【目的】以杉木幼叶为试验材料,通过克隆杉木ClWRKY44基因,分析其表达特性,为揭示杉木抗逆生理的分子机制提供理论依据。【方法】采用RT-PCR技术克隆杉木ClWRKY44 基因,对其进行生物信息学分析和亚细胞定位研究,并利用实时荧光定量PCR技术分析其表达情况。【结果】杉木ClWRKY44 基因的开放阅读框(ORF)为1 776 bp,编码591个氨基酸,具有WRKY结构域。系统进化树分析结果显示ClWRKY44 基因与拟南芥(Arabidopsis thaliana)ATWRKY44 的亲缘关系较密切,其在杉木的不同器官(叶、根、茎) 中均有表达,嫩叶中的表达量最高,茎中次之,根中最低。低温胁迫处理6 h,ClWRKY44 基因相对表达量最高,约是对照组的79.8倍。干旱胁迫处理24 h,ClWRKY44 基因在高浓度干旱胁迫处理下的相对表达量最高,约是对照组的46.6倍。【结论】杉木ClWRKY44基因编码的蛋白质序列与拟南芥ATWRKY44的同源性很高,在调控植物逆境胁迫应答过程中可以发挥功能,为杉木幼苗生长发育的适应性提供参考。

【Objective】The aim of this study is to clone the Cunninghamia lanceolata ClWRKY44 gene and analyze its expression characteristics, so as to provide a theoretical basis for revealing the molecular mechanism of stress resistance physiology of Cunninghamia lanceolata. 【Method】 We cloned the Chinese fir ClWRKY44 gene by using RT-PCR technology, conducted bioinformatics analysis and subcellular localization study, and used real-time fluorescent quantitative PCR technology to analyze its expression. 【Result】The open reading frame (ORF)of ClWRKY44 gene is 1 776 bp, which encodes 591 amino acids and has a WRKY domain. Phylogenetic tree analysis showed that ClWRKY44 gene was closely related to Arabidopsis thaliana ATWRKY44. It was expressed in different organs (leaves, roots, stems) of Cunninghamia lanceolata. After 6 h of low temperature stress treatment, the relative expression of ClWRKY44 gene was the highest, about 79.8 times that of the control group. After 24 h of drought stress treatment, the relative expression of ClWRKY44 gene was the highest under high concentration drought stress treatment, about 46.6 times that of the control group. 【Conclusion】The protein sequence encoded by Chinese fir ClWRKY44 gene has high homology with Arabidopsis ATWRKY44. It can play a role in regulating plant stress response, and provide a reference for the adaptability of Chinese fir seedling growth and development.