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以普通不结球白菜品种“中脚黑叶”和“矮脚黑叶”种子无菌苗的子叶为外植体材料 ,在附加 2 mg/LCPPU、0 .1m g/L NAA和 7.5 m g/L Ag NO3的稍作修改的 MS培养基上诱导不定芽。子叶 (柄 )和子叶切段诱导不定芽频率分别为 32 .5 2 %、4.35 %和 4.79%、11.5 5 %。应用农杆菌介导法将含有豇豆胰蛋白酶抑制剂基因 (Cp TI)和新霉素磷酸转移酶基因 (N PT )的重组质粒导入普通不结球白菜 ,获得具有卡那霉素抗性的再生植株。具有卡那霉素抗性的当代转基因植株 (T0 )及其自交后代 (T1 )植株经 PCR和 Southern blot分子检测 ,确认抗虫基因已整合到受体植株的基因组中 ,并通过有性生殖遗传给后代。体外生物学测定结果 ,显示抗虫性状在转基因植株自交后代植株中得到表达。
Cotyledons of common non-heading Chinese cabbage (Brassica campestris ssp.chinensis var. Chinensis) were used as explants with 2 mg / L CPPU, 0.1 mg / L NAA and 7.5 mg / Adventitious buds were induced on slightly modified MS medium of L Ag NO3. The frequency of adventitious bud induction by cotyledon (cotyledon) and cotyledon cuttings was 32.52%, 4.35% and 4.79%, 11.5% respectively. A recombinant plasmid containing the cowpea trypsin inhibitor gene (Cp TI) and the neomycin phosphotransferase gene (N PT) was introduced into common non-heading Chinese cabbage using Agrobacterium-mediated method to obtain kanamycin-resistant regeneration Plant. The present transgenic plants (T0) and their selfed offspring (T1) plants with kanamycin resistance were detected by PCR and Southern blot. It was confirmed that the insect-resistant genes had been integrated into the genome of the recipient plant, Inherited to future generations. In vitro biological assays showed that the insect-resistant traits were expressed in the selfing offspring of transgenic plants.