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Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia.

Batovska J, Blacket MJ, Brown K, Lynch SE - Ecol Evol (2016)

Bottom Line: The average conspecific and congeneric p-distance was 0.8% and 7.6%, respectively.In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport.The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance.

View Article: PubMed Central - PubMed

Affiliation: Department of Economic Development, Jobs, Transport and Resources (DEDJTR) BioSciences Research AgriBio Centre for AgriBioscience Bundoora Victoria 3083 Australia.

ABSTRACT
DNA barcoding is a modern species identification technique that can be used to distinguish morphologically similar species, and is particularly useful when using small amounts of starting material from partial specimens or from immature stages. In order to use DNA barcoding in a surveillance program, a database containing mosquito barcode sequences is required. This study obtained Cytochrome Oxidase I (COI) sequences for 113 morphologically identified specimens, representing 29 species, six tribes and 12 genera; 17 of these species have not been previously barcoded. Three of the 29 species ─ Culex palpalis, Macleaya macmillani, and an unknown species originally identified as Tripteroides atripes ─ were initially misidentified as they are difficult to separate morphologically, highlighting the utility of DNA barcoding. While most species grouped separately (reciprocally monophyletic), the Cx. pipiens subgroup could not be genetically separated using COI. The average conspecific and congeneric p-distance was 0.8% and 7.6%, respectively. In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport. The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance. The DNA barcodes produced by this study have been uploaded to the 'Mosquitoes of Australia-Victoria' project on the Barcode of Life Database (BOLD), which will serve as a resource for the Victorian Arbovirus Disease Control Program and other national and international mosquito surveillance programs.

No MeSH data available.


Related in: MedlinePlus

Comparison of primer locations within COI in different mosquito DNA barcoding studies. Mitochondrial gene positions sourced from (Hardy et al. 2014), GenBank accession number: NC_025473.
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ece32095-fig-0001: Comparison of primer locations within COI in different mosquito DNA barcoding studies. Mitochondrial gene positions sourced from (Hardy et al. 2014), GenBank accession number: NC_025473.

Mentions: Currently, the most commonly used barcode region for animals is a 5′‐segment of the mitochondrial gene Cytochrome Oxidase I (COI) called the ‘Universal’ or ‘Folmer’ region. This region is the standard marker chosen by the Barcode of Life Database (BOLD), which is an online platform for collating and curating DNA barcoding information from around the world (Ratnasingham and Hebert 2007). While the majority of mosquito barcoding studies use this region, some studies have used other areas of COI (Fig. 1). Often more than one marker will be used, with both mitochondrial and nuclear genes exhibiting utility in distinguishing species (Lin and Danforth 2004). In mosquito barcoding studies, a variety of nuclear markers have been used, including elongation factor‐1 alpha (EF‐1α), acetylcholinesterase 2 (ace‐2), alpha amylase, zinc finger, and internal transcribed spacer subunit 2 (ITS2) (Foley et al. 2007; Hasan et al. 2009; Hemmerter et al. 2009; Puslednik et al. 2012). Using multiple genes can help to distinguish members of species complexes and subgroups, which are closely related species that may not be genetically distinct when using just one barcoding region (Foster et al. 2013; Jiang et al. 2014).


Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia.

Batovska J, Blacket MJ, Brown K, Lynch SE - Ecol Evol (2016)

Comparison of primer locations within COI in different mosquito DNA barcoding studies. Mitochondrial gene positions sourced from (Hardy et al. 2014), GenBank accession number: NC_025473.
© Copyright Policy - creativeCommonsBy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4863023&req=5

ece32095-fig-0001: Comparison of primer locations within COI in different mosquito DNA barcoding studies. Mitochondrial gene positions sourced from (Hardy et al. 2014), GenBank accession number: NC_025473.
Mentions: Currently, the most commonly used barcode region for animals is a 5′‐segment of the mitochondrial gene Cytochrome Oxidase I (COI) called the ‘Universal’ or ‘Folmer’ region. This region is the standard marker chosen by the Barcode of Life Database (BOLD), which is an online platform for collating and curating DNA barcoding information from around the world (Ratnasingham and Hebert 2007). While the majority of mosquito barcoding studies use this region, some studies have used other areas of COI (Fig. 1). Often more than one marker will be used, with both mitochondrial and nuclear genes exhibiting utility in distinguishing species (Lin and Danforth 2004). In mosquito barcoding studies, a variety of nuclear markers have been used, including elongation factor‐1 alpha (EF‐1α), acetylcholinesterase 2 (ace‐2), alpha amylase, zinc finger, and internal transcribed spacer subunit 2 (ITS2) (Foley et al. 2007; Hasan et al. 2009; Hemmerter et al. 2009; Puslednik et al. 2012). Using multiple genes can help to distinguish members of species complexes and subgroups, which are closely related species that may not be genetically distinct when using just one barcoding region (Foster et al. 2013; Jiang et al. 2014).

Bottom Line: The average conspecific and congeneric p-distance was 0.8% and 7.6%, respectively.In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport.The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance.

View Article: PubMed Central - PubMed

Affiliation: Department of Economic Development, Jobs, Transport and Resources (DEDJTR) BioSciences Research AgriBio Centre for AgriBioscience Bundoora Victoria 3083 Australia.

ABSTRACT
DNA barcoding is a modern species identification technique that can be used to distinguish morphologically similar species, and is particularly useful when using small amounts of starting material from partial specimens or from immature stages. In order to use DNA barcoding in a surveillance program, a database containing mosquito barcode sequences is required. This study obtained Cytochrome Oxidase I (COI) sequences for 113 morphologically identified specimens, representing 29 species, six tribes and 12 genera; 17 of these species have not been previously barcoded. Three of the 29 species ─ Culex palpalis, Macleaya macmillani, and an unknown species originally identified as Tripteroides atripes ─ were initially misidentified as they are difficult to separate morphologically, highlighting the utility of DNA barcoding. While most species grouped separately (reciprocally monophyletic), the Cx. pipiens subgroup could not be genetically separated using COI. The average conspecific and congeneric p-distance was 0.8% and 7.6%, respectively. In our study, we also demonstrate the utility of DNA barcoding in distinguishing exotics from endemic mosquitoes by identifying a single intercepted Stegomyia aegypti egg at an international airport. The use of DNA barcoding dramatically reduced the identification time required compared with rearing specimens through to adults, thereby demonstrating the value of this technique in biosecurity surveillance. The DNA barcodes produced by this study have been uploaded to the 'Mosquitoes of Australia-Victoria' project on the Barcode of Life Database (BOLD), which will serve as a resource for the Victorian Arbovirus Disease Control Program and other national and international mosquito surveillance programs.

No MeSH data available.


Related in: MedlinePlus