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

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.