Isolation and characterization of Bacillus thuringiensis from soils in contrasting agroecological zones of Ethiopia

Abstract

Phenotypic and molecular methods were used to isolate and characterize B. thuringiensis from diverse agro-ecological zones of Ethiopia. Bioassays were used to test the insecticidal activity of B. thuringiensis strains against the major malaria vector, Anopheles arabiensis (Diptera). B. thuringiensis were isolated from 32% of the total 503 soil samples collected from the 16 agro-ecological zones. All sequenced isolates were 99%-100% identical to each other and to B. thuringiensis entries in Genbank. B. thuringiensis with similar 16S rRNA gene sequences from these different zones were characterized with regard to maximum growth rate and temperature optima for growth to test if there was local adaptation in these functional traits. The result showed a narrow temperature range around 30°C for maximal growth rate, and there were no significant differences between agro-ecological zones. Of 110 Bacillus thuringiensis isolates analyzed for the presence of crystal protein genes, 7 tested positive for cry 4, cry 11, and cyt toxin genes. Sequencing of these genes in positive strains demonstrated 99-100 % homology to known mosquitocidal cry and cyt genes in Bacillus thuringiensis subsp. israelensis. The present study shows that this biotechnologically important species is wide spread in Ethiopian soils and that it does not demonstrate local adaptation to temperature regimes, at least not for basic functions such as growth-temperature response. Our finding also pointed the potential for exploiting this species in vector control programs.