Transient down-regulation of the RNA silencing machinery increases efficiency of Agrobacterium-mediated transformation of Arabidopsis.
Identifieur interne : 000385 ( Main/Exploration ); précédent : 000384; suivant : 000386Transient down-regulation of the RNA silencing machinery increases efficiency of Agrobacterium-mediated transformation of Arabidopsis.
Auteurs : Andriy Bilichak [Canada] ; Youli Yao ; Igor KovalchukSource :
- Plant biotechnology journal [ 1467-7652 ] ; 2014.
Descripteurs français
- KwdFr :
- ADN bactérien (génétique), ARN messager (génétique), ARN messager (métabolisme), Agrobacterium (physiologie), Arabidopsis (génétique), Arabidopsis (microbiologie), Cassures double-brin de l'ADN (MeSH), DNA-directed RNA polymerases (métabolisme), Gènes de plante (MeSH), Génétique inverse (MeSH), Interférence par ARN (MeSH), Liaison aux protéines (MeSH), Locus génétiques (MeSH), Modèles génétiques (MeSH), Mutation (génétique), Méthylation de l'ADN (génétique), Petit ARN interférent (métabolisme), Protéines d'Arabidopsis (métabolisme), Régulation négative (MeSH), Sous-unités de protéines (métabolisme), Technique de Southern (MeSH), Transformation génétique (MeSH), Végétaux génétiquement modifiés (MeSH), Épigenèse génétique (MeSH).
- MESH :
- génétique : ADN bactérien, ARN messager, Arabidopsis, Mutation, Méthylation de l'ADN.
- microbiologie : Arabidopsis.
- métabolisme : ARN messager, DNA-directed RNA polymerases, Petit ARN interférent, Protéines d'Arabidopsis, Sous-unités de protéines.
- physiologie : Agrobacterium.
- Cassures double-brin de l'ADN, Gènes de plante, Génétique inverse, Interférence par ARN, Liaison aux protéines, Locus génétiques, Modèles génétiques, Régulation négative, Technique de Southern, Transformation génétique, Végétaux génétiquement modifiés, Épigenèse génétique.
English descriptors
- KwdEn :
- Agrobacterium (physiology), Arabidopsis (genetics), Arabidopsis (microbiology), Arabidopsis Proteins (metabolism), Blotting, Southern (MeSH), DNA Breaks, Double-Stranded (MeSH), DNA Methylation (genetics), DNA, Bacterial (genetics), DNA-Directed RNA Polymerases (metabolism), Down-Regulation (MeSH), Epigenesis, Genetic (MeSH), Genes, Plant (MeSH), Genetic Loci (MeSH), Models, Genetic (MeSH), Mutation (genetics), Plants, Genetically Modified (MeSH), Protein Binding (MeSH), Protein Subunits (metabolism), RNA Interference (MeSH), RNA, Messenger (genetics), RNA, Messenger (metabolism), RNA, Small Interfering (metabolism), Reverse Genetics (MeSH), Transformation, Genetic (MeSH).
- MESH :
- chemical , genetics : DNA, Bacterial, RNA, Messenger.
- chemical , metabolism : Arabidopsis Proteins, DNA-Directed RNA Polymerases, Protein Subunits, RNA, Messenger, RNA, Small Interfering.
- genetics : Arabidopsis, DNA Methylation, Mutation.
- microbiology : Arabidopsis.
- physiology : Agrobacterium.
- Blotting, Southern, DNA Breaks, Double-Stranded, Down-Regulation, Epigenesis, Genetic, Genes, Plant, Genetic Loci, Models, Genetic, Plants, Genetically Modified, Protein Binding, RNA Interference, Reverse Genetics, Transformation, Genetic.
Abstract
Agrobacterium tumefaciens is a plant pathogen that is widely used in plant transformation. As the process of transgenesis includes the delivery of single-stranded T-DNA molecule, we hypothesized that transformation rate may negatively correlate with the efficiency of the RNA-silencing machinery. Using mutants compromised in either the transcriptional or post-transcriptional gene-silencing pathways, two inhibitors of stable transformation were revealed-AGO2 and NRPD1a. Furthermore, an immunoprecipitation experiment has shown that NRPD1, a subunit of Pol IV, directly interacts with Agrobacterium T-DNA in planta. Using the Tobacco rattle virus (TRV)--based virus-induced gene silencing (VIGS) technique, we demonstrated that the transient down-regulation of the expression of either AGO2 or NRPD1a genes in reproductive organs of Arabidopsis, leads to an increase in transformation rate. We observed a 6.0- and 3.5-fold increase in transformation rate upon transient downregulation of either AGO2 or NRPD1a genes, respectively. This is the first report demonstrating the increase in the plant transformation rate via VIGS-mediated transient down-regulation of the components of epigenetic machinery in reproductive tissue.
