Rapid detection of pityophthorus juglandis (Blackman) (coleoptera, curculionidae) with the loop-mediated isothermal amplification (lamp) method
| Autor: | Umberto Bernardo, Giovanni Cappellini, Nicola Luchi, Chiara Salemi, Daniele Da Lio, Domenico Rizzo, Alessandra Benigno, Santa Olga Cacciola, Matteo Bracalini, Salvatore Moricca, Tiziana Panzavolta, Francesco Nugnes |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Bark beetle bark beetle invasive species molecular identification thousand canker disease walnut Loop-mediated isothermal amplification Plant Science 01 natural sciences Article 03 medical and health sciences Walnut twig beetle medicine Ecology Evolution Behavior and Systematics Phytosanitary certification Geosmithia morbida Ecology biology Frass Botany biology.organism_classification medicine.disease Horticulture 030104 developmental biology Curculionidae Thousand cankers disease QK1-989 010606 plant biology & botany |
| Zdroj: | Plants 10 (2021). doi:10.3390/plants10061048 info:cnr-pdr/source/autori:Rizzo D., Moricca S., Bracalini M., Benigno A., Bernardo U., Luchi N., Da Lio D., Nugnes F., Cappellini G., Salemi C., Cacciola S.O., Panzavolta T./titolo:Rapid detection of pityophthorus juglandis (Blackman) (coleoptera, curculionidae) with the loop-mediated isothermal amplification (lamp) method/doi:10.3390%2Fplants10061048/rivista:Plants/anno:2021/pagina_da:/pagina_a:/intervallo_pagine:/volume:10 Plants, Vol 10, Iss 1048, p 1048 (2021) Plants Volume 10 Issue 6 |
| DOI: | 10.3390/plants10061048 |
| Popis: | The walnut twig beetle Pityophthorus juglandis is a phloem-boring bark beetle responsible, in association with the ascomycete Geosmithia morbida, for the Thousand Cankers Disease (TCD) of walnut trees. The recent finding of TCD in Europe prompted the development of effective diagnostic protocols for the early detection of members of this insect/fungus complex. Here we report the development of a highly efficient, low-cost, and rapid method for detecting the beetle, or even just its biological traces, from environmental samples: the loop-mediated isothermal amplification (LAMP) assay. The method, designed on the 28S ribosomal RNA gene, showed high specificity and sensitivity, with no cross reactivity to other bark beetles and wood-boring insects. The test was successful even with very small amounts of the target insect’s nucleic acid, with limit values of 0.64 pg/µL and 3.2 pg/µL for WTB adults and frass, respectively. A comparison of the method (both in real time and visual) with conventional PCR did not display significant differences in terms of LoD. This LAMP protocol will enable quick, low-cost, and early detection of P. juglandis in areas with new infestations and for phytosanitary inspections at vulnerable sites (e.g., seaports, airports, loading stations, storage facilities, and wood processing companies). |
| Databáze: | OpenAIRE |
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