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Fast molecular assay to detect the rate of decay of Bactrocera oleae (Diptera: Tephritidae) DNA in Pterostichus melas (Coleoptera: Carabidae) gut contents

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Abstract

The molecular analysis of predation through specific DNA amplification has been utilized extensively over the last decade, and has been shown to be fast and effective. However, it is necessary to evaluate the prey detectability half-life if we are to correctly infer the relevance of particular predators to particular pests and to accurately model the effect of biocontrol. We present here the design and analysis of a set of primers to amplify olive fruit fly (Bactrocera oleae) DNA in predator gut contents, allowing fast evaluation of the digestion time. We modified the existing protocol by solubilizing the prey DNA directly from the gut, and we applied this modified protocol to demonstrate that Pterostichus melas, one of the most common carabids in olive groves in Italy, feeds on B. oleae pupae. After feeding carabids with a single pupa, traces of the pest were found to be detectable more than 20 h after ingestion. This method could also be applied to other predators to evaluate trophic interactions of the olive fruit fly. The relevance of generalist predation to the mortality of the pupal stage of B. oleae is of great economic interest since B. oleae causes serious damage during olive production, reducing the commercial value of olive oil and table olives.

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Acknowledgements

The work was supported by the Fondo di Finanziamento Ordinario of the Italian Ministry of Instruction, University and Research.

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Authors and Affiliations

  1. Department of Biology, Ecology and Earth Science, University of Calabria, Ponte Bucci 6C, 87036, Rende, CS, Italy

    Simona Panni & Roberto Pizzolotto

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  1. Simona Panni
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  2. Roberto Pizzolotto
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Contributions

SP and RP conceived the research and conducted the experiments. RP analyzed the data and conducted the statistical analyses. SP and RP wrote the manuscript. RP secured funding. All authors read and approved the manuscript.

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Correspondence to Simona Panni.

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Fig. S1

. Multiple sequence alignment of part of the cox1 gene. The sequence from Bactrocera oleae was compared with those of other carabids that live in olive groves and those of some potential prey species of Pterostichus (earthworm, springtail, annelid, moth, firefly, and pot worm). The positions of the primers p_CoxO and p_CoxD are indicated. Pterostichus melas: genebank KM452177.1; Bactrocera oleae: AY210703.1 Lumbricus terrestris NC_001673.1; Orchesella cincta (Collembola) AJ272261.1; Enchytraeidae KM206565.1; Calathus fuscipes KM451314.1; Calathus cinctus KM452419.1; Carabus coriaceus KP067574.1; Oniscus asellus KU955993.1; Lepidoptera JN297456.1; Luciola italica KM448530.1 (PDF 73 kb)

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Panni, S., Pizzolotto, R. Fast molecular assay to detect the rate of decay of Bactrocera oleae (Diptera: Tephritidae) DNA in Pterostichus melas (Coleoptera: Carabidae) gut contents. Appl Entomol Zool 53, 425–431 (2018). https://doi.org/10.1007/s13355-018-0564-x

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