Thermoregulation has been suggested to influence mitochondrial DNA (mtDNA) evolution. Previous studies revealed that the mitochondrial protein-coding genes of fish living in temperate climates have smaller dN/dS (Non-synonymous substitution rate/Synonymous substitution rate) than tropical species. However, it is unknown whether different geographic populations of one fish species experience stronger selective pressures between cold and warm climates. The biological characteristics of the Japanese sand lance, Ammodytes personatus in the North-western Pacific is well-suited for assessing the performance of mtDNA evolution among separate geographic populations. In this study, we focused on the mitochondrial ATP6 gene of A. personatus using 174 individuals from eight different sea temperature populations. Two distinct haplotype lineages and a significant population structure (P = 0.016) were found in this species. The frequencies of the two lineages varied with the changes of annual sea temperature. The southern lineage (lineage A, dN/dS = 0.0384) showed a larger dN/dS value than the northern lineage (lineage B, dN/dS = 0.0167), suggesting that sea temperature greatly influences the evolution of the two lineages. The result provides robust evidence of local adaptation between populations in A. personatus.