甘蓝型油菜抗虫转基因植株及其抗性分析

通过油菜子叶外植体－农杆菌共培养法将苏云金杆菌杀虫蛋白基因导入甘蓝型油菜，获得抗虫的转基因植株。带有１￣２ｍｍ子叶柄的油菜子叶经农杆菌感染后，共培养２￣３天，然后转移到附加１５ｍｇ／Ｌ卡那霉素的ＭＳ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２０￣５０ｍｇ／Ｌ卡那霉素的选择培养基上继代培养，再转移到含２５ｍｇ／Ｌ卡那霉素的生根培养基上诱导生根。以苏云金杆菌杀虫蛋白基因为探针，进行１

Insect-resistant Transgenic Plants of Brassica napus and Analysis of Resistance in the Plants

Cotyledons, each with a 1-2mm petiole at its base, were cut from axenic seedlings and infected with Arobacterium tumefaciens. After 2-3 days of cocultivation, the cotyledon explants were transferred to MS selection medium containing 15mg / L kanamycin and 4.5ing / L 6-BA to induce shoot differentiation. Kanamycin-resistant shoots were subeultured on selection medium with 20-50mg / L kanamycin for 3-6 months for eliminating escaped non-transformants, and then rooted on MS medium containing 25mg/ L kanamycin and 0.1mg/ L NAA. Whole Plants were transplanted into soil and grew in the field. DNA Southem 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 tolerant to pest insects such as lerhygma exigua and Pieris rapae 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.