Identification of Lactobacillus plantarum by molecular methods and genomic anti-phage systems

Lactic acid bacteria (LAB) are the most commonly used microorganisms in industrial food fermentations. The multidrug resistance, bacteriocin production, various enzyme activities and probiotic properties of LAB , as well as resistance against bacteriophages are also important in industrial use areas. Microorganisms have developed various survival strategies in the evolutionary process. One of these is the defense of the genomic anti-phage system CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats). CRISPR is defined as clustered short palindromic repeats at regular intervals. This is an RNA-based immune system that is involved in the collaboration of CRISPR-related (cas) genes with DNA sequences of foreign genetic elements inserted between repeat sequences. In this study, rapid diagnosis of Lactobacillus plantarum species from 40 Lactobacillus sp. isolates was carried out in Real-Time PCR with species-specific probes and primers. 25 isolates were identified as Lactobacillus plantarum. For the determined 25 Lactobacillus plantarum, the presence of CRISPR-Cas loci of genomic anti-phage system was investigated by molecular and bioinformatics methods. Specific primer design for L.plantarum crispr arrays was performed. Crispr loci was found in 14 of L. plantarum isolates with classical polymerase chain reaction experiments. The results were analyzed using NCBI-Blast, CRISPRFinder and CrisprTarget databases. At Lactobacillus plantarum subsp. plantarum nF1-FD, 6 repeat, 5 spacer arrays were found. Crispr-related proteins (Dead / Deah box helicases) were detected in other isolates. Key words: L. plantarum, anti-phage, CRISPR-Cas