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)

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 an...

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Published in3 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
LanguageEnglish
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
Online AccessGet full text
ISSN2190-572X
2190-5738
DOI10.1007/s13205-021-02786-9

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Abstract 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.
AbstractList 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.
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.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.
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.
Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus , and (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 , segments of galleries in thin infested twigs were cut and processed; in the case of , raw frass extruded from exit holes was used, while DNA of 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 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.
ArticleNumber 259
Author Rizzo, Domenico
Aronadio, Antonio
Pennacchio, Fabrizio
Bartolini, Linda
Binazzi, Francesco
Salemi, Chiara
Da Lio, Daniele
Del Nista, Dalia
Francardi, Valeria
Rossi, Elisabetta
Garonna, Antonio P.
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Issue 6
Keywords Black twig-borer
Molecular diagnostics
Granulate ambrosia beetle
Black timber bark beetle
Language English
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Snippet Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus Xylosandrus , X. compactus, X. crassiusculus and X. germanus...
Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus , and (Coleoptera Curculionidae Scolytinae). These ambrosia...
Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus Xylosandrus, X. compactus, X. crassiusculus and X. germanus...
SourceID pubmedcentral
proquest
pubmed
crossref
springer
SourceType Open Access Repository
Aggregation Database
Index Database
Enrichment Source
Publisher
StartPage 259
SubjectTerms 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
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Title 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)
URI https://link.springer.com/article/10.1007/s13205-021-02786-9
https://www.ncbi.nlm.nih.gov/pubmed/33996371
https://www.proquest.com/docview/2524919113
https://www.proquest.com/docview/2528434140
https://www.proquest.com/docview/2551959375
https://pubmed.ncbi.nlm.nih.gov/PMC8110672
Volume 11
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