DOI: 10.1111/pbi.12165
PubMed: 24472037
Affiliations:
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As the process of transgenesis includes the delivery of single-stranded T-DNA molecule, we hypothesized that transformation rate may negatively correlate with the efficiency of the RNA-silencing machinery. Using mutants compromised in either the transcriptional or post-transcriptional gene-silencing pathways, two inhibitors of stable transformation were revealed-AGO2 and NRPD1a. Furthermore, an immunoprecipitation experiment has shown that NRPD1, a subunit of Pol IV, directly interacts with Agrobacterium T-DNA in planta. Using the Tobacco rattle virus (TRV)--based virus-induced gene silencing (VIGS) technique, we demonstrated that the transient down-regulation of the expression of either AGO2 or NRPD1a genes in reproductive organs of Arabidopsis, leads to an increase in transformation rate. We observed a 6.0- and 3.5-fold increase in transformation rate upon transient downregulation of either AGO2 or NRPD1a genes, respectively. This is the first report demonstrating the increase in the plant transformation rate via VIGS-mediated transient down-regulation of the components of epigenetic machinery in reproductive tissue.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24472037</PMID><DateCompleted><Year>2015</Year><Month>01</Month><Day>22</Day></DateCompleted><DateRevised><Year>2014</Year><Month>06</Month><Day>03</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1467-7652</ISSN><JournalIssue CitedMedium="Internet"><Volume>12</Volume><Issue>5</Issue><PubDate><Year>2014</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Plant biotechnology journal</Title><ISOAbbreviation>Plant Biotechnol J</ISOAbbreviation></Journal><ArticleTitle>Transient down-regulation of the RNA silencing machinery increases efficiency of Agrobacterium-mediated transformation of Arabidopsis.</ArticleTitle><Pagination><MedlinePgn>590-600</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1111/pbi.12165</ELocationID><Abstract><AbstractText>Agrobacterium tumefaciens is a plant pathogen that is widely used in plant transformation. As the process of transgenesis includes the delivery of single-stranded T-DNA molecule, we hypothesized that transformation rate may negatively correlate with the efficiency of the RNA-silencing machinery. Using mutants compromised in either the transcriptional or post-transcriptional gene-silencing pathways, two inhibitors of stable transformation were revealed-AGO2 and NRPD1a. Furthermore, an immunoprecipitation experiment has shown that NRPD1, a subunit of Pol IV, directly interacts with Agrobacterium T-DNA in planta. Using the Tobacco rattle virus (TRV)--based virus-induced gene silencing (VIGS) technique, we demonstrated that the transient down-regulation of the expression of either AGO2 or NRPD1a genes in reproductive organs of Arabidopsis, leads to an increase in transformation rate. We observed a 6.0- and 3.5-fold increase in transformation rate upon transient downregulation of either AGO2 or NRPD1a genes, respectively. This is the first report demonstrating the increase in the plant transformation rate via VIGS-mediated transient down-regulation of the components of epigenetic machinery in reproductive tissue.</AbstractText><CopyrightInformation>© 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bilichak</LastName><ForeName>Andriy</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yao</LastName><ForeName>Youli</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Kovalchuk</LastName><ForeName>Igor</ForeName><Initials>I</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType 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IdType="doi">10.1111/pbi.12165</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Kovalchuk, Igor" sort="Kovalchuk, Igor" uniqKey="Kovalchuk I" first="Igor" last="Kovalchuk">Igor Kovalchuk</name><name sortKey="Yao, Youli" sort="Yao, Youli" uniqKey="Yao Y" first="Youli" last="Yao">Youli Yao</name></noCountry><country name="Canada"><noRegion><name sortKey="Bilichak, Andriy" sort="Bilichak, Andriy" uniqKey="Bilichak A" first="Andriy" last="Bilichak">Andriy Bilichak</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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