【摘要】 向作物中转多价抗虫基因是增强作物抗虫性、拓宽抗虫谱、延长抗虫时限的有效措施。从粳稻品种吉粳81、吉粳88和通887的成熟胚诱导出愈伤组织,以愈伤组织作为转化的受体,利用农杆菌介导法,转化苏云金杆菌毒蛋白基因CryIA(a)和半夏凝集素(Pinelliaternataagglutinin,PTA)基因。表达双价抗虫基因的载体p3300-bt-pta的bt和pta分别由Ubiquitin启动子和CaMV35S启动子驱动,宿主菌株为EHA105。用除草剂草丁膦(PPT)筛选得到转基因植株25株。以bt基因和pta基因两端序列为引物在一个反应体系同时做2个基因的PCR检测,检测到bt、pta基因的2条带,Bar基因的PCR检测也显阳性。转化株对PPT除草剂抗性鉴定呈抗性,证明外源基因已整合到水稻基因组中。水稻二化螟接虫鉴定结果表明,23株转基因植株抗虫性比对照明显提高,2株表现感虫。更多还原

【Abstract】 Now transformation of multi-genes of insect resistance into plants has been an efficiency measure. It can improve insect resistance, extend different insect resistance ranges and prolong insect resistance times. The pCAMBIA3300 vector carrying multi-genes (CryIA(a)、pta and bar) mediated by Agrobacterium tumefaciens were transferred into calli which derived from mature embryos of three japonica rice varieties (Jijing81?Jijing88 and Tong887). The CryIA(a) and pta genes of the expression vector were drived by Ubi and CaMV 35S promoters. EHA105 is the host bacterial strain. Twenty-five herbicide-resistant plants were regenerated through herbicide (PPT) selection. The three genes transgenic status of the herbicide-resistant plants was confirmed by PCR analysis. Insect resistance assay showed that twenty-three transgenic plants had enhanced resistance to rice stem borer.更多还原