节点文献
芥菜型油菜抗虫转基因植株及其后代株系的研究
Transgenic Plants of Brassica juncea with Insect resistance and Genetic Analysis of the ProgenyThe Project Was Supported by National Foundation of Sciences of China(No. 39670389) and Foundation of Sciences of Hubei Province.
【摘要】 带有1 ~2 m m 子叶柄的芥菜型油菜子叶经农杆菌感染后,培养在附加10 ~20 mg/ L卡那霉素的 M S 选择培养基上筛选转化愈伤组织及不定芽。卡那霉素抗性苗相继在含30 ~50 m g/ L 卡那霉素的选择培养基上继代培养,再转移到含20 mg/ L 卡那霉素的生根培养基上诱导生根。以苏云金杆菌杀虫晶体蛋白基因为探针,进行 Southern blot 分子杂交,得到阳性结果。 P C R 分析也证明外源基因整合到油菜基因组并稳定传递到后代。转基因植株的抗虫性和卡那霉素抗性在自交后代中得到保持,筛选得到纯合的转基因植株后代株系
【Abstract】 Cotyledons with the petioles were cut from axenic seedlings and infected with Agrobacterium tumefaciens . After 2 days of cultivation, the explants were transferred to MS selection medium with 10~20mg/L kanamycin and 4.5 mg/L 6 BA for shoot differentiantion. Shoots with kanamycin resistance were subcultured on selection medium with 30~50 mg/L kanamycin for eliminating escaped non transformants, and then rooted on MS medium containing 20mg/L kanamycin and 0.5 mg/L IBA. Transgenic plants were transplanted into soil and grew in the field. DNA Southern blot hybridization and polymerase chain reaction showed that some of the plants were positive when probed with the insecticidal crystal protein gene. The transgenic plants exhibited resisteance to pest insects such as Laphygma exigua in leaf feeding experiments. Kanamycin resistance and insect resistance were maintained in the progeny. The foreign genes were delivered to the progeny according to Mendelian Law of single gene segregation.
【Key words】 Brassica juncea; insecticidal crytal protein gene; genetic transformation; insect resisteance;
- 【文献出处】 生物工程学报 ,CHINESE JOURNAL OF BIOTECHNOLOGY , 编辑部邮箱 ,1999年04期
- 【分类号】Q789
- 【被引频次】53
- 【下载频次】193