Distribution and diversity of Dipteran-specific cry and cyt genes in native Bacillus thuringiensis strains obtained from different ecosystems of Iran

Abstract

One hundred and twenty-eight Bacillus thuringiensis isolates from Welds of diVerent ecological regions of Iran were collected to study the distribution and diversity of Dipteran-speciWc cry and cyt genes. The percentage of samples with Bt showed signiWcant diVerences between diVerent regions and also between diVerent Welds. The most Bt frequency was observed in the soil samples collected from Caspianic zone (7%) and soils of cotton (17%). Characterization of isolates was based on morphological characteristics of crystals, plasmid proWles and protein band patterns as well as PCR analysis using general and speciWc primers for 22 diVerent cry and cyt genes encoding proteins active against mosquitoes. Thirty-eight diVerent cry gene proWles were detected in this collection. Several of them were found to be diVerent from all previously published proWles and none of the previous researches reported these numbers of proWles. Strains containing cry2-type genes were the most abundant and represent 57.1% of the isolates. Strains harboring cry24 and cry10 genes were also highly abundant (38.7 and 32.8%, respectively). cry11, cry4, cry17, cry19, cry21, cry29, cyt1, and cry9 genes were less abundant, found in 25.7, 14.3, 11.4, 1.4, 4.3, 1.4, and 10% of the strains, respectively. Among the cry2 gene containing isolates, 37.5% strains harbored cry2Aa, 55% cry2Ab, 2.5% cry2Ac, and 5% other or novel cry2-type genes. Among the cry4 gene containing isolates, 0% strains harbored cry4A, 60% cry4B, and 40% cry4C, cry4D or novel cry4 type genes. Finally, based on crystal morphology, protein patterns and PCR, 21 strains were selected as potentially high Dipteran-active for bioassays. Also our results showed that some of the isolates may harbor minimum a putative novel cry gene.