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Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
Published on: January 14, 2016
The transcription factor DNA binding with one finger 27 suppresses chilling tolerance by negatively regulating
Wenjun Sun1, Haishen Xu2, Junyi Zhan3
1Key Laboratory of Coarse Cereal Processing, Ministry of Agriculture and Rural Affairs, Sichuan Engineering and Technology Research Center of Coarse Cereal Industrialization, School of Food and Biological Engineering, Chengdu University, Chengdu, Sichuan, China; College of Life Science, Sichuan Agricultural University, Ya'an, Sichuan, China.
Abstract:
Chilling stress significantly limits crop growth and productivity. Quinoa (Chenopodium quinoa), recognized for its high nutritional value and tolerance to diverse abiotic stresses, including chilling, remains poorly understood at the genetic level regarding its response to chilling treatment. In this study, we identified the DNA binding with one finger (DOF) transcription factor family in quinoa and characterized CqDOF27 as a chilling-inducible gene. Overexpressing of CqDOF27 in quinoa resulted in reduced levels of flavonoids and photosynthetic pigments, decreased antioxidant enzyme activity, and heightened sensitivity to chilling. Gene expression analysis of the flavonoid biosynthesis pathway showed that chilling treatment upregulated CqCHS27 and CqFLS1, while transient overexpression of CqDOF27 suppressed their expression. Yeast one-hybrid (Y1H) assays demonstrated that CqDOF27 directly binds to the promoters of CqCHS27 and CqFLS1. Furthermore, dual-luciferase reporter (Dual-Luc) assays confirmed that CqDOF27 represses the transcriptional activity of these genes. Collectively, these findings indicate that CqDOF27 functions as a negative regulator of chilling tolerance in quinoa by inhibiting flavonoid accumulation through direct repression of CqCHS27 and CqFLS1. This study provides a comprehensive analysis of the DOF transcription factor and flavonoid biosynthetic gene families in quinoa, offering valuable insights for the development of genetically engineered cultivars with enhanced chilling tolerance.
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