AgrobacTransV1, Main, Exploration, bibRecord, 000506

Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.

Identifieur interne : 000506 ( Main/Exploration ); précédent : 000505; suivant : 000507

Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.

Auteurs : Ayako Nishizawa-Yokoi [Japon] ; Satoko Nonaka ; Hiroaki Saika ; Yong-Ik Kwon ; Keishi Osakabe ; Seiichi Toki

Source :

The New phytologist [ 1469-8137 ] ; 2012.

RBID : pubmed:23050791

Descripteurs français

English descriptors

KwdEn :
- Agrobacterium (genetics), DNA Breaks, Double-Stranded (MeSH), DNA End-Joining Repair (genetics), DNA Helicases (genetics), DNA Helicases (metabolism), DNA Ligase ATP (MeSH), DNA Ligases (genetics), DNA Ligases (metabolism), DNA Repair (MeSH), DNA, Bacterial (MeSH), DNA-Binding Proteins (genetics), DNA-Binding Proteins (metabolism), Gene Expression Regulation, Plant (MeSH), Gene Knockdown Techniques (MeSH), Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Homologous Recombination (MeSH), Oryza (genetics), Oryza (metabolism), Plant Proteins (genetics), Plant Proteins (metabolism), Plants, Genetically Modified (MeSH), Transformation, Genetic (MeSH).
MESH :
- chemical , genetics : DNA Helicases, DNA Ligases, DNA-Binding Proteins, Green Fluorescent Proteins, Plant Proteins.
- genetics : Agrobacterium, DNA End-Joining Repair, Oryza.
- chemical , metabolism : DNA Helicases, DNA Ligases, DNA-Binding Proteins, Green Fluorescent Proteins, Oryza, Plant Proteins.
- DNA Breaks, Double-Stranded, DNA Ligase ATP, DNA Repair, DNA, Bacterial, Gene Expression Regulation, Plant, Gene Knockdown Techniques, Homologous Recombination, Plants, Genetically Modified, Transformation, Genetic.

Abstract

Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza sativa) using scutellum-derived calli, we examined here the involvement of the NHEJ pathway in Agrobacterium-mediated stable transformation in rice. Rice calli from OsKu70, OsKu80 and OsLig4 knockdown (KD) plants were infected with Agrobacterium harboring a sensitive emerald luciferase (LUC) reporter construct to evaluate stable expression and a green fluorescent protein (GFP) construct to monitor transient expression of T-DNA. Transient expression was not suppressed, but stable expression was reduced significantly, in KD plants. Furthermore, KD-Ku70 and KD-Lig4 calli exhibited an increase in the frequency of homologous recombination (HR) compared with control calli. In addition, suppression of OsKu70, OsKu80 and OsLig4 induced the expression of HR-related genes on treatment with DNA-damaging agents. Our findings suggest strongly that NHEJ is involved in Agrobacterium-mediated stable transformation in rice, and that there is a competitive and complementary relationship between the NHEJ and HR pathways for DNA double-strand break repair in rice.

DOI: 10.1111/j.1469-8137.2012.04350.x
PubMed: 23050791
PubMed Central: PMC3532656

Affiliations:

Japon

Links toward previous steps (curation, corpus...)

