iForest - Biogeosciences and Forestry


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.


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

Authors’ address

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)
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)
Fabrizio Pennacchio 0000-0001-6609-4386
CREA - Research Centre for Plant Protection and Certification, v. Lanciola 12/A, I-50125 Florence (Italy)
Antonio P Garonna 0000-0002-8441-5208
Department of Agricultural Sciences, University of Naples “Federico II”, v. Università 100, I-80055 Portici (Italy)

Corresponding author

Antonio P Garonna


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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Albo JE, Marelli JP, Puig AS (2020)
Rapid molecular identification of Scolytinae (Coleoptera: Curculionidae). International Journal of Molecular Science 20 (23): 5944.
CrossRef | Gscholar
Amini S, Nozari J, Hosseini R, Rahati R (2020)
Comparison of different DNA extraction methods for the molecular study of bark beetles (Coleoptera: Curculionidae, Scolytinae). Journal of Insect Biodiversity and Systematics 6 (1): 113-124.
CrossRef | Gscholar
Avtzis DN, Lakatos F (2021)
Bark and wood boring insects. past, present, and the future know ledge we need. Insects 12: 28.
CrossRef | Gscholar
Briski E, Ghabooli S, Bailey SA, Maclsaac HJ (2016)
Are genetic databases sufficiently populated to detect non-indigenous species? Biological Invasions 18: 1911-1922.
CrossRef | Gscholar
Bucini D, Balestra GM, Pucci C, Paparatti B (2005)
Bio-ethology of Anisandrus dispar F. and its possible involvement in dieback (Moria) diseases of hazelnut (Corylus avellana L.) plants in Central Italy. Acta Horticulturae 686: 435-444.
CrossRef | Gscholar
Cognato AI, Sari G, Smith SM, Beaver RA, Li Y, Hulcr J, Jordal BH, Kajimura H, Lin CS, Pham TH, Singh S, Sittichaya W (2020)
The essential role of taxonomic expertise in the creation of DNA databases for the identification and delimitation of Southeast Asian ambrosia beetle species (Curculionidae: Scolytinae: Xyleborini). Frontiers in Ecology and Evolution 8: 27.
CrossRef | Gscholar
Colombari F, Martinez Sañudo I, Battisti A (2022)
First report of the alien ambrosia beetle Cnestus mutilatus and further finding of Anisandrus maiche in the EPPO region (Coleoptera: Curculionidae: Scolytinae: Xyleborini). EPPO Bulletin 00: 1-5.
CrossRef | Gscholar
EPPO (2021)
PM 7/98 (5) Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity. Bulletin OEPP/EPPO 51: 468-498.
CrossRef | Gscholar
EPPO (2022)
First record of the ambrosia beetle Anisandrus maiche in Switzerland. EPPO Reporting Service no. 07, pp. 148.
Online | Gscholar
French JRJ, Roeper RA (1975)
Studies on the biology of the ambrosia beetle Xyleborus dispar (F.) (Coleoptera: Scolytidae). Zeitschrift für Angewandte Entomologie 78: 241-247.
CrossRef | Gscholar
Galko J, Nikolov C, Kimoto T, Kunca A, Gubka A, Vakula J, Zúbrik M, Ostrihon M (2014)
Attraction of ambrosia beetles to ethanol baited traps in a Slovakian oak forest. Biologia 69: 1376-1383.
CrossRef | Gscholar
Gohli J, Selvarajah T, Kirkendall LR, Jordal BL (2016)
Globally distributed Xyleborus species reveal recurrent intercontinental dispersal in a landscape of ancient worldwide distributions. BMC Evolutionary Biology 16: 37.
CrossRef | Gscholar
Gomez DF, Rabaglia RJ, Fairbanks KEO, Hulcr J (2018)
