Analysis of RNA Silencing in Agroinfiltrated Leaves of Nicotiana Benthamiana and Nicotiana Tabacum
Identifieur interne : 003021 ( Main/Exploration ); précédent : 003020; suivant : 003022Analysis of RNA Silencing in Agroinfiltrated Leaves of Nicotiana Benthamiana and Nicotiana Tabacum
Auteurs : Edyta Ko Cia Ska [Grèce, Pologne] ; Kriton Kalantidis [Grèce] ; Krzysztof Wypijewski [Pologne, Royaume-Uni] ; Jan Sadowski [Pologne] ; Martin Tabler [Grèce]Source :
- Plant Molecular Biology [ 0167-4412 ] ; 2005-11-01.
Descripteurs français
- KwdFr :
- ARN des plantes (génétique), ARN des plantes (métabolisme), ARN double brin (génétique), ARN double brin (métabolisme), Agrobacterium tumefaciens (génétique), Feuilles de plante (génétique), Génie génétique, Interférence par ARN, Petit ARN interférent (génétique), Petit ARN interférent (métabolisme), Protéines à fluorescence verte (génétique), Protéines à fluorescence verte (métabolisme), Régulation de l'expression des gènes végétaux, Tabac (), Tabac (génétique), Transgènes (génétique), Végétaux génétiquement modifiés.
- MESH :
- génétique : ARN des plantes, ARN double brin, Agrobacterium tumefaciens, Feuilles de plante, Petit ARN interférent, Protéines à fluorescence verte, Tabac, Transgènes.
- métabolisme : ARN des plantes, ARN double brin, Petit ARN interférent, Protéines à fluorescence verte.
- Génie génétique, Interférence par ARN, Régulation de l'expression des gènes végétaux, Tabac, Végétaux génétiquement modifiés.
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (genetics), Gene Expression Regulation, Plant, Genetic Engineering, Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Plant Leaves (genetics), Plants, Genetically Modified, RNA Interference, RNA interference, RNA silencing, RNA, Double-Stranded (genetics), RNA, Double-Stranded (metabolism), RNA, Plant (genetics), RNA, Plant (metabolism), RNA, Small Interfering (genetics), RNA, Small Interfering (metabolism), Tobacco (classification), Tobacco (genetics), Transgenes (genetics), agroinfiltration, microRNA, post-transcriptional gene silencing, short interfering RNA.
- MESH :
- chemical , genetics : Green Fluorescent Proteins, RNA, Double-Stranded, RNA, Plant, RNA, Small Interfering.
- classification : Tobacco.
- genetics : Agrobacterium tumefaciens, Plant Leaves, Tobacco, Transgenes.
- chemical , metabolism : Green Fluorescent Proteins, RNA, Double-Stranded, RNA, Plant, RNA, Small Interfering.
- Gene Expression Regulation, Plant, Genetic Engineering, Plants, Genetically Modified, RNA Interference.
Abstract
Abstract: In this study we analyse several aspects of cytoplasmic RNA silencing by agroinfiltration of DNA constructs encoding single- and double-stranded RNAs derived from a GFP transgene and from the endogenous Virp1 gene. Both types of inductors resulted after 2–4 days in much higher concentration of siRNAs in the agroinfiltrated zone than normally seen during systemic silencing. More specifically, infiltration of two transgene hairpin constructs resulted in elevated levels of siRNAs. However, differences between the two constructs were observed: the antisense–sense arrangement was more effective than the sense–antisense order. For both double-stranded forms, we observed a relative increase of the 24-mer size class of siRNAs. When a comparable hairpin construct of the endogenous Virp1 gene was assayed, the portion of the 24-mer siRNA class remained low as observed for all kinds of single-stranded inducers. The lack of increase of Virp1-derived 24-mers was independent of the expression level, as demonstrated by agroinfiltration into a transgenic plant that overexpressed Virp1 and showed the same pattern. Using transducer constructs, we could detect within a week transitive silencing from GFP to GUS sequences in the infiltrated zone and in either direction 5′–3′ and 3′–5′. Conversely, for the endogenous Virp1 gene neither transitive silencing nor the induction of systemic silencing could be observed. These results are discussed in view of the current models of RNA silencing.