to stream Main, to step Corpus: 000490
to stream Main, to step Curation: 000490

Le document en format XML

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<term>DNA Breaks, Double-Stranded (MeSH)</term>
<term>DNA End-Joining Repair (genetics)</term>
<term>DNA Helicases (genetics)</term>
<term>DNA Helicases (metabolism)</term>
<term>DNA Ligase ATP (MeSH)</term>
<term>DNA Ligases (genetics)</term>
<term>DNA Ligases (metabolism)</term>
<term>DNA Repair (MeSH)</term>
<term>DNA, Bacterial (MeSH)</term>
<term>DNA-Binding Proteins (genetics)</term>
<term>DNA-Binding Proteins (metabolism)</term>
<term>Gene Expression Regulation, Plant (MeSH)</term>
<term>Gene Knockdown Techniques (MeSH)</term>
<term>Green Fluorescent Proteins (genetics)</term>
<term>Green Fluorescent Proteins (metabolism)</term>
<term>Homologous Recombination (MeSH)</term>
<term>Oryza (genetics)</term>
<term>Oryza (metabolism)</term>
<term>Plant Proteins (genetics)</term>
<term>Plant Proteins (metabolism)</term>
<term>Plants, Genetically Modified (MeSH)</term>
<term>Transformation, Genetic (MeSH)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN bactérien (MeSH)</term>
<term>Agrobacterium (génétique)</term>
<term>Cassures double-brin de l'ADN (MeSH)</term>
<term>DNA ligase ATP (MeSH)</term>
<term>DNA ligases (génétique)</term>
<term>DNA ligases (métabolisme)</term>
<term>Helicase (génétique)</term>
<term>Helicase (métabolisme)</term>
<term>Oryza (génétique)</term>
<term>Oryza (métabolisme)</term>
<term>Protéines de liaison à l'ADN (génétique)</term>
<term>Protéines de liaison à l'ADN (métabolisme)</term>
<term>Protéines végétales (génétique)</term>
<term>Protéines végétales (métabolisme)</term>
<term>Protéines à fluorescence verte (génétique)</term>
<term>Protéines à fluorescence verte (métabolisme)</term>
<term>Recombinaison homologue (MeSH)</term>
<term>Régulation de l'expression des gènes végétaux (MeSH)</term>
<term>Réparation de l'ADN (MeSH)</term>
<term>Réparation de l'ADN par jonction d'extrémités (génétique)</term>
<term>Techniques de knock-down de gènes (MeSH)</term>
<term>Transformation génétique (MeSH)</term>
<term>Végétaux génétiquement modifiés (MeSH)</term>
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<term>DNA Ligases</term>
<term>DNA-Binding Proteins</term>
<term>Green Fluorescent Proteins</term>
<term>Plant Proteins</term>
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<term>DNA ligases</term>
<term>Helicase</term>
<term>Oryza</term>
<term>Protéines de liaison à l'ADN</term>
<term>Protéines végétales</term>
<term>Protéines à fluorescence verte</term>
<term>Réparation de l'ADN par jonction d'extrémités</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA Helicases</term>
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<term>DNA-Binding Proteins</term>
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<term>Protéines végétales</term>
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<term>DNA Ligase ATP</term>
<term>DNA Repair</term>
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<term>Gene Knockdown Techniques</term>
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<term>DNA ligase ATP</term>
<term>Recombinaison homologue</term>
<term>Régulation de l'expression des gènes végétaux</term>
<term>Réparation de l'ADN</term>
<term>Techniques de knock-down de gènes</term>
<term>Transformation génétique</term>
<term>Végétaux génétiquement modifiés</term>
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<front><div type="abstract" xml:lang="en">Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza sativa) using scutellum-derived calli, we examined here the involvement of the NHEJ pathway in Agrobacterium-mediated stable transformation in rice. Rice calli from OsKu70, OsKu80 and OsLig4 knockdown (KD) plants were infected with Agrobacterium harboring a sensitive emerald luciferase (LUC) reporter construct to evaluate stable expression and a green fluorescent protein (GFP) construct to monitor transient expression of T-DNA. Transient expression was not suppressed, but stable expression was reduced significantly, in KD plants. Furthermore, KD-Ku70 and KD-Lig4 calli exhibited an increase in the frequency of homologous recombination (HR) compared with control calli. In addition, suppression of OsKu70, OsKu80 and OsLig4 induced the expression of HR-related genes on treatment with DNA-damaging agents. Our findings suggest strongly that NHEJ is involved in Agrobacterium-mediated stable transformation in rice, and that there is a competitive and complementary relationship between the NHEJ and HR pathways for DNA double-strand break repair in rice.</div>
</front>
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<Abstract><AbstractText>Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza sativa) using scutellum-derived calli, we examined here the involvement of the NHEJ pathway in Agrobacterium-mediated stable transformation in rice. Rice calli from OsKu70, OsKu80 and OsLig4 knockdown (KD) plants were infected with Agrobacterium harboring a sensitive emerald luciferase (LUC) reporter construct to evaluate stable expression and a green fluorescent protein (GFP) construct to monitor transient expression of T-DNA. Transient expression was not suppressed, but stable expression was reduced significantly, in KD plants. Furthermore, KD-Ku70 and KD-Lig4 calli exhibited an increase in the frequency of homologous recombination (HR) compared with control calli. In addition, suppression of OsKu70, OsKu80 and OsLig4 induced the expression of HR-related genes on treatment with DNA-damaging agents. Our findings suggest strongly that NHEJ is involved in Agrobacterium-mediated stable transformation in rice, and that there is a competitive and complementary relationship between the NHEJ and HR pathways for DNA double-strand break repair in rice.</AbstractText>
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Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.

Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.

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