Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all st...

Autores
Quadrana, Leandro Daniel; Rodriguez, Maria Cecilia; Bermudez Salazar, Luisa; Nunes Nesi, Adriano; Fernie, Alisdair R.; Descalzo, Adriana Maria; Asis, Ramón; Rossi, Magdalena; Asurmendi, Sebastian; Carrari, Fernando
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Since the advent of the postgenomic era, efforts have focused on the development of rapid strategies for annotating plant genes of unknown function. Given its simplicity and rapidity, virus-induced gene silencing (VIGS) has become one of the preeminent approaches for functional analyses. However, several problems remain intrinsic to the use of such a strategy in the study of both metabolic and developmental processes. The most prominent of these is the commonly observed phenomenon of “sectoring” the tissue regions that are not effectively targeted by VIGS. To better discriminate these sectors, an effective marker system displaying minimal secondary effects is a prerequisite. Utilizing a VIGS system based on the tobacco rattle virus vector, we here studied the effect of silencing the endogenous phytoene desaturase gene (pds) and the expression and subsequent silencing of the exogenous green fluorescence protein (gfp) on the metabolism of Arabidopsis (Arabidopsis thaliana) leaves and tomato (Solanum lycopersicum) fruits. In leaves, we observed dramatic effects on primary carbon and pigment metabolism associated with the photobleached phenotype following the silencing of the endogenous pds gene. However, relatively few pleiotropic effects on carbon metabolism were observed in tomato fruits when pds expression was inhibited. VIGS coupled to gfp constitutive expression revealed no significant metabolic alterations after triggering of silencing in Arabidopsis leaves and a mild effect in mature green tomato fruits. By contrast, a wider impact on metabolism was observed in ripe fruits. Silencing experiments with an endogenous target gene of interest clearly demonstrated the feasibility of cosilencing in this system; however, carefully constructed control experiments are a prerequisite to prevent erroneous interpretation.
Fil: Quadrana, Leandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Rodriguez, Maria Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bermudez Salazar, Luisa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Nunes Nesi, Adriano. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; Alemania
Fil: Fernie, Alisdair R. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; Alemania
Fil: Descalzo, Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Tecnología de Alimentos; Argentina
Fil: Asis, Ramón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas, Universidad Nacional de Córdoba; Argentina
Fil: Rossi, Magdalena. Universidade de São Paulo. Instituto de Biociências. Departamento de Botânica; Brasil
Fil: Asurmendi, Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Carrari, Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Cátedra de Genética; Argentina
Fuente
Plant physiology 156 (3) : 1278–1291. (July 2011)
Materia
Tomate
Solanum Lycopersicum
Virus de las Plantas
Arabidopsis
Etapas de Desarrollo de la Planta
Proteínas Virales
Genomas
Fruto
Fruit
Genomes
Viral Proteins
Plant Developmental Stages
Plant Viruses
Tomatoes
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/2186

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oai_identifier_str oai:localhost:20.500.12123/2186
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit developmentQuadrana, Leandro DanielRodriguez, Maria CeciliaBermudez Salazar, LuisaNunes Nesi, AdrianoFernie, Alisdair R.Descalzo, Adriana MariaAsis, RamónRossi, MagdalenaAsurmendi, SebastianCarrari, FernandoTomateSolanum LycopersicumVirus de las PlantasArabidopsisEtapas de Desarrollo de la PlantaProteínas ViralesGenomasFrutoFruitGenomesViral ProteinsPlant Developmental StagesPlant VirusesTomatoesSince the advent of the postgenomic era, efforts have focused on the development of rapid strategies for annotating plant genes of unknown function. Given its simplicity and rapidity, virus-induced gene silencing (VIGS) has become one of the preeminent approaches for functional analyses. However, several problems remain intrinsic to