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Position: Home > Articles > Construction of marker gene-free RNAi vector and genetic transformation of rice mediated resistance to Ricestripevirus Plant Protection 2013,39 (6) 14-20

抗水稻条纹病毒安全表达载体的构建及遗传转化

作  者：
雷阳;李羽;王彪;李红伟;李莉;王锡锋
单  位：
植物病虫害生物学国家重点实验室;中国农业科学院植物保护研究所
关键词：
水稻条纹病毒;RNA干涉载体;去除标记基因;共转化
摘  要：
构建了水稻条纹病毒(RSV)外壳蛋白(CP)基因RNAi载体pCMBIA1301+/-R,该载体在理论上可以获得无标记基因的转基因后代植株。在双菌双质粒体系中,比较了不同品种、重组农杆菌浓度配比对遗传转化效率的影响。结果表明:改进的转化体系最适合品种‘爱知旭’的转化,共转化效率达9.07%;推广品种‘武育粳3号’上也转化成功获得了转基因阳性植株(6.59%)。以‘爱知旭’为受体转化时,重组农杆菌E13R和E1301浓度比为3∶1时,共转化率最高(16.33%),其次分别是2∶1(13.56%)、1∶1(9.09%),以1∶2时最低(5.97%)。通过T1代自交分离和抗病性筛选,成功获得了无选择标记基因的转基因抗病毒植株16株,为抗RSV转基因水稻的安全释放奠定了基础。
译  名：
Construction of marker gene-free RNAi vector and genetic transformation of rice mediated resistance to Ricestripevirus
作  者：
Lei Yang;Li Yu;Wang Biao;Li Hongwei;Li Li;Wang Xifeng;State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences;
关键词：
Rice stripe virus;;RNA interference vector;;marker gene-free;;co-transformation
摘  要：
In this study,a marker gene-free RNA interference(RNAi)vector pCMBIA1301+/-R was constructed with a partial coat protein(CP)gene of RSV.The vectors pCMBIA1301+/-R and pCMBIA1301 were transformed into Agrobacterium tumefaciens"shooter"strain EHA105 by electroporation,respectively.We also attempted to improve the efficiency of genetic transformation in the test.The effects of both rice varieties and density ratio of two recombinant agrobacterium strains on the co-transformation efficiency were investigated.The results showed that the rice variety‘Aichiasahi'(9.07%)was more adaptive to co-transformation system by RNAi vector than that of the variety‘Wuyujing3'(6.59%)and‘Huaidao5'(0%).Additionally,when the density of recombinant agrobacterium E13R and E1301 was3∶1,the co-transformation efficiency was 16.33%,higher than those of the other density ratios,including 2∶1(13.56%),1∶1(9.09%)and1∶2(5.97%).Sixteen marker gene-free plants were successfully obtained in this study.

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