Diabrotica virgifera zeae Krysan and Smith, (MCR) and D. virgifera virgifera LeConte (WCR) are currently classified as subspecies; both are key pests of corn. MCR is distributed from Central America to Oklahoma in the United States. WCR is found from Northern Mexico up into the Unites States Corn Belt and into Canada. The latter subspecies has been recently introduced into Europe. This study was conducted to examine genetic variability within and among populations of the two subspecies throughout much of the geographical range of the species in North and Central America. The analysis of COI mitochondrial DNA sequence data from individuals revealed the presence of two phylogroups within Diabrotica virgifera, one present primarily to the North and the other to the South of the species range. However, there is not a good correlation between this geographic subdivision and that dictated by external morphological characters used to separate the subspecies. Results of phylogeographical analyses are used to revisit current hypotheses pertaining to historical corn-corn rootworm associations and the relatively recent changes in geographic distribution of the WCR.