A cotton (Gossypium hirsutum) DRE-binding transcription factor gene, GhDREB, confers enhanced tolerance to drought, high salt, and freezing stresses in transgenic wheat
A cotton (Gossypium hirsutum) DRE-binding transcription factor gene, GhDREB, confers enhanced tolerance to drought, high salt, and freezing stresses in transgenic wheat
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棉花（Gossypium hirsutum）DRE 结合转录因子基因 GhDREB 可增强转基因小麦对干旱、高盐和冷冻胁迫的耐受性
DOI：
10.1007/s00299-008-0623-9
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发表时间：
2009-02-01
影响因子：
6.2
通讯作者：
Ma, You-Zhi
中科院分区：
文献类型：
--
作者：
Gao, Shi-Qing;Chen, Ming;Ma, You-Zhi
从棉花 cv. 的 cDNA 文库中分离得到棉花 (G. hirsutum L.) 脱水反应元件结合蛋白基因 GhDREB，该基因编码含有保守 AP2/EREBP 结构域的 153 个氨基酸的蛋白质。 Simian 3 由酵母单杂交系统产生。 RNA印迹分析表明，GhDREB基因在棉苗中受到干旱、高盐和冷胁迫的诱导。电泳迁移率变动分析 (EMSA) 表明 GhDREB 蛋白在体外与 DRE 核心元件 (A/GCCGAC) 特异性结合。构建了两个含有带有泛素或rd29A启动子的GhDREB基因的表达载体，并通过轰击将其转移到小麦（Triticum aestivum L.）中。通过PCR分析分别鉴定了阳麦（36株）和鲁麦（39株）的58株Ubi::GhDREB和17株rd29A::GhDREB T-0植物。 Southern blot和RT-PCR分析表明，2或3个拷贝的GhDREB整合到扬麦10基因组中并在转录水平表达，3或4个拷贝整合到鲁麦23基因组中。功能分析表明，转基因植物在胁迫处理后通过在叶片中积累更高水平的可溶性糖和叶绿素来提高对干旱、高盐和冷冻胁迫的耐受性。转基因植物与其非转基因对照之间没有观察到表型差异。这些结果表明 GhDREB 可能有助于通过基因工程提高小麦的胁迫耐受性。
A cotton (G. hirsutum L.) dehydration responsive element binding protein gene, GhDREB, which encodes a 153 amino acid protein containing a conserved AP2/EREBP domain, was isolated from the cDNA library of cotton cv. Simian 3 by a yeast one-hybrid system. RNA blot analysis showed that the GhDREB gene was induced in cotton seedlings by drought, high salt and cold stresses. An electrophoretic mobility shift assay (EMSA) indicated that the GhDREB protein bound specifically to the DRE core element (A/GCCGAC) in vitro. Two expression vectors containing the GhDREB gene with either of the Ubiqutin or rd29A promoters were constructed and transferred into wheat (Triticum aestivum L.) by bombardment. Fifty-eight Ubi::GhDREB and 17 rd29A::GhDREB T-0 plants of Yangmai (36 plants) and Lumai (39 plants) were identified by PCR analysis, respectively. Southern blot and RT-PCR analyses showed that two or three copies of the GhDREB were integrated into the Yangmai 10 genome and were expressed at the transcriptional level, and three or four copies were integrated into the Lumai 23 genome. Functional analysis indicated that the transgenic plants had improved tolerance to drought, high salt, and freezing stresses through accumulating higher levels of soluble sugar and chlorophyll in leaves after stress treatments. No phenotype differences were observed between transgenic plants and their non-transgenic controls. These results indicated that GhDREB might be useful in improving wheat stress tolerance through genetic engineering.