Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus

[EN] Citrus tristeza virus (CTV), the causal agent of the most devastating viral disease of citrus, has evolved three silencing suppressor proteins acting at intra- (p23 and p20) and/or intercellular level (p20 and p25) to overcome host antiviral defence. Previously, we showed that Mexican lime tran...

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Authors: Soler, Nuria, Plomer Sáez, Montserrat, Fagoaga García, Carmen Concepción, Moreno, Pedro, Navarro, Luis, Flores Pedauye, Ricardo, PEÑA GARCIA, LEANDRO|||0000-0002-9853-366X
Format: article
Publication Date:2012
Country:España
Institution:Universitat Politècnica de València (UPV)
Repository:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Language:English
OAI Identifier:oai:riunet.upv.es:10251/85385
Online Access:https://riunet.upv.es/handle/10251/85385
Access Level:Open access
Keyword:Antiviral defence
Citrus
RNA interference
Silencing suppressor
Transgenic citrus
Virus resistance
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spelling Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virusSoler, NuriaPlomer Sáez, MontserratFagoaga García, Carmen ConcepciónMoreno, PedroNavarro, LuisFlores Pedauye, RicardoPEÑA GARCIA, LEANDRO|||0000-0002-9853-366XAntiviral defenceCitrusRNA interferenceSilencing suppressorTransgenic citrusVirus resistance[EN] Citrus tristeza virus (CTV), the causal agent of the most devastating viral disease of citrus, has evolved three silencing suppressor proteins acting at intra- (p23 and p20) and/or intercellular level (p20 and p25) to overcome host antiviral defence. Previously, we showed that Mexican lime transformed with an intron-hairpin construct including part of the gene p23 and the adjacent 3' untranslated region displays partial resistance to CTV, with a fraction of the propagations from some transgenic lines remaining uninfected. Here, we transformed Mexican lime with an intron-hairpin vector carrying full-length, untranslatable versions of the genes p25, p20 and p23 from CTV strain T36 to silence the expression of these critical genes in CTV-infected cells. Three transgenic lines presented complete resistance to viral infection, with all their propagations remaining symptomless and virus-free after graft inoculation with CTV-T36, either in the nontransgenic rootstock or in the transgenic scion. Accumulation of transgene-derived siRNAs was necessary but not sufficient for CTV resistance. Inoculation with a divergent CTV strain led to partially breaking the resistance, thus showing the role of sequence identity in the underlying mechanism. Our results are a step forward to developing transgenic resistance to CTV and also show that targeting simultaneously by RNA interference (RNAi) the three viral silencing suppressors appears critical for this purpose, although the involvement of concurrent RNAi mechanisms cannot be excluded.We thank J.E. Peris for his excellent technical assistance and Dr. W.O. Dawson (University of Florida, C.R.E.C., Lake Alfred, FL, USA) for providing the GFP-tagged CTV strain CTV947R-GFP. N.S. was supported by a PhD fellowship from the IVIA. C.F. is recipient of a postdoctoral Ramon y Cajal contract from the Ministerio de Ciencia e Innovacion (MICINN). This research was supported by grants AGL2009-08052, co-financed by Fondo Europeo de Desarrollo Regional-MICINN, and Prometeo/2008/121 from the Generalitat Valenciana.Wiley Open AccessInstituto Universitario Mixto de Biología Molecular y Celular de PlantasMinisterio de Ciencia e InnovaciónGeneralitat ValencianaRepositorio Institucional de la Universitat Politècnica de València Riunet20122012-06-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://riunet.upv.es/handle/10251/85385reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengMinisterio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 AGL2009-08052 Mejora Genetica De La Calidad Y De La Respuesta A Estreses Bioticos De Los Citricos Mediante Ingenieria GeneticaGeneralitat Valenciana https://doi.org/10.13039/501100003359 PROMETEO08%2F2008%2F121 Biotecnología de cítricosopen accesshttp://purl.org/coar/access_right/c_abf2Reserva de todos los derechoshttp://rightsstatements.org/vocab/InC/1.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/853852026-06-13T07:49:27Z
dc.title.none.fl_str_mv Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
title Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
