For the study of the polymorphism and evolutionary characteristics of MYB1 and its allele MYB10 involved in the regulation of anthocyanin biosynthesis in genus Malus. The MYB1 and MYB10 gene sequences amplified by RT-PCR technology in several Malus plants were sequenced and predicted for their protein structures to detect the differences. Six MYB1 sequences were obtained and the full-length CDS were 1 239 bp except for those of Malus cv. Eleyi and Malus sieversii (1 209 bp). While nine MYB10 were sequenced and the full-length CDS were 732 bp (Malus micromalus was 717 bp). The result from the comparison of protein sequences of MYB1 and MYB10 showed that the polymorphism of MYB1 was significantly higher than MYB10 and the polymorphism difference between the alleles may be related to the differentiation of gene function. Two polymorphism sites, the interchange of alkaline arginine and non-polar leucine, located on functional domain which may affect the function of MYB1 in different Malus plants. The result suggested that sequence conservatism of the allele MYB10 and MYB1 significantly differed from each other, and the genetic function responsible for evolution was differentiated. The result can provide further theoretical basis for the study of the gene function and regulation mechanism of apple fruit color.