Agrobacterium-mediated Genetic Transformation of Bentazon Resistant Gene (Cyp81A6) in Cotton

Muhammad Daud Khan, Lei Mei, Irum Raza, Saeed Hassan, Shui Jin Zhu

Abstract


Present study reports the Agrobacterium-mediated genetic transformation of bentazon resistant gene (Cyp81A6) in cotton genotypes (Coker 312 and Coker 201). Main aims were to genetically transform Cyp81A6 in cotton genome and to perform non-PCR and PCR-based confirmation studies regarding its stable transformation. The embryogeniccalli (EC) were first inoculated with the Agrobacterium tumefaciensstrain LBA4404 carrying foreign gene (Cyp81A6) and were transferred to the selection medium having kanamycin (100 mg/L) for 4 weeks. Embryogeniccalli were stunted in growth in both cultivars when transferred to selective MS medium. The kanamycin resistant (Knr) calli grew well after few sub-culturings. Knr and benr assays showed greater percentage of Knrembryogeniccalli. Also EC of Coker 312 showed greater resistance than Coker 201. Both genotypes profusely produced tulip shaped somatic embryos; however, their number was more in Coker 312 as compared with Coker 201. Molecular analysis of putative transgenic calli confirmed the presence of Cyp81A6 in the genomes of Coker 312 and Coker 201. Present study reveals that both cotton genotypes were efficient in integration of the gene and grew well on normal and selective MS media.


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ISSN: 2311-4630

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