spellingShingle Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
Soler, Nuria
Antiviral defence
Citrus
RNA interference
Silencing suppressor
Transgenic citrus
Virus resistance
title_short Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
title_full Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
title_fullStr Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
title_full_unstemmed Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
title_sort Transformation of Mexican lime with an intron-hairpin construct expressing untranslatable versions of the genes coding for the three silencing suppressors of Citrus tristeza virus confers complete resistance to the virus
dc.creator.none.fl_str_mv Soler, Nuria
Plomer Sáez, Montserrat
Fagoaga García, Carmen Concepción
Moreno, Pedro
Navarro, Luis
Flores Pedauye, Ricardo
PEÑA GARCIA, LEANDRO|||0000-0002-9853-366X
author Soler, Nuria
author_facet Soler, Nuria
Plomer Sáez, Montserrat
Fagoaga García, Carmen Concepción
Moreno, Pedro
Navarro, Luis
Flores Pedauye, Ricardo
PEÑA GARCIA, LEANDRO|||0000-0002-9853-366X
author_role author
author2 Plomer Sáez, Montserrat
Fagoaga García, Carmen Concepción
Moreno, Pedro
Navarro, Luis
Flores Pedauye, Ricardo
PEÑA GARCIA, LEANDRO|||0000-0002-9853-366X
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto Universitario Mixto de Biología Molecular y Celular de Plantas
Ministerio de Ciencia e Innovación
Generalitat Valenciana
Repositorio Institucional de la Universitat Politècnica de València Riunet
dc.subject.none.fl_str_mv Antiviral defence
Citrus
RNA interference
Silencing suppressor
Transgenic citrus
Virus resistance
topic Antiviral defence
Citrus
RNA interference
Silencing suppressor
Transgenic citrus
Virus resistance
description [EN] Citrus tristeza virus (CTV), the causal agent of the most devastating viral disease of citrus, has evolved three silencing suppressor proteins acting at intra- (p23 and p20) and/or intercellular level (p20 and p25) to overcome host antiviral defence. Previously, we showed that Mexican lime transformed with an intron-hairpin construct including part of the gene p23 and the adjacent 3' untranslated region displays partial resistance to CTV, with a fraction of the propagations from some transgenic lines remaining uninfected. Here, we transformed Mexican lime with an intron-hairpin vector carrying full-length, untranslatable versions of the genes p25, p20 and p23 from CTV strain T36 to silence the expression of these critical genes in CTV-infected cells. Three transgenic lines presented complete resistance to viral infection, with all their propagations remaining symptomless and virus-free after graft inoculation with CTV-T36, either in the nontransgenic rootstock or in the transgenic scion. Accumulation of transgene-derived siRNAs was necessary but not sufficient for CTV resistance. Inoculation with a divergent CTV strain led to partially breaking the resistance, thus showing the role of sequence identity in the underlying mechanism. Our results are a step forward to developing transgenic resistance to CTV and also show that targeting simultaneously by RNA interference (RNAi) the three viral silencing suppressors appears critical for this purpose, although the involvement of concurrent RNAi mechanisms cannot be excluded.
publishDate 2012
dc.date.none.fl_str_mv 2012
2012-06-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://riunet.upv.es/handle/10251/85385
url https://riunet.upv.es/handle/10251/85385
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 AGL2009-08052 Mejora Genetica De La Calidad Y De La Respuesta A Estreses Bioticos De Los Citricos Mediante Ingenieria Genetica
Generalitat Valenciana https://doi.org/10.13039/501100003359 PROMETEO08%2F2008%2F121 Biotecnología de cítricos
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reserva de todos los derechos
http://rightsstatements.org/vocab/InC/1.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reserva de todos los derechos
http://rightsstatements.org/vocab/InC/1.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley Open Access
publisher.none.fl_str_mv Wiley Open Access
dc.source.none.fl_str_mv reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
instname:Universitat Politècnica de València (UPV)
instname_str Universitat Politècnica de València (UPV)
reponame_str RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
collection RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
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