Tamyb10 genes, a kind of transcription factors of the MYB family, determine the grain color of bread wheat and also have certain influence upon pre-harvest sprouting in bread wheat. In this study, the Tamyb10-1 functional markers reported previously were used to identify allelic variations on the Tamyb10-A1, Tamyb10-B1 and Tamyb10-D1 loci in 100 landrace wheat varietiesfrom Huanghuai Wheat Region. The results indicated that there were two types of alleles on each Tamyb10-1 locus, which resulted in 7 allelic combinations of Tamyb10-1 genes, i.e., Tamyb10-A1a/Tamyb10-B1a/Tamyb10-D1a, Tamyb10-A1a/Tamyb10-B1a/Tamyb10-D1b, Tamyb10-A1a/Tamyb10-B1b/Tamyb10-D1a, Tamyb10-A1b/Tamyb10-B1a/Tamyb10-D1a, Tamyb10-A1b/Tamyb10-B1b/Tamyb10-D1a, Tamyb10-A1b/Tamyb10-B1a/Tamyb10-D1b and Tamyb10-A1b/Tamyb10-B1b/Tamyb10-D1b. Their distribution percentage were 38.0%, 15.0%, 1.0%, 8.0%, 1.0%, 33.0% and 4.0%, respectively. Further analysis suggested that wheat cultivars with wild-type Tamyb10-1 at three loci showed white grains, whereas wheat cultivars with at least one mutant of Tamyb10-1 at three loci showed red grains. In addition, the Tamyb10-1 genes also affect pre-harvest sprouting and grain color is closely related with antioxidant capacity. Therefore, this study could provide certain useful information for the purpose of screening relatively superior wheat germplasms.