Url:
DOI: 10.1007/s11103-005-0668-x
Affiliations:
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Box 1527, GR-71110, Crete, Heraklion</wicri:regionArea><wicri:noRegion>Heraklion</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Grèce</country></affiliation></author><author><name sortKey="Wypijewski, Krzysztof" sort="Wypijewski, Krzysztof" uniqKey="Wypijewski K" first="Krzysztof" last="Wypijewski">Krzysztof Wypijewski</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Biotechnology, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Midzychodzka 5, 60-371, Poznań</wicri:regionArea><wicri:noRegion>Poznań</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Scottish Crop Research Institute, DD2 5DA, Dundee, Invergowrie</wicri:regionArea><wicri:noRegion>Invergowrie</wicri:noRegion></affiliation></author><author><name sortKey="Sadowski, Jan" sort="Sadowski, Jan" uniqKey="Sadowski J" first="Jan" last="Sadowski">Jan Sadowski</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Biotechnology, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Midzychodzka 5, 60-371, Poznań</wicri:regionArea><wicri:noRegion>Poznań</wicri:noRegion></affiliation></author><author><name sortKey="Tabler, Martin" sort="Tabler, Martin" uniqKey="Tabler M" first="Martin" last="Tabler">Martin Tabler</name><affiliation wicri:level="1"><country xml:lang="fr">Grèce</country><wicri:regionArea>Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology – Hellas, P.O. Box 1527, GR-71110, Crete, Heraklion</wicri:regionArea><wicri:noRegion>Heraklion</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Plant Molecular Biology</title><title level="j" type="sub">An International Journal on Molecular Biology, Molecular Genetics and Biochemistry</title><title level="j" type="abbrev">Plant Mol Biol</title><idno type="ISSN">0167-4412</idno><idno type="eISSN">1573-5028</idno><imprint><publisher>Kluwer Academic Publishers</publisher><pubPlace>Dordrecht</pubPlace><date type="published" when="2005-11-01">2005-11-01</date><biblScope unit="volume">59</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="647">647</biblScope><biblScope unit="page" to="661">661</biblScope></imprint><idno type="ISSN">0167-4412</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4412</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>Gene Expression Regulation, Plant</term><term>Genetic Engineering</term><term>Green Fluorescent Proteins (genetics)</term><term>Green Fluorescent Proteins (metabolism)</term><term>Plant Leaves (genetics)</term><term>Plants, Genetically Modified</term><term>RNA Interference</term><term>RNA interference</term><term>RNA silencing</term><term>RNA, Double-Stranded (genetics)</term><term>RNA, Double-Stranded (metabolism)</term><term>RNA, Plant (genetics)</term><term>RNA, Plant (metabolism)</term><term>RNA, Small Interfering (genetics)</term><term>RNA, Small Interfering (metabolism)</term><term>Tobacco (classification)</term><term>Tobacco (genetics)</term><term>Transgenes (genetics)</term><term>agroinfiltration</term><term>microRNA</term><term>post-transcriptional gene silencing</term><term>short interfering RNA</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN des plantes (génétique)</term><term>ARN des plantes (métabolisme)</term><term>ARN double brin (génétique)</term><term>ARN double brin (métabolisme)</term><term>Agrobacterium tumefaciens (génétique)</term><term>Feuilles de plante (génétique)</term><term>Génie génétique</term><term>Interférence par ARN</term><term>Petit ARN interférent (génétique)</term><term>Petit ARN interférent (métabolisme)</term><term>Protéines à fluorescence verte (génétique)</term><term>Protéines à fluorescence verte (métabolisme)</term><term>Régulation de l'expression des gènes végétaux</term><term>Tabac ()</term><term>Tabac (génétique)</term><term>Transgènes (génétique)</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>RNA, Double-Stranded</term><term>RNA, Plant</term><term>RNA, Small Interfering</term></keywords><keywords scheme="MESH" qualifier="classification" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Plant