Abstract: 5 restriction endonucleases, namely BamHⅠ, TaqⅠ, HaeⅢ, RsaⅠand HincⅡ, were used to analyze the polymorphism of mtDNA Cytb gene part sequences of 256 horses from 6 types including importing breed, cultivating breed and local breed. The products of endonuclease cutting were detected by 8 percent non-denatured polyacry lamide gel electrophoresis and showing in silver staining protocol. The result indicated BamHⅠand TaqⅠshowed polymorphism, 7 restriction morph were defected and 3 haplotypes sorted, in which haplotypeⅠand Ⅲ were the basic haplotypes, but we guessed that these horses came of one female ancestor analyzed thorough one especial morph, namely BamHⅠ-B.