Construction of expression vectors of male sterility and its fertility restoration in rice (Oryza sativa L.)

The 1.7kb promoter 6B of tapetum-specific genes from Oryza sativa L. by PCR was connected to the 270 bp NOS terminator from pROKII to generate pGEM7Z-6BNOS. The genes for Barnase and Barstar from Bacillus amylolique faciens were inserted between the BamH I and Nco I sites of pGEM7Z-6BNOS to give pGEM7Z-6BBNNOS and pGEM7Z-6BBNOS, respectively. The fragments of 2.3kb 6BBNNOS and 2.2kb 6BBSNOS were separated from pGEM7Z-6BBNNOS and pGEM7Z-6BBSNOS with the Sal I and Xho I digestion, and cloned into the Xho I sites of pDM302 conferred resistance to phosphinothricin (PPT), respectively. The orientation of the 6BBNNOS and 6BBSNOS fragments was diverse in pDM302. Therefore, the plant expression vectors of male sterility and its fertility restoration were successfully constructed. The immatured embryos from rice were transformed by particle bombardment. The rice transgenic plants with resistance to PPT were generated. The superior male sterile line and its restorer are being expected.