【Background】Dendroctonus spp., the noxious forest pests, have been repeatedly intercepted in many ports in China. For custom officers, the main challenge remains to determine the provenance of these insects. 【Method】Using nested PCR, the cytochrome C oxidase subunit I gene (COⅠ) in mtDNA was examined for 12 samples from Dendroctonus valens, D.rufipennis, D.ponderosae, D.simplex and D.adjunctus. 【Result】The nested PCR showed variation between and within groups for a 530 portion of COⅠ. Molecular phylogenetic analysis based on COⅠ gene showed that Dendroctonus spp. could be divided into two clades. Clade 1 included D.adjunctus, and Clade 2, D.valens, D.rufipennis, D.ponderosae and D.simplex. The provenance of the specimens analyzed in this study can be identified within the evolutionary tree. 【Conclusion and significance】The results suggest that COⅠ may help determine the provenance of specimens intercepted in ports thus providing more adequate information to custom officers.