农杆菌介导的花魔芋遗传转化体系的优化  

Optimized Genetic Transformation System of Amorphophallus konjac Mediated by Agrobacterium tumfaciens

作　　者：陈磊[1] 郭政宏[1] 程海丽[1] 乐超银[1] 

机构地区：[1]三峡大学生物技术研究中心,湖北宜昌443002

出　　处：《安徽农业科学》2015年第5期21-24,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30871579)

摘　　要：[目的]探讨魔芋愈伤组织培养基、不同转化条件(菌液浓度、侵染时间、预培养时间及共培养时间)及转化植株的筛选条件(抗生素浓度和乙酰丁香酮(AS)浓度)等因素对转化效率的影响。[方法]采用卡那霉素抗性基因为选择标记,对农杆菌介导的花魔芋遗传转化体系进行优化。[结果]在预培养2 d,用OD600为0.6的菌液侵染30 min、共培养3 d,卡那霉素100 mg/L,羧苄青霉素250 mg/L,AS浓度100μmol/L的条件下能有效提高转化效率。再生花魔芋植株经GUS染色及PCR检测,结果表明外源目的基因已经整合到花魔芋基因组中。[结论]为魔芋抗病品种的转基因培育,丰富其种质资源,寻找抗病新途径提供理论依据和试验基础。[ Objective ] Factors that influence Agrobacterium tumfaciens-mediated transformation of Amorphophallus konjac, including culture medium, transformation conditions and selection reagents were investigated. [ Method ] An transformation system mediated by Agrobacterium tumfaciens has been optimized to obtain transgenic plants of Amorphophallus konjac by using kanamyein resistance gene as marker gene. [ Result ] The results showed that precuhure for 2 days before infection by Agrobacterium （ OD6oo = 0.6, 30 min） and then coeuhure for 3 days yielded the highest rate of antibiotic resistant callus formation with the addition of kanamycin （ 100 mg/L）, carbenieillin （250 mg/L） and acetosyrlngone（ 100 μmol/L）. The transgenic plants were screened and re-clarlfied by GUS staining and PCR analysis that the exogenous gene was integrated into Amorphophallus konjac genome. [ Conclusion] The study provided theoretical basis and experimental basis for cultivating genetically resistant varieties, riching germplasm resources and finding new ways for disease resistance.

关 键 词：花魔芋 农杆菌 遗传转化体系 

分 类 号：S188[农业科学—农业基础科学]