the use of such a strategy in the study of both metabolic and developmental processes. The most prominent of these is the commonly observed phenomenon of “sectoring” the tissue regions that are not effectively targeted by VIGS. To better discriminate these sectors, an effective marker system displaying minimal secondary effects is a prerequisite. Utilizing a VIGS system based on the tobacco rattle virus vector, we here studied the effect of silencing the endogenous phytoene desaturase gene (pds) and the expression and subsequent silencing of the exogenous green fluorescence protein (gfp) on the metabolism of Arabidopsis (Arabidopsis thaliana) leaves and tomato (Solanum lycopersicum) fruits. In leaves, we observed dramatic effects on primary carbon and pigment metabolism associated with the photobleached phenotype following the silencing of the endogenous pds gene. However, relatively few pleiotropic effects on carbon metabolism were observed in tomato fruits when pds expression was inhibited. VIGS coupled to gfp constitutive expression revealed no significant metabolic alterations after triggering of silencing in Arabidopsis leaves and a mild effect in mature green tomato fruits. By contrast, a wider impact on metabolism was observed in ripe fruits. Silencing experiments with an endogenous target gene of interest clearly demonstrated the feasibility of cosilencing in this system; however, carefully constructed control experiments are a prerequisite to prevent erroneous interpretation.Fil: Quadrana, Leandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Rodriguez, Maria Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bermudez Salazar, Luisa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Nunes Nesi, Adriano. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; AlemaniaFil: Fernie, Alisdair R. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; AlemaniaFil: Descalzo, Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Tecnología de Alimentos; ArgentinaFil: Asis, Ramón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas, Universidad Nacional de Córdoba; ArgentinaFil: Rossi, Magdalena. Universidade de São Paulo. Instituto de Biociências. Departamento de Botânica; BrasilFil: Asurmendi, Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Carrari, Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Cátedra de Genética; Argentina2018-04-06T12:43:01Z2018-04-06T12:43:01Z2011-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/2186http://www.plantphysiol.org/content/156/3/12781532-2548https://doi.org/10.1104/pp.111.177345Plant physiology 156 (3) : 1278–1291. (July 2011)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:44:17Zoai:localhost:20.500.12123/2186instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:44:17.411INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
title Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
spellingShingle Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
Quadrana, Leandro Daniel
Tomate
Solanum Lycopersicum
Virus de las Plantas
Arabidopsis
Etapas de Desarrollo de la Planta
Proteínas Virales
Genomas
Fruto
Fruit
Genomes
Viral Proteins
Plant Developmental Stages
Plant Viruses
Tomatoes
title_short Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
title_full Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
title_fullStr Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
title_full_unstemmed Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
title_sort Coupling virus-induced gene silencing to exogenous green fluorescence protein expression provides a highly efficient system for functional genomics in arabidopsis and across all stages of tomato fruit development
dc.creator.none.fl_str_mv Quadrana, Leandro Daniel
Rodriguez, Maria Cecilia
Bermudez Salazar, Luisa
Nunes Nesi, Adriano
Fernie, Alisdair R.
Descalzo, Adriana Maria
Asis, Ramón
Rossi, Magdalena
Asurmendi, Sebastian
Carrari, Fernando
author Quadrana, Leandro Daniel
author_facet Quadrana, Leandro Daniel
Rodriguez, Maria Cecilia
Bermudez Salazar, Luisa
Nunes Nesi, Adriano
Fernie, Alisdair R.
Descalzo, Adriana Maria
Asis, Ramón
Rossi, Magdalena
Asurmendi, Sebastian
Carrari, Fernando
author_role author
author2 Rodriguez, Maria Cecilia
Bermudez Salazar, Luisa
Nunes Nesi, Adriano
Fernie, Alisdair R.
