CryIA(c)转基因结球甘蓝的抗虫性研究  

Inheritance and Resistance to Insect in CryIA(c) Transgenic Cabbage

作　　者：李汉霞[1,2] 尹若贺[1,2] 陆芽春[1,2] 张俊红[1,2] 

机构地区：[1]华中农业大学园艺林学学院 [2]国家蔬菜改良中心华中分中心,武汉430070

出　　处：《农业生物技术学报》2006年第4期546-550,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.AA212221)资助。

摘　　要：以结球甘蓝(Brassicaoleraceavar.capitata)下胚轴为外植体,通过根癌农杆菌(Agrobacteriumtumefaciecin)介导法将苏云金芽孢杆菌(Bacillusthuringiensis,Bt)杀虫蛋白基因CryIA(c)导入结球甘蓝栽培品种中,共获得56株卡那霉素抗性植株。分别以CryIA(c)和NPTⅡ基因引物进行PCR扩增,结果43株均扩增出了预期的目的带。以CryIA(c)基因为探针,进行Southernblot分析,证明外源基因已经整合到甘蓝基因组中,且多数转基因植株只含有单拷贝的外源基因。离体叶片抗虫实验表明,转基因植株幼虫校正死亡率为40%,平均单虫重达到极显著水平,叶片损害程度差异明显,转基因植株抗性较对照显著提高。通过卡那霉素涂抹叶片和PCR扩增两种方法鉴定,表明外源基因在转基因甘蓝自交后代呈孟德尔单基因遗传。Using hypocotyl segments of aseptic seedlings of cabbage （Brassica oleracea var. capitata） as explants, regenerated plants with kanamycin resistance were obtained mediated by Agrobacterium tumefaciecin （sWain LBA4404）. The transformed plants with CryIA（c） （Bt） gene were confirmed by Southern blotting analysis. And the results showed that CryIA（c） gene had been integrated into the cabbage genome. And majority of the transgenic plants had only one single copy of the inserted CryIA （c） gene. Leaf section bioassays of insect resistance showed that CryIA（c） transgenic cabbage significantly enhanced the level of resistance against the larvae of insect diamondback moth （DBM）. The inheritance patterns of the transgene in T1 offspring of transgenic cabbage were investigated using PCR with NPT Ⅱ primers and kanamycin resistance test on young seedling leaves. The results showed that dominant gene locus, CryIA（c） gene and NPTⅡ gene followed the Mendel＇s fashion with a ratio of 3：1 segregation in T1 populations.

关 键 词：甘蓝 BT转基因 抗虫性 

分 类 号：S635[农业科学—蔬菜学]