Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen
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...
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| Formato: | article |
| Lenguaje: | Inglés |
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MDPI
2020
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/6594 https://www.mdpi.com/2223-7747/9/9/1151 |
| _version_ | 1855032454826426368 |
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| author | Morán, Félix Sassalou, Chrysoula-Lito Canales, Celia Maliogka, Varvara Olmos, Antonio Ruiz-García, Ana B. |
| author_browse | Canales, Celia Maliogka, Varvara Morán, Félix Olmos, Antonio Ruiz-García, Ana B. Sassalou, Chrysoula-Lito |
| author_facet | Morán, Félix Sassalou, Chrysoula-Lito Canales, Celia Maliogka, Varvara Olmos, Antonio Ruiz-García, Ana B. |
| author_sort | Morán, Félix |
| collection | ReDivia |
| description | 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. |
| format | article |
| id | ReDivia6594 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | ReDivia65942025-04-25T14:47:26Z Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen Morán, Félix Sassalou, Chrysoula-Lito Canales, Celia Maliogka, Varvara Olmos, Antonio Ruiz-García, Ana B. GRLDaV plant virus diagnostics grapevine real-time PCR Detection H20 Plant diseases Grapevines Pathogenic viruses Plant pathology 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. 2020-09-09T12:50:12Z 2020-09-09T12:50:12Z 2020 article publishedVersion 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 2223-7747 http://hdl.handle.net/20.500.11939/6594 10.3390/plants9091151 https://www.mdpi.com/2223-7747/9/9/1151 en info:eu-repo/grantAgreement/MINECO/Programa Estatal de I+D+i Orientada a los Retos de la Sociedad/RTA-2014-00061 Prevention and control of emerging grapevine viruses in Spain. RTA 2014-00061 Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess MDPI electronico |
| spellingShingle | GRLDaV plant virus diagnostics grapevine real-time PCR Detection H20 Plant diseases Grapevines Pathogenic viruses Plant pathology Morán, Félix Sassalou, Chrysoula-Lito Canales, Celia Maliogka, Varvara Olmos, Antonio Ruiz-García, Ana B. Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title | Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title_full | Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title_fullStr | Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title_full_unstemmed | Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title_short | Specific Real-Time PCR for the Detection and Absolute Quantitation of Grapevine Roditis Leaf Discoloration-Associated Virus, an EPPO Alert Pathogen |
| title_sort | specific real time pcr for the detection and absolute quantitation of grapevine roditis leaf discoloration associated virus an eppo alert pathogen |
| topic | GRLDaV plant virus diagnostics grapevine real-time PCR Detection H20 Plant diseases Grapevines Pathogenic viruses Plant pathology |
| url | http://hdl.handle.net/20.500.11939/6594 https://www.mdpi.com/2223-7747/9/9/1151 |
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