中国农业科学2016，Vol.49Issue(12)：2245-2254,10.DOI:10.3864/j.issn.0578-1752.2016.12.001
普通小麦转录因子基因TaWRKY35的克隆及功能分析
Cloning and Characterization of Transcription Factor TaWRKY35 in Wheat (Triticum aestivum)
摘要
Abstract
Objective]Abiotic stresses are major limitations to wheat production worldwide. Transcription factors play crucial roles in abiotic stress signaling in plants. It is predicted that there are at least 200 WRKY genes in common wheat genomes, yet only a few of them have been functionally characterized. The aim of this study is to decipher the roles of WRKY transcription factors in abiotic stress signaling and facilitate the utilization of WRKY genes in the improvement of abiotic stress tolerance in wheat.[Method]A wheat WRKY gene designatedTaWRKY35was cloned via the wheat full-length cDNA libraries. Quantitative real-time PCR was performed to identify the dynamic expression of target gene in different tissues at various developmental stages, and to characterize the transcriptional patterns responding to ABA, PEG, NaCl and low temperature treatments in wheat. To probe the subcellular location of TaWRKY35, the construct encoding TaWRYK35::GFP fusion protein was transferred into wheat protoplast by PEG mediated method. To characterize the function of TaWRYK35, target gene driven by the 35S promoter was delivered into Arabidopsis byAgrobacterium mediated method.[Result]The cDNA ofTaWRYK35 contains an 1 134 bp open reading frame, encoding a 377- amino acid protein. TaWRYK35 possesses a typical WRKY domain in the N-terminal and a C2HC type zinc finger domain in the C-terminal, belonging to group IIIof WRKY family. In 32 hexaploid wheat materials of highly polymorphic, TaWRKY35 coding region sequence is very conservative. The dynamic expression ofTaWRKY35 was identified in different tissues at various developmental stages, and the highest expression occurred in the root base of seedling, while the lowest expression was observed in seedling leaf. Furthermore, its transcript was inducible by ABA, PEG, NaCl and low temperature treatments, yet the expression patterns to difference stress varied significantly. Subcellular localization indicated that TaWRKY35 specifically located in the nucleus. Overexpression ofTaWRKY35resulted in enhanced tolerance to high salinity, supported by improved cell membrane stability and survival rate relative to wild typeArabidopsis.[Conclusion]TaWRKY35 transcription factor contains a WRKY and a C2HC type zinc finger domain, belonging to subgroup III of the WRKY family. The expression ofTaWRKY35occurs in different tissues at various developmental stages, andTaWRKY35is an abiotic stress responsive gene. Overexpression ofTaWRKY35 confers remarkably enhanced tolerance to high salinity.关键词
小麦/WRKY转录因子基因/非生物胁迫/耐盐性Key words
wheat/WRKY transcription factor gene/abiotic stress/salt tolerance引用本文复制引用
刘自成,苗丽丽,王景一,杨德龙,毛新国,景蕊莲..普通小麦转录因子基因TaWRKY35的克隆及功能分析[J].中国农业科学,2016,49(12):2245-2254,10.基金项目
国家高技术研究发展计划（“863”计划）（2011AA100501）、北京市自然科学基金 （）
