长寿沙田柚Ri T-DNA转型根优化培养体系研究.pdfVIP

中国农业科学 2005,38(12):2496-2501 Scientia Agricultura Sinica 长寿沙田柚Ri T-DNA转型根优化培养体系研究 1,2 1 2 杨晓红 ，孙中海 ，仝瑞建 1 2 ( 华中农业大学柑桔研究所/教育部园艺植物生物学重点实验室，武汉 430070 ； 西南大学园艺园林学院，重庆 400716) 摘要：设计了长寿沙田柚不同外植体在光、暗条件下进行发根农杆菌遗传转化试验，建立了长寿沙田柚 Ri T-DNA转型根优化培养体系，该体系为：用对数期的发根农杆菌A4菌株感染活化胚中单片子叶的创伤腹面，25℃ ±2℃暗培养 25～30 d，多数子叶上发出转型根（毛状根），遗传转化率达（83±11）%。继代脱菌 5 代后获得无 菌转型根。外植体活化采用 MT 培养基，继代脱菌培养采用 MS 培养基。采用这个体系获得了大量激素自主的长寿 沙田柚Ri T-DNA转型根。获得的新鲜转型根白色，长1.14±0.07 cm，粗0.73±0.04 mm，负向地性生长。 关键词：长寿沙田柚；发根农杆菌；Ri T-DNA转型根；优化培养体系 Optimized Culture System for Ri T-DNA Transformed Roots of Citrus grandis cv. Changshou Shatian You 1, 2 1 2 YANG Xiao-hong , SUN Zhong-hai , TONG Rui-jian (1 Key Laboratory of Horticultural Plant Biology, Ministry of Education / Institute of Citrus, Huazhong Agricultural University, Wuhan 430070; 2College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400716) Abstract: Genetic transformation experiments of the different exploids from Citrus grandis cv. Changshou Shatian You infected with Agrobacterium rhizogenes were carried out under dark and light. The optimized culture system for Ri T-DNA transformed roots of C. grandis cv. Changshou Shatian You was constructed as follows: After the ventral wounded striations on the single activation cotyledon were inoculated by A. rhizogenes A4 (logarithmic period), they were cultured at the temperature of 25±2 ℃ in the dark for 25-30 days, some transformed roots were generated