Correlation of mitochondrial haplotypes with cuticular hydrocarbon phenotypes of sympatric Reticulitermes species from the southeastern United States

Three sympatric Reticuelitermes species have been identified in Georgia, US A, based on morphological characters from alates and soldiers: R. flavipes, R. virginicus, and R. hageni, but species identification at individual col lection sites is often difficult because alate production is seasonal and s oldiers comprise 1-3% of the colony. We therefore set up an experiment to d etermine if chemical phenotypes and mtDNA haplotypes can be used together t o separate species of subterranean termites. Subterranean termites of the o rder Reticulitermes (Isoptera: Rhinotermitidae) were collected from 20 insp ection ports across Four soil provinces in Georgia. Each collection was ide ntified to species using dichotomous keys. Two collections, HH11 and BH25, however, could not be unequivocally keyed to species and were classified as unknown. The mitochondrial cytochrome oxidase II (COII) gene was sequenced from individual members of each collection and the variation in cuticular hydrocarbon phenotypes from these same collections was characterized. The c uticular hydrocarbon and mtDNA phylogenetic analyses show agreement with bo th unknown collections falling out in a separate clade. Specimens from HH11 and BH25 are different morphologically, chemically, and genetically from t he three known sympatric species in Georgia. Our results suggest that these two collections may represent at least one new taxon in Reticulitermes. Fu rthermore, the association of cuticular hydrocarbon phenotypes and mtDNA ha plotypes demonstrates that. when combined with morphological characters, th ey are useful in separating known species, determining new species, and und erstanding termite evolution.