基因枪法和农杆菌介导的Bt抗虫基因转化芥蓝  

Pyramiding of Bt Gene into Brassica alboglabra L. H. Bailey by Biolistic and Agrobacterium-mediated Transformation Methods

作　　者：陈长明[1] 赵祥明[1] 雷建军[1] 曹必好[1] 陈国菊[1] 

机构地区：[1]华南农业大学园艺学院,广东广州510642

出　　处：《中国蔬菜》2016年第8期21-28,共8页China Vegetables

基　　金：广东省教育厅特色创新项目(2014KTSCX026);广东省教育厅青年人才项目(2014KQNCX035);华南农业大学校长基金项目(K13012)

摘　　要：以芥蓝品种小香菇为试材,分别采用基因枪法和农杆菌介导转化法将Bt抗虫基因Cry2Aa2转化到芥蓝中,并对两种转化方法进行了优化和比较。结果表明:基因枪法在轰击距离为6cm、轰击次数为1次时转化效率最高,为0.43%;农杆菌介导转化法以带子叶下胚轴为外植体、预培养2d、侵染10min、共培养2d、抑菌培养5d后转入筛选培养基为最佳参数,转化效率为0.37%。经PCR和PCR-Southern鉴定,基因枪法和农杆菌介导转化法均成功将目的基因导入芥蓝基因组中。经抗虫性表型鉴定,T_0代转基因芥蓝植株对甘蓝夜蛾幼虫的抗性高于非转基因植株。The Bt gene,Cry2Aa2,were transferred into Chinese kale（Brassica alboglabra L. H.Bailey）by biolistic and agrobacterium-mediated transformation methods and these 2 transformation systems wereoptimized and compared.High transformation rate was obtained by bombarding Chinese kale callus with a distanceof 6 cm and for one time.The transformation efficiency was 0.43%. The suitable procedure for agrobacteriummediatedtransformation method in Chinese kale was using hypocotyl as material,pre-culture for 2 days,infectedby agrobacterium for 10 minutes,co-culture for 2 days,antibacterial-culture for 5 days and then screened inselective culture with 5 mg·L-1 kanamycin.The transformation efficiency was 0.37%.Identified by PCR,PCRSouthernblot analyses,the target gene were transferred into the Chinese kale by both biolistic and agrobacteriummediatedtransformation methods.Insect resistance identification proved that the positive transgenic Chinese kaleplants had stronger resistance to Mamestra brassicae larva than the non-transgenic plants.

关 键 词：芥蓝 BT基因 Cry2Aa2 基因枪法 农杆菌介导转化法 

分 类 号：S635.9[农业科学—蔬菜学]