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Identification of morphologically challenging Delia (Diptera: Anthomyiidae) species from field vegetable crops using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP)

Published online by Cambridge University Press:  02 June 2021

Julia J. Mlynarek Open the ORCID record for Julia J. Mlynarek [Opens in a new window]  and
Kathrin A. Sim
Julia J. Mlynarek*
Affiliation:
Montreal Insectarium, 4581 Sherbrooke Street East, Montréal, Québec, H1X 2B2, Canada Agriculture and Agri-Food Canada, Harrow Research and Development Centre, 2585 County Road 20, Harrow, Ontario, N0R 1G0, Canada
Kathrin A. Sim
Affiliation:
Agriculture and Agri-Food Canada, Harrow Research and Development Centre, 2585 County Road 20, Harrow, Ontario, N0R 1G0, Canada
*
*Corresponding author. Email: [email protected]
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Abstract

Root feeding by the larvae of multiple Delia species can lead to economic loss in many agricultural crops. Field vegetables are subject to infestations by a species complex composed of Delia radicum (Linnaeus) (Diptera: Anthomyiidae), a pest in brassica crops (Brassicaceae), Delia antiqua (Meigen), believed to cause the majority of crop damage in onions, and the generalists Delia florilega (Zetterstedt), Delia platura (Meigen), and Botanophila fugax (Meigen) (Diptera: Anthomyiidae). Correct species identification is necessary to implement field management strategies, but these species are challenging to identify morphologically. We propose a polymerase chain reaction–restriction fragment length polymorphism method as a molecular tool to distinguish between five species of Delia and between two genetic lines of D. platura. The mitochondrial DNA cytochrome c oxidase subunit 1 barcode fragment is targeted, then the polymerase chain reaction product digested with four different restriction enzymes (AccI, BsrI, MlyI, and StyI). The BsrI enzyme distinguishes the two genetic lines of D. platura and D. florilega. The MlyI enzyme identifies B. fugax from the Delia species. Combining BsrI, StyI, AccI, and MlyI into double digestion reactions allows for rapid diagnostics among the species tested. Our method was validated using DNA from specimens collected in eastern Canada. This method provides tools in ecological and environmental studies where these species are of interest.

Type
Research Papers
Information
The Canadian Entomologist , Volume 153 , Issue 5 , October 2021 , pp. 630 - 639
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Copyright
© The Author(s) and her Majesty, the Queen, in right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada, 2021. Published by Cambridge University Press on behalf of the Entomological Society of Canada

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Footnotes

Subject editor: Hervé Colinet

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