普通烟草NtWRKY53基因克隆、鉴定及表达分析  

Cloning,characterization and expression pattern analysis of NtWRKY53 gene in Nicotiana tabacum

作　　者：刘万峰 蒲文宣 杨爱国[2] 高军平 王奇 孙明铭 张新要 孙楠 陈凯 王国平[2,3] 李晓旭 LIU Wanfeng;PU Wenxuan;YANG Aiguo;GAO Junping;WANG Qi;SUN Mingming;ZHANG Xinyao;SUN Nan;CHEN Kai;WANG Guoping;LI Xiaoxu(Technology Center,China Tobacco Hunan Industrial Co.,Ltd.,Changsha 410007,China;Key Laboratory of Tobacco Gene Resources,Tobacco Research Institute,Chinese Academy of Agricultural Sciences,Qingdao 266101,Shandong,China;Yuxi Zhongyan Tobacco Seed Co.,Ltd.,Yuxi 653100,Yunnan,China)

机构地区：[1]湖南中烟工业有限责任公司技术中心,长沙市410007 [2]中国农业科学院烟草研究所烟草行业烟草基因资源利用重点实验室,山东省青岛市266101 [3]玉溪中烟种子有限责任公司,云南省玉溪市653100

出　　处：《烟草科技》2021年第1期1-9,共9页Tobacco Science & Technology

基　　金：国家烟草专卖局科技重大专项项目“基于K326品种的工厂化基因组定向编辑与育种素材创制”[110201801030(JY-07)];国家烟草专卖局科技重大专项项目“烟草核心基因库编辑素材的脂类、色素类相关基因功能鉴定研究”[110202001020(JY-12)];湖南中烟工业有限责任公司科技项目“高抗病毒病K326品种的生态适应性和工业可用性研究”(KY2017YC0002)。

摘　　要：WRKY转录因子家族是一类植物特有的转录因子家族,其成员WRKY53在生物及非生物胁迫和叶片衰老过程中发挥着重要的调控作用。为明确WRKY在烟草中的相关功能,克隆了普通烟草NtWRKY53基因。结构域分析表明,NtWRKY53基因编码典型的WRKY转录因子并具有高度保守且典型的WRKY结构域;进化和共线性分析发现,所鉴定的普通烟草NtWRKY53基因与拟南芥AtWRKY53基因为同源基因;亚细胞定位结果表明,NtWRKY53转录因子定位于细胞核中;转录激活试验发现,NtWRKY53转录因子具有转录激活活性;表达模式分析表明,NtWRKY53基因在衰老的叶片中表达量较高且能够被盐、干旱胁迫,以及CMV和黑胫病所诱导。因此,普通烟草NtWRKY53基因编码WRKY型核定位转录因子,在落黄的烟草叶片中表达量较高,受多种生物、非生物逆境诱导,并参与相应的生物和非生物逆境胁迫响应。WRKY transcription factor family is plant-specific and WRKY53,a member of this gene family,plays a significant role in plant biotic and abiotic stress response and leaf senescence.To explore the biological functions of NtWRKY53 gene in Nicotiana tabacum,the NtWRKY53 gene of tobacco was cloned.WRKY transcription factor encoded by NtWRKY53 gene was proved to possess a highly conservative and typical WRKY domain by domain analysis.Phylogenetic and collinearity analyses revealed that NtWRKY53 gene of Nicotiana tabacum and AtWRKY53 gene of Arabidopsis were paralogous genes.Subcellular localization assay indicated that NtWRKY53 transcription factor was localized in cell nucleus.Transactivation analysis implied that NtWRKY53 transcription factor displayed transcriptional activation activity.In addition,the results of expression pattern analysis showed that the NtWRKY53 gene was up-regulated in the senescent leaves and could be induced by multiple stress treatments,such as salt and drought stresses,CMV and black shank disease.Therefore,NtWRKY53 gene encoded a WRKY transcription factor while localized in the nucleus and had a high expression level in yellowing tobacco leaves.It was induced by biotic and abiotic stresses,thus taking part in the biotic and abiotic stress responses.

关 键 词：烟草 WRKY转录因子 生物逆境胁迫 非生物逆境胁迫 叶片衰老 

分 类 号：TS413[农业科学—烟草工业]