The New phytologist, 2012-12, Vol.196 (4), p.1048-1059
2012
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- Autor(en) / Beteiligte
- Titel
- Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice
- Ist Teil von
- The New phytologist, 2012-12, Vol.196 (4), p.1048-1059
- Ort / Verlag
- England: New Phytologist Trust
- Erscheinungsjahr
- 2012
- Quelle
- Wiley-Blackwell Journals
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0028-646XeISSN: 1469-8137DOI: 10.1111/j.1469-8137.2012.04350.xTitel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3532656
- Format
- –
- Schlagworte
- Agrobacterium, Agrobacterium - genetics, Arabidopsis, Beryl, Deoxyribonucleic acid, DNA, DNA Breaks, Double-Stranded, DNA damage, DNA double‐strand breaks (DSBs), DNA End-Joining Repair - genetics, DNA Helicases - genetics, DNA Helicases - metabolism, DNA ligase 4 (Lig4), DNA Ligase ATP, DNA Ligases - genetics, DNA Ligases - metabolism, DNA Repair, DNA, Bacterial, DNA-Binding Proteins - genetics, DNA-Binding Proteins - metabolism, Fluorescence, Gene expression, Gene Expression Regulation, Plant, Gene Knockdown Techniques, Genes, Genetic transformation, Genomes, Green fluorescent protein, Green Fluorescent Proteins - genetics, Green Fluorescent Proteins - metabolism, Homologous Recombination, homologous recombination (HR), Homology, Ku70, Ku80, LIG4 protein, Luminescence, Non-homologous end joining, nonhomologous end joining (NHEJ), Oryza - genetics, Oryza - metabolism, Oryza sativa, Oryza sativa (rice), Plant bacterial diseases, Plant Proteins - genetics, Plant Proteins - metabolism, Plants, Plants, Genetically Modified, Recombination, Rice, transferred DNA (T‐DNA), Transformation, Genetic, Transformations, Transgenic plants