干旱响应的大豆G-box结合因子GmDIBL1的克隆和功能鉴定  

Cloning and Functional Identification of Drought Responsive G-box Binding Factor Gm DIBL1 in Soybean

作　　者：乔志清 邱宇姗 高映慧 黎烊村 刘菲 张宏菊 田惠丹 王巧玲 柳宁 谭娉婷 刘晨 罗明[1,2] QIAO Zhiqing;QIU Yushan;GAO Yinghui;LI Yangcun;LIU Fei;ZHANG Hongju;TIAN Huidan;WANG Qiaoling;LIU Ning;TAN Pingting;LIU Chen;LUO Ming(College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China;Engineering Research Center of South Upland Agriculture,Ministry of Education,Chongqing 400715,China;College of Life Sciences,Southwest University,Chongqing 400715,China)

机构地区：[1]西南大学农学与生物科技学院 [2]南方山地农业教育部工程研究中心 [3]西南大学生命科学学院

出　　处：《西南大学学报（自然科学版）》2024年第9期35-44,共10页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(31971984);重庆市技术创新与应用发展专项—重点项目(cstc2021jscx-gksbx0011);重庆市科企联合体种质资源收集利用与品种试验项目(cqnyncw-kqlhtxm)。

摘　　要：干旱是典型的非生物逆境之一,严重影响作物的产量品质.鉴定作物抗/耐旱关键基因功能,解析其调控机制,对于培育抗/耐旱作物新种质、保障粮食安全具有重要意义.大豆是对干旱逆境较敏感的作物,挖掘寻找大豆抗/耐旱基因、鉴定其功能、明确其调控机制是大豆抗逆研究领域的热点.通过分析干旱处理的大豆根转录组数据,获得一个受干旱诱导的G-box结合蛋白基因GmDIBL1(Drought Induced B12D Like 1),克隆的cDNA序列包含1个长261 bp的开放阅读框,编码86个氨基酸残基,推导的分子量为10.1 kD,等电点为9.647.GmDIBL1蛋白质序列与其他物种的G-box结合蛋白之间具有较高的保守性,包含B12D结构域(6~73个氨基酸残基)和跨膜螺旋,N-端具有信号肽,C-端构成非胞质区域(34~86氨基酸残基),该蛋白定位于细胞核.GmDIBL1基因在根瘤中的表达水平最高,根中次之.干旱处理12 h后,其表达水平显著升高.在耐旱性较强的‘晋豆21’根中的表达水平显著高于干旱敏感的‘绥农26’中的表达水平.进一步通过在大豆毛状根中超量表达和敲除GmDIBL1基因,鉴定了转基因材料对干旱的响应,证明超量表达GmDIBL1基因显著提高毛状根复合体的抗旱性.这些结果说明GmDIBL1基因是大豆中一个响应干旱胁迫的基因,在大豆抗/耐干旱逆境中具有重要作用.Drought is one of the typical abiotic stresses that seriously affect the yield and quality of crops.Identifying the functions of key genes for the drought resistance/tolerance of crop and analyzing their regulatory mechanisms are of great significance for developing new drought resistant germplasm of crops and ensuring food security.Soybean is a crop that is more sensitive to drought stress.Exploring and searching for soybean drought resistance/tolerance genes,identifying their functions,and revealing their regulatory mechanisms are hot topics in the field of soybean stress resistance research.This study analyzed transcriptome data of drought treated soybean roots and obtained a drought induced G-box binding protein gene GmDIBL1(Drought Induced B12D Like 1).The cloned cDNA contained an open reading frame of 261 bp length,which encoded 86 amino acid residues.The molecular weight and isoelectric point of GmDIBL1 was 10.1 kD and 9.647,respectively.The GmDIBL1 protein sequence exhibits a high conservation with G-box binding proteins from other species,including the B12D domain(6-73 amino acid residues) and transmembrane helix.The N-terminus has a signal peptide,while the C-terminus forms a non-cytoplasmic region(34-86 amino acid residues).GmDIBL1is located in the nucleus.The expression level of GmDIBL1 gene in root nodules is the highest,followed by roots.After drought treatment for 12 h,GmDIBL1 expression level significantly increased.The expression level of GmDIBL1 was much higher in the root of 'Jindou 21'(a drought resistant variety) than in that of 'Suinong 26'(a drought sensitive variety).Furthermore,the response of this gene to drought was identified by overexpressing and knocking out the GmDIBL1 gene in soybean.Overexpressing GmDIBL1 gene significantly enhanced the drought resistance of the hairy roots complex.These results indicated that the GmDIBL1 gene is a drought stress responsive gene in soybeans and plays an important role in resistance/tolerance to drought stress.

关 键 词：干旱响应 大豆 G-box结合蛋白 GmDIBL1基因 克隆 毛状根 基因编辑 

分 类 号：S565.1[农业科学—作物学]