To reveal the relationship and genetic diversity of colored-grain wheat, we analyzed 18 varieties (lines) with red, purple, blue, and black seed coats using 305 polymorphic SSR markers. A total of 1084 alleles were detected with 3.6 alleles per locus. The allele numbers in seven homoeologous groups (HG) were aligned as HG3>HG2>HG1>HG4>HG5>HG7>HG6 in red and purple wheat and HG3>HG2>HG1 and HG 4>HG5>HG7>HG6 in black and blue wheat. The polymorphic information content (PIC) values in the seven homoeologous groups were similar, which were 0.525-0.543 in red and purple wheat and 0.517-0.545 in black and blue wheat. Among the 18 colored-grain varieties (lines), genetic difference was significant (P = 0.0001), and the genetic similarity coefficients ranged from 0.41 to 0.76. According to UPGMA analysis, the 18 varieties (lines) were classified into six groups, which were in relation to variety kindred and geographic distance.