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Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
Published on: January 14, 2016
A nuclear-localized protein, KOLD SENSITIV-1, affects the expression of cold-responsive genes during prolonged
Sarah J Purdy1, John D Bussell, David C Nelson
1Australian Research Council Centre of Excellence in Plant Energy Biology, The University of Western Australia, Stirling Highway, Crawley, Perth, Western Australia, 6009, Australia.
Abstract:
Plants respond to cold by transcriptional and metabolic responses which underlie tolerance and acclimation mechanisms, but details at the molecular level are incomplete. Here we describe KOLD SENSITIV-1 (KOS1), a new gene required for responses to cold. KOS1 protein is predicted to have coiled-coil, Structural Maintenance of Chromosomes and nuclear-targeting domains. GFP-labeled KOS1 localizes to the nucleus. Null mutants could not be isolated but two independent knockdown T-DNA mutants were obtained. Growth and development of kos1 knockdown mutant plants was comparable to wild type when grown at 21°C. However, when grown at 4°C these mutants exhibited accelerated leaf yellowing and smaller rosette size than wild type. Quantitative RT-PCR revealed that in the cold kos1 mutants had reduced expression of cold-responsive transcripts COR15A, COR15B, BAM3 and AMY3. Metabolite profiling revealed that ascorbate levels were lower in the mutants in the cold relative to wild type. KOS1 therefore represents a new gene that influences the regulation of transcript and metabolite levels in response to prolonged chilling temperatures.
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