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Position: Home > Articles > Optimized Genetic Transformation System of Amorphophallus konjac Mediated by Agrobacterium tumfaciens Journal of Anhui Agricultural Sciences 2015 (5) 21-24

农杆菌介导的花魔芋遗传转化体系的优化

作  者:
陈磊;郭政宏;程海丽;乐超银
单  位:
三峡大学生物技术研究中心
关键词:
花魔芋;农杆菌;遗传转化体系
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。
译  名:
Optimized Genetic Transformation System of Amorphophallus konjac Mediated by Agrobacterium tumfaciens
作  者:
CHEN Lei;GUO Zheng-hong;CHENG Hai-li;YUE Chao-yin;Biotechnology Research Center,Three Gorges University;
关键词:
Amorphophallus konjac;;Agrobacterium tumfaciens;;Genetic transformation system
摘  要:
[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 kanamycin resistance gene as marker gene. [Result]The results showed that preculture for 2 days before infection by Agrobacterium( OD600= 0. 6,30 min) and then coculture for 3 days yielded the highest rate of antibiotic resistant callus formation with the addition of kanamycin( 100 mg / L),carbenicillin( 250 mg / L) and acetosyringone( 100 μmol / L). The transgenic plants were screened and re-clarified 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.

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