利用共转化和花药培养技术快速获得无选择标记的三价转基因水稻  

Rapidly Obtaining the Marker-Free Transgenic Rice with Three Target Genes by Co-Transformation and Anther Culture

作　　者：朱丽[1] 傅亚萍[1] 刘文真[1] 胡国成[1] 斯华敏[1] 唐克轩[2] 孙宗修[1] 

机构地区：[1]中国水稻研究所水稻生物学国家重点实验室,浙江杭州310006 [2]上海交通大学农业与生物学院植物生物技术研究中心,上海200030

出　　处：《中国水稻科学》2007年第5期475-481,共7页Chinese Journal of Rice Science

基　　金：国家转基因植物研究与产业化专项资助项目(FY03-B-07);国家973计划资助项目(G19990116-1;2005CB120801)

摘　　要：采用农杆菌介导的水稻转化体系,将包含选择标记潮霉素磷酸转移酶基因(hpt)和目的基因人乳铁蛋白(hLF)、高赖氨酸(SB401)、高甲硫氨酸(RZ10)基因的双T-DNA表达载体p13HSR转化水稻。筛选潮霉素磷酸转移酶基因和目的基因都是阳性的转基因植株,按单株进行花药培养,快速得到无选择标记而目的基因阳性的转基因纯合植株,得率为9.87%。RT-PCR检测结果显示外源基因已整合到转基因水稻基因组中并转录。同时观测到三价表达载体在转化过程中部分目的基因丢失。The ＂double T DNA＂ binary vector pl3HSR which carried two independent T-DNAs, containing hygromycin phosphotransferase gene （hpt） in one T DNA region and three target genes （hLF ,SB401 , RZ10） in the other T-DNA region, was used to generate selectable rnarker-free transgenic rice by Agrobacterium-mediated transformation. To plants with the three target genes and hpt gene we：e selected for anther culture. It took only one year to obtain double haploid selectable marker-free transgenic plants containing the three target genes through co-transformation followed bY anther culture, with an efficiency of 9.87%. RT PCR analysis indicated that the target genes were inserted into rice genomic DNA and successfully transcripted. It was noted that one or two target genes derived from the binary vector were lost in some transgenic rice plants.

关 键 词：花药培养 共转化 无选择标记 转基因水稻 双T-DNA结构 

分 类 号：S511[农业科学—作物学]