Liu, Ting Li, Qi Song, Junlin and Yu, Hong 2017. Development of genomic microsatellite multiplex PCR using dye-labeled universal primer and its validation in pedigree analysis of Pacific oyster (Crassostrea gigas). Journal of Ocean University of China, Vol. 16, Issue. 1, p. 151.
Benefer, Carly M. D'Ahmed, Karzan S. Murray, Philip J. and Blackshaw, Rod P. 2017. Molecular identification and distribution of leatherjackets (Diptera: Tipulidae) in U.K. agricultural grassland. Agricultural and Forest Entomology, Vol. 19, Issue. 4, p. 400.
Milosavljević, Ivan Esser, Aaron D. and Crowder, David W. 2016. Effects of environmental and agronomic factors on soil-dwelling pest communities in cereal crops. Agriculture, Ecosystems & Environment, Vol. 225, p. 192.
Ritter, Claudia de Mol, Friederike Richter, Ellen Struck, Christine and Katroschan, Kai-Uwe 2016. Antipredator Behavioral Traits of some Agriotes Wireworms (Coleoptera: Elateridae) and their Potential Implications for Species Identification. Journal of Insect Behavior, Vol. 29, Issue. 2, p. 214.
Han, Taeman Lee, Wonhoon Lee, Seunghwan Park, In Gyun Park, Haechul and Fontaneto, Diego 2016. Reassessment of Species Diversity of the Subfamily Denticollinae (Coleoptera: Elateridae) through DNA Barcoding. PLOS ONE, Vol. 11, Issue. 2, p. e0148602.
Furlan, Lorenzo and Kreutzweiser, David 2015. Alternatives to neonicotinoid insecticides for pest control: case studies in agriculture and forestry. Environmental Science and Pollution Research, Vol. 22, Issue. 1, p. 135.
Oba, Yuichi Ôhira, Hitoo Murase, Yukio Moriyama, Akihiko Kumazawa, Yoshinori and Mans, Ben J 2015. DNA Barcoding of Japanese Click Beetles (Coleoptera, Elateridae). PLOS ONE, Vol. 10, Issue. 1, p. e0116612.
G van Herk, Willem Vernon, Robert S. Vojtko, Bobbi Snow, Shelby Fortier, Jacqueline and Fortin, Chantelle 2015. Contact behaviour and mortality of wireworms exposed to six classes of insecticide applied to wheat seed. Journal of Pest Science, Vol. 88, Issue. 4, p. 717.
Traugott, Michael Benefer, Carly M. Blackshaw, Rod P. van Herk, Willem G. and Vernon, Robert S. 2015. Biology, Ecology, and Control of Elaterid Beetles in Agricultural Land. Annual Review of Entomology, Vol. 60, Issue. 1, p. 313.
Gomez-Polo, Priscila Traugott, Michael Alomar, Oscar Castañé, Cristina Rojo, Santos and Agustí, Nuria 2014. Identification of the most common predatory hoverflies of Mediterranean vegetable crops and their parasitism using multiplex PCR. Journal of Pest Science, Vol. 87, Issue. 2, p. 371.
Raso, Lorna Sint, Daniela Rief, Alexander Kaufmann, Rüdiger Traugott, Michael and Lin, Baochuan 2014. Molecular Identification of Adult and Juvenile Linyphiid and Theridiid Spiders in Alpine Glacier Foreland Communities. PLoS ONE, Vol. 9, Issue. 7, p. e101755.
Furlan, Lorenzo 2014. IPM thresholds for Agriotes wireworm species in maize in Southern Europe. Journal of Pest Science, Vol. 87, Issue. 4, p. 609.
Serjeant, Anthony F. and Beebee, Trevor J. C. 2013. Identification of Dytiscus larvae by cytochrome oxidase I (COI) partial sequencing. Journal of Insect Conservation, Vol. 17, Issue. 6, p. 1191.
Barsics, Fanny Haubruge, Eric and Verheggen, François 2013. Wireworms’ Management: An Overview of the Existing Methods, with Particular Regards to Agriotes spp. (Coleoptera: Elateridae). Insects, Vol. 4, Issue. 4, p. 117.
Ritter, Claudia and Richter, Ellen 2013. Control methods and monitoring of Agriotes wireworms (Coleoptera: Elateridae). Journal of Plant Diseases and Protection, Vol. 120, Issue. 1, p. 4.
Benefer, Carly M. van Herk, W. G. Ellis, J. S. Blackshaw, R. P. Vernon, R. S. and Knight, M. E. 2013. The molecular identification and genetic diversity of economically important wireworm species (Coleoptera: Elateridae) in Canada. Journal of Pest Science, Vol. 86, Issue. 1, p. 19.
Kleespies, Regina G. Ritter, Claudia Zimmermann, Gisbert Burghause, Frank Feiertag, Simon and Leclerque, Andreas 2013. A survey of microbial antagonists of Agriotes wireworms from Germany and Italy. Journal of Pest Science, Vol. 86, Issue. 1, p. 99.
Traugott, Michael Schallhart, Nikolaus Staudacher, Karin and Wallinger, Corinna 2013. Understanding the ecology of wireworms and improving their control: a special issue. Journal of Pest Science, Vol. 86, Issue. 1, p. 1.
Staudacher, Karin Schallhart, Nikolaus Pitterl, Peter Wallinger, Corinna Brunner, Nina Landl, Marion Kromp, Bernhard Glauninger, Johann and Traugott, Michael 2013. Occurrence of Agriotes wireworms in Austrian agricultural land. Journal of Pest Science, Vol. 86, Issue. 1, p. 33.
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.
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