A total of 135 EST, 27 STS, and 253 SSR primer pairs located in 7 homoeologous groups of wheat and barly were used for amplification of 24 possible Wheat-R. ciliaris disomic addition lines. Fifty-five primer pairs could amplified polymorphic bands between common wheat variety "Inayama Komugi" and the Inayama Komugi- R. ciliaris amphiploid, and 31 of the 55 could amplify the specific bands of R. ciliaris in the addition lines. According to the PCR (polymerase Chain Reaction) results, the added R. ciliaris chromosomes in lines 07K02, 07K06, 07K39, 07K201, 07K202, 07K255, and 07K256 belong to homoeologous group 1 of wheat; the added R. ciliaris chromosomes in lines 07K07, 07K08, 07K09, 07K11, 07K14, and 07K17 belong to homoeologous group 2 of wheat; the added R. ciliaris chromosomes in lines 07K15, 07K16, 07K21, and 07K47 belong to homoeologous group 6 of wheat.