On the Evolution of Saline Tolerance in the Larvae of Mosquitoes in the Genus Ochlerotatus

We examined physiological and morphological parameters associated with saline tolerance in the larvae of 11 species of mosquito in the genus Ochlerotatus that were collected from the wild in a variety of sites around North America. Saline tolerance was assayed, and all of the species were osmoregulators. Six of the species examined were found to be physiologically restricted to freshwater habitats, while the other five could successfully osmoregulate in both freshwater and saline water, including seawater. All larvae that were obligate freshwater forms had only one rectal segment, while all of the euryhaline osmoregulators had two. We were interested in the evolutionary pathway by which saline tolerance arose in this lineage. DNA sequence data were obtained by polymerase chain reaction amplification and sequencing of the D2 region of the 28s rDNA gene in all of the freshwater and saline-tolerant Ochlerotatus species we studied. When the morphological and physiological characters were mapped on the resultant cladogram, they revealed a complex pattern, with freshwater and saline-water forms being adjacent and interspersed through the tree. The data also demonstrate that saline tolerance has been gained and then lost at least once in this lineage. Two possible evolutionary scenarios are presented, but the one we favor is that saline tolerance arose one time in this lineage and repeated reversions to the freshwater condition have occurred.