PtrWRKY51基因的克隆及抗旱表达特性分析  

Cloning and Drought Tolerance Expression Analysis of PtrWRKY51 Gene in Populus trichocarpa

作　　者：黄安瀛 夏德安[3] 张洋[3] 那冬晨[4] 燕青 魏志刚 HUANG Anying;XIA Dean;ZHANG Yang;NA Dongchen;YAN Qing;WEI Zhigang(Research Center for Saline-alkali Land,National Forestry and Grassland Administration,Beijing 100091;China Eucalypt Research Centre,Chinese Academy of Forestry(CAF),Zhanjiang 524022;State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040;College of Life Science,Shanxi Normal University,Linfen 041000)

机构地区：[1]国家林业和草原局盐碱地研究中心,北京100091 [2]国家林业和草原局桉树研究开发中心,湛江524022 [3]东北林业大学林木遗传育种国家重点实验室,哈尔滨150040 [4]山西师范大学生命科学学院,临汾041000

出　　处：《植物研究》2022年第6期1005-1013,共9页Bulletin of Botanical Research

基　　金：中央级公益性科研院所基本科研业务费专项资金(CAFYBB2020ZA005);国家自然科学基金(31770640)。

摘　　要：为明确毛果杨WRKY家族成员PtrWRKY51基因功能,以Nisqually-1株系毛果杨为模板,克隆得到PtrWRKY51基因CDS序列。通过生物信息学分析,结合酵母自激活验证、亚细胞定位及模拟干旱胁迫下的实时荧光定量PCR(qRT-PCR)对PtrWRKY51基因功能进行初步研究。结果表明:PtrWRKY51全长579 bp,编码192 aa。生物信息学分析及亚细胞定位试验结果表明,PtrWRKY51蛋白为非跨膜碱性不稳定亲水蛋白,定位于细胞核,含有WRKY家族特有的保守结构域,是第IIc类WRKY转录因子;酵母自激活验证试验表明PtrWRKY51基因具有自激活活性;qRT-PCR分析表明,8%PEG6000模拟干旱胁迫下,该基因在胁迫12 h后茎部与叶部相对表达量达到最大值,根部则出现在24 h,研究可为PtrWRKY51抗逆及生物学功能进一步研究提供参考。In order to clarify the function of PtrWRKY51 gene of Populus trichocarpa,the CDS of PtrWRKY51 gene was cloned using P.trichocarpa from Nisqually-1 strain as material.The function of PtrWRKY51 gene was studied by bioinformatics analysis,and assayed by yeast self-activation verification,and subcellular localization and simulated drought stress by real-time qPCR.The results showed that the CDS of PtrWRKY51was 579 bp and encoded 192 aa.Bioinformatics analysis and subcellular localization experiments showed that PtrWRKY51 was a non-transmembrane alkaline unstable hydrophilic protein,located in the nucleus and contained a conserved domain unique to WRKY family,it was a WRKY transcription factor classified to class IIc The yeast self-activation verification experiment showed that PtrWRKY51 gene could self-activation activity qRT-PCR showed that the expression of this gene was significantly induced by 8% PEG6000 and reached the highest expression in stem and leaf after 12 h stress,while the roots appeared after 24 h stress.This study could provide reference for further study on stress tolerance and biological function of PtrWRKY51.

关 键 词：毛果杨 PtrWRKY51 生物信息学 酵母自激活 亚细胞定位 抗旱表达分析 

分 类 号：S792.11[农业科学—林木遗传育种]