A phylogeographic analysis inferred from the partial mitochondrial cytochrome oxidase subunit I gene (433 bp) was performed with 22 populations of Diaphorina citri Kuwayama collected in the Americas and one in the Pacific. Eight populations from four countries in South America, 14 from four countries in North America, and one from Hawaii were analyzed. Twenty-three haplotypes (hp) were identified and they fell into two groups: hp1–8 were identified in South America (group 1) and hp9–23 were identified in North America and Hawaii (group 2). Hp1 and nine were present in the highest frequencies within each population and within their group, 81 and 85% for group 1 and group 2, respectively. A diagnostic nucleotide at position 48 was identified that allowed for the discrimination of the two groups; in addition, no haplotypes were shared between the two groups. An analysis of molecular variance uncovered significant genetic structure (φ;CT = 0.733; P < 0.001) between the two groups of the Americas. Two haplotype networks (ParsimonySplits and Statistical Parsimony) discriminated the two groups and both networks identified hp1 and nine as the predicted ancestral or founding haplotypes within their respective group. The data suggest that two separate introductions or founding events of D. citri occurred in the Americas, one in South America and one in North America. Furthermore, North America and Hawaii appear to share a similar source of invasion. These data may be important to the development of biological control programs against D. citri in the Americas.