tailieunhanh - báo cáo khoa học: " Agrobacterium-mediated genetic transformation of Coffea arabica (L.) is greatly enhanced by using established embryogenic callus cultures"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Agrobacterium-mediated genetic transformation of Coffea arabica (L.) is greatly enhanced by using established embryogenic callus cultures | Ribas et al. BMC Plant Biology 2011 11 92 http 1471-2229 11 92 BMC Plant Biology RESEARCH ARTICLE Open Access Agrobacterium-mediated genetic transformation of Coffea arabica L. is greatly enhanced by using established embryogenic callus cultures Alessandra F Ribas1 Eveline Dechamp1 Anthony Champion2 Benoit Bertrand1 Marie-Christine Combes2 Jean-Luc Verdeil3 Fabienne Lapeyre3 Philippe Lashermes2 and Hervé Etienne1 Abstract Background Following genome sequencing of crop plants one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures co-cultivation with LBA1119 OD600 for five days at 20 C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures particularly a high auxin to cytokinin ratio the age of the culture optimum for 7-10 months of proliferation and the use of a yellow callus phenotype were the most important factors for achieving highly efficient transformation 90 . At the histological level successful transformation was related to the number of proembryogenic masses present. All the selected plants .