Construction of RYMV-based vectors for rice functional genomic
Rice is a major crop and a plant model for monocotyledons genomic, and especially for cereals. A new field has to be developed with the functional genomic, to take advantage of the growing number of genomic sequences. The potential of plant virus-based transient expression vectors is reported in fundamental virology but also for plant biology. At present plant virus vectors offer number of benefits for the expression of foreign genes, by overexpress (gain of function mutant) or suppress (loss-of-function mutant) gene expression. The potential of such vectors for plant molecular biology is substantial particularly to identify previous unknown genes. Rice yellow mottle virus (RYMV) is a single-stranded-positivesense RNA virus that specifically infects rice leaves and causes serious disease in irrigated rice systems in East and West Africa. The purpose of this work is to develop for rice a transitory expression vector based on an infectious full-length cDNA clone of RYMV to assess gene function. Three main RYMV-based vector constructs is being generated. The first approach consists in producing a fusion between a reporter gene (encoding GUS or GFP) and the viral coat protein. With such construct, gene expression can be monitored in planta. In a second approach, to over-express a foreign gene, we consider duplicating sub-genomic mRNA promoter of the coat protein in order to improve gene expression thanks to an additional stem-loop structure. The third approach is the construction of an interesting virus-based vector for gene complementation, a construct devoid of its infectious elements like genes encoding viral coat protein and/or movement protein. Accumulation of recombinant virus is evaluated by RT-PCR in electroporated rice protoplasts and mechanically inoculated plant. Then, recombinant vectors will be challenged for the ability to express levels of viral proteins, GUS and GFP expression in protoplasts and in planta. To improve gene expression all these constructs will be cloned under the control of constitutive promoters. In parallel, as we also want to better understand molecular mechanisms involved in gene silencing induced by RYMV, we are studying inhibition of gene silencing by P1 protein of different RYMV-isolates. The purpose of the study is to estimate how the inhibition of gene silencing by RYMV occurs, in order to improve vectors expression. With this aim in view, we will inoculate these transgenic rice lines with different RYMV-isolates, to show variability on the suppressor of silencing expression and thus, highlight isolates that generates the main effect on silencing inhibition. (Texte intégral)
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Subjects: | F30 - Génétique et amélioration des plantes, H20 - Maladies des plantes, sobemovirus marbrure jaune riz, Oryza, riz irrigué, virus des végétaux, résistance aux maladies, génie génétique, vecteur génétique, expression des gènes, arn, http://aims.fao.org/aos/agrovoc/c_34331, http://aims.fao.org/aos/agrovoc/c_5435, http://aims.fao.org/aos/agrovoc/c_15724, http://aims.fao.org/aos/agrovoc/c_5985, http://aims.fao.org/aos/agrovoc/c_2328, http://aims.fao.org/aos/agrovoc/c_15974, http://aims.fao.org/aos/agrovoc/c_27532, http://aims.fao.org/aos/agrovoc/c_27527, http://aims.fao.org/aos/agrovoc/c_6618, http://aims.fao.org/aos/agrovoc/c_8355, http://aims.fao.org/aos/agrovoc/c_2442, |
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F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 |
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F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 Sire, Christelle Bangratz, Martine Sallaud, Christophe Guiderdoni, Emmanuel Fargette, Denis Ghesquière, Alain Brugidou, Christophe Construction of RYMV-based vectors for rice functional genomic |
description |
Rice is a major crop and a plant model for monocotyledons genomic, and especially for cereals. A new field has to be developed with the functional genomic, to take advantage of the growing number of genomic sequences. The potential of plant virus-based transient expression vectors is reported in fundamental virology but also for plant biology. At present plant virus vectors offer number of benefits for the expression of foreign genes, by overexpress (gain of function mutant) or suppress (loss-of-function mutant) gene expression. The potential of such vectors for plant molecular biology is substantial particularly to identify previous unknown genes. Rice yellow mottle virus (RYMV) is a single-stranded-positivesense RNA virus that specifically infects rice leaves and causes serious disease in irrigated rice systems in East and West Africa. The purpose of this work is to develop for rice a transitory expression vector based on an infectious full-length cDNA clone of RYMV to assess gene function. Three main RYMV-based vector constructs is being generated. The first approach consists in producing a fusion between a reporter gene (encoding GUS or GFP) and the viral coat protein. With such construct, gene expression can be monitored in planta. In a second approach, to over-express a foreign gene, we consider duplicating sub-genomic mRNA promoter of the coat protein in order to improve gene expression thanks to an additional stem-loop structure. The third approach is the construction of an interesting virus-based vector for gene complementation, a construct devoid of its infectious elements like genes encoding viral coat protein and/or movement protein. Accumulation of recombinant virus is evaluated by RT-PCR in electroporated rice protoplasts and mechanically inoculated plant. Then, recombinant vectors will be challenged for the ability to express levels of viral proteins, GUS and GFP expression in protoplasts and in planta. To improve gene expression all these constructs will be cloned under the control of constitutive promoters. In parallel, as we also want to better understand molecular mechanisms involved in gene silencing induced by RYMV, we are studying inhibition of gene silencing by P1 protein of different RYMV-isolates. The purpose of the study is to estimate how the inhibition of gene silencing by RYMV occurs, in order to improve vectors expression. With this aim in view, we will inoculate these transgenic rice lines with different RYMV-isolates, to show variability on the suppressor of silencing expression and thus, highlight isolates that generates the main effect on silencing inhibition. (Texte intégral) |
