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DREB/CBF(dehydration-responsive element binding protein/C-repeat binding factor)转录因子能特异的与DRE/CRT(dehydration-responsive element/C-repeat)顺式作用元件结合,在植物干旱、高盐和低温等非生物胁迫应答中起着重要的调控作用。本研究从山葡萄中克隆得到了一个VaDREB转录因子基因(登录号XM_002283076.1),并对其进行了序列分析、亚细胞定位分析以及酵母和拟南芥中的功能鉴定。序列分析表明,VaDREB开放阅读框全长459 bp,编码152个氨基酸,预测蛋白质分子量为17 kDa,等电点为8.67,包含一个AP2/ERF结合域。氨基酸序列比对和系统进化分析结果表明,该基因属于DREB/CBF转录因子A-5亚类。亚细胞定位结果表明VaDREB蛋白定位于细胞核中。重组酵母菌株与表达VaDREB基因的转基因拟南芥株系均表现出明显增强的盐、干旱、低温和高温胁迫耐受性表型,基因表达分析结果表明,转基因拟南芥株系中抗逆相关功能基因的表达量出现了明显上调,这些结果证明VaDREB转录因子在植物抗逆调控过程中起着重要作用。
DREB / CBF (dehydration-responsive element binding protein / C-repeat binding factor) transcription factors can specifically bind to cis-acting elements of DRE / CRT (dehydration-responsive element / C-repeat) And other non-biological stress response plays an important regulatory role. In this study, a VaDREB transcription factor gene (accession number XM_002283076.1) was cloned from the grapevine, and its sequence analysis, subcellular localization analysis and functional identification in yeast and Arabidopsis were carried out. Sequence analysis showed that the open reading frame of VaDREB was 459 bp in length and encoded a protein of 152 amino acids. The predicted protein was 17 kDa with an isoelectric point of 8.67 and contained an AP2 / ERF binding domain. Amino acid sequence alignment and phylogenetic analysis indicated that the gene belonged to the DREB / CBF transcription factor A-5 subtype. Subcellular localization results indicate that the VaDREB protein is localized in the nucleus. The recombinant yeast strain and the transgenic Arabidopsis thaliana strain expressing VaDREB gene all showed significantly enhanced salt, drought, hypothermia and high temperature stress tolerance phenotypes. The gene expression analysis indicated that the anti-retrogradation in transgenic Arabidopsis lines The expression of functional genes showed a significant increase, these results demonstrate that VaDREB transcription factor plays an important role in the plant anti-retrograde regulation.