TILLING (Targeting Induced local Lesions IN Genomes) is known to be an excellent methodology for reverse genetics approach. About 15,000 M3 TILLING lines have been developed after gamma-ray irradiation to the rice seeds of Donganbye. In order to assess genetic diversity of the TILLING population. we have employed a multiple dominant marker technique, such as AFLP. A total of 96 (0.64%) lines including Dongganbye were randomly selected and their genetic diversity was assessed on the basis of AFLP marker polymorphism by using 5 primer combinations. An average of 100.4 loci with a range of 97 to 106 were detected by using the primer combinations, resulting in 173 (34.6%) polymorphic loci among 96 lines. A broad range of similarities with 80% to 96% was evidenced between Donganbye and each of 96 TILLING lines, reflecting genetic diversity of the TILLING population. About 30 polymorphic loci have been cloned and their sequences have been blasted against rice whole genome sequences. The sequences evidenced highly significant matches to each of genes including exons and introns, upstream sequences and downstream of genes, and intergeneic sequences. Therefore, the TILLING rice population would be valuable genetic sources for rice functional genomics.