Descalzo, Adriana Maria
Asis, Ramón
Rossi, Magdalena
Asurmendi, Sebastian
Carrari, Fernando
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Tomate
Solanum Lycopersicum
Virus de las Plantas
Arabidopsis
Etapas de Desarrollo de la Planta
Proteínas Virales
Genomas
Fruto
Fruit
Genomes
Viral Proteins
Plant Developmental Stages
Plant Viruses
Tomatoes
topic Tomate
Solanum Lycopersicum
Virus de las Plantas
Arabidopsis
Etapas de Desarrollo de la Planta
Proteínas Virales
Genomas
Fruto
Fruit
Genomes
Viral Proteins
Plant Developmental Stages
Plant Viruses
Tomatoes
dc.description.none.fl_txt_mv Since the advent of the postgenomic era, efforts have focused on the development of rapid strategies for annotating plant genes of unknown function. Given its simplicity and rapidity, virus-induced gene silencing (VIGS) has become one of the preeminent approaches for functional analyses. However, several problems remain intrinsic to the use of such a strategy in the study of both metabolic and developmental processes. The most prominent of these is the commonly observed phenomenon of “sectoring” the tissue regions that are not effectively targeted by VIGS. To better discriminate these sectors, an effective marker system displaying minimal secondary effects is a prerequisite. Utilizing a VIGS system based on the tobacco rattle virus vector, we here studied the effect of silencing the endogenous phytoene desaturase gene (pds) and the expression and subsequent silencing of the exogenous green fluorescence protein (gfp) on the metabolism of Arabidopsis (Arabidopsis thaliana) leaves and tomato (Solanum lycopersicum) fruits. In leaves, we observed dramatic effects on primary carbon and pigment metabolism associated with the photobleached phenotype following the silencing of the endogenous pds gene. However, relatively few pleiotropic effects on carbon metabolism were observed in tomato fruits when pds expression was inhibited. VIGS coupled to gfp constitutive expression revealed no significant metabolic alterations after triggering of silencing in Arabidopsis leaves and a mild effect in mature green tomato fruits. By contrast, a wider impact on metabolism was observed in ripe fruits. Silencing experiments with an endogenous target gene of interest clearly demonstrated the feasibility of cosilencing in this system; however, carefully constructed control experiments are a prerequisite to prevent erroneous interpretation.
Fil: Quadrana, Leandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Rodriguez, Maria Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bermudez Salazar, Luisa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Nunes Nesi, Adriano. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; Alemania
Fil: Fernie, Alisdair R. Max-Planck-Institut fur Molekulare Pflanzenphysiologie; Alemania
Fil: Descalzo, Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Tecnología de Alimentos; Argentina
Fil: Asis, Ramón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas, Universidad Nacional de Córdoba; Argentina
Fil: Rossi, Magdalena. Universidade de São Paulo. Instituto de Biociências. Departamento de Botânica; Brasil
Fil: Asurmendi, Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Carrari, Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Cátedra de Genética; Argentina
description Since the advent of the postgenomic era, efforts have focused on the development of rapid strategies for annotating plant genes of unknown function. Given its simplicity and rapidity, virus-induced gene silencing (VIGS) has become one of the preeminent approaches for functional analyses. However, several problems remain intrinsic to the use of such a strategy in the study of both metabolic and developmental processes. The most prominent of these is the commonly observed phenomenon of “sectoring” the tissue regions that are not effectively targeted by VIGS. To better discriminate these sectors, an effective marker system displaying minimal secondary effects is a prerequisite. Utilizing a VIGS system based on the tobacco rattle virus vector, we here studied the effect of silencing the endogenous phytoene desaturase gene (pds) and the expression and subsequent silencing of the exogenous green fluorescence protein (gfp) on the metabolism of Arabidopsis (Arabidopsis thaliana) leaves and tomato (Solanum lycopersicum) fruits. In leaves, we observed dramatic effects on primary carbon and pigment metabolism associated with the photobleached phenotype following the silencing of the endogenous pds gene. However, relatively few pleiotropic effects on carbon metabolism were observed in tomato fruits when pds expression was inhibited. VIGS coupled to gfp constitutive expression revealed no significant metabolic alterations after triggering of silencing in Arabidopsis leaves and a mild effect in mature green tomato fruits. By contrast, a wider impact on metabolism was observed in ripe fruits. Silencing experiments with an endogenous target gene of interest clearly demonstrated the feasibility of cosilencing in this system; however, carefully constructed control experiments are a prerequisite to prevent erroneous interpretation.
publishDate 2011
dc.date.none.fl_str_mv 2011-07
2018-04-06T12:43:01Z
2018-04-06T12:43:01Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/2186
http://www.plantphysiol.org/content/156/3/1278
1532-2548
https://doi.org/10.1104/pp.111.177345
url http://hdl.handle.net/20.500.12123/2186
http://www.plantphysiol.org/content/156/3/1278
https://doi.org/10.1104/pp.111.177345
identifier_str_mv 1532-2548
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Plant physiology 156 (3) : 1278–1291. (July 2011)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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