Molecular identification of chlorpyrifos and Malathion degrading bacteria

Abstract

Fifteen strains of bacteria were successfully isolated from agriculture soil samples contains pesticides and from soil moisture around them. The isolates were capable of utilizing chlorpyrifos (Cp) and malathion as the sole source of carbon, phosphorus and energy. High yielding isolate KSCM-08 was identified based on16SrRNA sequencing. Genomic DNA was successfully extracted from bacterial isolate KSCM-08, and the total DNA content was determined to be 734±23 µg/gm. The absorbance ratio of extracted DNA at A260/280 has been found to be 1.96±0.2. 1381 nucleotide sequence of the PCR amplified 16s rRNA gene from bacterial isolate KSCM-08. The 16s rRNA gene sequence comprises 344 bp of adenine (A), 280 bp of thymine (T), 430 bp of guanine (G), and 327 bp of cytosine (C). The G+C percentage was determined to be 54.8%. The BLAST report demonstrates the similar sequences with the KSCM-08 16s rRNA sequence, along with their percentage of identity. The BLASTn analysis of the 1381 bp length 16s rRNA gene sequence of KSCM-08 against the GenBank database revealed that KSCM-08 exhibits similarity with Bacillus species. The evolutionary divergence of bacterial isolate KSCM-08 with its relative members was determined. A satisfactory result was established by the use of the 16S rRNA gene as a marker to evaluate the phylogenetic relationship and the bacteria was confirmed as Bacillus safensis.