Fingerprinting and identification of closely related wheat (Triticum aestivum L.) cultivars using ISSR and fluorescence-labeled TP-M13-SSR markers

DNA based molecular markers allow precise, objective and rapid cultivar identification. In this study, five ISSR and two sets of fluorescence-labeled TP-M13-SSR markers were applied for discrimination of eight closely related wheat cultivars. Five ISSR primers revealed a total of 43 distinct reproducible bands, 29 (67.44%) of the 43 bands were polymorphic. The number of polymorphic bands detected by each ISSR primer ranged from 3 to 8 with an average of 4.8 per primer. One ISSR primer UBC 849 was able to identify all the eight wheat cultivars. The two sets of fluorescence-labeled TP-M13-SSR markers produced a total of 29 alleles with an average of 14.5 per locus in the eight wheat cultivars. All of the 29 alleles were polymorphic, and the numbers and sizes of alleles revealed were used for cultivar discrimination. Combination of the two sets of fluorescence-labeled TP-M13-SSR primers could distinguish all the eight closely related wheat cultivars. Finally, two fluorescence-labeled TP-M13-SSR markers successfully fingerprinted the eight closely related wheat cultivars. The result suggested that fluorescence-labeled TP-M13-SSR markers could be used as a potential and valuable method for fingerprinting and identification of wheat cultivars, and fluorescence-labeled TP-M13-SSR markers should be given priority in fingerprinting and identification of wheat cultivars compared with ISSR for its high-throughput and high accuracy. DNA fingerprinting of wheat cultivars can be expanded by additional cultivars and markers, and the data obtained can be used for cultivar protection.