format |
conference_item |
topic_facet |
F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 |
author |
Sire, Christelle Bangratz, Martine Sallaud, Christophe Guiderdoni, Emmanuel Fargette, Denis Ghesquière, Alain Brugidou, Christophe |
author_facet |
Sire, Christelle Bangratz, Martine Sallaud, Christophe Guiderdoni, Emmanuel Fargette, Denis Ghesquière, Alain Brugidou, Christophe |
author_sort |
Sire, Christelle |
title |
Construction of RYMV-based vectors for rice functional genomic |
title_short |
Construction of RYMV-based vectors for rice functional genomic |
title_full |
Construction of RYMV-based vectors for rice functional genomic |
title_fullStr |
Construction of RYMV-based vectors for rice functional genomic |
title_full_unstemmed |
Construction of RYMV-based vectors for rice functional genomic |
title_sort |
construction of rymv-based vectors for rice functional genomic |
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AAB |
url |
http://agritrop.cirad.fr/516968/ |
work_keys_str_mv |
AT sirechristelle constructionofrymvbasedvectorsforricefunctionalgenomic AT bangratzmartine constructionofrymvbasedvectorsforricefunctionalgenomic AT sallaudchristophe constructionofrymvbasedvectorsforricefunctionalgenomic AT guiderdoniemmanuel constructionofrymvbasedvectorsforricefunctionalgenomic AT fargettedenis constructionofrymvbasedvectorsforricefunctionalgenomic AT ghesquierealain constructionofrymvbasedvectorsforricefunctionalgenomic AT brugidouchristophe constructionofrymvbasedvectorsforricefunctionalgenomic |
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dig-cirad-fr-5169682024-01-28T11:58:08Z http://agritrop.cirad.fr/516968/ http://agritrop.cirad.fr/516968/ Construction of RYMV-based vectors for rice functional genomic. Sire Christelle, Bangratz Martine, Sallaud Christophe, Guiderdoni Emmanuel, Fargette Denis, Ghesquière Alain, Brugidou Christophe. 2003. In : Advances in plant virology : A three day International conference at CIRAD, Montpellier, France on 29 September - 1 October 2003. AAB, CIRAD-AMIS, INRA, ENSAM, IRD. Warwick : AAB, Résumé, 1 p. International Conference : Advances in Plant Virology, Montpellier, France, 29 Septembre 2003/1 Octobre 2003. Construction of RYMV-based vectors for rice functional genomic Sire, Christelle Bangratz, Martine Sallaud, Christophe Guiderdoni, Emmanuel Fargette, Denis Ghesquière, Alain Brugidou, Christophe eng 2003 AAB Advances in plant virology : A three day International conference at CIRAD, Montpellier, France on 29 September - 1 October 2003 F30 - Génétique et amélioration des plantes H20 - Maladies des plantes sobemovirus marbrure jaune riz Oryza riz irrigué virus des végétaux résistance aux maladies génie génétique vecteur génétique expression des gènes arn http://aims.fao.org/aos/agrovoc/c_34331 http://aims.fao.org/aos/agrovoc/c_5435 http://aims.fao.org/aos/agrovoc/c_15724 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_2328 http://aims.fao.org/aos/agrovoc/c_15974 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_6618 Afrique occidentale Afrique orientale http://aims.fao.org/aos/agrovoc/c_8355 http://aims.fao.org/aos/agrovoc/c_2442 Rice is a major crop and a plant model for monocotyledons genomic, and especially for cereals. A new field has to be developed with the functional genomic, to take advantage of the growing number of genomic sequences. The potential of plant virus-based transient expression vectors is reported in fundamental virology but also for plant biology. At present plant virus vectors offer number of benefits for the expression of foreign genes, by overexpress (gain of function mutant) or suppress (loss-of-function mutant) gene expression. The potential of such vectors for plant molecular biology is substantial particularly to identify previous unknown genes. Rice yellow mottle virus (RYMV) is a single-stranded-positivesense RNA virus that specifically infects rice leaves and causes serious disease in irrigated rice systems in East and West Africa. The purpose of this work is to develop for rice a transitory expression vector based on an infectious full-length cDNA clone of RYMV to assess gene function. Three main RYMV-based vector constructs is being generated. The first approach consists in producing a fusion between a reporter gene (encoding GUS or GFP) and the viral coat protein. With such construct, gene expression can be monitored in planta. In a second approach, to over-express a foreign gene, we consider duplicating sub-genomic mRNA promoter of the coat protein in order to improve gene expression thanks to an additional stem-loop structure. The third approach is the construction of an interesting virus-based vector for gene complementation, a construct devoid of its infectious elements like genes encoding viral coat protein and/or movement protein. Accumulation of recombinant virus is evaluated by RT-PCR in electroporated rice protoplasts and mechanically inoculated plant. Then, recombinant vectors will be challenged for the ability to express levels of viral proteins, GUS and GFP expression in protoplasts and in planta. To improve gene expression all these constructs will be cloned under the control of constitutive promoters. In parallel, as we also want to better understand molecular mechanisms involved in gene silencing induced by RYMV, we are studying inhibition of gene silencing by P1 protein of different RYMV-isolates. The purpose of the study is to estimate how the inhibition of gene silencing by RYMV occurs, in order to improve vectors expression. With this aim in view, we will inoculate these transgenic rice lines with different RYMV-isolates, to show variability on the suppressor of silencing expression and thus, highlight isolates that generates the main effect on silencing inhibition. (Texte intégral) conference_item info:eu-repo/semantics/conferenceObject Conference info:eu-repo/semantics/closedAccess http://agritrop.cirad.fr/516952/ |