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Pak. J. Bot., 42(4): 2269-2279, 2010. |
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Updated: 09-12-10 |
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AGROBACTERIUM MEDIATED TRANSFORMATION OF SOYBEAN (GLYCINE MAX L.): SOME CONDITIONS STANDARDIZATION
MUHAMMAD ZIA*, ZARRIN FATIMA RIZVI, RIAZ-UR-REHMAN AND M FAYYAZ CHAUDHARY
Abstract: Present study was aimed to standardize some transformation conditions for soybean cultivar NARC-4 using Agrobacterium tumefaciens strain EHA 101 harboring pGUSintNPTII. Soybean half seed cotyledonary node method was opted. It was found that explant preparation in Agro-suspension culture resulted in highest transformation efficiency (48.3%) than in infection medium and water. One hour infection time was found optimum (55.9% transformation efficiency) in culture OD600 1.0. Co-cultivation of soybean half seed explants with Agrobacterium for five days showed better results as compared with three and four days. Two hr washing in washing medium containing 1g/L cefotaxime controlled prevalence of Agrobacterium in further steps. It was found that varying kanamycin concentration in selection medium resulted in high survival rate of transformed shoots. NARC-4 soybean cultivar showed better transformation efficiency than NARC-7 when genotype dependency was examined using Agrobacterium strain EHA101.
Department of Biotechnology, Quaid-i-Azam University, Islamabad, Pakistan 45320 *Correspondence E-mail: ziachaudhary@gmail.com; Ph: +92-51-9064-3011 |
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