Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe
Watermelon mosaic virus (WMV) causes serious damage to several crops worldwide, mainly cucurbits. Disease control is based on preventing spread and search for natural resistances for plant breeding, which requires tools for sensitive detection and precise quantitation. We developed a procedure based...
| Main Authors: | , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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Elsevier
2022
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.11939/8168 https://www.sciencedirect.com/science/article/abs/pii/S0166093421003554?via%3Dihub |
| _version_ | 1855492438389424128 |
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| author | Rubio, Luis Giménez, Karen Romero, Juan Font-San-Ambrosio, Isabel Alfaro-Fernández, Ana Galipienso, Luis |
| author_browse | Alfaro-Fernández, Ana Font-San-Ambrosio, Isabel Galipienso, Luis Giménez, Karen Romero, Juan Rubio, Luis |
| author_facet | Rubio, Luis Giménez, Karen Romero, Juan Font-San-Ambrosio, Isabel Alfaro-Fernández, Ana Galipienso, Luis |
| author_sort | Rubio, Luis |
| collection | ReDivia |
| description | Watermelon mosaic virus (WMV) causes serious damage to several crops worldwide, mainly cucurbits. Disease control is based on preventing spread and search for natural resistances for plant breeding, which requires tools for sensitive detection and precise quantitation. We developed a procedure based on reverse transcription followed by real-time quantitative polymerase chain reaction (RT-qPCR) with a primer pair and a TaqMan® probe specific for WMV. The primers and probe were designed from conserved sequence stretches to target a wide range of WMV isolates. A standard curve performed with transcripts enabled estimation of WMV RNA copies per ng of total RNA, with a wide dynamic range and sensitivity (104 to 1011). This RT-qPCR was assayed with field samples from different cucurbits and used to evaluate the temporal accumulation in pumpkin plants. |
| format | Artículo |
| id | ReDivia8168 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | ReDivia81682025-04-25T14:48:49Z Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe Rubio, Luis Giménez, Karen Romero, Juan Font-San-Ambrosio, Isabel Alfaro-Fernández, Ana Galipienso, Luis Watermelon mosaic virus Cucurbits Viral accumulations Diagnostics Real time-PCR H20 Plant diseases U40 Surveying methods Watermelon mosaic virus (WMV) causes serious damage to several crops worldwide, mainly cucurbits. Disease control is based on preventing spread and search for natural resistances for plant breeding, which requires tools for sensitive detection and precise quantitation. We developed a procedure based on reverse transcription followed by real-time quantitative polymerase chain reaction (RT-qPCR) with a primer pair and a TaqMan® probe specific for WMV. The primers and probe were designed from conserved sequence stretches to target a wide range of WMV isolates. A standard curve performed with transcripts enabled estimation of WMV RNA copies per ng of total RNA, with a wide dynamic range and sensitivity (104 to 1011). This RT-qPCR was assayed with field samples from different cucurbits and used to evaluate the temporal accumulation in pumpkin plants. 2022-05-27T17:04:55Z 2022-05-27T17:04:55Z 2022 article acceptedVersion Rubio, L., Giménez, K., Romero, J., Font-San-Ambrosio, M. I., Alfaro-Fernández, A. & Galipienso, L. (2022). Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe. Journal of Virological Methods, 300, 114416. 0166-0934 http://hdl.handle.net/20.500.11939/8168 10.1016/j.jviromet.2021.114416 https://www.sciencedirect.com/science/article/abs/pii/S0166093421003554?via%3Dihub en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess Elsevier electronico |
| spellingShingle | Watermelon mosaic virus Cucurbits Viral accumulations Diagnostics Real time-PCR H20 Plant diseases U40 Surveying methods Rubio, Luis Giménez, Karen Romero, Juan Font-San-Ambrosio, Isabel Alfaro-Fernández, Ana Galipienso, Luis Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title | Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title_full | Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title_fullStr | Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title_full_unstemmed | Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title_short | Detection and absolute quantitation of watermelon mosaic virus by real-time RT-PCR with a TaqMan probe |
| title_sort | detection and absolute quantitation of watermelon mosaic virus by real time rt pcr with a taqman probe |
| topic | Watermelon mosaic virus Cucurbits Viral accumulations Diagnostics Real time-PCR H20 Plant diseases U40 Surveying methods |
| url | http://hdl.handle.net/20.500.11939/8168 https://www.sciencedirect.com/science/article/abs/pii/S0166093421003554?via%3Dihub |
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