Abstract: In this study,15 restriction enzymes were employed to investigate the mitochondrial DNA polymorphism among 93 individuals from 4 Guizhou goat breeds.Mitochondrial DNA polymorphism was detected by 3 enzymes (BamHⅠ,HindⅢ and SalⅠ).18 restriction morphs,which could be sorted into 3 mtDNA haplotypes,were found.Haplotype Ⅰ and Ⅱ are distributed in every breed with relatively high frequency,I.e.77.42% and 21.50%.The frequency of haplotype Ⅲ is lower (1.08%).The relationship between white goat and black goat is the closest,and they are related to ma goat.Our results suggest that haplotype Ⅰ and Ⅱ of Guizhou goats mtDNA might represent two maternal origin,which might separate about 190 thousand years ago.