The plumage on the dorsal trunk of normal quail embryos exhibits longitudinal black and brown stripes of pigments produced by melanocytes. However, this pigmentation pattern disappeared in Bh (black at hatch) heterozygous and homozygous embryos because of overall black and brown pigmentation of plumages, respectively. To investigate the mechanisms of the pigment pattern formation of plumage and clarify the roles of the Bh locus in the pattern formation, we examined the expression pattern of genes relating to melanocyte development (Mitf, MelEM antigen, Kitl, Kit and EdnrB2) and melanin pigment production (Dct, Tyrp1, Tyr and Mmp115) in Bh mutant and wild-type embryos throughout development. As a result, we found that MelEM antigen was expressed in melanoblasts committed to produce black pigment before apparent melanogenic gene expression, and that Bh heterozygotes and homozygotes showed abnormal expression patterns of the MelEM antigen. These results indicate that MelEM antigen is a good marker for melanoblasts committed to produce black pigment, and suggests that the Bh locus directs melanocytes to produce eumelanin in proper positions.