The Analysis Binding Specificity Of Taobf1and Expression Pattern Of Seven Kinds Transcription Factors In Dongnongdongmai1Under Cold Stress

Cold, drough, salt and other abiotic stress has a great impact on plant growth and development, Cold stress is one of the main affect factors on plant growth and distribution. Recent evidence indicates that the regulation of gene expression, especially transcription factors play an important role in plant responses to cold stress. Winter wheat Dongnongdongmai1is cultivated by the Northeast Agricultural University, field observations show that the varieties can be resistance to-30℃Cold damage. In this study, known in the wheat bZIP family of seven kinds of transcription factors by the Extremely low temperature stress conditions (-25℃) expression, in order to find the Dongnongdongmai1, and cold stress-related transcription factor, and its regulatory mechanism. TaOBF1transcription factor is a wheat OBF1transcription factor, while the TaOBF1and DNA binding specificity has not been reported.In this study by gel retardation assay proved the TaOBF1transcription factors with the G-box binding specificity, to reveal the law of TaOBF1transcription factors regulate gene expression, and cold stress under winter wheat gene expression and regulation mechanism of reference. Experimental results show that:1.real-time quantitative PCR showed that as the temperature decreases in the1st Dongnongdongmai transcription factor TaOBF1, MYB3and HvWRKY6transcription factor gene expression first increased and then decreased, and the change of magnitude greater than than the Jimai22.These three kinds of transcription factors and winter wheat response to cold stress is relevant. While the regularity of the transcription the factor DRFL2, FLbaf113d20, HvWRKY6and NAC69-1gene expression compared to less significant, indicating that the four kinds of transcription factors and winter wheat response to cold stress is not clear.2.Cloned full-length TaOBFl transcription factor gene construct pET-32b-TaOBF1prokaryotic expression vector and optimization of expression conditions of fusion protein, and obtained by further separation and purification of high purity TaOBFl transcription factor protein.3.EMS A show that TaOBF with the components of the G-box binding transcription factor protein, and can not be combined with the G-box element mutant mG,-box, therefore proved TaOBF transcription factor protein to combine with the G-box components, and This combined with specificity.