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Article

Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen

1
Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera km 4.5, Moncada, 46113 Valencia, Spain
2
Plant Pathology Laboratory, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
*
Author to whom correspondence should be addressed.
Plants 2020, 9(9), 1151; https://doi.org/10.3390/plants9091151
Received: 10 August 2020 / Revised: 1 September 2020 / Accepted: 3 September 2020 / Published: 4 September 2020
(This article belongs to the Special Issue Diagnosis and Control of Plant Viral Diseases)
Grapevine Roditis leaf discoloration-associated virus (GRLDaV) is an emerging grapevine pathogen included in the European and Mediterranean Plant Protection Organization (EPPO) alert list due to its ability to damage grapevine crops and cause production losses. This work aimed to develop a specific and reliable diagnostic tool that would contribute to preventing the spread of this pathogen. Therefore, a TaqMan real-time quantitative PCR was developed. The method was validated according to EPPO guidelines showing a high degree of analytical sensitivity, analytical specificity, selectivity, and repeatability and reproducibility. The sensitivity of this method is much higher than the sensitivity reached by previously reported methods even when tested in crude extracts, which could allow rapid testing by avoiding nucleic acid extraction steps. The method was also able to detect GRLDaV isolates from all the geographic origins reported so far, despite their high degree of genetic diversity. In addition, this new technique has been successfully applied for the quantitative detection of GRLDaV in plant material and two mealybug species, Planococcus citri and Pseudococcus viburni. In conclusion, the methodology developed herein represents a significant contribution to the diagnosis and control of this emerging pathogen in grapevine. View Full-Text
Keywords: GRLDaV; plant virus diagnostics; grapevine; real-time PCR GRLDaV; plant virus diagnostics; grapevine; real-time PCR
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MDPI and ACS Style

Morán, F.; Sassalou, C.-L.; Canales, C.; Maliogka, V.I.; Olmos, A.; Ruiz-García, A.B. Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen. Plants 2020, 9, 1151. https://doi.org/10.3390/plants9091151

AMA Style

Morán F, Sassalou C-L, Canales C, Maliogka VI, Olmos A, Ruiz-García AB. Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen. Plants. 2020; 9(9):1151. https://doi.org/10.3390/plants9091151

Chicago/Turabian Style

Morán, Félix, Chrysoula-Lito Sassalou, Celia Canales, Varvara I. Maliogka, Antonio Olmos, and Ana Belén Ruiz-García. 2020. "Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen" Plants 9, no. 9: 1151. https://doi.org/10.3390/plants9091151

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