Cry1 Ac基因组织特异表达载体构建及水稻遗传转化

Construction of Tissue-specific Expression Vectors of Cry1Ac and Rice Genetic Transformation

为了提高转基因抗虫水稻的生物安全性,构建了由组织特异性启动子序列RSP1、RSP2及组成型启动子35S带动Bt基因Cry1Ac的双元载体pCAM RSP1 Cry1A(c)、pCAM RSP2 Cry1A(c)及pCAM 35S Cry1A(c),并通过农杆菌EHA105介导对两个水稻品种"秀水11"和"秀水63"进行了遗传转化。"秀水11"和"秀水63"的平均转化率分别为72 9%和83 2%,经抗性愈伤组织的分化培养两个品种分别获得346和448株再生植株,其中PCR检测阳性株数分别为295和414。

In order to improve the biosafety of insect-resistant transgenic rice plants, three Agrobacterium binary vectors,pCAM-RSP1-Cry1A(c), pCAM-RSP2-Cry1A(c) and pCAM-35S-Cry1A(c) were constructed, in which the insecticidal gene Cry1Ac was driven by tissue-specific promoter fragments RSP1, RSP2 and the constitutive promoter 35S respectively.Two rice cultivars Xiushui 11' and Xiushui 63' were transformed by Agrobacterium EHA105 containing the constructed vectors.The results showed that the average transformation ratio of the two cultivars are 72.9% and 83.2 respectively. After differentiation culturing of Hygromycin resistant callus, 346 plantlets of Xiushui 11' and 448 plantlets of Xiushui 63' were regenerated in total and the PCR assay of them indicated that the number of Cry1Ac positive plantlets were 295 and 414 respectively.