In this study, we use three mitochondrial markers, cytochrome b gene (Cyt b), NADH dehydrogenase subunit 4 (ND4) and control region (D-loop) to investigate the phylogenetic relationships of extant zokor species in Mysopalacinae. The phylogenetic tree constructed based on Cyt b strongly supports the monophyly genera Eospalax and Myospalax with E. fontanierii being the most ancient species in Eospalax. Further phylogenetic analyses of four species of Eospalax based on ND4 and D-loop sequences revealed two clades that correspond to two geographical distributions. The basal clade includes E. cansus which is mainly found on Loess Plateau (LP) and another clade including E. baileyi, E. smithii and E. rufescens that inhabits areas above 2000 m on Qinghai-Tibetan Plateau (QTP) and Qinling Mountains. Geographical events of QTP and LP may have played a major role in the diversification and evolution of Mysopalacinae.