The application discloses a CYP703A3 gene based cultivation method for a transgenic rice sterile line, which comprises the steps as follows: A, acquisition of a rice gene cassette CYP703A3; B, acquisition of an sfGFP gene cassette; C, amplification of a wheat pollen lethal gene Ki; D, amplification of a wheat pollen specific promoter Pg47; and E, connection of each gene expression element. The step E of connection of each gene expression element comprises: firstly, introducing a complete rice CYP703A3 gene into a pCAMBIA1300 vector; and then connecting the sfGFP gene cassette, the wheat pollenlethal gene Ki and the wheat pollen specific promoter pPg47 to the vector. According to the invention, a F1-generation heterozygote follows the Mendel segregation law in the self fruitfulness process, and generated progenies not only contain heterozygotes which can keep a triple linked gene, but also contain the sterile line without fertility.