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PCR-based species identification of Agriotes larvae

Published online by Cambridge University Press:  01 November 2010

K. Staudacher ,
P. Pitterl ,
L. Furlan ,
P.C. Cate  and
M. Traugott
K. Staudacher*
Affiliation:
Institute of Ecology, Mountain Agriculture Research Unit, University of Innsbruck, Technikerstraße 25, 6020 Innsbruck, Austria
P. Pitterl
Affiliation:
Institute of Ecology, Mountain Agriculture Research Unit, University of Innsbruck, Technikerstraße 25, 6020 Innsbruck, Austria
L. Furlan
Affiliation:
Veneto Agricoltura, Legnaro, viale dell'Università 14, Agripolis, 35020 Legnaro PD, Italy
P.C. Cate
Affiliation:
Hebragasse 4/18, 1090 Vienna, Austria
M. Traugott*
Affiliation:
Institute of Ecology, Mountain Agriculture Research Unit, University of Innsbruck, Technikerstraße 25, 6020 Innsbruck, Austria
*
*Authors for correspondence Fax: +43(0)512 507 6190 E-mail: Michael.Traugott@uibk.ac.at; Karin.Staudacher@uibk.ac.at
*Authors for correspondence Fax: +43(0)512 507 6190 E-mail: Michael.Traugott@uibk.ac.at; Karin.Staudacher@uibk.ac.at
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Abstract

Click beetle larvae within the genus Agriotes (Coleoptera: Elateridae), commonly known as wireworms, are abundant ground-dwelling herbivores which can inflict considerable damage to field crops. In Central Europe up to 20 species, which differ in their distribution, ecology and pest status, occur in arable land. However, the identification of these larvae based on morphological characters is difficult or impossible. This hampers progress towards controlling these pests. Here, we present a polymerase chain reaction (PCR)-based approach to identify, for the first time, 17 Agriotes species typically found in Central Europe. Diagnostic sequence information was generated and submitted to GenBank, allowing the identification of these species via DNA barcoding. Moreover, multiplex PCR assays were developed to identify the nine most abundant species rapidly within a single-step reaction: Agriotes brevis, A. litigiosus, A. obscurus, A. rufipalpis, A. sordidus, A. sputator, A. ustulatus, A. lineatus and A. proximus. The latter two species remain molecularly indistinguishable, questioning their species status. The multiplex PCR assays proved to be highly specific against non-agrioted elaterid beetles and other non-target soil invertebrates. By testing the molecular identification system with over 900 field-collected larvae, our protocol proved to be a reliable, cheap and quick method to routinely identify Central European Agriotes species.

Keywords

barcodingmultiplex PCRwirewormsElateridae

Information

Type
Research Paper
Information
Bulletin of Entomological Research , Volume 101 , Issue 2 , April 2011 , pp. 201 - 210
Copyright
Copyright © Cambridge University Press 2010

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