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...
| Authors: | , , , , , , |
|---|---|
| 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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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 |
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open access http://purl.org/coar/access_right/c_abf2 Reserva de todos los derechos http://rightsstatements.org/vocab/InC/1.0/ |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 Reserva de todos los derechos http://rightsstatements.org/vocab/InC/1.0/ |
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openAccess |
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application/pdf |
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Wiley Open Access |
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Wiley Open Access |
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