The nucleotide sequence variations of mitochondrial DNA were investigated to understand genetic differentiation for five different Pungitius kaibarae populations at five study sites from selected four streams flowing into the East Sea. The complete sequences of mtDNA control regions of them were determined the constant sites 342 bp and variable sites 183 bp including parsimony infromative sites 122 bp. Based on the phylogenetic tree, five populations were monophyletic unit (97% MPand 100% NJ) and separated two groups (Myongpa-Songhuen stream group and others group). The population of Baebong stream (82nd bp, G-A) consistently formed them of the Jasan stream into a monophyletic unit even though it is closer then Myongpa stream (99th bp, T-C) from the Baebong stream. Further studies on the molecular phylogeny for the primary freshwater fish are needed to establish for the fish biodiversity conservation in mountainous and upland streams.