Cloning and Characterization of a Putative CTR1 Gene from Wheat
Cloning and Characterization of a Putative CTR1 Gene from Wheat
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小麦中假定的 CTR1 基因的克隆和表征
DOI：
10.1016/s1671-2927(09)60213-3
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发表时间：
2010-09-20
影响因子：
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通讯作者：
Liu Xu
中科院分区：
文献类型：
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作者：
Bi Cai-Li;Wen Xiao-Jie;Liu Xu
CTR 1是乙烯信号转导中一个关键的负调控因子。从小麦中克隆了一个盐诱导的类CTR 1基因（TaCTR 1），研究了该基因在非生物胁迫下的表达、亚细胞定位以及过量表达TaCTR 1对烟草耐盐性的影响。利用RACE和RT-PCR技术从小麦中克隆了一个CTRI基因。利用半定量RT-PCR分析了TaCTR 1在逆境下的表达，并研究了TaCTR 1过表达对烟草耐盐性的影响。TaCTR 1的cDNA全长为2 635 bp，编码759个氨基酸。在羧基末端有一个保守的丝氨酸/苏氨酸蛋白激酶结构域，含有ATP结合位点。Southern杂交分析表明，TaCTR 1在小麦中属于一个基因家族。不同生物间CTR 1的氨基酸同源性较高。表达分析表明，TaCTR 1诱导NaCl和干旱胁迫，但抑制阿坝处理。TaCTR 1-GFP在洋葱表皮细胞中的瞬时表达表明TaCTR 1可能靶向于质膜。过量表达TaCTR 1降低了转基因烟草的耐盐性。与对照组相比。据我们所知，TaCTR 1是第一个在小麦中克隆的CTR 1基因，可能与多种非生物胁迫有关。过量表达TaCTR 1降低了烟草的耐盐性，表明TaCTR 1可能是植物盐胁迫的负调节因子。
CTR1 is a key negative regulator in ethylene signal transduction. A salt-induced CTR1 like gene (TaCTR1) was cloned from wheat, its expression under abiotic stresses, subcellular localization and the effect of overexpression of TaCTR1 on salt tolerance in tobacco was studied. A putative CTRI gene was cloned and characterized from wheat via rapid amplification of cDNA ends (RACE) and RT-PCR. TaCTR1 expression under stresses was analyzed using semi-quantitative RT-PCR and the effect of overexpression of TaCTR1 on salt tolerance was conducted in tobacco. The full-length cDNA of TaCTR1 is 2 635 bp which codes for a polypeptide of 759 amino acids. There is a conserved serine/threonine protein kinase domain at the carboxyl terminus containing an ATP-binding site. Southern blot analysis revealed that TaCTR1 consisted of a gene family in wheat. The amino acid homologies of CTR1 among different organisms share higher similarities. Expression analysis revealed that TaCTR1 was induced by NaCl and drought stress but inhibited by ABA treatment. Transient expression of TaCTR1-GFP in the onion epidermal cells indicated that TaCTR1 was probably targeted to the plasma membrane. Overexpression of TaCTR1 decreased salt tolerance in transgenic tobacco (Nicotiana tabacum L.) plants compared with the control. To our knowledge, TaCTR1 is the first CTR1 gene cloned in wheat and may be involved in various abiotic stresses. Overexpression of TaCTR1 decreased the salt tolerance in tobacco suggested that TaCTR1 may act as a negative regulator of salt stress in plants.