Isolation and functional characterization of the JcERF gene, a putative AP2/EREBP domain-containing transcription factor, in the woody oil plant Jatropha curcas
Isolation and functional characterization of the JcERF gene, a putative AP2/EREBP domain-containing transcription factor, in the woody oil plant Jatropha curcas
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DOI：
10.1007/s11103-006-9098-7
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发表时间：
2007-02
影响因子：
5.1
通讯作者：
Mingjuan Tang;Jingwen Sun;Yun Liu;Fan Chen;S. Shen
中科院分区：
文献类型：
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作者：
Mingjuan Tang;Jingwen Sun;Yun Liu;Fan Chen;S. Shen
从木本油料植物麻疯树幼苗中分离到一个cDNA克隆，命名为JcERF。根据多个序列比对和系统发育特征，该基因被归类为ERF亚家族成员。JcERF克隆推导的氨基酸序列除了保守的AP2/EREBP DNA结合区外，与其他已知的ERF蛋白没有明显的序列相似性。JcERF基因在盐、干旱、乙烯和机械伤害处理下迅速诱导表达。在ABA胁迫下，JcERF的表达没有明显变化。凝胶滞留实验表明，JcERF蛋白能与GCC盒和C/DRE基序特异结合。此外，从凝胶位移可以推断，GCC盒的邻近序列可能对DNA结合活性有重要影响。在酵母中，JcERF蛋白特异性地与DRE序列结合，并激活由DRE序列驱动的两个报告基因His3和LacZ的转录。当与LexA DNA结合域融合时，全长JcERF在酵母单杂交试验中有效地作为反式激活剂发挥作用。JcERFcDNA在转基因拟南芥中的过表达提高了植株的耐盐性和抗冻性。同时，不同浓度的ABA对转JcERF基因植株的种子萌发没有影响。综上所述，结果表明JcERF是一种新的转录因子，与热带木本植物中存在的其他AP2/EREBP调节子一样，表现出植物对环境因子的反应机制。
A cDNA clone, namedJcERF, was isolated fromJatropha curcasseedlings (a woody oil plant). It was classified as an ERF subfamily member based on multiple sequence alignment and phylogenetic characterization. The deduced amino acid sequences of theJcERFclone showed no significant sequence similarity with other known ERF proteins except for the conserved AP2/EREBP DNA-binding domain. Expression of theJcERFgene was rapidly induced upon salinity, drought, ethylene and mechanical wounding treatments. No significant changes in theJcERFexpression were observed under ABA stress. Gel retardation assay revealed that the JcERF protein could bind specifically to the GCC box as well as to the C/DRE motif. Also it can be inferred from the gel-shift that there is a possibility that the near sequence of the GCC box has an important effect on the DNA-binding activity. In yeast, the JcERF protein specifically bound to the DRE sequence and activated the transcription of two reporter genesHis3andLacZdriven by the DRE sequence. When fused to the LexA DNA-binding domain, the full-lengthJcERFfunctioned effectively as a trans-activator in the yeast one-hybrid assay. Overexpression ofJcERFcDNA in transgenicArabidopsisenhanced the salt and freezing tolerance. Meanwhile the seed germination ofJcERFtransgenic plants was not affected by various concentrations ABA in MS medium. Taken together, the results showed that JcERF functioned as a novel transcription factor and it exhibited a mechanism of plant response to environmental factors like the other AP2/EREBP regulons that also exist in tropical woody plants.