农杆菌介导转化甜菜影响因素的研究

[目的]以甜菜无菌苗的叶柄为外植体材料,对农杆菌转化条件和方法进行较为系统地比较研究,以期建立一种转化频率高、稳定性好、通用性较强的甜菜遗传转化技术体系.[方法]对影响农杆菌介导的不同因素分别进行比较试验.[结果]对甜菜叶柄进行培养后用不同菌液浓度OD600=0.3～0.4的农杆菌感染8～10 min,脱菌处理后接入卡那霉素浓度为100 mg/L、头孢霉素浓度为500 mg/L的选择培养基中进行筛选,获得的抗性芽较多,抗性芽分化频率可达到34.3;.[结论]初步建立了一种转化频率高、稳定性好、通用性较强的甜菜遗传转化技术体系,通过植物转基因技术为选育高产、优质、广谱抗病的甜菜品种奠定基础.

Factors Influencing the Sugarbeet Transformation Mediated by Agrobacterium

【Objective and Method】Using sugarbeet petioles as explants,the factors that affected transformation condition and method were investigated,such as preculture time,concentration of Kanamycin(Kan),concentration of Cefazolin sodium(Cef),infection time and infection concentration of Agrobacterium,the purpose is to establish a stable and efficient Agrobacterium mediated transformation system.【Result】When the sugarbeet petioles were infected with infection concentration of OD600=0.3-0.4 and infection time of 8-10 minutes it was then added into the seclective medium with the concentration of 100 mg/L Kan and 500 mg/L Cef.The resistant buds obtained were more and the highest differentiation frequency could reach 34.3%.【Conclusion】A stable and efficient Agrobacterium mediated genetic transformation technology system for sugarbeet has been established,which offers a base for the selective breeding of high yield,high quality and broad resistance of disease sugarbeet variety.