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《ACTA AGRICUL TURAE SHANGHAI》 1998-02
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HIGH EFFICIENCY REGENERATION AND Agrobacterium MEDIATED TRANSFORMATION OF HYPOCOTYL IN Brassica oleracea var.capitata WITH B.t.GENE

Wei Zhiming Huang Jiangqiu Xu Shuping Xue Hongwei(Shanghai Institute of Plant Physiology,Chinese Academy of Sciences,Shanghai 200032)Zhong Zhongxian Gu Zhenxiang Xu Tiwei Li Xian(Horticultural Research Institute,Shanghai Academy of Agricultural Sciences,S  
We have developed a high efficiency regeneration system of hypocotyl explants in Brassica oleracea var. capitata based on the study of the time of sampling and hormone composition in medium. The results indicated that when hypocotyls of 6~7 days wereplaced on the bud differentiation medium MSB+BA 2. 5 mg/L +ZT 1. 0 mg/L +IBA 0. 1mg/L(with 3 % sucrose and 0. 75 % agar,pH5. 8),supplemented with AgNO3 7. 5 mg/L,thehighest differentiation frequencies of 81. 67 % (Female parent "103")and 78. 56 % (male parent "60d Zaoyecai") could be obtained from hypocotyl explants, respectively. Furthermore,some factors affecting the transformation frequency,i. e. the kinds and concentrations of antibiotics for inhibiting the growth of Agrobacterium,preculture of hypocotyl explants,concentration of Agrobacterium and infection time,co-infection time,the time for supplementing theselecting antibiotic were also discussed. More than one hundred kanamycin-resistant plantshave been regenerated. All of them survived after transplantation and grew well in pots.Southern blotting analysis of plant DNA confirmed that nptⅡgenes have been introduced into the plant genome.
【Fund】: 上海市科委应用基因工程项目
【CateGory Index】: Q943
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