Abstract: In order to evaluate the influence of the extension (E) locus on the color of cattle′s coat, the genetic characteristic and variations of the encoding region for melanocortin-1 receptor (MC1R) gene in four cattle populations sampled in Yunnan Province, including three indigenous populations and an imported breed, were assayed and analyzed by using DNA sequencing and PCRRFLP techniques. The alleles E+, e and genotypes E+/E+, E+/e, e/e were identified in Wenshan Yellow cattle, Zhaotong Yellow cattle and Shorthorn cattle. The frequencies of E+ and e alleles were 0.54 and 0.46 in Wenshan Yellow cattle, 0.70 and 0.30 in Zhaotong Yellow cattle. The alleles E+, ED and e were identified in Diqing Yellow cattle with the frequencies of 0.76, 0.14 and 0.10, respectively. The genotypes E+/E+，ED/ED, E+/ED, E+/e and ED/e were identified in Diqing Yellow cattle, with the only exception of e/e. In contact with the coat color of the selected samples, it was found that E locus had an important influence on determinating the color of the cattle′s coat. E+ and ED alleles were associated with black coat, and e allele was connected to red coat. Whereas yellow, brown and red fur were also influenced by other loci.