Abstract： Totally, 24 three-line parents of indica hybrid rice (Oryza sativa L.) were used to analyze their genetic diversity and genetic relationship by using 26 polymorphous primer pairs selected from 110 SSR (Simple sequence repeats) which dispersed on 12 chromosomes in rice. Those primers detected 116 alleles from all the 24 parents. The alleles number might range from 2 to 7 with an average of 4.4. The PIC (Polymorphism index content) value of the polymorphic markers ranged from 0.068 9 to 0.803 6, with average of 0.603 4. The expected heterzygosity (H_e) ranged from 0.081 6 to 0.829 8 and the average was 0.616 9. However, Shannon's information index (Ⅰ) ranged from 0.173 2 to 1.791 3 with average of 1.160 0. Furthermore, according to the results of cluster analysis by UPGMA method, the 24 parents materials were classified into three groups by microsatellite markers, and the parents among the groups showed relatively large genetic variation, which could be used to identify the relationship among the parents, and also to predict the heterosis of new materials by molecule markers assisted selection. The results provide a good reference to the choice of higher heterosis hybrid combinations in rice breeding. Fig 2, Tab 3, Ref 24