普通烟草Nt WRKY40基因克隆及其抗旱功能  

Cloning of NtWRKY40 gene in Nicotiana tabacum and analysis of its drought resistance functions

作　　者：耿少武 柴云霞 丁以纾 林云红 布云虹 王磊 熊茜 艾可彪 孙嘉贤 李晓燕 周贵贤 贺丽 刘正杰[2] GENG Shaowu;CHAI Yunxia;DING Yishu;LIN Yunhong;BU Yunhong;WANG Lei;XIONG Qian;AI Kebiao;SUN Jiaxian;LI Xiaoyan;ZHOU Guixian;HE Li;LIU Zhengjie(Chuxiong Branch of Yunnan Provincial Tobacco Company,Chuxiong 675099,Yunnan,China;College of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南省烟草公司楚雄州公司,云南省675099 [2]云南农业大学农学与生物技术学院,昆明市650201

出　　处：《烟草科技》2025年第3期60-69,共10页Tobacco Science & Technology

基　　金：中国烟草总公司云南省公司科技重大项目“楚雄K326植烟土壤精准管理及烟叶品质调控关键技术研究”(2023530000241019)。

摘　　要：从普通烟草K326中克隆WRKY转录因子编码基因NtWRKY40,对该基因进行亚细胞定位并在烟草K326中过表达,通过对比转基因和对照植株的抗旱能力,检测抗旱生理指标和干旱胁迫响应基因的表达量,解析NtWRKY40基因的抗旱功能。结果表明,烟草NtWRKY40基因核心编码区(CDS)全长777bp,编码258个氨基酸,亚细胞定位显示该基因主要在细胞核中表达;NtWRKY40基因过表达植株的抗旱性显著高于对照;在聚乙二醇(PEG)模拟干旱胁迫处理下,转基因植株中游离脯氨酸(Pro)含量、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均高于对照,而丙二醛(MDA)含量低于对照,转基因植株中NtSOD、NtAPX、NtP5CS和NtNCED14个干旱胁迫响应基因的表达量显著高于对照。初步验证烟草NtWRKY40基因能够有效提高烟草抗旱性,可为该基因在植物抗逆中的应用提供理论依据。The NtWRKY40 gene encoding a WRKY transcription factor was cloned from tobacco cv.K326 and the gene was subcellularly mapped and overexpressed in tobacco.By comparing the drought resistance of transgenic and control(wild-type)plants,the physiological indexes of drought resistance and the expression of drought stress response genes were determined,and the drought resistance function of the NtWRKY40 gene was analyzed.The results showed that the coding sequence(CDS)of the tobacco NtWRKY40 gene was 777 bp in length,encoding 258 amino acids.Subcellular localization showed that the gene was mainly expressed in the nucleus.Compared with the control plants,the overexpressed transgenic plants of the NtWRKY40 gene exhibited significantly higher drought resistance,the free proline(Pro)content and the superoxide dismutase(SOD)and peroxidase(POD)activities in the transgenic plants were higher,while the malondialdehyde(MDA)content was lower;and the expression levels of NtSOD,NtAPX,NtP5CS and NtNCED1 in transgenic plants were significantly higher under polyethylene glycol(PEG)-simulated drought stress.This study provided preliminary evidence that the NtWRKY40 gene in tobacco could effectively improve drought resistance and provides a research basis for the application of this gene in plant stress resistance genetic engineering.

关 键 词：普通烟草 NtWRKY40基因 亚细胞定位 干旱胁迫 

分 类 号：S572.034[农业科学—烟草工业]