A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites
Grapevine Pinot gris virus (GPGV) is a widely distributed grapevine pathogen that has been associated to the grapevine leaf mottling and deformation disease. With the aim of better understanding the disease epidemiology and providing efficient control strategies a specific and quantitative duplex Ta...
| Autores principales: | , , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
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PLOS ONE
2018
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.11939/6143 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0197237 |
| _version_ | 1855032374362898432 |
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| author | Morán, Félix Olmos, Antonio Lotos, Leonidas Katis, Nikolaos Glasa, Miroslav Predajna, Lukas Ruiz-García, Ana B. |
| author_browse | Glasa, Miroslav Katis, Nikolaos Lotos, Leonidas Morán, Félix Olmos, Antonio Predajna, Lukas Ruiz-García, Ana B. |
| author_facet | Morán, Félix Olmos, Antonio Lotos, Leonidas Katis, Nikolaos Glasa, Miroslav Predajna, Lukas Ruiz-García, Ana B. |
| author_sort | Morán, Félix |
| collection | ReDivia |
| description | Grapevine Pinot gris virus (GPGV) is a widely distributed grapevine pathogen that has been associated to the grapevine leaf mottling and deformation disease. With the aim of better understanding the disease epidemiology and providing efficient control strategies a specific and quantitative duplex TaqMan real-time RT-PCR assay has been developed. This method has allowed reliable quantitation of the GPGV titer ranging from 30 up to 3 x 108 transcript copies, with a detection limit of 70 viral copies in plant material. The assay targets a grapevine internal control that reduces the occurrence of false negative results, thus increasing the diagnostic sensitivity of the technique. Viral isolates both associated and non-associated to symptoms from Greece, Slovakia and Spain have been successfully detected. The method has also been applied to the absolute quantitation of GPGV in its putative transmission vector Colomerus vitis. Moreover, the viral titer present in single mites has been determined. In addition, in the current study a new polymorphism in the GPGV genome responsible for a shorter movement protein has been found. A phylogenetic study based on this genomic region has shown a high variability among Spanish isolates and points to a different evolutionary origin of this new polymorphism. The methodology here developed opens new possibilities for basic and epidemiological studies as well as for the establishment of efficient control strategies. |
| format | article |
| id | ReDivia6143 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | PLOS ONE |
| publisherStr | PLOS ONE |
| record_format | dspace |
| spelling | ReDivia61432025-04-25T14:46:18Z A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites Morán, Félix Olmos, Antonio Lotos, Leonidas Katis, Nikolaos Glasa, Miroslav Predajna, Lukas Ruiz-García, Ana B. Detection, vector, grapevine, quantification, variability Trichovirus H20 Plant diseases Disease prevention Grapevines Trichovirus Grapevine Pinot gris virus (GPGV) is a widely distributed grapevine pathogen that has been associated to the grapevine leaf mottling and deformation disease. With the aim of better understanding the disease epidemiology and providing efficient control strategies a specific and quantitative duplex TaqMan real-time RT-PCR assay has been developed. This method has allowed reliable quantitation of the GPGV titer ranging from 30 up to 3 x 108 transcript copies, with a detection limit of 70 viral copies in plant material. The assay targets a grapevine internal control that reduces the occurrence of false negative results, thus increasing the diagnostic sensitivity of the technique. Viral isolates both associated and non-associated to symptoms from Greece, Slovakia and Spain have been successfully detected. The method has also been applied to the absolute quantitation of GPGV in its putative transmission vector Colomerus vitis. Moreover, the viral titer present in single mites has been determined. In addition, in the current study a new polymorphism in the GPGV genome responsible for a shorter movement protein has been found. A phylogenetic study based on this genomic region has shown a high variability among Spanish isolates and points to a different evolutionary origin of this new polymorphism. The methodology here developed opens new possibilities for basic and epidemiological studies as well as for the establishment of efficient control strategies. 2018-12-11T08:15:00Z 2018-12-11T08:15:00Z 2018 article publishedVersion Morán F, Olmos A, Lotos L, Predajňa L, Katis N, Glasa M, et al. (2018) A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites. PLoS ONE 13(5): e0197237. https://doi.org/10.1371/journal.pone.0197237 http://hdl.handle.net/20.500.11939/6143 10.1371/journal.pone.0197237 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0197237 en info:eu-repo/grantAgreement/EC/H2020/734736 info:eu-repo/grantAgreement/AEI/Programa Estatal de I+D+i Orientada a los Retos de la Sociedad/E-RTA2017-00009 INIA RTA-2014-0006, INIA E-RTA2017-00009, European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 734736 y COST Action FA1407 INIA RTA-2014-0006, INIA E-RTA2017-00009, European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 734736 y COST Action FA1407 PLOS ONE electronico |
| spellingShingle | Detection, vector, grapevine, quantification, variability Trichovirus H20 Plant diseases Disease prevention Grapevines Trichovirus Morán, Félix Olmos, Antonio Lotos, Leonidas Katis, Nikolaos Glasa, Miroslav Predajna, Lukas Ruiz-García, Ana B. A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title | A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title_full | A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title_fullStr | A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title_full_unstemmed | A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title_short | A novel specific duplex real-time RT-PCR method for absolute quantitation of Grapevine Pinot gris virus in plant material and single mites |
| title_sort | novel specific duplex real time rt pcr method for absolute quantitation of grapevine pinot gris virus in plant material and single mites |
| topic | Detection, vector, grapevine, quantification, variability Trichovirus H20 Plant diseases Disease prevention Grapevines Trichovirus |
| url | http://hdl.handle.net/20.500.11939/6143 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0197237 |
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