The cultivation of walnuts (Juglans sp.) in Europe retains high economic, social, and environmental value. The recent reporting of the Thousand Cankers Disease (TCD) fungus, Geosmithia morbida, and of its vector, Pityophthorus juglandis, in walnut trees in Italy is alarming the whole of Europe. Although Italy is at present the only foothold of the disease outside North America, given the difficulties inherent in traditional identification of both members of this beetle/fungus complex, a rapid and effective protocol for the early detection and identification of TCD organisms is an absolute priority for Europe. Here we report the development of an effective and sensitive molecular tool based on simplex/duplex qPCR assays for the rapid, accurate and highly specific detection of both the bionectriaceous fungal pathogen and its bark-beetle vector. Our assay performed excellently, detecting minute amounts of target DNA without any non-specific amplification. Detection limits from various and heterogeneous matrices were lower than other reported assays. Our molecular protocol could assist in TCD organism interception at entry points, territory monitoring for the early identification and eradication of outbreaks, delineation of quarantine areas, and tracing back TCD entry and dispersal pathways.

A duplex real-time PCR with probe for simultaneous detection of Geosmithia morbida and its vector Pityophthorus juglandis / Rizzo, Domenico; Da Lio, Daniele; Bartolini, Linda; Cappellini, Giovanni; Bruscoli, Tommaso; Bracalini, Matteo; Benigno, Alessandra; Salemi, Chiara; Del Nista, Dalia; Aronadio, Antonio; Panzavolta, Tiziana; Moricca, Salvatore. - In: PLOS ONE. - ISSN 1932-6203. - STAMPA. - 15:(2020), pp. e0241109-16. [10.1371/journal.pone.0241109]

A duplex real-time PCR with probe for simultaneous detection of Geosmithia morbida and its vector Pityophthorus juglandis

Cappellini, Giovanni;Bruscoli, Tommaso;Bracalini, Matteo;Benigno, Alessandra;Panzavolta, Tiziana
;
Moricca, Salvatore
2020

Abstract

The cultivation of walnuts (Juglans sp.) in Europe retains high economic, social, and environmental value. The recent reporting of the Thousand Cankers Disease (TCD) fungus, Geosmithia morbida, and of its vector, Pityophthorus juglandis, in walnut trees in Italy is alarming the whole of Europe. Although Italy is at present the only foothold of the disease outside North America, given the difficulties inherent in traditional identification of both members of this beetle/fungus complex, a rapid and effective protocol for the early detection and identification of TCD organisms is an absolute priority for Europe. Here we report the development of an effective and sensitive molecular tool based on simplex/duplex qPCR assays for the rapid, accurate and highly specific detection of both the bionectriaceous fungal pathogen and its bark-beetle vector. Our assay performed excellently, detecting minute amounts of target DNA without any non-specific amplification. Detection limits from various and heterogeneous matrices were lower than other reported assays. Our molecular protocol could assist in TCD organism interception at entry points, territory monitoring for the early identification and eradication of outbreaks, delineation of quarantine areas, and tracing back TCD entry and dispersal pathways.
2020
15
e0241109
16
Goal 3: Good health and well-being for people
Rizzo, Domenico; Da Lio, Daniele; Bartolini, Linda; Cappellini, Giovanni; Bruscoli, Tommaso; Bracalini, Matteo; Benigno, Alessandra; Salemi, Chiara; Del Nista, Dalia; Aronadio, Antonio; Panzavolta, Tiziana; Moricca, Salvatore
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Descrizione: Sono stati messi a punto dei saggi molecolari rapidi, accurati e ripetibili per la pronta identificazione dell'agente della malattia dei "Mille cancri del Noce" Geosmithia morbida e del suo insetto vettore Pityophthorus juglandis
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1213774
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