TILLING (Targeting Induced Local Lesions IN Genomes) is broadly regarded as an excellent methodology for reverse genetics applications. Approximately 15,000 M3 TILLING lines have been developed via the application of gamma-ray irradiation to rice seeds (cv. Donganbyeo), followed by subsequent selections. In an effort to evaluate the genetic diversity of the TILLING population, we have employed the AFLP multiple dominant marker technique. A total of 96 (0.64%) TILLING lines as well as Donganbyeo were selected randomly and their genetic diversity was assessed based on AFLP marker polymorphisms using 5 primer combinations. An average of 100.4 loci in a range of 97 to 106 was detected using these primer combinations, yielding a total of 158 (31.4%) polymorphic loci between Donganbyeo and each of the 96 lines. A broad range of similarity from 80% to 96% with an average of 89.4% between Donganbyeo and each of the 96 lines was also observed, reflecting the genetic diversity of the TILLING population. Approximately 28 polymorphic loci have been cloned and their sequences were BLAST-searched against rice whole genome sequences, resulting in 20 matches to each of the gene bodies including exon, intron, 1 kb upstream and 1 kb downstream regions. Six polymorphic loci evidenced changes in the coding regions of genes as compared to the rice pseudomolecules, 4 loci of which exhibited missense mutations and 2 loci of which exhibited silent mutations. Therefore, the results of our study show that the TILLING rice population should prove to be a useful genetic material pool for functional genomics as well as mutation breeding applications.

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.