拟南芥中WRKY31转录因子的转录活性与互作蛋白分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0918

摘要/Abstract

摘要： 为深化对AtWRKY基因家族中功能尚未被报道成员的认识,探究拟南芥中WRKY转录因子基因在非生物逆境响应中的作用与可能的机制,通过对AtWRKY31进行亚细胞定位以及转录活性分析,并利用qRT-PCR检测AtWRKY31在不同逆境处理1、3和12 h后的表达变化,通过酵母双杂交（Y2H）和双分子荧光互补（BiFC）对其互作蛋白进行了筛选与验证。发现AtWRKY31定位于细胞核,具有转录抑制作用,且表达受热害、低钾、低氮等处理显著的抑制。此外,发现WRKY31可以与WRKY31、WRKY6和WRKY42在体内发生互作。拟南芥WRKY31作为一个转录抑制子,可能通过与WRKY6和WRKY42转录因子互作来参与调控对一些非生物逆境的应答。

关键词: 拟南芥, WRKY31, 亚细胞定位, 酵母双杂交, 双分子荧光互补

Abstract: In order to deepen our understanding on functionally-unknown members in AtWRKY gene family,and to explore the roles and regulatory mechanisms of WRKY genes in Arabidopsis thaliana in response to abiotic stresses,we chose to study AtWRKY31 gene. First,we performed the subcellular localization assay of AtWRKY31 and analyzed its transcriptional activity. Then,we examined the expression changes of AtWRKY31 under different stress treatments for 1 h,3 h and 12 h through qRT-PCR. At last,we used yeast two-hybrid（Y2H）and bimolecular fluorescence complementation（BiFC）assays to screen and validate the interacting partners of WRKY31. The results showed that AtWRKY31 protein was located in the nucleus and acted as a transcriptional repressor. qRT-PCR showed that the expression of WRKY31 was significantly inhibited under heat,low potassium and low nitrogen treatments. Through yeast two-hybrid and BiFC assays,WRKY31 protein was identified to interact with itself,WRKY6 and WRKY42,respectively. Conclusively,WRKY31 in Arabidopsis thaliana,as a transcriptional repressor,may be involved in the regulation of some abiotic stress responses by the interactions with transcriptional factor WRKY6 and WRKY42.

Key words: Arabidopsis thaliana, WRKY31, subcellular localization, yeast two-hybrid, bimolecular fluorescence complementation

王艺桥,赵新杰,牛芳芳,郭小华,杨博,江元清. 拟南芥中WRKY31转录因子的转录活性与互作蛋白分析[J]. 生物技术通报, 2018, 34(5): 101-109.

WANG Yi-qiao, ZHAO Xin-jie, NIU Fang-fang, GUO Xiao-hua, YANG Bo, JIANG Yuan-qing. Analysis of Transcriptional Activity and Interaction Proteins of WRKY31 Transcription Factor in Arabidopsis thaliana[J]. Biotechnology Bulletin, 2018, 34(5): 101-109.

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