Leaves</term><term>Tobacco</term><term>Transgenes</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN des plantes</term><term>ARN double brin</term><term>Agrobacterium tumefaciens</term><term>Feuilles de plante</term><term>Petit ARN interférent</term><term>Protéines à fluorescence verte</term><term>Tabac</term><term>Transgènes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Green Fluorescent Proteins</term><term>RNA, Double-Stranded</term><term>RNA, Plant</term><term>RNA, Small Interfering</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN des plantes</term><term>ARN double brin</term><term>Petit ARN interférent</term><term>Protéines à fluorescence verte</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression Regulation, Plant</term><term>Genetic Engineering</term><term>Plants, Genetically Modified</term><term>RNA Interference</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Génie génétique</term><term>Interférence par ARN</term><term>Régulation de l'expression des gènes végétaux</term><term>Tabac</term><term>Végétaux génétiquement modifiés</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: In this study we analyse several aspects of cytoplasmic RNA silencing by agroinfiltration of DNA constructs encoding single- and double-stranded RNAs derived from a GFP transgene and from the endogenous Virp1 gene. Both types of inductors resulted after 2–4 days in much higher concentration of siRNAs in the agroinfiltrated zone than normally seen during systemic silencing. More specifically, infiltration of two transgene hairpin constructs resulted in elevated levels of siRNAs. However, differences between the two constructs were observed: the antisense–sense arrangement was more effective than the sense–antisense order. For both double-stranded forms, we observed a relative increase of the 24-mer size class of siRNAs. When a comparable hairpin construct of the endogenous Virp1 gene was assayed, the portion of the 24-mer siRNA class remained low as observed for all kinds of single-stranded inducers. The lack of increase of Virp1-derived 24-mers was independent of the expression level, as demonstrated by agroinfiltration into a transgenic plant that overexpressed Virp1 and showed the same pattern. Using transducer constructs, we could detect within a week transitive silencing from GFP to GUS sequences in the infiltrated zone and in either direction 5′–3′ and 3′–5′. Conversely, for the endogenous Virp1 gene neither transitive silencing nor the induction of systemic silencing could be observed. These results are discussed in view of the current models of RNA silencing.</div></front></TEI><affiliations><list><country><li>Grèce</li><li>Pologne</li><li>Royaume-Uni</li></country></list><tree><country name="Grèce"><noRegion><name sortKey="Ko Cia Ska, Edyta" sort="Ko Cia Ska, Edyta" uniqKey="Ko Cia Ska E" first="Edyta" last="Ko Cia Ska">Edyta Ko Cia Ska</name></noRegion><name sortKey="Kalantidis, Kriton" sort="Kalantidis, Kriton" uniqKey="Kalantidis K" first="Kriton" last="Kalantidis">Kriton Kalantidis</name><name sortKey="Kalantidis, Kriton" sort="Kalantidis, Kriton" uniqKey="Kalantidis K" first="Kriton" last="Kalantidis">Kriton Kalantidis</name><name sortKey="Tabler, Martin" sort="Tabler, Martin" uniqKey="Tabler M" first="Martin" last="Tabler">Martin Tabler</name></country><country name="Pologne"><noRegion><name sortKey="Ko Cia Ska, Edyta" sort="Ko Cia Ska, Edyta" uniqKey="Ko Cia Ska E" first="Edyta" last="Ko Cia Ska">Edyta Ko Cia Ska</name></noRegion><name sortKey="Sadowski, Jan" sort="Sadowski, Jan" uniqKey="Sadowski J" first="Jan" last="Sadowski">Jan Sadowski</name><name sortKey="Wypijewski, Krzysztof" sort="Wypijewski, Krzysztof" uniqKey="Wypijewski K" first="Krzysztof" last="Wypijewski">Krzysztof Wypijewski</name></country><country name="Royaume-Uni"><noRegion><name sortKey="Wypijewski, Krzysztof" sort="Wypijewski, Krzysztof" uniqKey="Wypijewski K" first="Krzysztof" last="Wypijewski">Krzysztof Wypijewski</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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