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水通道蛋白不仅控制着水分和一些小分子溶质的跨膜运输，也是植物体应对非生物胁迫的重要组成部分。为了进一步研究小麦水通道蛋白的功能，本研究以NCBI和Ensemble Plant数据库中查找出的已报道的小麦水通道蛋白基因序列为模板，针对其保守区设计了19对引物，应用PCR的方法从5个小麦品种中克隆出19个基因片段。通过比对鉴定到一个此前未曾报道的新基因，并对其进行生物信息学分析，将其命名为 TaNIP4-1。理化特性分析表明，该基因位于3A染色体短臂，基因全长1 315 bp，CDS序列长度864 bp，含有3个外显子和2个内含子，编码含有288个氨基酸的多肽，含有保守的沙漏模型，有6个跨膜区和2个保守的NPA基序。亚细胞定位预测结果表明该基因位于质膜上。该基因启动子序列含有与逆境响应相关的顺式调控元件，而对7日龄的麦苗进行干旱和盐胁迫处理后，其qRT-PCR结果显示，干旱和盐处理5 h后， TaNIP4-1基因在小麦叶片中的表达量明显上调；盐处理7 d后， TaNIP4-1基因在小麦根部的表达量显著上调。由此可见， TaNIP4-1基因参与了小麦应对逆境的过程。

英文摘要:

Aquaporins can not only transport water and some small molecules through the membrane, but also have an effect on abiotic stress. In order to study the function of aquaporins involved in drought resistance mechanism of wheat, reported wheat aquaporin gene sequences were selected from the NCBI and Ensemble Plant databases. A total of 19 pairs of primers were designed, and 19 gene fragments were cloned from five wheat cultivars’s genome using PCR. An aquaporin gene, which has not been identified in the previous bioinformatics analysis, was found via BLAST analysis and some bioinformatics analysis was further carried out. The results showed that one of the 19 genes was not yet reported and it was named as TaNIP4-1 according to the phylogenetic tree and conserved domain analysis. The physical and chemical properties analysis showed that the gene is located on the chromosome 3AS,DNA sequence length is 1315 bp,coding sequence length is 864 bp, containing three exons and two introns, encoding 288 amino acids and the protein is hydrophobic. In addition, the transmembrane domain analysis and 3D model indicated that the protein contains a conservative hourglass model with six transmembrane regions and two conserved NPA motif. Subcellular localization prediction results indicated that the gene is localized on the plasma membrane, while promoter analysis revealed this gene contained cis-acting regulatory elements which were related to drought.The qRT-PCR results showed that the expression level of TaNIP4-1 in wheat leaves was obviously increased after 5 h both under drought and salt treatments,and the expression level of TaNIP4-1 in wheat roots was significantly up-regulated after 7 days under salt treatment. Thus, these results suggested that this gene was response to the abiotic stress.

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