Isolation and characterization of Bacillus thuringiensis strains from olive‐related habitats in Turkey

• Published in: Journal of applied microbiology Vol. 104; no. 2; pp. 515 - 525
• Main Authors: Cinar, C., Apaydin, O., Yenidunya, A.F., Harsa, S., Gunes, H.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.02.2008; Blackwell Science
• Subjects: 16S‐internal transcribed spacer rDNA restriction fragment length polymorphism; Bacillus thuringiensis; Bacillus thuringiensis - genetics; Bacillus thuringiensis - isolation & purification; Bacillus thuringiensis - ultrastructure; Bacterial Proteins - genetics; Biological and medical sciences; characterization; crystal protein; Crystallization; DNA, Ribosomal - analysis; Environmental Microbiology; Fundamental and applied biological sciences. Psychology; Genetic Variation; Genome, Bacterial; Insecticides; Microbiology; Microscopy, Electron, Scanning; Microscopy, Phase-Contrast; Olea - microbiology; olive‐related habitats; polymerase chain reaction; Polymorphism, Restriction Fragment Length; Spores, Bacterial; Turkey; Asia; Turkey; 16S-internal transcribed spacer rDNA restriction fragment length polymorphism; Applied microbiology; Bacillus thuringiensis; Crystals; Ribosomal DNA; Spacer DNA; olive-related habitats; Bacillaceae; Protein; Characterization; Olive; Polymerase chain reaction; Restriction fragment length polymorphism; Bacillales; 16S-RNA; Bacteria; Isolation; crystal protein
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2007.03571.x

Aims:  To isolate Bacillus thuringiensis strains from different olive‐related habitats (olive groves and olive oil factories) in Turkey and to characterize these strains by molecular methods. Methods and Results:  A total of 150 samples, consisting of olive grove soil, green olive leaves, olive leaf residues, animal faeces, olive pomace and dust, were examined for the presence of B. thuringiensis. One hundred B. thuringiensis strains were isolated from 54 environmental samples (36%) and characterized in terms of crystal morphology, cry and cyt gene content by polymerase chain reaction, plasmid profiles and 16S‐internal transcribed spacer ribosomal DNA restriction fragment length polymorphism (16S‐ITS rDNA RFLP). The highest percentage of samples containing B. thuringiensis was found in 38 out of 54 total soil samples (70%). Of the 100 B. thuringiensis isolates, the most frequent crystal shapes were irregularly shaped (24%), spherical‐irregular pointed (19%), cuboidal (17%) and spherical (16%). The cry1 plus cry4 genotype was the most abundant genotype in our collection (21%). RFLP analysis of the amplified 16S‐ITS rDNA revealed 11 distinct patterns for the isolates and 10 reference strains. Conclusions:  Bacillus thuringiensis isolates showed a great genetic diversity and crystal shape heterogeneity. Significance and Impact of the Study:  This is the first study on the isolation and characterization of B. thuringiensis from olive‐related habitats in Turkey. No correlation was observed between the cry genotypes and insecticidal crystal shapes of the isolates. Restriction profiles of 23% of the isolates were found to be different from those of the 10 reference strains used.