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dc.contributor.authorAmbrós, Silvia
dc.contributor.authorRuiz-Ruiz, Susana
dc.contributor.authorPena, Leandro
dc.contributor.authorMoreno, Pedro
dc.date.accessioned2020-05-21T16:28:03Z
dc.date.available2020-05-21T16:28:03Z
dc.date.issued2013es
dc.identifier.citationAmbrós, S., Ruiz-Ruiz, S., Peña, L., & Moreno, P. (2013). A genetic system for citrus tristeza virus using the non-natural host Nicotiana benthamiana: an update. Frontiers in microbiology, 4, 165.es
dc.identifier.issn1664-302X
dc.identifier.urihttp://hdl.handle.net/20.500.11939/6455
dc.description.abstractIn nature Citrus tristeza virus (CTV), genus Closterovirus, infects only the phloem cells of species of Citrus and related genera. Finding that the CTV T36 strain replicated in Nicotiana benthamiana (NB) protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with RNA transcribed from a full-genome cDNA clone, a laborious and uncertain system requiring several months for each experiment. We developed a more efficient system based on agroinfiltration of NB leaves with CTV-T36-based binary plasmids, which caused systemic infection in this non-natural host within a few weeks yielding in the upper leaves enough CTV virions to readily infect citrus by slash inoculation. Stem agroinoculation of citrus and NB plants with oncogenic strains of Agrobacterium tumefaciens carrying a CTV-T36 binary vector with a GUS marker, induced GUS positive galls in both species. However, while most NB tumors were CTV positive and many plants became systemically infected, no coat protein or viral RNA was detected in citrus tumors, even though CTV cDNA was readily detected by PCR in the same galls. This finding suggests (1) strong silencing or CTV RNA processing in transformed cells impairing infection progress, and (2) the need for using NB as an intermediate host in the genetic system. To maintain CTV-T36 in NB or assay other CTV genotypes in this host, we also tried to graft-transmit the virus from infected to healthy NB, or to mechanically inoculate NB leaves with virion extracts. While these trials were mostly unsuccessful on non-treated NB plants, agroinfiltration with silencing suppressors enabled for the first time infecting NB plants by side-grafting and by mechanical inoculation with virions, indicating that previous failure to infect NB was likely due to virus silencing in early infection steps. Using NB as a CTV host provides new possibilities to study virus-host interactions with a simple and reliable system.es
dc.language.isoenes
dc.publisherFrontiers Media S.A.es
dc.rightsAtribución-NoComercial-SinDerivadas 3.0 España*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/es/*
dc.subjectInfectious cDNA cloneses
dc.subjectAgroinoculationes
dc.subjectOncogenic Strainses
dc.subjectGraft transmissiones
dc.subjectNicotiana Benthamiana protoplastses
dc.subjectRNA silencing suppressores
dc.titleA genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an updateen
dc.typearticlees
dc.authorAddressInstituto Valenciano de Investigaciones Agrarias (IVIA), Carretera CV-315, Km. 10’7, 46113 Moncada (Valencia), Españaes
dc.entidadIVIACentro de Protección Vegetal y Biotecnologíaes
dc.identifier.doi10.3389/fmicb.2013.00165es
dc.identifier.urlhttps://www.frontiersin.org/articles/10.3389/fmicb.2013.00165/fulles
dc.journal.issueNumber4es
dc.journal.titleFrontiers in Microbiologyes
dc.page.final165es
dc.page.initial165es
dc.source.typeelectronicoes
dc.subject.agrovocCitrus tristeza viruses
dc.subject.agrovocAgrobacterium tumefacienses
dc.type.hasVersionpublishedVersiones


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Except where otherwise noted, this item's license is described as Atribución-NoComercial-SinDerivadas 3.0 España