Comparation Of Ability Of Somatic Embryogenesis Of Cotton Genotypes And Production Of IAP-P35 (the Anti-apoptotic Baculovirus) Gene Transgenic Plants

In this study, we developed a plant regeneration system of upland cotton through somatic embryogenesis. The regeneration of transgenic plants of XLZ15 with iap-p35 gene is developed through transformation mediated by Agrobacterium tumefaciens.the main results obtained are listed below:1.Callus is initiated from hypocotyls on modified MSB medium with 0.1mg/LKT and 1.0mg/LIBA, the callus induction rates is reached to 90%. The optimum medium for callus differentiation is MSB medim with 0.1mg/L IBA and double amount of KNO3, the differentiation rates reached to 60%.2.The optimum medium of somatic embryogenesis is MSB medium with double amount of KNO3, without NH4NO3, 0.5g/L asparagines, 1.0g/L glutamine, 0.05mg/L IBA and 2.0g/L Gelrite. We obtained somatic embryo and regenerated plantlets of six upland cotton lines (ZM35, ZM44, XZL15, XLZ7, X9). The duatation of regeneration is reached about 6 to 7months. The two upland cotton lines (yellow river valleys and Xin Jiang cotton) showed the similar regeneable abilities, except a relative longer time taken during the somatic embryogenesis of cultivars of the Xing Jiang lines.3.Agrobacterium-mediated transformation system of upland cotton embryogenic calli were developed and obtained the transgenic plant of IAP-P35 genes. The transformation condition was enhanced by ten days of pre-culture of embryogenic calli, optimum concentration of Agrobacterium with OD value of 0.5 and co-culture for 24 hour with no antibiotics. The cocultrued embryonic callus is transferred to the selection medium applied with 50mg/L HyB and 200 mg/L cef. The rate of transgenic embryogenic callus is about 15%. Among them, about 5% callus can be developed to the normal plants. The putative transgenic plants were tested by PCR, the results showed that the transgenic has inserted into cotton genome.