A PCR-based diagnostic assay for detecting DNA of the olive fruit fly, Bactrocera oleae, in the gut of soil-living arthropods

• Published in: Bulletin of entomological research 2016-10, Vol.106 (5), p.695-699
• Main Authors: Rejili, Mokhtar, Fernandes, Telma, Dinis, A. M., Pereira, J. A., Baptista, P., Santos, S. A. P., Lino-Neto, T.
• Format: Article
• Language: English
• Subjects: Animals; Bactrocera oleae; Biological; Ciências Biológicas; Ciências Naturais; Coleoptera; diagnostic PCR; DNA; DNA Primers; feeding assay; Feeding Behavior; Mitochondrial; Olea; Pest Control; Polymerase Chain Reaction; Predatory Behavior; Reproducibility of Results; Science & Technology; Soil; Species Specificity; specific primers; Tephritidae
• ISSN: 1475-2670
• DOI: 10.1017/S000748531600050X

Bactrocera oleae (Rossi) (Diptera: Tephritidae) is considered the most devastating pest of the olive tree worldwide. In an effort to develop management and biological control strategies against this pest, new molecular tools are urgently needed. In this study, we present the design of B. oleae-specific primers based on mitochondrial DNA sequences of cytochrome oxidase subunit I (COI) gene. Two pairs of B. oleae-specific primers were successfully designed and named as SBo1-F/SBo1-R and SBo2-F/SBo1-R, being able to amplify 108 and 214 bp COI fragments, respectively. The specificity of designed primers was tested by amplifying DNA from phylogenetically related (i.e. Diptera order) and other non-pest insects living in olive groves from the Mediterranean region. When using these primers on a PCR-based diagnostic assay, B. oleae DNA was detected in the gut content of a soil-living insect, Pterostichus globosus (Fabricius) (Coleoptera: Carabidae). The detection of B. oleae DNA in the guts of arthropods was further optimized by adding bovine serum albumin enhancer to the PCR reaction, in order to get a fast, reproducible and sensitive tool for detecting B. oleae remains in the guts of soil-living arthropods. This molecular tool could be useful for understanding pest-predator relationships and establishing future biological control strategies for this pest. This work was supported by FEDER Funds throughout Programa Operacional Factores de Competitividade – COMPETE and National Funds throughout FCT – Fundação para a Ciência e Tecnologia, within the project EXCL/ AGR-PRO/0591/2012: Olive crop protection in sustainable production under global climatic changes: linking ecological infrastructures to ecosystem functions. M. Rejili was supported by a fellowship on the frame of Erasmus Mundus EU MARE NOSTRUM.