In this study, an efficient plant regeneration protocol in vitro and transformation by Agrobacterium-mediated method of Camellia sinensis was achieved, which would lay the foundation for genetic improvement of tea pla...In this study, an efficient plant regeneration protocol in vitro and transformation by Agrobacterium-mediated method of Camellia sinensis was achieved, which would lay the foundation for genetic improvement of tea plant by genetic engineering technology. The cotyledon callus of C.sinensis were used as the receptors for transformation by Agrobacterium tumefaciens EHA105 containing PS1aG-3. Some factors which affected the result of Agrobacterium-mediated transformation of C. sinensis were studied on the basis of GUS transient expression system. The optimum system of Agrobacterium-mediated transformation was that the cotyledon callus were pre-cultured for 3 d, and then infected by EHA105 for 15 min followed by 3 d co-culture in the dark on the YEB medium containing 150 μmol·L^(-1) acetosyringone(AS). The transient expression rate of GUS gene was 62.6%. After being delayed selective culture for 3 d, infected callus were transferred into the differentiation medium and the root induction medium both of which were supplemented with 100 mg·L^(-1) spectinomycin, and then resistant seedlings of C. sinensis were obtained. The conversion rate was 3.6%.展开更多
基金supported financially by the China Earmarked Fund for Modern Agro-industry Technology Research System (CARS-23)Jiangsu Provincial Agricultural Project (SXGC[2015]018)+3 种基金Fudiyingcai Talent Project of Jurong (2014)the Priority Academic Program Development of Jiangsu Higher Education InstitutionsJiangsu Provincial Project of Research and Practice on the Teaching Reform of Postgraduate Education (JGLX15_111)the Teaching Research Project of College of Horticulture of Nanjing Agricultural University
文摘In this study, an efficient plant regeneration protocol in vitro and transformation by Agrobacterium-mediated method of Camellia sinensis was achieved, which would lay the foundation for genetic improvement of tea plant by genetic engineering technology. The cotyledon callus of C.sinensis were used as the receptors for transformation by Agrobacterium tumefaciens EHA105 containing PS1aG-3. Some factors which affected the result of Agrobacterium-mediated transformation of C. sinensis were studied on the basis of GUS transient expression system. The optimum system of Agrobacterium-mediated transformation was that the cotyledon callus were pre-cultured for 3 d, and then infected by EHA105 for 15 min followed by 3 d co-culture in the dark on the YEB medium containing 150 μmol·L^(-1) acetosyringone(AS). The transient expression rate of GUS gene was 62.6%. After being delayed selective culture for 3 d, infected callus were transferred into the differentiation medium and the root induction medium both of which were supplemented with 100 mg·L^(-1) spectinomycin, and then resistant seedlings of C. sinensis were obtained. The conversion rate was 3.6%.
