Duplex Real-Time PCR Assays for the Simultaneous Detection and Quantification of Botryosphaeriaceae Species Causing Canker Diseases in Woody Crops

Woody canker diseases caused by fungi of the family are producing increasing losses in many economically important woody crops, including almond. To develop a molecular tool for the detection and quantification of the most aggressive and threatening species is of main importance. This will help to p...

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Bibliographic Details
Published in:Plants (Basel) 2023-06, Vol.12 (11), p.2205
Main Authors: Romero-Cuadrado, Laura, López-Herrera, Carlos José, Aguado, Ana, Capote, Nieves
Format: Article
Language:English
Subjects:
almond
Botryosphaeria
Botryosphaeria dothidea
Botryosphaeriaceae
Canker
Crop diseases
Crops
Dieback
DNA probes
Economic importance
Fungi
Infections
Latent infection
multiplex qPCR
Neofusicoccum parvum
Orchards
Pathogens
plant crude extracts
Sample preparation
Target detection
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Summary:Woody canker diseases caused by fungi of the family are producing increasing losses in many economically important woody crops, including almond. To develop a molecular tool for the detection and quantification of the most aggressive and threatening species is of main importance. This will help to prevent the introduction of these pathogens in new orchards and to conveniently apply the appropriate control measures. Three reliable, sensitive and specific duplex qPCR assays using TaqMan probes have been designed for the detection and quantification of (a) and the genus, (b) and the family and (c) and the family. The multiplex qPCR protocols have been validated on artificially and naturally infected plants. Direct systems to process plant materials, without DNA purification, allowed high-throughput detection of targets even in asymptomatic tissues. These results validate the qPCR using the direct sample preparation method as a valuable tool for Botryosphaeria dieback diagnosis allowing a large-scale analysis and the preventive detection of latent infection.
ISSN:2223-7747
2223-7747
DOI:10.3390/plants12112205