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iForest - Biogeosciences and Forestry

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Identification of the ambrosia beetle Anisandrus dispar (Fabricius) (Coleoptera Curculionidae Scolytinae) using TaqMan™ probe assay on biological samples

Domenico Rizzo (1), Andrea D’Agostino (2), Igor Stabile (2), Chiara Ranaldi (2), Andrea Marrucci (2), Claudia Gabriela Zubieta (2), Daniele Da Lio (2), Linda Bartolini (1), Fabrizio Pennacchio (3), Elisabetta Rossi (2), Antonio P Garonna (4)   

iForest - Biogeosciences and Forestry, Volume 16, Issue 3, Pages 182-187 (2023)
doi: https://doi.org/10.3832/ifor4287-016
Published: Jun 30, 2023 - Copyright © 2023 SISEF

Research Articles


The European shot-hole borer Anisandrus dispar (Fabricius) (Coleoptera Curculionidae Scolytinae) is a well-known ambrosia beetle living on shrubs and several ornamental and fruiting trees where it can cause heavy damages. Like other harmful xyleborine species, A. dispar can represent a potential threat outside its native region. Molecular diagnostic tools can lead to accurate identification of xylophagous species hidden in wooden matrix in phytosanitary surveys at entry points. A molecular assay based on qPCR TaqMan™ Probes was developed for the identification of A. dispar from different matrices. To setup and perform the test, DNA extraction was carried out from adults, larvae, and artificial samples of wood chips from oak healthy plants whose lysates were contaminated with a known amount of DNA of A. dispar adults. The assay has proven inclusive for A. dispar, and exclusive towards the non-target organisms, showing 100% analytical specificity. The limit of detection was 0.32 pg µL-1 for the samples of insect adult DNA of A. dispar, and at 0.8 pg µL-1 for the samples containing lysates of Quercus spp. and 0.1 ng µL-1 of A. dispar adult DNA. Repeatability and reproducibility showed low values independently from the matrix used for DNA extraction, confirming the possible use in diagnostics of biological samples even if not directly related to the presence of A. dispar developmental stages. The presented approach may be adjusted and applied for phytosanitary purposes to other quarantine pests and rapidly detect possible infestations in vegetal matrices globally traded.

  Keywords


European Shot-hole Borer, Xyleborini, Quarantine Species, Molecular Diagnostics, Biological Traces

Authors’ address

(1)
Domenico Rizzo 0000-0003-2834-4684
Linda Bartolini
Laboratory of Phytopathological Diagnostics and Molecular Biology, Plant Protection Service of Tuscany, v. Ciliegiole 99, I-51100 Pistoia (Italy)
(2)
Andrea D’Agostino
Igor Stabile
Chiara Ranaldi
Andrea Marrucci
Claudia Gabriela Zubieta
Daniele Da Lio 0000-0002-9222-9501
Elisabetta Rossi 0000-0002-6073-1046
Department of Agricultural, Food and Agro-Environmental Sciences, University of Pisa, v. del Borghetto 80, I-56124 Pisa (Italy)
(3)
Fabrizio Pennacchio 0000-0001-6609-4386
CREA - Research Centre for Plant Protection and Certification, v. Lanciola 12/A, I-50125 Florence (Italy)
(4)
Antonio P Garonna 0000-0002-8441-5208
Department of Agricultural Sciences, University of Naples “Federico II”, v. Università 100, I-80055 Portici (Italy)
(5)
retired.

Corresponding author

 
Antonio P Garonna
garonna@unina.it

Citation

Rizzo D, D’Agostino A, Stabile I, Ranaldi C, Marrucci A, Zubieta CG, Da Lio D, Bartolini L, Pennacchio F, Rossi E, Garonna AP (2023). Identification of the ambrosia beetle Anisandrus dispar (Fabricius) (Coleoptera Curculionidae Scolytinae) using TaqMan™ probe assay on biological samples. iForest 16: 182-187. - doi: 10.3832/ifor4287-016

Academic Editor

Matteo Marchioro

Paper history

Received: Dec 09, 2022
Accepted: Mar 28, 2023

First online: Jun 30, 2023
Publication Date: Jun 30, 2023
Publication Time: 3.13 months

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