Genetic Diversity Study of Locally Isolated Bacillus thuringiensis Strains from Kuwait Using Random Amplified Polymorphic DNA Analysis

Abstract

Introduction: Bacillus thuringiensis (Bt) is significant bacteria in the field of insect biological control due to their insecticidal properties and its importance in agriculture. Aim: The objective of this study was to analyze genetic variation of 15 native isolates of Bacillus thuringiensis and four reference strains (total 19) using the random amplified polymorphic DNA (RAPD) technique, this allowed the analysis the genetic diversity of this species in the microbial populations from different soil samples of Kuwait. Methodology: We isolated 109 Bacillus thuringiensis strains, out of which, 15 strains were subspeciesthuringiensis, using culture and serological method. A rapid typing method of Bacillus thuringiensis local isolates from Kuwait soil was established using the RAPD technique. A single decamer primer was used to study and characterize Bacillus Spp, Bacillus thuringiensis var. thuringiensis, to differentiate the isolated species. Based on RAPD pattern, data were subjected to cluster analysis using Alpha Ease software. Results: We found three groups each, with two strains that had a similar pattern of DNA and one group that had three subspecies that have a similar DNA pattern. The rest of the eight isolates each had a unique pattern of DNA. These isolates were classified according to the same chemical and physical characterization, but were different genetically. Conclusion: In conclusion, using molecular methods for comparison of genomic DNA between different bacterial species of the same genus is a good measurement of genetic relations between different species, which could lead to discovery of new species unique to the local environment.