DNA barcoding: complementing morphological identification of mosquito species in Singapore

Abstract

BackgroundTaxonomy that utilizes morphological characteristics has been the gold standard method to identify mosquito species. However, morphological identification is challenging when the expertise is limited and external characters are damaged because of improper specimen handling. Therefore, we explored the applicability of mitochondrial cytochrome C oxidase subunit 1 (COI) gene-based DNA barcoding as an alternative tool to identify mosquito species. In the present study, we compared the morphological identification of mosquito specimens with their differentiation based on COI barcode, in order to establish a more reliable identification system for mosquito species found in Singapore.MethodsWe analysed 128 adult mosquito specimens, belonging to 45 species of 13 genera. Phylogenetic trees were constructed for Aedes, Anopheles, Culex and other genera of mosquitoes and the distinctive clustering of different species was compared with their taxonomic identity.ResultsThe COI-based DNA barcoding achieved a 100% success rate in identifying the mosquito species. We also report COI barcode sequences of 16 mosquito species which were not available previously in sequence databases.ConclusionsOur study utilised for the first time DNA barcoding to identify mosquito species in Singapore. COI-based DNA barcoding is a useful tool to complement taxonomy-based identification of mosquito species.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-014-0569-4) contains supplementary material, which is available to authorized users.

The mitochondrial genomes of twelve Anopheles mosquitoes (Diptera: Culicidae) and their phylogenetic implications

“…Given that distinct species normally differ in their resistance to insecticides, accurate species identification has become especially crucial for implementing biological control and disease prevention (Bickford et al 2007). For such purposes, mitochondrial DNA markers have proven powerful (Ashfaq et al 2014;Chan et al 2014;Ruiz-Lopez et al 2012). In this study, we assembled mitochondrial genomes for 12 Anopheles mosquitoes from Illumina sequencing reads (Table 1).…”

Phylogenetic Analysis of Aedes aegypti Based on Mitochondrial ND4 Gene Sequences in Almadinah, Saudi Arabia

“…COI-based DNA barcoding has been also used by Abigail Chan and co-workers to indentify mosquitoes in Singapore (15). This study has demonstrated that COI-based DNA barcoding can be also a good molecular marker for mosquitos' taxonomy.…”

Aedes nigrinus (Eckstein, 1918) (Diptera, Culicidae), a new country record for England, contrasted with Aedes sticticus (Meigen, 1838)

“…nigrinus using the DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) in accordance to the manufacturer's instructions. Amplification of the mitochondrial cytochrome oxidase subunit I (COI) gene and the nuclear internal transcriber spacer 2 (ITS2) region of ribosomal DNA was carried out using the following primers: 5'-GGATTTGGAAATTGATT-AGTTCCTT-3' (COIF) and 5'-AAAAATTTTAATTCCAGTTGGAACAGC-3' (COIR) (Chan et al 2014), 5'-TGTGAACTGCAGGACACATG-3' (ITS2F) and 5'-ATGCTTAAATTTAGGGGGTA-3' (ITS2R) (Walton et al 2007). PCR was Recorded by Medlock and Vaux (2009).…”