农杆菌介导的Btcry1Ⅰa基因转化花椰菜的研究  

Studies on Genetic Transformation Bt cry1Ⅰa Gene into Cauliflower by Agrobacterium-mediated Transformation

作　　者：张永侠[1,2] 郎志宏[1] 朱莉[1] 杨丽梅[3] 张杰[4] 苍晶[2] 黄大昉[1] 

机构地区：[1]中国农业科学院生物技术研究所,北京100081 [2]东北农业大学生命科学学院,哈尔滨150030 [3]中国农业科学院蔬菜花卉研究所,北京100081 [4]中国农业科学院植物保护研究所,北京100193

出　　处：《中国农业科技导报》2011年第1期15-19,共5页Journal of Agricultural Science and Technology

基　　金：国家863计划项目(2008AA10Z155)资助

摘　　要：利用苏云金芽胞杆菌(Bacillus thuringiensis,简称Bt)cry1Ⅰa基因构建了双T-DNA边界的植物表达载体pCS1Ⅰa-LR,以瑞士雪球花椰菜具柄子叶和下胚轴为外植体,通过农杆菌介导法将cry1Ⅰa基因导入花椰菜中,共获得30株转化植株,经PCR检测,其中18株为PCR阳性植株,PCR阳性率为60%;RT-PCR和Western杂交检测结果表明,cry1Ⅰa基因在转基因花椰菜中可以正常转录和正确表达。转基因植株用小菜蛾幼虫进行饲虫试验,结果表明,转cry1Ⅰa基因花椰菜对小菜蛾具有很强的毒杀作用,昆虫幼虫的校正死亡率高达100%。获得的转基因花椰菜可用于下一步的抗虫花椰菜的培育。The double T-DNA border expression vector pCS1Ⅰa-LR harboring Bacillus thuringiensis cry1 Ⅰ a gene was introduced into the cotyledon petioles and hypocotyls of cauliflower（Brassica oleracea var.botrytis） mediated by Agrobacterium.Thirty regenerated plants were obtained,18 of them were positive by PCR detection,which demonstrated that cry1 Ⅰ a gene was integrated into the cauliflower genome.The PCR positive rate is 60%.Moreover,RT-PCR and Western blot analysis showed that cry1 Ⅰ a gene were transcripted and translated,respectively in transgenic cauliflower.Bioassay results showed that the transgenic cauliflowers harboring cry1 Ⅰ a gene had the high toxicity to diamondback moth and the mortality of insect larvae was accounted for 100%.The transgenic cauliflower can be used for further insect resistant cauliflower breeding.

关 键 词：花椰菜 农杆菌介导转化 cry1Ⅰa基因 转基因植株 

分 类 号：Q789[生物学—分子生物学]