Djemal Rania, Khoudi Habib
Laboratory of Plant Protection and Improvement, Center of Biotechnology of Sfax, University of Sfax, B.P' 1177, Route Sidi Mansour Km 6, 3018, Sfax, Tunisia.
Protoplasma. 2015 Nov;252(6):1461-73. doi: 10.1007/s00709-015-0775-8. Epub 2015 Feb 17.
Over the last decade, APETALA2/Ethylene Responsive Factor (AP2/ERF) proteins have become the subject of intensive research activity due to their involvement in a variety of biological processes. This research led to the identification of AP2/ERF genes in many species; however, little is known about these genes in durum wheat, one of the most important cereal crops in the world. In this study, a new member of the AP2/ERF transcription factor family, designated TdSHN1, was isolated from durum wheat using thermal asymetric interlaced PCR (TAIL-PCR) method. Protein sequence analysis showed that TdSHN1 contained an AP2/ERF domain of 63 amino acids and a putative nuclear localization signal (NLS). Phylogenetic analysis showed that TdSHN1 belongs to a group Va protein in the ERF subfamily which contains the Arabidopsis ERF proteins (SHN1, SHN2, and SHN3). Expression of TdSHN1 was strongly induced by salt, drought, abscisic acid (ABA), and cold. In planta, TdSHN1 protein was able to activate the transcription of GUS reporter gene driven by the GCC box and DRE element sequences. In addition, TdSHN1 was targeted to the nucleus when transiently expressed in tobacco epidermal cells. In transgenic yeast, overexpression of TdSHN1 increased tolerance to multiple abiotic stresses. Taken together, the results showed that TdSHN1 encodes an abiotic stress-inducible, transcription factor which confers abiotic stress tolerance in yeast. TdSHN1 is therefore a promising candidate for improvement of biotic and abiotic stress tolerance in wheat as well as other crops.
在过去十年中,APETALA2/乙烯响应因子(AP2/ERF)蛋白因其参与多种生物学过程而成为深入研究的对象。这项研究促使人们在许多物种中鉴定出了AP2/ERF基因;然而,对于硬粒小麦(世界上最重要的谷类作物之一)中的这些基因却知之甚少。在本研究中,利用热不对称交错PCR(TAIL-PCR)方法从硬粒小麦中分离出了AP2/ERF转录因子家族的一个新成员,命名为TdSHN1。蛋白质序列分析表明,TdSHN1包含一个由63个氨基酸组成的AP2/ERF结构域和一个假定的核定位信号(NLS)。系统发育分析表明,TdSHN1属于ERF亚家族中的Va组蛋白,该组包含拟南芥ERF蛋白(SHN1、SHN2和SHN3)。盐、干旱、脱落酸(ABA)和低温强烈诱导TdSHN1的表达。在植物体内,TdSHN1蛋白能够激活由GCC盒和DRE元件序列驱动的GUS报告基因的转录。此外,当在烟草表皮细胞中瞬时表达时,TdSHN1定位于细胞核。在转基因酵母中,TdSHN1的过表达提高了对多种非生物胁迫的耐受性。综上所述,结果表明TdSHN1编码一种非生物胁迫诱导型转录因子,该因子赋予酵母非生物胁迫耐受性。因此,TdSHN1有望成为提高小麦以及其他作物对生物和非生物胁迫耐受性的候选基因。