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
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, s...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Artículo |
| Language: | Inglés |
| Published: |
2018
|
| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/2186 http://www.plantphysiol.org/content/156/3/1278 https://doi.org/10.1104/pp.111.177345 |
| _version_ | 1855483009454571520 |
|---|---|
| author | 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_browse | Asis, Ramón Asurmendi, Sebastian Bermudez Salazar, Luisa Carrari, Fernando Descalzo, Adriana Maria Fernie, Alisdair R. Nunes Nesi, Adriano Quadrana, Leandro Daniel Rodriguez, Maria Cecilia Rossi, Magdalena |
| 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_sort | Quadrana, Leandro Daniel |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA2186 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| record_format | dspace |
| spelling | INTA21862019-03-13T11:09:36Z 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 Rodriguez, Maria Cecilia Bermudez Salazar, Luisa Nunes Nesi, Adriano Fernie, Alisdair R. Descalzo, Adriana Maria Asis, Ramón Rossi, Magdalena Asurmendi, Sebastian Carrari, Fernando 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 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 2018-04-06T12:43:01Z 2018-04-06T12:43:01Z 2011-07 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf Plant physiology 156 (3) : 1278–1291. (July 2011) |
| spellingShingle | 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 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 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 |
| 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_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_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 |
| 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 |
| 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 |
| work_keys_str_mv | AT quadranaleandrodaniel couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT rodriguezmariacecilia couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT bermudezsalazarluisa couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT nunesnesiadriano couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT ferniealisdairr couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT descalzoadrianamaria couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT asisramon couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT rossimagdalena couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT asurmendisebastian couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment AT carrarifernando couplingvirusinducedgenesilencingtoexogenousgreenfluorescenceproteinexpressionprovidesahighlyefficientsystemforfunctionalgenomicsinarabidopsisandacrossallstagesoftomatofruitdevelopment |