平榛ChWRKY28基因克隆及表达模式分析

目的]研究平榛ChWRKY28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ChWRKY28.构建的系统发育树表明:该序列与拟南芥AtWRKY28及杨树PtrWRKY93的关系最近,相似性分别为49%和60%.基因表达分析表明:ChWRKY28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ChWRKY28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:ChWRKY28蛋白分布在细胞核内,是一个核蛋白.结论]推测ChWRKY28基因可能参与植物响应非生物胁迫的信号转导过程.

Cloning and Expression Analysis of ChWRKY28 from Corylus heterophylla Fisch

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 cDNA 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 ChWRKY28. Phylogeny tree results showed ChWRKY28 was much closer to AtWRKY28 from Arabidopsis thaliana and PtrWRKY93 from Populus trichocarpa, generated 49% and 60% amino acids similarity. Spatial expression analyses demonstrated that the expression level of ChWRKY28 was higher in stem than that in male anthotaxy and floral buds which indicating tissue-specific expression. ChWRKY28 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 ChWRKY28 protein was targeted to the nucleus. Conclusion]This study indicated that ChWRKY28 gene may be involved in response to abiotic stress signal transduction pathway.