Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.
Identifieur interne : 000861 ( Main/Exploration ); précédent : 000860; suivant : 000862Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.
Auteurs : L K Johansen [États-Unis] ; J C CarringtonSource :
- Plant physiology [ 0032-0889 ] ; 2001.
Descripteurs français
- KwdFr :
- ARN des plantes (génétique), Clonage moléculaire (MeSH), Cysteine endopeptidases (génétique), Extinction de l'expression des gènes (MeSH), Feuilles de plante (métabolisme), Potyvirus (génétique), Protéines luminescentes (génétique), Protéines virales (génétique), Protéines virales non structurales (génétique), Protéines à fluorescence verte (MeSH), Rhizobium (génétique), Régulation de l'expression des gènes végétaux (MeSH), Solanaceae (génétique), Solanaceae (métabolisme), Tabac (MeSH), Végétaux toxiques (MeSH).
- MESH :
- génétique : ARN des plantes, Cysteine endopeptidases, Potyvirus, Protéines luminescentes, Protéines virales, Protéines virales non structurales, Rhizobium, Solanaceae.
- métabolisme : Feuilles de plante, Solanaceae.
- Clonage moléculaire, Extinction de l'expression des gènes, Protéines à fluorescence verte, Régulation de l'expression des gènes végétaux, Tabac, Végétaux toxiques.
English descriptors
- KwdEn :
- Cloning, Molecular (MeSH), Cysteine Endopeptidases (genetics), Gene Expression Regulation, Plant (MeSH), Gene Silencing (MeSH), Green Fluorescent Proteins (MeSH), Luminescent Proteins (genetics), Plant Leaves (metabolism), Plants, Toxic (MeSH), Potyvirus (genetics), RNA, Plant (genetics), Rhizobium (genetics), Solanaceae (genetics), Solanaceae (metabolism), Tobacco (MeSH), Viral Nonstructural Proteins (genetics), Viral Proteins (genetics).
- MESH :
- chemical , genetics : Cysteine Endopeptidases, Luminescent Proteins, RNA, Plant, Viral Nonstructural Proteins, Viral Proteins.
- genetics : Potyvirus, Rhizobium, Solanaceae.
- metabolism : Plant Leaves, Solanaceae.
- Cloning, Molecular, Gene Expression Regulation, Plant, Gene Silencing, Green Fluorescent Proteins, Plants, Toxic, Tobacco.
Abstract
The Agrobacterium-mediated transient expression assay in intact tissues has emerged as a rapid and useful method to analyze genes and gene products in plants. In many cases, high levels of active protein can be produced without the need to produce transgenic plants. In this study, a series of tools were developed to enable strong or weak induction of RNA silencing and to suppress RNA silencing in the absence of stable transgenes. Transient delivery of a gene directing production of a double-stranded green fluorescent protein (GFP) transcript rapidly induced RNA silencing of a codelivered GFP reporter gene, effectively preventing accumulation of GFP protein and mRNA. RNA silencing triggered by the strong dsGFP inducer was partially inhibited by the tobacco etch virus silencing suppressor, P1/HC-Pro. In the absence of the strong double-stranded GFP inducer, the functional GFP gene served as a weak RNA silencing inducer in the transient assay, severely limiting accumulation of the GFP mRNA over time. The weak silencing induced by the GFP gene was suppressed by P1/HC-Pro. These results indicate RNA silencing can be triggered by a variety of inducers and analyzed entirely using transient gene delivery systems. They also indicate that RNA silencing may be a significant limitation to expression of genes in the Agrobacterium-mediated transient assay but that this limitation can be overcome by using RNA silencing suppressors.
