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目的:运用根癌农杆菌介导的方法,成功建立神农香菊遗传转化体系。为对神农香菊性状进行定性改良奠定基础。方法:以神农香菊无菌苗幼嫩叶片为受体材料,通过根癌农杆菌介导的遗传转化方法,建立了神农香菊遗传转化体系。结果:神农香菊遗传转化的卡那霉素最适分化临界耐受浓度为45 mg/L,最适生根临界耐受浓度为60 mg/L。头孢霉素抑菌浓度在延迟培养阶段选用200 mg/L最佳,筛选培养阶段选用100 mg/L最佳。预培养3d,侵染时间30 min,共培养3 d,延迟培养3 d为神农香菊叶盘最优遗传转化体系。结论:经PCR检测,初步获得转基因植株19株。成功建立了神农香菊的遗传转化体系。
Objective: The Agrobacterium tumefaciens genetic transformation system was successfully established by Agrobacterium tumefaciens-mediated method. For the Shennongxiang traits of qualitative improvement and lay the foundation. Methods: The young leaves of A. sinensis germplasm were used as the receptor materials. The Agrobacterium tumefaciens genetic transformation system was established by Agrobacterium tumefaciens-mediated genetic transformation. Results: The critical tolerance concentration of kanamycin genetically transformed by Shennongxiangju was 45 mg / L, and the critical tolerable rooting concentration was 60 mg / L. Cefotaxime antibacterial concentration in the delay of incubation phase selection of 200 mg / L best screening stage selection of 100 mg / L best. Pre-culture 3d, infection time 30 min, co-culture for 3 days, delayed culture for 3 days for Agrocybe media optimal genetic transformation system. Conclusion: 19 transgenic plants were obtained by PCR. Successfully established Shennongxiangju genetic transformation system.