Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/91696
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | Real-time PCR quantification of Peronospora arborescens, the opium poppy downy mildew pathogen, in seed stocks and symptomless infected plants |
Autor: | Montes Borrego, Miguel CSIC; Muñoz Ledesma, Francisco Javier; Jiménez-Díaz, Rafael M. CSIC ; Landa, Blanca B. CSIC ORCID | Fecha de publicación: | feb-2011 | Editor: | American Phytopathological Society | Citación: | Plant Disease 95(2): 143-152 (2011) | Resumen: | In this study, we developed a reliable, quick, and accurate quantitative polymerase chain reaction (qPCR) assay based on the MIQE (Minimum Information for publication of Quantitative Real-Time PCR Experiments) guidelines for the quantification of Peronospora arborescens in infected downy mildew-symptomless opium poppy (Papaver somniferum) tissues and commercial seed stocks. The protocol was highly reproducible and allowed accurate quantification of pathogen DNA up to 10 fg in different plant DNA backgrounds without losing specificity and efficiency. Moreover, to further overcome difficulties conferred by the strict biotrophy of this pathogen, we developed dilution series of DNA extracted from a plasmid with the target pathogen DNA as a cloned insert. This facilitated the demonstration of the robustness of the protocol in different laboratories with different qPCR equipment and reagents, which may help in its use on a broad scale. Finally, we validated the usefulness of the qPCR protocol for quarantine purposes and downy mildew resistance screening by quantifying P. arborescens in complex, naturally infested opium poppy samples. Thus, a pathogen biomass of 0.0003 to 0.007%o or of 0.110 to 5,557 ppm was quantified in symptomless capsules in commercial seed stocks, or in stem samples from symptomless opium poppy plants systemically infected by the pathogen, respectively. © 2011 The American Phytopathological Society. | URI: | http://hdl.handle.net/10261/91696 | DOI: | 10.1094/PDIS-07-10-0499 | Identificadores: | doi: 10.1094/PDIS-07-10-0499 issn: 0191-2917 |
Aparece en las colecciones: | (IAS) Artículos |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
---|---|---|---|---|
accesoRestringido.pdf | 15,38 kB | Adobe PDF | Visualizar/Abrir |
CORE Recommender
SCOPUSTM
Citations
33
checked on 10-abr-2024
WEB OF SCIENCETM
Citations
30
checked on 25-feb-2024
Page view(s)
265
checked on 19-abr-2024
Download(s)
114
checked on 19-abr-2024
Google ScholarTM
Check
Altmetric
Altmetric
NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.