Phylogenetics and molecular identification of theOchlerotatus communiscomplex (Diptera: Culicidae) using DNA barcoding and polymerase chain reaction-restriction fragment length polymorphism

Abstract

AbstractTheOchlerotatus communis(De Geer, 1776) complex consists of four cryptic mosquito species in North America, including:O. communis,Ochlerotatus churchillensis(Ellis and Brust, 1973),Ochlerotatus nevadensis(Chapman and Barr, 1964), andOchlerotatus tahoensis(Dyar, 1916). Most of the morphological characters used for the identification of these species are quantitative and overlap across species. Here we evaluated the efficacy of DNA barcoding for identification of three members of thecommuniscomplex (O. nevadensisis not included in this study) and developed diagnostic restriction fragment length polymorphism (RFLP) patterns forO. communisandO. churchillensis. A phylogeny of 23OchlerotatusLynch Arribálzaga, 1891 species was inferred using mitochondrial cytochromecoxidase subunit I gene sequences. All species included in our analysis within theO. communiscomplex were delineated using cytochromecoxidase subunit I barcodes. However, this complex was recovered as paraphyletic with respect toOchlerotatus abserratus(Felt and Young, 1904) andOchlerotatus implicatus(Vockeroth, 1954), indicating the need for increased genetic and taxonomic sampling to infer the phylogenetic relationships of these taxa. The RFLP profile for multiple field specimens ofO. communiswas distinct from all RFLP patterns forO. churchillensis, and this method can be used as an efficient molecular method for the identification these species.