平榛ChWRKY28基因克隆及表达模式分析  

Cloning and Expression Analysis of Ch WRKY28 from Corylus heterophylla Fisch

作　　者：赵天田[1] 梁丽松[1] 马庆华[1] 王贵禧[1] 

机构地区：[1]中国林业科学研究院林业研究所,国家林业局林木培育重点实验室,林木遗传育种国家重点实验室,北京100091

出　　处：《林业科学研究》2016年第2期250-255,共6页Forest Research

基　　金：国家自然基金(31500555);林业公益性行业科研专项(201304710);林业科学技术推广项目[2014]-3号

摘　　要：[目的]研究平榛Ch WRKY28基因序列特征及其在不同非生物胁迫下的表达规律。[方法]以平榛为试材,采用RACE-PCR方法进行基因克隆;利用实时荧光定量PCR方法检测基因在不同组织及不同非生物胁迫下的表达模式。[结果]表明:克隆得到的WRKY基因,全长1 342 bp,基因内部包含1个长963 bp的完整开放阅读框,编码320个氨基酸残基,命名为Ch WRKY28。构建的系统发育树表明:该序列与拟南芥At WRKY28及杨树Ptr WRKY93的关系最近,相似性分别为49%和60%。基因表达分析表明:Ch WRKY28在雄花序、雌花芽及茎中均有表达,但在茎部(皮)中的表达量高于雄花序和雌花芽中的表达量,具有组织表达特异性;低温、干旱及盐胁迫均能诱导Ch WRKY28基因的表达,但受诱导程度存在差异。亚细胞定位分析结果表明:Ch WRKY28蛋白分布在细胞核内,是一个核蛋白。[结论]推测Ch WRKY28基因可能参与植物响应非生物胁迫的信号转导过程。[Objective]To analyze the sequence features and expression rules of WRKY gene from Corylus heterophylla Fisch. [Method]The gene was cloned by RACE-PCR. Quantitative real-time PCR was used in analyzing gene expression in various tissues and different abiotic stresses,including cold,high salinity and drought. [Result]The c DNA of WRKY is 1 342 bp in length,including a complete open reading frame（ ORF） of 963 bp encoding a protein of 320 amino acids,designated as Ch WRKY28. Phylogeny tree results showed Ch WRKY28 was much closer to At WRKY28 from Arabidopsis thaliana and Ptr WRKY93 from Populus trichocarpa,generated 49% and 60% amino acids similarity. Spatial expression analyses demonstrated that the expression level of Ch WRKY28 was higher in stem than that in male anthotaxy and floral buds which indicating tissue-specific expression. Ch WRKY28 was clearly induced by cold,high salinity and drought,but that the expression tendency were evidently different of this gene. The subcellular localization analysis showd that Ch WRKY28 protein was targeted to the nucleus. [Conclusion]This study indicated that Ch WRKY28 gene may be involved in response to abiotic stress signal transduction pathway.

关 键 词：平榛 转录因子 克隆 亚细胞定位 表达 

分 类 号：S718.46[农业科学—林学]