寒地冬小麦膨胀素基因TaEXPB12同源基因的克隆及功能分析  

Cloning and Function Verification of TaEXPB12 Homologous Genes in Frigid Region Winter Wheat

作　　者：冯珊珊 徐永清[1] 赵梓颐 李凤兰[1] 胡宝忠[1] FENG Shanshan;XU Yongqing;ZHAO Ziyi;LI Fenglan;HU Baozhong(College of Life Science,Northeast Agricultural University,Harbin 150038,China)

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150038

出　　处：《华北农学报》2020年第6期74-80,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家基础科学人才培养基金子课题资助项目(J1210069);国际科技合作资助项目(2013DFR30270);伊春市科技计划推广与产业化项目(T2016-2);哈尔滨市应用科技研究与开发项目(2015RAQXJ021)。

摘　　要：为了进一步研究冬小麦的根系建成,并为今后高寒地区的冬小麦育种奠定基础。以东农冬麦2号为材料,利用PCR技术得到TaEXPB12-A/B/D 3个同源基因的CDS全长。其核酸序列长度均为846 bp,编码281个氨基酸,同源性达到98.46%,分别定位在2AL、2BL、2DL染色体上,蛋白均具备膨胀素基因序列特有的结构域DPBB_1,分别位于142-810 bp,142-834 bp,142-834 bp。启动子分析结果显示,这3段序列中均含有多个与根特异表达相关的作用元件,以及多个激素响应元件和非生物胁迫响应元件。qRT-PCR分析发现TaEXPB12-A/B/D基因在根中特异性表达,低温会抑制TaEXPB12-A/B/D 3个同源基因的表达,而干旱、脱落酸、茉莉酸甲酯及水杨酸等处理会促进它们的表达。洋葱表皮亚细胞定位结果显示,3个基因均定位于细胞壁。TaEXPB12-A/B/D蛋白质在冬小麦根的细胞壁中特异性分布,并参与冬小麦抵御外界多种非生物胁迫的过程,在3个基因响应外界胁迫的过程中,TaEXPB12-A占据表达优势。结果说明,TaEXPB12-A/B/D在冬小麦根系建成和抵御多种非生物胁迫过程中具有重要作用。In order to further study the establishment of the root system of winter wheat,and lay the foundation for winter wheat breeding in alpine regions in the future.Dongnongdongmai 2 was used as the material,and the full-length CDS of three homologous genes of TaEXPB12-A/B/D was obtained by PCR technology.The nucleic acid sequence length was 846 bp,encoding 281 amino acids,homology reached 98.46%,the three genes located on the 2 AL,2 BL,2 DL chromosomes respectively,their protein had the unique domain DPBB_1 of the expand in gene sequence,locating at 142-810 bp,142-834 bp,142-834 bp,respectively.Promoter analysis results showed that all three sequences contained multiple action elements related to root-specific expression,as well as multiple hormone response elements and abiotic stress response elements.qRT-PCR analysis found that TaEXPB12-A/B/D genes were specifically expressed in roots.Low temperature would inhibit the expression of three homologous genes of TaEXPB12-A/B/D,while drought,abscisic acid,methyl jasmonate and salicylic acid would promote their expression.Onion epidermal subcellular localization results showed that all three genes were localized on the cell wall.TaEXPB12-A/B/D protein was specifically distributed in the cell wall of winter wheat roots,and participated in the process of winter wheat resisting a variety of abiotic stresses.In the process of three genes responding to external stress,TaEXPB12-A had an expression advantage.These results indicated that TaEXPB12-A/B/D played an important role in the establishment of winter wheat roots and resistance to various abiotic stresses.

关 键 词：寒地冬小麦 膨胀素 TaEXPB12-A/B/D 同源基因 表达特性分析 

分 类 号：Q78[生物学—分子生物学] S512.03[农业科学—作物学]