Abstract: The "double T-DNA" binary vector p13HSR 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 marker-free transgenic rice by Agrobacterium-mediated transformation.T0 plants with the three target genes and hpt gene were 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.

Key words: anther culture, co-transformation, selectable marker-free, transgenic rice, double T-DNAs

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。

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