Lan H Y, Wang C H, Zhang L H, Liu G Z, Wan L L, Chen Z H, Tian Y C
Institute of Genetics, Chinese Academy of Sciences, Beijing.
Sheng Wu Gong Cheng Xue Bao. 2000 Mar;16(2):142-6.
By Agrobacterium-mediated method, the cotyledonary petiole of good quality rape variety H165 was transformed with plant expression vector pBLGC which constitutively express beta-1,3-glucanase and chitinase genes. We obtained some Kanamycin(Kan)-resistant regenerational green shoots, with these shoots, PCR identification was conducted. Results showed that 30% green shoots which grew in medium of Kan 15 mg/L and 53% green shoots in Kan 25 mg/L had positive reaction. We also made dot blot analysis with those green shoots, some of them gave positive signal, indicating that the foreign genes had integrated into rape genome. Fungal challenge of these transgenic plants showed that some plants were much more resistant to Sclerotinia sclerotiorium than non-transgenic control plants.
通过农杆菌介导法，用组成型表达β-1,3-葡聚糖酶和几丁质酶基因的植物表达载体pBLGC转化优质油菜品种H165的子叶柄。我们获得了一些抗卡那霉素（Kan）的再生绿苗，并对这些绿苗进行了PCR鉴定。结果表明，在含15 mg/L Kan的培养基中生长的绿苗有30%呈阳性反应，在含25 mg/L Kan的培养基中生长的绿苗有53%呈阳性反应。我们还对这些绿苗进行了斑点杂交分析，其中一些给出了阳性信号，表明外源基因已整合到油菜基因组中。对这些转基因植株进行真菌挑战试验表明，一些植株对核盘菌的抗性比非转基因对照植株强得多。
