油莎豆抗旱基因CeDREB2.1的克隆与功能分析  

Cloning,structure and function analysis of CeDREB2.1 genein Cyperus esculentus

作　　者：柴乖强 秦媛媛 康雨欣 黄凡 段义忠[1,2] 亢福仁 CHAI Guaiqiang;QIN Yuanyuan;KANG Yuxin;HUANG Fan;DUAN Yizhong;KANG Furen(College of Life Science,Yulin University, Yulin, Shaanxi 719000, China;Shaanxi Key Laboratory of Ecological Restoration in Shanbei Mining Area, Yulin, Shaanxi 719000, China)

机构地区：[1]榆林学院生命科学学院,陕西榆林719000 [2]陕西省陕北矿区生态修复重点实验室,陕西榆林719000

出　　处：《干旱地区农业研究》2022年第3期11-19,共9页Agricultural Research in the Arid Areas

基　　金：陕西省教育厅科研计划项目(21JS042);榆林市科技局产学研合作项目(2019-107-2,CXY2020-009-02);榆林市高新区科技局项目(CXY-2020-18);榆林学院博士科研启动基金项目(18GK11)。

摘　　要：干旱应答元件结合蛋白(DREB)能够激活逆境相关基因的表达,提高作物的抗性。沙地植物油莎豆(Cyperus esculentus)具有耐旱、耐瘠等特性,在抗旱节水和生态保护过程中发挥着重要作用。本研究在前期转录组测序中发现了一个DREB转录因子,进而采用PCR扩增、序列分析、表达载体构建和非生物胁迫表达等方法,对油莎豆CeDREB2.1的功能进行初步分析。结果表明,从油莎豆中克隆的CeDREB2.1基因(登录号MZ713253),其开放阅读框为1191 bp,编码397个氨基酸;序列分析表明,CeDREB2.1具有典型的AP2/ERFBP结构域,为亲水性蛋白,α螺旋、延伸链、β折叠和无规卷曲所占比重分别为26.01%、6.57%、2.78%和64.65%;系统进化分析表明,CeDREB2.1与其他物种DREB间序列相似性较高,与胡杨、杨树和苹果的DREB蛋白亲缘关系较近;同时将CeDREB2.1连接到pCSXN上,构建了植物超表达载体并转化拟南芥,转基因拟南芥具有抵抗干旱胁迫的作用;表达分析表明,油莎豆CeDREB2.1能够被干旱、高温、低温、盐和ABA等非生物胁迫诱导表达。Drought response element binding protein(DREB)can activate the expression of stress related genes and improve crop resistance.Cyperus esculentus plays an important role in drought resistance,water saving and ecological protection due to its drought tolerance and barren resistance.In this study,based on the previous transcription alsequencing,a DREB-related transcription factor was found,and its function was preliminarily analyzed by PCR amplification,bioinformatics analysis,expression vector construction and abiotic stress expression.The results showed that a CeDREB2.1(accession number:MZ713253)had an open reading frame of 1191bp and encoding 397 amino acids.Bioinformatics analysis showed that CeDREB2.1 had a typical AP2/ERFBP domain and was a hydrophilic protein.The proportion ofαhelix,extended chain,βfold and random curl were 26.01%,6.57%,2.78%and 64.65%,respectively.Phylogenetic analysis showed that CeDREB2.1 had high sequence similarity with DREB of other species,and it was closely related to DREB proteins of Populus euphrila,poplar and apple.Meanwhile,CeDREB2.1 was connected to pCSXN,and the plant over expression vector was successfully constructed,genetic transformation in Arabidopsis indicated CeDREB2.1 has the biological function of drought resistance.Furthermore,expression analysis in Cyperus esculentus showed that CeDREB2.1 could be induced by abiotic stresses such as drought,high temperature,low temperature,salt and ABA.This study provides theoretical reference for further explaining the biological function of CeDREB2.1 and mechanism of action involved in stress response.

关 键 词：油莎豆 DREB转录因子 基因克隆 干旱胁迫 表达模式 

分 类 号：S548[农业科学—作物学]