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

Abstract

TheOchlerotatus 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.

Integrative taxonomy improves understanding of native beneficial fauna: revision of the Nearctic Peristenus pallipes complex (Hymenoptera: Braconidae) and implications for release of exotic biocontrol agents

“…Integrative taxonomy combines information from multiple sources, such as morphology, DNA, ecology and behaviour (Dayrat, ; Schlick‐Steiner et al ., ). This provides a more holistic taxonomic approach using multiple independent lines of evidence and has largely improved cryptic species delimitation (Boring et al ., ; Ceccarelli et al ., ; Gebiola et al ., ; Baur et al ., ; Grossi et al ., ; Namin et al ., ; Schwarzfeld & Sperling, ; Zhang et al ., ).…”

When taxonomy and biological control researchers unite: Species delimitation of Eadya parasitoids (Braconidae) and consequences for classical biological control of invasive paropsine pests of Eucalyptus

“…Three gene regions were amplified, including two mitochondrial genes (Cytochrome oxidase I [ COI ] and Cytochrome b [ Cytb ]) and one nuclear gene (28S rDNA regions D1-D3 [ 28S ]). COI has long been the standard for species delimitation in insects [ 45 , 16 , 46 , 18 , 47 ] including Braconidae [ 48 , 49 , 17 , 19 ]. Cytb is generally more conserved but can help provide an independent test to prevent overestimations of species [ 50 ].…”

“…As the majority of taxonomic works have been based on only morphological characters, cryptic species complexes typically include a set of undescribed species [ 12 ]. Molecular taxonomy has revealed numerous cryptic species complexes, particularly within insects [ 13 – 18 ]. If a prospective biological control agent is a part of a cryptic species complex, multiple species may end up being released causing non-target effects, or if the wrong species is released, control of the target pest may not occur [ 11 , 7 , 19 ].…”

Novel Mitochondrial DNA Lineage Found among Ochlerotatus communis (De Geer, 1776) of the Nordic-Baltic Region

“…Morphology-based delimitation of these species is highly problematic due to a lack of reliable distinguishing traits, especially in adults [ 2 ]. Thus, researchers have employed both DNA sequencing, using mainly the mitochondrial cytochrome c oxidase subunit I (COI or COX1), and restriction fragment length polymorphism (RFLP) patterns to help with differentiation [ 3 ]. The namesake of the group, Oc.…”

“…Because of these factors, it has been theorized that the sister species comprising Oc. communis complex may have derived from allopatric as well as sympatric speciation [ 3 ].…”