Purple pericarp is an interesting and useful trait in Triticum aestivum, but the molecular mechanism behind this phenotype remains unclear. The allelic variation in the MYB transcriptors is associated with the phenotype of pigmented organs in many plants. In this study, a MYB transcription factor gene, TaMYB3, was isolated using homology-based cloning and a differentially expressed gene mining approach, to verify the function of the MYB transcriptor in the purple pericarp. The coding sequence of TaMYB3 in cultivar Gy115 was the same as that in cultivar Opata. TaMYB3 was localized to FL0.62–0.95 on chromosome 4BL. The TaMYB3 protein contains DNA-binding and transcription-activation domains, and clustered on a phylogenetic tree with the MYB proteins that regulates anthocyanin and proanthocyanin biosynthesis. TaMYB3 localized in the nuclei of Arabidopsis thaliana and wheat protoplasts after it was transiently expressed with PEG transformation. TaMYB3 induced anthocyanin synthesis in the pericarp cells of Opata in the dark in collaboration with the basic helix–loop–helix protein ZmR, which is also the function of ZmC1. However, TaMYB3 alone did not induce anthocyanin biosynthesis in the pericarp cells of the white grain wheat cultivar Opata in the light after bombardment, whereas the single protein ZmR did. Light increased the expression of TaMYB3 in the pericarp of Gy115 and Opata, but only induced anthocyanin biosynthesis in the grains of Gy115. Our results extend our understanding of the molecular mechanism of the purple pericarp trait in T. aestivum.