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.

作　　者：李学宝[1] 秦明辉 施荣华[1] 陈光荣[1] 白永延[2] 

机构地区：[1]华中师范大学生命科学学院,武汉430079 [2]中国科学院上海植物生理研究所,上海200032

出　　处：《生物工程学报》1999年第4期482-488,共7页Chinese Journal of Biotechnology

基　　金：国家自然科学基金;湖北省自然科学基金

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。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.

关 键 词：芥菜型油菜 杀虫晶体蛋白 基因 遗传转化 抗虫性 

分 类 号：S565.403.4[农业科学—作物学] Q784[生物学—分子生物学]