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dc.contributor.authorPalacio-Bielsa, Ana
dc.contributor.authorCubero, Jaime
dc.contributor.authorCambra, Miguel A.
dc.contributor.authorCollados, Raquel
dc.contributor.authorBerruete, Isabel M.
dc.contributor.authorLópez, María M.
dc.date.accessioned2017-06-01T10:09:47Z
dc.date.available2017-06-01T10:09:47Z
dc.date.issued2011
dc.identifier.citationPalacio-Bielsa, Ana, Cubero, Jaime, Cambra, M.A., Collados, Raquel, Berruete, I.M., Lopez, M.M. (2011). Development of an Efficient Real-Time Quantitative PCR Protocol for Detection of Xanthomonas arboricola pv. pruni in Prunus Species. Applied and Environmental Microbiology, 77(1), 89-97.
dc.identifier.issn0099-2240
dc.identifier.urihttp://hdl.handle.net/20.500.11939/4286
dc.description.abstractXanthomonas arboricola pv. pruni, the causal agent of bacterial spot disease of stone fruit, is considered a quarantine organism by the European Union and the European and Mediterranean Plant Protection Organization (EPPO). The bacterium can undergo an epiphytic phase and/or be latent and can be transmitted by plant material, but currently, only visual inspections are used to certify plants as being X. arboricola pv. pruni free. A novel and highly sensitive real-time TaqMan PCR detection protocol was designed based on a sequence of a gene for a putative protein related to an ABC transporter ATP-binding system in X. arboricola pv. pruni. Pathogen detection can be completed within a few hours with a sensitivity of 10(2) CFU ml(-1), thus surpassing the sensitivity of the existing conventional PCR. Specificity was assessed for X. arboricola pv. pruni strains from different origins as well as for closely related Xanthomonas species, non-Xanthomonas species, saprophytic bacteria, and healthy Prunus samples. The efficiency of the developed protocol was evaluated with field samples of 14 Prunus species and rootstocks. For symptomatic leaf samples, the protocol was very efficient even when washed tissues of the leaves were directly amplified without any previous DNA extraction. For samples of 117 asymptomatic leaves and 285 buds, the protocol was more efficient after a simple DNA extraction, and X. arboricola pv. pruni was detected in 9.4% and 9.1% of the 402 samples analyzed, respectively, demonstrating its frequent epiphytic or endophytic phase. This newly developed real-time PCR protocol can be used as a quantitative assay, offers a reliable and sensitive test for X. arboricola pv. pruni, and is suitable as a screening test for symptomatic as well as asymptomatic plant material.
dc.language.isoen
dc.titleDevelopment of an Efficient Real-Time Quantitative PCR Protocol for Detection of Xanthomonas arboricola pv. pruni in Prunus Species
dc.typearticle
dc.date.issuedFreeFormJAN 2011
dc.identifier.doi10.1128/AEM.01593-10
dc.journal.abbreviatedTitleAppl.Environ.Microbiol.
dc.journal.issueNumber1
dc.journal.titleApplied and Environmental Microbiology
dc.journal.volumeNumber77
dc.page.final97
dc.page.initial89
dc.rights.accessRightsopenAccess
dc.source.typeImpreso


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