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Differential RNAi responses of Nicotiana benthamiana individuals transformed with a hairpin-inducing construct during Plum pox virus challenge

hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorMONTÈS, Christian
hal.structure.identifierBiotechnology Laboratory
hal.structure.identifierUniversidad de Santiago de Chile [Santiago] [USACH]
dc.contributor.authorCASTRO, Alvaro
hal.structure.identifierDepartment of Plant Breeding and Genetics
dc.contributor.authorBARBA, Paola
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorRUBIO, Julia
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorSÁNCHEZ, Evelyn
hal.structure.identifierUniversidad de Chile = University of Chile [Santiago] [UCHILE]
dc.contributor.authorCARVAJAL, Denisse
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorAGUIRRE, Carlos
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorTAPIA, Eduardo
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorDELLORTO, Paola
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorDECROOCQ, Véronique
hal.structure.identifierBiotechnology Laboratory
dc.contributor.authorPRIETO, Humberto
dc.date.issued2014
dc.identifier.issn0920-8569
dc.description.abstractEnGene silencing and large-scale small RNA analysis can be used to develop RNA interference (RNAi)-based resistance strategies for Plum pox virus (PPV), a high impact disease of Prunus spp. In this study, a pPPViRNA hairpin-inducing vector harboring two silencing motif-rich regions of the PPV coat protein (CP) gene was evaluated in transgenic Nicotiana benthamiana (NB) plants. Wild-type NB plants infected with a chimeric PPV virus (PPV:: GFP) exhibited affected leaves with mosaic chlorosis congruent to GFP fluorescence at 21 day post-inoculation; transgenic lines depicted a range of phenotypes from fully resistant to susceptible. ELISA values and GFP fluorescence intensities were used to select transgenic-resistant (TG-R) and transgenic-susceptible (TG-S) lines for further characterization of small interfering RNAs (siRNAs) by large-scale small RNA sequencing. In infected TG-S and untransformed (WT) plants, the observed siRNAs were nearly exclusively 21- and 22-nt siRNAs that targeted the whole PPV:: GFP genome; 24-nt siRNAs were absent in these individuals. Challenged TG-R plants accumulated a full set of 21- to 24-nt siRNAs that were primarily associated with the selected motif-rich regions, indicating that a trans-acting siRNAs process prevented viral multiplication. BLAST analysis identified 13 common siRNA clusters targeting the CP gene. 21-nt siRNA sequences were associated with the 22-nt siRNAs and the scarce 23- and 24-nt molecules in TG-S plants and with most of the observed 22-, 23-, and 24-nt siRNAs in TG-R individuals. These results validate the use of a multi-hot spot silencing vector against PPV and elucidate the molecules by which hairpin-inducing vectors initiate RNAi in vivo.
dc.language.isoen
dc.publisherSpringer Verlag
dc.subjectsiRNA
dc.subject.enPlum pox virus
dc.subject.enRNA interference
dc.subject.enNicotiana benthamiana
dc.subject.enMassive sequencing
dc.title.enDifferential RNAi responses of Nicotiana benthamiana individuals transformed with a hairpin-inducing construct during Plum pox virus challenge
dc.typeArticle de revue
dc.identifier.doi10.1007/s11262-014-1093-5
dc.subject.halSciences du Vivant [q-bio]
dc.subject.halSciences du Vivant [q-bio]/Biologie végétale
bordeaux.journalVirus Genes
bordeaux.page325-338
bordeaux.volume49
bordeaux.issue2
bordeaux.peerReviewedoui
hal.identifierhal-02640010
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-02640010v1
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