DOI: 10.1104/pp.126.3.930
PubMed: 11457942
PubMed Central: PMC1540124
Affiliations:
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Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.</title><author><name sortKey="Johansen, L K" sort="Johansen, L K" uniqKey="Johansen L" first="L K" last="Johansen">L K Johansen</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340</wicri:regionArea><wicri:noRegion>Washington 99164-6340</wicri:noRegion></affiliation></author><author><name sortKey="Carrington, J C" sort="Carrington, J C" uniqKey="Carrington J" first="J C" last="Carrington">J C Carrington</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2001">2001</date><idno type="RBID">pubmed:11457942</idno><idno type="pmid">11457942</idno><idno type="pmc">PMC1540124</idno><idno type="doi">10.1104/pp.126.3.930</idno><idno type="wicri:Area/Main/Corpus">000867</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000867</idno><idno type="wicri:Area/Main/Curation">000867</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000867</idno><idno type="wicri:Area/Main/Exploration">000867</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.</title><author><name sortKey="Johansen, L K" sort="Johansen, L K" uniqKey="Johansen L" first="L K" last="Johansen">L K Johansen</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340</wicri:regionArea><wicri:noRegion>Washington 99164-6340</wicri:noRegion></affiliation></author><author><name sortKey="Carrington, J C" sort="Carrington, J C" uniqKey="Carrington J" first="J C" last="Carrington">J C Carrington</name></author></analytic><series><title level="j">Plant physiology</title><idno type="ISSN">0032-0889</idno><imprint><date when="2001" type="published">2001</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cloning, Molecular (MeSH)</term><term>Cysteine Endopeptidases (genetics)</term><term>Gene Expression Regulation, Plant (MeSH)</term><term>Gene Silencing (MeSH)</term><term>Green Fluorescent Proteins (MeSH)</term><term>Luminescent Proteins (genetics)</term><term>Plant Leaves (metabolism)</term><term>Plants, Toxic (MeSH)</term><term>Potyvirus (genetics)</term><term>RNA, Plant (genetics)</term><term>Rhizobium (genetics)</term><term>Solanaceae (genetics)</term><term>Solanaceae (metabolism)</term><term>Tobacco (MeSH)</term><term>Viral Nonstructural Proteins (genetics)</term><term>Viral Proteins (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN des plantes (génétique)</term><term>Clonage moléculaire (MeSH)</term><term>Cysteine endopeptidases (génétique)</term><term>Extinction de l'expression des gènes (MeSH)</term><term>Feuilles de plante (métabolisme)</term><term>Potyvirus (génétique)</term><term>Protéines luminescentes (génétique)</term><term>Protéines virales (génétique)</term><term>Protéines virales non structurales (génétique)</term><term>Protéines à fluorescence verte (MeSH)</term><term>Rhizobium (génétique)</term><term>Régulation de l'expression des gènes végétaux (MeSH)</term><term>Solanaceae (génétique)</term><term>Solanaceae (métabolisme)</term><term>Tabac (MeSH)</term><term>Végétaux toxiques (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Cysteine Endopeptidases</term><term>Luminescent Proteins</term><term>RNA, Plant</term><term>Viral Nonstructural Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Potyvirus</term><term>Rhizobium</term><term>Solanaceae</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN des plantes</term><term>Cysteine endopeptidases</term><term>Potyvirus</term><term>Protéines luminescentes</term><term>Protéines virales</term><term>Protéines virales non structurales</term><term>Rhizobium</term><term>Solanaceae</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Plant Leaves</term><term>Solanaceae</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Feuilles de plante</term><term>Solanaceae</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>Gene Expression Regulation, Plant</term><term>Gene Silencing</term><term>Green Fluorescent Proteins</term><term>Plants, Toxic</term><term>Tobacco</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Clonage moléculaire</term><term>Extinction de l'expression des gènes</term><term>Protéines à fluorescence verte</term><term>Régulation de l'expression des gènes végétaux</term><term>Tabac</term><term>Végétaux toxiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The Agrobacterium-mediated transient expression assay in intact tissues has emerged as a rapid and useful method to analyze genes and gene products in plants. In many cases, high levels of active protein can be produced without the need to produce transgenic plants. In this study, a series of tools were developed to enable strong or weak induction of RNA silencing and to suppress RNA silencing in the absence of stable transgenes. Transient delivery of a gene directing production of a double-stranded green fluorescent protein (GFP) transcript rapidly induced RNA silencing of a codelivered GFP reporter gene, effectively preventing accumulation of GFP protein and mRNA. RNA silencing triggered by the strong dsGFP inducer was partially inhibited by the tobacco etch virus silencing suppressor, P1/HC-Pro. In the absence of the strong double-stranded GFP inducer, the functional GFP gene served as a weak RNA silencing inducer in the transient assay, severely limiting accumulation of the GFP mRNA over time. The weak silencing induced by the GFP gene was suppressed by P1/HC-Pro. These results indicate RNA silencing can be triggered by a variety of inducers and analyzed entirely using transient gene delivery systems. They also indicate that RNA silencing may be a significant limitation to expression of genes in the Agrobacterium-mediated transient assay but that this limitation can be overcome by using RNA silencing suppressors.