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

The 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.