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 (φCT = 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.

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Hypogeococcus pungens , a mealybug native of southern South America, is devastating native cacti in Puerto Rico and threatening cactus diversity in the Caribbean, and potentially in Central and North America. The taxonomic status of H . pungens is controversial since it has been reported feeding not only on Cactaceae but also on other plant families throughout its distribution range. However, in Australia, where the species had been exported from Argentina to control weedy American cacti, it was never found on host plants other than Cactaceae. These conflicting pieces of evidence not only cast doubt on the species identity that invaded Puerto Rico, but also have a negative impact on the search for natural enemies to be used in biological control programs against this pest. Here we present reproductive incompatibility and phylogenetic evidences that give support to the hypothesis that H . pungens is a species complex in which divergence appears to be driven by the host plants. The nuclear EF1α and 18S and the mitochondrial COI genes were used as markers to evaluate the phylogenetic relationships among H . pungens populations collected in Argentina, Australia and Puerto Rico feeding on Cactaceae and/or Amaranthaceae. Additionally, we conducted reciprocal crosses between mealybugs from both hosts. Species delimitation analysis revealed two well-supported putative species within H . pungens , one including mealybugs feeding on Amaranthaceae ( H . pungens sensu stricto ), and a new undescribed species using Cactaceae as hosts. Additionally, we found asymmetric reproductive incompatibility between these putative species suggesting recent reproductive isolation. The Bayesian species delimitation also suggested that the Australian mealybug population may derive from another undescribed species. Overall, the patterns of genetic differentiation may be interpreted as the result of recent speciation events prompted by host plant shifts. Finally, the finding of a single haplotype in the Puerto Rico population suggests only one invasive event. We still need to identify the geographical origin of the pest in order to enable the use of biological control to reduce the threat to cacti diversity in the Caribbean.

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