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">11457942</PMID><DateCompleted><Year>2001</Year><Month>09</Month><Day>13</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>14</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0032-0889</ISSN><JournalIssue CitedMedium="Print"><Volume>126</Volume><Issue>3</Issue><PubDate><Year>2001</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Plant physiology</Title><ISOAbbreviation>Plant Physiol</ISOAbbreviation></Journal><ArticleTitle>Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.</ArticleTitle><Pagination><MedlinePgn>930-8</MedlinePgn></Pagination><Abstract><AbstractText>The Agrobacterium-mediated transient expression assay in intact tissues has emerged as a rapid and useful method to analyze genes and gene products in plants. In many cases, high levels of active protein can be produced without the need to produce transgenic plants. In this study, a series of tools were developed to enable strong or weak induction of RNA silencing and to suppress RNA silencing in the absence of stable transgenes. Transient delivery of a gene directing production of a double-stranded green fluorescent protein (GFP) transcript rapidly induced RNA silencing of a codelivered GFP reporter gene, effectively preventing accumulation of GFP protein and mRNA. RNA silencing triggered by the strong dsGFP inducer was partially inhibited by the tobacco etch virus silencing suppressor, P1/HC-Pro. In the absence of the strong double-stranded GFP inducer, the functional GFP gene served as a weak RNA silencing inducer in the transient assay, severely limiting accumulation of the GFP mRNA over time. The weak silencing induced by the GFP gene was suppressed by P1/HC-Pro. These results indicate RNA silencing can be triggered by a variety of inducers and analyzed entirely using transient gene delivery systems. They also indicate that RNA silencing may be a significant limitation to expression of genes in the Agrobacterium-mediated transient assay but that this limitation can be overcome by using RNA silencing suppressors.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Johansen</LastName><ForeName>L K</ForeName><Initials>LK</Initials><AffiliationInfo><Affiliation>Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Carrington</LastName><ForeName>J C</ForeName><Initials>JC</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>AI27832</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>AI43288</GrantID><Acronym>AI</Acronym><Agency>NIAID NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, Non-P.H.S.</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Plant Physiol</MedlineTA><NlmUniqueID>0401224</NlmUniqueID><ISSNLinking>0032-0889</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008164">Luminescent Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018749">RNA, Plant</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017361">Viral Nonstructural Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014764">Viral Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C402506">pI protein, Filamentous phage phiLf</NameOfSubstance></Chemical><Chemical><RegistryNumber>147336-22-9</RegistryNumber><NameOfSubstance UI="D049452">Green Fluorescent Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.4.22.-</RegistryNumber><NameOfSubstance UI="D003546">Cysteine Endopeptidases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.4.22.-</RegistryNumber><NameOfSubstance UI="C072775">HC-Pro protein, potyvirus</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003546" MajorTopicYN="N">Cysteine Endopeptidases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="Y">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020868" MajorTopicYN="Y">Gene Silencing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D049452" MajorTopicYN="N">Green Fluorescent Proteins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008164" MajorTopicYN="N">Luminescent Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010947" MajorTopicYN="N">Plants, Toxic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017800" MajorTopicYN="N">Potyvirus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018749" MajorTopicYN="N">RNA, Plant</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012231" MajorTopicYN="N">Rhizobium</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019657" MajorTopicYN="N">Solanaceae</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017361" MajorTopicYN="N">Viral Nonstructural Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014764" MajorTopicYN="N">Viral Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2001</Year><Month>7</Month><Day>18</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2001</Year><Month>9</Month><Day>14</Day><Hour>10</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2001</Year><Month>7</Month><Day>18</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">11457942</ArticleId><ArticleId IdType="pmc">PMC1540124</ArticleId><ArticleId IdType="doi">10.1104/pp.126.3.930</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Carrington, J C" sort="Carrington, J C" uniqKey="Carrington J" first="J C" last="Carrington">J C Carrington</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Johansen, L K" sort="Johansen, L K" uniqKey="Johansen L" first="L K" last="Johansen">L K Johansen</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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