苦荞麦FtWRKY6基因的克隆及其在低磷与激素诱导下根中的表达分析  

The Cloning and Expression Analysis of FtWRKY6 Gene in Roots of Fagopyrum tataricum Treated by Low Phosphorus and Hormones

作　　者：田建红 彭喜旭[1,2] 邬清韬 文碧瑶 邓楚楚 唐新科 王海华[1,2,3] TIAN Jianhong;PENG Xixu;WU Qingtao;WEN Biyao;DENG Chuchu;TANG Xinke;WANG Haihua(School of Life and Health Sciences,Hunan University of Science and Technology,Xiangtan 411201,China;Key Laboratory of Genetic Improvement and Multiple Utilization of Economic Crops in Hunan Province,Xiangtan 411201,China;Key Laboratory of Ecological Remediation and Safe Utilization of Heavy Metal-polluted Soils,College of Hunan Province,Xiangtan 411201,China)

机构地区：[1]湖南科技大学生命科学与健康学院,湖南湘潭411201 [2]经济作物遗传改良与综合利用湖南省重点实验室,湖南湘潭411201 [3]重金属污染生态土壤修复与安全利用湖南省高校重点实验室,湖南湘潭411201

出　　处：《华北农学报》2023年第1期32-39,共8页Acta Agriculturae Boreali-Sinica

基　　金：科技部国家重点研发计划(2017YFE0117600);湖南省教育厅重点科研项目(19A176);湖南省研究生科研创新项目(CX20211005)。

摘　　要：WRKY转录因子是植物非生物胁迫反应中的重要调节子。荞麦耐贫瘠,磷利用率较高。为了探究WRKY基因在荞麦磷饥饿反应中可能的调节作用,以苦荞麦为材料,采用逆转录PCR方法,从低磷处理的根RNA样品中获得了FtWRKY6基因的编码序列。FtWRKY6 cDNA长1 572 bp,编码524个氨基酸,含有2个典型的WRKY保守结构域,锌指类型为C_(2)H_(2),归属于第Ⅰ类WRKY,与绿茶CsWRKY24在氨基酸水平上的同一性较高(55.5%)。拟南芥原生质体瞬时表达分析表明,FtWRKY6定位于细胞核中;酵母单杂交试验表明,FtWRKY6具有转录激活活性。实时荧光定量PCR分析表明,在根中FtWRKY6的表达受低磷和3种重要的低磷反应相关激素—吲哚乙酸(IAA)、赤霉素(GA)和细胞分裂素(CTK)显著诱导。上述结果提示,FtWRKY6具有转录因子的基本结构与生化特征,在根中可能通过IAA、GA、CTK信号通路的交互作用介导苦荞麦在低磷条件下的响应过程。WRKY transcription factors act important regulators in plant response to low phosphorus.Buckwheat performs well in under-fertilized soils with higher phosphorus use efficiency.Taking tartary buckwheat as experimental materials, this study aims to explore the possible regulatory roles of WRKY genes in phosphorus starvation response of buckwheat.The entire coding sequence(CDS)of FtWRKY6 gene was cloned from RNA samples generated from roots treated by low phosphorus using reverse transcription PCR.The obtained CDS of FtWRKY6 was 1 572 bp in length, encoded a polypeptide of 524 amino acid residues which consists of two conserved WRKY domain each with a zinc finger motif of CCHH,and belonged to the WRKY group Ⅰ.FtWRKY6 shared the highest identity(55.5%)at the amino acid level with Camellia sinensis CsWRKY24.Transient expression assay in protoplasts showed that FtWRKY6 protein was localized in nucleus.Yeast one-hybrid assay revealed that FtWRKY6 had transcription-activating activity.Real-time fluorescence quantitative PCR analysis showed that the expression of FtWRKY6 in roots was significantly induced by low phosphorus and three related hormones such as indole acetic acid(IAA),gibberellin(GA)and cytokinin(CTK).Taken together, FtWRKY6 possesses basic structural and biochemical characteristics as a putative transcription factor, and may be involved in low phosphorus response in roots possibly by crosstalk of IAA,GA and CTK signaling pathways.

关 键 词：苦荞麦 低磷胁迫 WRKY基因 基因表达 转录激活活性 亚细胞定位 

分 类 号：S517[农业科学—作物学] Q943.2[生物学—植物学]