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
Institución: Instituto Nacional de Tecnología Agropecuaria
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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)2025-10-23T11:16:29Zoai: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:l2025-10-23 11:16:30.083INTA 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)
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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)
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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)
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repository.mail.fl_str_mv	tripaldi.nicolas@inta.gob.ar
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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...

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