| 刘 燕,赵 毅,吕 千,张 丽,李立群,李学军.小麦TaHTAS-5A基因的克隆、表达分析及亚细胞定位[J].麦类作物学报,2018,(10):1137 |
| 小麦TaHTAS-5A基因的克隆、表达分析及亚细胞定位 |
| Cloning, Expression Analysis and Subcellular Localization of TaHTAS-5A Gene from Wheat(Triticum aestivum) |
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| DOI：10.7606/j.issn.1009-1041.2018.10.01 |
| 中文关键词: 小麦 TaHTAS-5A 非生物胁迫 表达分析 亚细胞定位 |
| 英文关键词:Wheat TaHTAS-5A Abiotic stress Expression analysis Subcellular localization |
| 基金项目:国家重点研发计划项目（2017YFD0100704）；农业部高产转基因小麦新品种培育科技重大专项（2016ZX08002003）；杨凌示范区产学研用协同创新重大项目（2016CXY-01) |
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| 中文摘要: |
| 非生物胁迫对小麦的生长发育具有不良影响，克隆与非生物胁迫相关的基因，并对其结构及表达特性进行分析，可以为进一步探索小麦的抗逆机制和抗逆育种奠定基础。本研究利用同源克隆技术从普通小麦品种中国春中克隆了水稻E3泛素连接酶基因OsHTAS的同源基因TaHTAS-5A（GeneBank登录号：MF967573），并利用生物信息学方法对其基因序列特征进行分析；利用qRT-PCR技术对该基因在小麦不同组织及不同逆境胁迫下的表达模式进行分析；同时，对该基因的表达产物进行了亚细胞定位。结果表明，小麦TaHTAS-5A基因含有220 bp的5′UTR、1 242 bp的ORF以及126 bp的3′UTR，共编码413个氨基酸；基因组分析表明，该基因全长3 819 bp，共包含4个外显子和3个内含子；缺体-四体定位表明该基因位于小麦5A染色体上；蛋白结构分析显示，TaHTAS-5A蛋白的N端包含四个跨膜结构域，C端包含一个E3泛素连接酶特有的RING finger保守结构域，具有植物E3泛素连接酶的结构特征；qRT-PCR结果显示，TaHTAS-5A对高温、低温和ABA都有响应，对干旱和盐响应不敏感；TaHTAS-5A在小麦的根、茎、叶和幼穗等不同组织中均有表达，但表达量有差异，在叶和穗中的表达量明显高于根部和茎部；亚细胞定位结果表明， TaHTAS-5A位于细胞膜上。本研究为进一步探究小麦TaHTAS-5A基因的功能奠定了基础，也为小麦非生物胁迫抗性改良提供了一定的理论依据。 |
| 英文摘要: |
| Abiotic stress has adverse effects on plant growth and development. Cloning the abiotic stress related genes laid the foundation for further exploring the mechanism of plant stress resistance and provided a theoretical basis for the stress resistance breeding of wheat through the analysis of the gene structure and expression of the target gene. In this study, we cloned the gene TaHTAS-5A(GeneBank accession number:MF967573) homologous to rice E3 ubiquitin ligase gene OsHTAS from Chinese spring in Triticum aestivum. We analyzed its sequence features by bioinformatic techniques. The expression pattern of TaHTAS-5A in different tissues and under different stresses of wheat was analyzed by qRT-PCR. At the same time, the expression products of this gene were analyzed by subcellular localization. The result show that the TaHTAS-5A gene contained 220 bp 5′UTR, 1 242 bp intact CDS and 126 bp 3′UTR, encoding 413 amino acids. Genomic analysis showed that the full length of TaHTAS-5A in genome is 3819 bp, containing four exons and three introns. Nullisomic localization indicated that the gene is located on wheat 5A chromosome. Protein structure analysis showed that the N-terminus of TaHTAS-5A protein included four transmembrane domains and C-terminal contained an E3 ubiquitin ligase-specific conservative RING finger domain, which has the structural characteristics of plant E3 ubiquitin ligase. qRT-PCR results demonstrated that TaHTAS-5A was responsive to high temperature, low temperature and ABA, but insensitive to drought and salt stress. Tissue expression analysis showed that TaHTAS-5A expressed in roots, stems, leaves and young spike, where it was least expressed in root and stem, but highly expressed in leaves and spikes. Subcellular localization showed that TaHTAS-5A is located on the cell membrane. The results laid a foundation for further exploring the function of wheat TaHTAS-5A gene and provided some theoretical basis for improving resistance to abiotic stress in wheat. |
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