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.
出处
《生物工程学报》
CAS
CSCD
北大核心
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
关键词
芥菜型油菜
杀虫晶体蛋白
基因
遗传转化
抗虫性
Brassica juncea, insecticidal crytal protein gene, genetic transformation, insect resisteance
