摘要： 为了揭示SlWRKY6基因与植物重金属胁迫的相关性，通过RT-PCR的方法从番茄中克隆得到SlWRKY6基因，该基因序列号为：NM_001365762.1。生物信息学分析结果表明，该基因含有一个长度为1 342 bp的完整开放阅读框，编码447个氨基酸残基。该基因编码的蛋白含有一个WRKY结构域，属于典型的Group Ⅱ亚家族成员。系统进化分析表明，与番茄SpWRKY31氨基酸序列之间的亲缘关系最近。组织特异性分析表明，SlWRKY6基因在番茄的不同组织中均有表达，在老叶中表达量最高。实时荧光定量PCR分析结果表明，SlWRKY6基因在3种重金属（CdCl₂、CuCl₂、HgSO₄）胁迫下均上调表达，且在Cd和Cu胁迫下番茄根部的表达量较高。CdCl₂胁迫下，SlWRKY6基因在番茄根中受胁迫诱导显著高于叶中；CuCl₂胁迫下，番茄叶中SlWRKY6基因表达量在3 h时达到最大值后降低，而根中的SlWRKY6基因水平随着浓度增加而下降；HgSO₄胁迫下，根部和叶片该基因的表达量显著高于对照（0 h），并随着HgSO₄胁迫时间延长，SlWRKY6基因相对表达量在番茄根和叶中呈现相反的趋势。综上，本研究获得了SlWRKY6基因的编码区序列，并初步阐述了番茄SlWRKY6基因在3种重金属胁迫下的表达情况，为筛选番茄中响应重金属胁迫功能基因的研究提供了研究基础。

关键词: 番茄, SlWRKY6 基因克隆, 生物信息学, 重金属胁迫, 表达分析

Abstract: In order to reveal the correlation between the SlWRKY6 gene and heavy metal stress in plants, the SlWRKY6 gene was cloned from tomato by RT-PCR, the gene sequence number was NM_001365762.1. Bioinformatics analysis showed that the gene contained a complete open reading frame of 1 342 bp, encoding 447 amino acid residues.The protein encoded by this gene contained a WRKY domain structure and belonged to a typical Group Ⅱ subfamily member. Phylogenetic analysis showed that the amino acid sequence of tomato SlWRKY6 and tomato SpWRKY31 was closely related. Tissue-specific analysis showed that the SlWRKY6 gene was expressed in different tissues of tomato, with the highest expression in old leaves. The results of Real-time fluorescence quantitative PCR analysis showed that the SlWRKY6 gene was up-regulated under the stress of three heavy metals(CdCl₂, CuCl₂, HgSO₄), and the expression level of tomato roots was higher under Cd and Cu stress. Under CdCl₂ stress, SlWRKY6 gene was induced significantly higher in tomato roots than in leaves;the expression level of SlWRKY6 gene decreased with the increase of CuCl₂ stress concentration;Under HgSO₄ stress, the expression of this gene in roots and leaves was significantly higher than that in the control(0 h), with the prolongation of HgSO₄ stress, the relative expression of SlWRKY6 gene showed a reverse trend in tomato roots and leaves. In summary, this study obtained the coding region sequence of the SlWRKY6 gene, and preliminarily described the expression of the tomato SlWRKY6 gene under three heavy metal stresses, which provided a research basis for the screening of functional genes in response to heavy metal stress in tomatoes.

Key words: Tomato, SlWRKY6 gene cloning, Bioinformatics, Heavy metals stress, Expression analysis

中图分类号: 

• Q78

• S641.03