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)

• Published in: 3 Biotech Vol. 11; no. 6; p. 259
• Main Authors: Rizzo, Domenico, Da Lio, Daniele, Bartolini, Linda, Salemi, Chiara, Del Nista, Dalia, Aronadio, Antonio, Pennacchio, Fabrizio, Binazzi, Francesco, Francardi, Valeria, Garonna, Antonio P., Rossi, Elisabetta
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.06.2021; Springer Nature B.V
• Subjects: Adults; Agriculture; ambrosia beetles; Assaying; Beetles; Bioinformatics; Biological properties; Biomaterials; Biotechnology; Cancer Research; Chemistry; Chemistry and Materials Science; Coleoptera; Curculionidae; Deoxyribonucleic acid; DNA; DNA probes; Europe; Extrusion; frass; Fruits; genus; Insects; Larvae; nontarget organisms; Original; Original Article; Reproducibility; Scolytinae; Shrubs; Species; Stem Cells; wood; Wood chips; Xylosandrus compactus; Xylosandrus germanus; Europe; Black twig-borer; Molecular diagnostics; Granulate ambrosia beetle; Black timber bark beetle
• ISSN: 2190-572X; 2190-5738
• DOI: 10.1007/s13205-021-02786-9

Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus Xylosandrus , X. compactus, X. crassiusculus and X. germanus (Coleoptera Curculionidae Scolytinae). These ambrosia beetles are xylophagous species alien to Europe, causing damages to many ornamental and fruiting trees as well as shrubs. DNA extraction was carried out from adults, larvae and biological samples derived from insect damages on infested plants. For X. compactus , segments of galleries in thin infested twigs were cut and processed; in the case of X. crassiusculus , raw frass extruded from exit holes was used, while DNA of X. germanus was extracted from small wood chips removed around insect exit holes. The assays were inclusive for the target species and exclusive for all the non-target species tested. The LoD was 3.2 pg/µL for the frass of X. crassiusculus and 0.016 ng/µL for the woody matrices of the other two species. Both repeatability and reproducibility were estimated on adults and woody samples, showing very low values ranging between 0.00 and 4.11. Thus, the proposed diagnostic assays resulted to be very efficient also on the woody matrices used for DNA extraction, demonstrating the applicability of the protocol in the absence of dead specimens or living stages.