North American Xyleborini north of Mexico: a review and key to genera and species (Coleoptera, Curculionidae, Scolytinae). ZooKeys 768: 19-68.
CrossRef | Gscholar
Ide T, Kanzaki N, Ohmura W, Okabe K (2016a)
Molecular identification of the western drywood termite (Isoptera: Kalotermitidae) by loop-mediated isothermal amplification of DNA from fecal pellets. Journal of Economic Entomology 109 (3): 2234-2237.
CrossRef | Gscholar
Ide T, Kanzaki N, Ohmura W, Okabe K (2016b)
Molecular identification of an invasive wood-boring insect Lyctus brunneus (Coleoptera: Bostrichidae: Lyctinae) using frass by loop-mediated isothermal amplification and nested PCR assays. Journal of Economic Entomology 109 (3): 1410-1414.
CrossRef | Gscholar
Ioos R, Fourrier C, Iancu G, Gordon TR (2009)
Sensitive detection of Fusarium circinatum in pine seed by combining an enrichment procedure with a real-time polymerase chain reaction using dual-labeled probe chemistry. Phytopathology 99: 582-590.
CrossRef | Gscholar
Katoh K, Standley DM (2013)
MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30 (4): 772-780.
CrossRef | Gscholar
Li R, Mocka R, Huang Q, Abad J, Hartung J, Kinarda G (2008)
A reliable and inexpensive method of nucleic acid extraction for the PCR-based detection of diverse plant pathogens. Journal of Virological Methods 154: 48-55.
CrossRef | Gscholar
Mandelshtam MY, Yakushkin EA, Petrov AV (2018)
Oriental ambrosia beetles (Coleoptera: Curculionidae: Scolytinae): new inhabitants of Primorsky krai in Russia. Russian Journal of Biological Invasions 9: 355-365.
CrossRef | Gscholar
Meurisse N, Rassati D, Hurley BP, Brockerhoff EG, Haack RA (2018)
Common pathways by which non-native forest insects move internationally and domestically. Journal of Pest Science 92: 13-27.
CrossRef | Gscholar
Pennacchio F, Sabbatini PG, Jucker C, Allegro G, Roversi PF (2012)
A key for the identification of larvae of Anoplophora chinensis, Anoplophora glabripennis and Psacothea hilaris (Coleoptera Cerambycidae Lamiinae) in Europe. REDIA - Journal of Zoology 95: 57-65.
Poland TM, Rassati D (2019)
Improved biosecurity surveillance of non-native forest insects: a review of current methods. Journal of Pest Science 92: 37-49.
CrossRef | Gscholar
Rabaglia RJ, Dole SA, Cognato AI (2006)
Review of American Xyleborina (Coleoptera: Curculionidae: Scolyinae) occurring North of Mexico, with an illustrated key. Annals of the Entomological Society of America 99 (6): 1034-1056.
CrossRef | Gscholar
Rabaglia RJ, Cognato AI, Hoebeke ER, Johnson CW, Labonte JR, Carter ME, Vlack JJ (2019)
Early detection and rapid response: a 10-year summary of the USDA Forest Service program of surveillance for non-native bark and Ambrosia beetles. American Entomologist 65 (1): 29-42.
CrossRef | Gscholar
Rassati D, Faccoli M, Petrucco Toffolo E, Battisti A, Marini L (2015)
Improving the early detection of alien wood-boring beetles in ports and surrounding forests. Journal of Applied Ecology 52: 50-58.
CrossRef | Gscholar
Rizzo D, Taddei F, Da Lio D, Nugnes F, Barra E, Stefani L, Bartolini L, Griffo RV, Spigno P, Cozzolino L, Rossi E, Garonna AP (2020a)
Identification of the red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) with Real-Time PCR on frass. Sustainability 12: 6041.
CrossRef | Gscholar
Rizzo D, Taddei A, Da Lio D, Bruscoli T, Cappellini G, Bartolini L, Salemi C, Luchi N, Pennacchio F, Rossi E (2020b)
Molecular identification of Anoplophora glabripennis (Coleoptera: Cerambycidae) from frass by loop-mediated isothermal amplification. Journal of Economic Entomology 113 (6): 2911-2919.
