PCR Based Amplification of Chitinase Gene from Native Isolates of Beauveria Bassiana


Abstract

Chitinase gene has been widely studied as one of the pest control agents because of its characteristic of degrading chitin. Chitinase is found in insects, nematodes, fungi, some algae and yeast. <i>Beauveria bassiana</i> is an insect pathogenic fungus successfully used as an insect pest control agent worldwide. In the present study, the entomopathogenic fungal isolates, <i>Beauveria</i> were isolated from the soil samples and insect cadavers collected from different places of Tamil Nadu. CTAB method was used for the isolation of DNA from entomopathogenic fungus. The ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and PN16 (5'-TCCCTTTCAACAATTTCACG-3') primers amplified a fragment of 930bp corresponding to the ITS1 and PN16 regions of the rDNA for the <i>Beauveria</i> isolates examined. Totally, 15 isolates were examined for the amplification of ITS region and all these isolates showed amplified product with size range of 930 bp which showed these isolates were belongs to <i>Beauveria</i> spp. A chitinase gene (1047 bp) was amplified from isolates of <i>B. bassiana</i> genomic DNA by PCR technique using forward primer 523ChitEcoRI (5'-ACATAGGAATTCATGGCTCCTTTTCTTCAAAC-3') and reverse primer 325ChitHindIII (5'-TACCTAACATGAACATTTAAGCTTTT-3'). All the isolates of <i>Beauveria</i> showed amplification at 1047bp. Chitinase has been considered as one of the most important factor for the pathogenicity of <i>B. bassiana</i> to various insect pests.

Keywords

Beauveria Bassiana, Chitinase Gene, Bbchit1 Gene

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