*3.7. DNA Isolation and Amplification and Phylogenetic Analysis of 16S rDNA Gene*

Genomic DNAs from bacterial isolates were prepared using NucleoSpin kit (Macherey-Nagel, Germany) according to the recommendation provided by the manufacturer. PCR amplification of 16S rDNA gene from all the isolates was performed according to [27], using primers BF-20 (5 -ATCACGCGTAAAAATCT-3 ) and BR2-22 (5 -CCGCAATATCATTGGTGGT-3 ), resulting in about a 1500 bp length PCR product. The purified PCR fragments were sequenced using the ABI PRISM 3130xl Genetic Analyzer (Applied Biosystems) and by the BigDye v.3.1 sequencing kit (Applied Biosystems) (see Table S1 in Supplementary Materials). Obtained sequences were analyzed on the highest percentage of similarities using the Ez-taxon database [8] and the MEGA program version 7 [28]. The 16S rRNA phylogenetic tree was constructed using the maximum likelihood (ML) method based on the Tamura 3-parameter model [29], with 1000 bootstrap replications in the MEGA program.
