大豆转录因子GmWRKY58亚细胞定位及在非生物胁迫下的表达分析

Subcellular Localization and Expression Analysis of a Soybean Transcription Factor GmWRKY58 in Response to Abiotic Stresses

WRKY转录因子在植物响应非生物胁迫,调控抗逆基因的表达方面具有重要作用。为了解析大豆转录因子GmWRKY58基因在非生物胁迫中的功能,从大豆中克隆了GmWRKY58 c DNA全长,推导其编码的氨基酸序列、生理生化特征与进化关系,并研究了其亚细胞定位和在不同组织及非生物胁迫下基因的表达变化。结果表明,GmWRKY58基因c DNA ORF全长为954 bp,编码317个氨基酸。亚细胞定位结果显示,GmWRKY58蛋白质定位于细胞核中。实时荧光定量PCR基因表达分析表明,GmWRKY58在大豆根、茎、叶、花和荚等组织均有表达,根、茎、叶中的表达量明显高于花和荚。在大豆根中,GmWRKY58基因受高盐、干旱、低N和缺Fe等非生物胁迫因子强烈诱导,在高盐胁迫下,基因表达量增加187.4倍;GmWRKY58基因受水杨酸(SA)、低温及低P和低K等诱导轻微,差异表达不显著。由此推测,GmWRKY58转录因子在大豆抗盐、抗旱、低N和缺Fe等非生物胁迫过程中起到重要的调控作用。

WRKY transcription factor plays an important role in plant response to abiotic stress and regulation of resistance gene expression.In order to reveal the role of a soybean transcription factor GmWRKY58 in response to abiotic stresses.The full length of GmWRKY58 cDNA was cloned from soybean,and the amino acid sequence,physiological and biochemical characteristics and evolutionary relationship were deduced.Subcellular localization and the changes of gene expression under different tissues and abiotic stress were studied.The results showed that GmWRKY58 had a 954 bp length open reading frame and encoded a total of 317 amino acids.Subcellular localization results showed that GFP signals combined GmWRKY58 was only detected in the nucleus.Real-time RT-PCR analysis showed that GmWRKY58 was expressed in soybean roots,stems,leave,flowers and pods,and the expression in each tissue of roots,stems and leave was significantly higher than flowers and pods.In soybean roots,GmWRKY58 gene can be significantly induced by abiotic stress factors such as high salinity,drought,low nitrogen and iron deficiency.Under high salt stress,GmWRKY58 gene expression was increased by more than 187.4 times.Moreover,this gene just slightly induced by other abiotic stresses shch as SA,low temperature,low phosphorus and low potassium.The results indicate that GmWRKY58 plays an important regulatory role in abiotic stress process of salt,drought,low nitrogen and iron deficiency in soybean.