. Results: Based on the antibiogram test, it was found that all the studied bacteria were resistant to kanamycin and amikacin antibiotics and sensitive to gentamicin, vancomycin and tetracycline antibiotics. Bacteria isolated from tarkhineh with 97% similarity and bacteria isolated from yogurt with 89% similarity were placed in the same group. Two bacterial strains isolated from yogurt were 94% similar to L.rhamnosus bacteria. In relation to the two bacterial strains isolated from tarkhineh sample, the results showed that these two bacteria were 98% similar to L. pentosus bacteria. Based on the results of grouping by RAPD-PCR marker, the studied bacteria formed 6 groups with a similarity coefficient of 57%, and one genotype alone formed one group. Regarding the geographical distances and the grouping obtained, the bacteria samples isolated from the traditional products of similar geographical areas were placed in close groups. Conclusion: The results generally showed that the use of genetic markers can be very effective in identifying the species and strains of lactic acid bacteria in dairy and fermented products.
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