Fluorescence in situ hybridization (FISH) can reveal minor structural differences of chromosomes. The karyotype of common wheat (Triticum aestivum L.) based on FISH pattern is seldom reported. In this study, non-denaturing FISH (ND-FISH) using Oligo-pSc119.2-1, Oligo-pTa535-1 and (AAG)6 as probes was used to investigate the chromosomal structure of 85 common wheat including 83 wheat-rye 1RS.1BL translocation cultivars/lines, a wheatrye 1RS.1AL translocation cultivar Amigo and Chinese Spring (CS). Two, three, two, three, six, three and four structural types respectively for 1A, 2A, 3A, 4A, 5A, 6A and 7A chromosomes were observed. Two, eight, two, two, four and six types of chromosome for 2B, 3B, 4B, 5B, 6B and 7B were respectively detected. The structure of 1B chromosomes in Amigo and CS is different. Five, two, two and two types of chromosomal structure respectively for 1D, 2D, 3D and 5D were distinguished. Polymorphisms of 1RS.1BL, 4D, 6D and 7D chromosomes were not detected. Chromosomes 1AI, 2AI, 3AI, 4AI, 5AIII, 6AI, 7AIII, 2BI, 3BV, 4BI, 5BII, 6BIII, 7BI, 1DIV, 2DI, 3DI and 5DII appeared in these 85 wheat cultivars/lines at high frequency. Each of the 85 wheat cultivars/lines has a unique karyotype. Amigo is a complex translocation cultivar. The FISH karyotype of wheat chromosomes built in this study provide a reference for the future analyzing wheat genetic stocks and help to learn structural variations of wheat chromosomes. In addition, the results in this study indicate that oligonucleotide probes and ND-FISH technology can be used to identify individual wheat cultivar.