CrossRef | Gscholar
Rizzo D, Da Lio D, Bartolini L, Cappellini G, Bruscoli T, Bracalini M, Benigno A, Salemi C, Del Nista D, Aronadio A, Panzavolta T, Moricca S (2020c)
A duplex real-time PCR with probe for simultaneous detection of Geosmithia morbida and its vector Pityophthorus juglandis. PLoS One 15 (10): e0241109.
CrossRef | Gscholar
Rizzo D, Da Lio D, Bartolini L, Salemi C, Del Nista D, Aronadio A, Pennacchio F, Binazzi F, Francardi V, Garonna AP, Rossi E (2021)
TaqMan probe assays on different biological samples for the identification of three ambrosia beetle species, Xylosandrus compactus (Eichoff), X. crassiusculus (Motschulsky) and X. germanus (Blandford) (Coleoptera Curculionidae Scolytinae). 3 Biotech 11: 259.
CrossRef | Gscholar
Ruzzier E, Bani L, Cavaletto G, Faccoli M, Rassati D (2022)
Anisandrus maiche Kurentzov (Curculionidae: Scolytinae), an Asian species recently introduced and now widely established in Northern Italy. BioInvasions Records 11 (3): 652-658.
CrossRef | Gscholar
Saruhan H, Akyol I (2012)
Monitoring population density and fluctuations of Anisandrus dispar and Xyleborinus saxesenii (Coleoptera: Scolytinae, Curculionidae) in hazelnut orchards. African Journal of Biotechnology 11: 4202-4207.
CrossRef | Gscholar
Sauvard D (2004)
General biology of bark beetles. Chapter 7. In: “Bark and Wood Boring Insects in Living Trees in Europe: A Synthesis” (Lieutier F, Day K, Battisti A, Gregorie J-C, Evans H eds). Springer, Dordrecht, Netherlands, pp. 63-88.
CrossRef | Gscholar
Smith SM, Beaver RA, Cognato AI (2020)
A monograph of the Xyleborini (Coleoptera, Curculionidae, Scolytinae) of the Indochinese Peninsula (except Malaysia) and China. Zookeys 983: 1-442.
CrossRef | Gscholar
Speranza S, Bucini D, Paparatti B (2009)
New observation on biology of European Shot-Hole Borer (Xyleborus dispar (F.)) on hazel in Northern Latium (Central Italy). Acta Horticolturae 845: 539-542.
CrossRef | Gscholar
Strangi A, Sabbatini Peverieri G, Roversi PF (2013)
Managing outbreaks of the citrus long-horned beetle Anoplophora chinensis (Forster) in Europe: molecular diagnosis of plant infestation. Pest Management Science 69: 627-634.
CrossRef | Gscholar
Taddei A, Becker M, Berger B, Da Lio D, Feltgen S, König S, Hoppe B, Rizzo D (2021)
Molecular identification of Anoplophora glabripennis (Coleoptera: Cerambycidae) and detection from frass samples based on real-time quantitative PCR. Journal of Plant Diseases and Protection 128: 1587-1601.
CrossRef | Gscholar
Tanasković S, Marjanović M, Gvozdenac S, Popović N, Drašković G (2016)
Sudden occurrence and harmfulness of Xyleborus dispar (Fabricius) on pear. Contemporary Agriculture 65 (3-4): 57-62.
CrossRef | Gscholar
Vega FE, Hofstetter RWE (2014)
Bark beetles, biology and ecology of native and invasive species. Academic Press, London, UK, pp. 620.
CrossRef | Gscholar
Wood SL, Bright DE (1992)
A catalog of Scolytinae and Platypotidae (Coleoptera). Part 2: Taxonomic Index, Volume A. Great Basin Naturalist Memories, pp. 1-833.
Wu Y, Trepanowski NF, Molongoski JJ, Reagel PF, Lingafelter SW, Nadel H, Myers SW, Ray AM (2017)
Identification of wood-boring beetles (Cerambycidae and Buprestidae) intercepted in trade associated solid wood packaging material using DNA barcoding and morphology. Scientific Reports 7: 40316.
CrossRef | Gscholar

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