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dc.contributor.authorRuiz-Ruiz, Susana
dc.contributor.authorMoreno, Pedro
dc.contributor.authorGuerri, José
dc.contributor.authorAmbrós, Silvia
dc.date.accessioned2017-06-01T10:10:10Z
dc.date.available2017-06-01T10:10:10Z
dc.date.issued2009
dc.identifier.citationRuiz-Ruiz, S., Moreno, P., Guerri, J., Ambros, S. (2009). Discrimination Between Mild and Severe Citrus tristeza virus Isolates with a Rapid and Highly Specific Real-Time Reverse Transcription-Polymerase Chain Reaction Method Using TaqMan LNA Probes. Phytopathology, 99(3), 307-315.
dc.identifier.issn0031-949X
dc.identifier.urihttp://hdl.handle.net/20.500.11939/4453
dc.description.abstractSevere isolates of Citrus tristeza virus (CTV) inducing seedling yellows (SY) and/or stem pitting (SP) in grapefruit or sweet orange are a major threat for the citrus industry worldwide. Identification of these CTV variants was achieved by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) using a general primer set and three TaqMan locked nucleic acids (LNA) probes targeting sequences characteristic of severe, mild (non-SY, non-SP), and T36-like isolates. Successful amplification was achieved from fresh or silica-desiccated CTV-infected samples and all isolates but one reacted with one or more probes. Standard curves using RNA transcripts homologous to the three probes allowed a reproducible quantitative assay, with a wide dynamic range of detection starting with 102 copies. RT-PCR assays with homologous and heterologous transcript RNA mixes demonstrated that each probe reacted only with its cognate sequence which was detected even at ratios below 2.5%. Analysis of 56 pathogenically distinct CTV isolates from 20 countries showed that mild isolates reacted only with the mild probe, whereas severe SP and SY isolates reacted with the severe-SP or the T36-like probes, respectively, and often with a second probe. This procedure can be useful to identify and control potentially dangerous CTV isolates in areas affected only by mild isolates.
dc.language.isoen
dc.titleDiscrimination Between Mild and Severe Citrus tristeza virus Isolates with a Rapid and Highly Specific Real-Time Reverse Transcription-Polymerase Chain Reaction Method Using TaqMan LNA Probes
dc.typearticle
dc.authorAddressInstituto Valenciano de Investigaciones Agrarias, Carretera CV-315, Km. 10,7 - 46113 Moncada (València)
dc.date.issuedFreeFormMAR 2009
dc.entidadIVIACentro de Protección Vegetal y Biotecnología
dc.identifier.doi10.1094/PHYTO-99-3-0307
dc.journal.abbreviatedTitlePhytopathology
dc.journal.issueNumber3
dc.journal.titlePhytopathology
dc.journal.volumeNumber99
dc.page.final315
dc.page.initial307
dc.rights.accessRightsopenAccess
dc.source.typeImpreso
dc.type.hasVersionpublishedVersion


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