Mitochondrial DNA(mtDNA) variations were investigated among fifteen strains in Drosophila kikkawai. Seven genotypes in total were revealed in fifteen strains of D. kikkawai by the use of ten restriction endonucleases. There were eleven variable sites among seven genotypes. Substitution rates of seven genotypes ranged from 0.0032 to 0.0706, with an average of 0.0174. In addition to the site variation, the length variation of mt genomes was also observed. Based on the restriction map, this length variation was occurred within the adenine and thymine(A+T) rich region. Geographic distribution of seven genotypes and length variations were not shown any geographic structuring. The total number of base pair of the D. kikkawai mtDNA molecule was estimated as 16.45±0.08 Kbp.