Provided is a breeding method for two lines hybrid rice based on the rice osms4 gene mutant comprising mutagenizing seeds of japonica cultivar 9522 with Co gamma ray backcrossing one male sterile mutant of mutagenized F2 progeny for three generations obtaining the rice plants having the osms4 gene mutant with stable inheritance which are regulated by recessive single gene then using the rice plants as male sterile line and maintainer line for two lines hybrid rice breeding under different illumination conditions.
