Contribution
A multiplex PCR assay for detecting slug species common in European arable land in the diet of carabid beetles
Guenay-Greunke, Yasemin; Bohan, David A.; Traugott, Michael; Wallinger, Corinna
Entomologia Generalis Volume 42 Number 1 (2022), p. 117 - 126
publié: Feb 11, 2022
publication en ligne: Jun 15, 2021
manuscrit accepté: Mar 24, 2021
révision final du manuscrit reçu: Feb 8, 2021
révision du manuscrit demandée: Feb 4, 2021
manuscrit reçu: Dec 15, 2020
DOI: 10.1127/entomologia/2021/1248
Open Access (article peut être télechargé gratuitement)
Abstract
DNA-based diet analysis of natural enemies is a valuable tool for unravelling the food choice of predators in agroecosystems. It enables the rapid identification of potential biocontrol agents of invertebrate pests. Here, we present a new multiplex PCR system for the identification of pest slug species in the diet of their natural enemies such as carabid beetles. It comprises three species-specific primers targeting the mitochondrial cytochrome c oxidase subunit I (COI) gene to detect DNA of the common garden slug, Arion distinctus (Stylommatophora: Arionidae), the Iberian slug, Arion lusitanicus (Stylommatophora: Arionidae) and the grey field slug Deroceras reticulatum (Stylommatophora: Agriolimacidae). We also include (super)family-specific primers for Arionidae and Limacoidea, which amplify parts of the 28S gene for ribosomal RNA (rRNA) in order to identify a wider range of slugs. The amplicons for Arionidae can be assigned to a total of seven Central European slug species of this family and the amplicons for Limacoidea to ten species. The multiplex assay showed high specificity against DNA extracts of field-collected slugs and co-occurring invertebrates. The assay also exhibited high sensitivity, which was confirmed by testing it with 223 dietary samples from field-collected carabids as potential natural enemies of slugs in agroecosystems. This methodology represents a new, cost-effective, highly sensitive and specific approach for the identification of common Central European slug species as well as for analysing trophic interactions to identify natural enemies for further biological control development. It can also be applied in any study where a rapid and reliable identification of slugs is needed.
Mots-clefs
28S rRNA •
Arion distinctus
•
Arion lusitanicus
•
Arionidae •
trophic interactions •
COI •
Deroceras reticulatum
•
diagnostic PCR •
Limacoidea •
slug primers