Abstract: In this research, DNA fingerprints of 1 male sterile lines and 1 restorer lines of two-line hybrid rice were constructed by using 270 SRAP (Sequence related amplified polymorphism) primers. The 54 SARP primers could amplify 136 polymorphic bands totally with a mean of 2. 5 bands per primer. The 200 single plants of Luliangyou 996 were tested in laboratory by a specific SRAP marker(Me3Em8)whose bands were complement in hybrid combination seeds. The seed purity was 97.50%, which was same to the field purity (97.50% in Hainan). The results of this research suggested that SRAP technique could be used in seed purity identification of two-line hybrid rice.