Bulletin of Entomological Research

Research Paper

PCR-based species identification of Agriotes larvae

K. Staudachera1 c1, P. Pitterla1, L. Furlana2, P.C. Catea3 and M. Traugotta1 c1

a1 Institute of Ecology, Mountain Agriculture Research Unit, University of Innsbruck, Technikerstraße 25, 6020 Innsbruck, Austria

a2 Veneto Agricoltura, Legnaro, viale dell'Università 14, Agripolis, 35020 Legnaro PD, Italy

a3 Hebragasse 4/18, 1090 Vienna, Austria

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.

(Accepted July 01 2010)

(Online publication November 01 2010)

Correspondence:

c1 Authors for correspondence Fax: +43(0)512 507 6190 E-mail: Michael.Traugott@uibk.ac.at; Karin.Staudacher@uibk.ac.at