Coleopteran-specific and putative novel cry genes in Iranian native Bacillus thuringiensis collection |
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Authors: | Amin Nazarian Gholamreza Salehi Jouzani Ali Seifinejad Omolbanin Bagheri Khalil Alamisaeid |
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Affiliation: | a Department of Microbial Biotechnology and Biosafety, Agricultural Biotechnology Research Institute of Iran (ABRII), Mahdasht Road, P.O. Box 31535-1897, Karaj, Iran b Seed and Plant Improvement Institute, Shahid Fahmideh Blvd., P.O. Box 4119-31585, Karaj, IR Iran c Department of Agronomy and Plant Breeding, Agricultural and Natural Resources University of Ramin, Molasani, Ahvaz, Iran |
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Abstract: | The characterization of the strains containing Coleopteran-specific and also putative novel cry genes in Iranian native Bacillus thuringiensis collection is presented. Characterization was based on PCR analysis using 31 general and specific primers for cry1B, cry1I, cry3A, cry3B, cry3C, cry7A, cry8A, cry8B, cry8C, cry14, cry18, cry26, cry28, cry34 and cry35 genes, protein band patterns as well as their insecticidal activity on Xanthogaleruca luteola Mull. larvae. Forty six isolates (65.7%) contained minimum one Coleopteran-active cry gene. Based on universal primers, strains containing cry18 and cry26 genes were the most abundant and represent 27.1% and 24% of the isolates, respectively, whereas cry14, cry3, cry28, cry34, cry35, cry7, cry8 genes were less abundant, found in 14.2, 12.5, 10, 7, 7 and 5.6% of the strains, respectively. Based on specific primers, isolates containing cry1I were the most abundant (48.5%). Two strains containing Coleopteran-active cry genes showed higher activity against X. luteola larvae than B. thuringiensis subsp. morrisoni pathovar tenebrionis. Thirty isolates, when assayed for cry1C, cry5, cry6, cry8b, cry9, cry10, cry11, cry18, cry24 and cry35 genes, showed unexpected size bands. Cloning and sequencing of the amplicons allowed both the identification of known cry genes and the detection of putative novel cry1C sequences. |
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Keywords: | Bacillus thuringiensis Coleoptera Iran Molecular characterization Novel cry genes PCR Xanthogaleruca luteola Mull |
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