Analysis of RNA silencing in agroinfiltrated leaves of Nicotiana benthamiana and Nicotiana tabacum.
Identifieur interne : 000377 ( Ncbi/Curation ); précédent : 000376; suivant : 000378Analysis of RNA silencing in agroinfiltrated leaves of Nicotiana benthamiana and Nicotiana tabacum.
Auteurs : Edyta Ko Cia Ska [Grèce] ; Kriton Kalantidis ; Krzysztof Wypijewski ; Jan Sadowski ; Martin TablerSource :
- Plant molecular biology [ 0167-4412 ] ; 2005.
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, 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).
- 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
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.
DOI: 10.1007/s11103-005-0668-x
PubMed: 16244913
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pubmed:16244913Le document en format XML
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<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, 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>
</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>
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<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>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term>
<term>Plant Leaves</term>
<term>Tobacco</term>
<term>Transgenes</term>
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<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>
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<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>
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<term>ARN double brin</term>
<term>Petit ARN interférent</term>
<term>Protéines à fluorescence verte</term>
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<term>Genetic Engineering</term>
<term>Plants, Genetically Modified</term>
<term>RNA Interference</term>
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<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>
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<front><div type="abstract" xml:lang="en">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>
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