Establishment Of The Regeneration System Of Island Cotton(Gossypium Barbadense L.) And The Genetic Transformation Of SATI Gene

In this study, we developed the plant regeneration system of island cotton (Gossypium barbadense L.) by the way of somatic embryogenesis and the Agrobacterium mediated transformation of XinHai 16 embryogenic calli by which way transgenic plant containing SATI was obtained.1 .The The Optimum medium for callus induction of island cotton hypocotyls is MSB medium applied 0.1mg/L KT and 0.1mg/L 2,4-D, the induction rates of callus reaches to 95%. The best medium for callus differentiation is based on 1/3MS and the differentiation rates is about 60%.The optimum condition of callus proliferation is add the two times of KNO3, 0.01mg/L KT and 0.3mg/L NAA to MSB medium.2. The optimum medium of somatic embryogenesis is add two times of KNO3, 0.1g/L asparagines, 0.1g/L glutamine, 0.05mg/L IBA and 2.0g/L Gelrite to MSB medium. The rate of plant regeneration is 10% and the duatation of regeneration is about 6 to 7 months.3. Agrobacterium-mediated transformation system of island cotton (Gossypium barbadense) embryogenic calli was developed. And the transgenic plant of SATI gene was regenerated. The transformation condition was enhanced by ten days of pre-culture of embryogenic calli, optimum concentration of Agrobacterium with OD value of 0.5～0.6 and co-culture for 24 hour with no antibiotics. Then the cocultrued embryonic callus is transferred to the selection medium applied with 50mg/L Kan.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 rate of positive plantlets is about 19 % by PCR analysis. The surviving rate of graft was about 90%.