Amplification profiles of the SSR markers in cultivars of hexaploid wheat and their relatives

The National Bioresource Project-WHEAT (NBRPWHEAT), launched by the Japanese government in 2002, aims to maintain and distribute seed stocks and DNA clones of “Wheat”. The second-term NBRPWHEAT started in 2007. Additionally to its primary roles in handling seed stocks and DNA clones, the second-term NBRP-WHEAT features the collection and characterization of DNA markers, which will make the seed stocks of NBRP-WHEAT more valuable for molecular works, such as, isolation and characterization of genes in wheat. We started to survey amplification profiles of the publically available SSR markers (barc, cfa, cfd, cfe, cft, gdm, gwm, hbg, hbe, hbd, wmc, and STM markers) in cultivars of hexaploid wheat and their relatives. The lines to be tested include; eight Aegilops species with representative diploid genomes, Triticum monococcum, T. boeoticum, T. urartu, T. durum, T. spelta and 31 hexaploid wheat accessions. We also took samples of Hordeum vulgare, H. spontaneum, and Secale cereale as outgroup species. The PCR products using template DNA from these lines are separated on a capillaryelectrophoresis machine and the amplification profiles are documented as chromatograph of fluorescence intensity and pseudo-electrophoretic-patterns. As pilot experiments, we tested 464 markers on 12 lines thus far and established the methodology for stable amplification of SSRs and reproducible separation of PCR fragments.