Abstract: Extensive polymorphism within D.virilis species has been studied in this paper.We analyzed physical maps of mitochondrial DNA (mtDNA) from five populations distributing in Chinese continent using nine restriction endonucleases (XbaⅠ,EcoRⅠ,PstⅠ,HindⅢ,BglⅡ,SacⅠ,ScaⅠ,EcoRⅤ and PvuⅡ).Five genotypes of mtDNA are found in all the five populations.The genetic difference π between the five genotypes is 0.46%-1.76%.Genetic variation within population πij is 0.00%-0.33%.Genetic variation between populations dxy is 0.00%-0.82%.The γst of D.virilis species in Chinese continent is 24.62%.We have found the life environment of D.virilis has an effect on genetic variations of mtDNA,but not found the relationship between geographic distance (latitute degree) and inter-populational genetic difference (dxy).