番茄金属硫蛋白基因家族鉴定、SlMT3基因表达及启动子活性分析  

Identifcation of the metallothionein gene family and expression and promoter activity analysis of SlMT3 from Solanum lycopersicum

作　　者：李文静 王秋平 黄丽婕 刘文杰 冯珂 刘亚雯 朱姝 王聪艳[1,2] 王晓铮 蒋秀虹 张新业 LI Wenjing;WANG Qiuping;HUANG Lijie;LIU Wenjie;FENG Ke;LIU Yawen;ZHU Shu;WANG Congyan;WANG Xiaozheng;JIANG Xiuhong;ZHANG Xinye(College of Life Science,Langfang Normal University,Langfang,Hebei 065000,China;Hebei Key Laboratory of Animal Diversity,Langfang,Hebei 065000,China;Langfang Key Laboratory of Cell Engineering and Application,Langfang,Hebei 065000,China)

机构地区：[1]廊坊师范学院生命科学学院,河北廊坊065000 [2]河北省动物多样性重点实验室,河北廊坊065000 [3]廊坊市细胞工程与应用研究重点实验室,河北廊坊065000

出　　处：《植物生理学报》2021年第9期1755-1766,共12页Plant Physiology Journal

基　　金：河北省高等学校科学技术研究项目(ZD2020122);河北省省属高等学校基本科研业务费研究项目(JYQ202101);廊坊师范学院大学生创新创业训练计划项目资金(S202010100016)。

摘　　要：金属硫蛋白(metallothioneins,MTs)是一类低分子量、富含半胱氨酸的金属结合蛋白,在维持金属稳态和去除金属毒性方面具有重要作用。本研究通过生物信息学方法对番茄(Solanum lycopersicum)MT家族基因(SlMT)进行鉴定,利用荧光定量PCR技术分析家族成员SlMT3基因表达模式,并利用GUS报告基因对SlMT3启动子活性进行分析。结果表明番茄基因组中共鉴定到5个MT基因,主要定位于细胞质,蛋白序列长度在72~112 aa,等电点介于4.49~5.71。基因结构分析表明,该基因家族由2~3个外显子构成。染色体分布结果显示,SlMT基因分布于3条染色体上。系统进化和保守结构域分析显示,SlMT分别归属于Ⅰ、Ⅱ和Ⅲ三个亚族,具有3种保守基序,蛋白保守性很高。荧光定量PCR结果表明,SlMT3在番茄根中特异性表达。启动子顺式作用元件及GUS组织化学染色分析结果表明,SlMT3基因启动子驱动GUS基因在拟南芥根部特异性表达,与荧光定量PCR结果一致。本研究为后续开展SlMT基因的功能研究和番茄遗传改良提供理论依据。Metallothionein(MT) proteins are low-molecular-weight, cysteine-rich and metal-binding proteins, which play important roles in the maintenance of metal homeostasis and detoxifcation. In this study, we performed a comprehensive bioinformatic analysis of MT gene family of tomato(Solanum lycopersicum), and detected the tissue expression pattern of Sl MT3 by quantitative real-time PCR(qRT-PCR). Furthernmore, we conducted promoter activity analysis using GUS reporter gene. The bioinformatic analysis showed that 5 Sl MTs were detected in the tomato genome, which mainly located in cytoplasm with protein length ranged from 72 to 112 aa and pI value distributed from 4.49 to 5.71. Gene structure analysis showed that the number of exons within SlMT genes was 2-3. Chromosome mapping analysis revealed that the 5 SlMTs were distributed on 3 chromosomes. The results of phylogenetic and conserved motif analysis showed that SlMT gene family was divided into Ⅰ, Ⅱ and Ⅲ subfamilies, and the protein domain of SlMT family is conserved. Quantitative RT-PCR results demonstrated that SlMT3 was specifically expressed in tomato roots. Analysis of cis-acting elements in promoter and histochemical staining revealed that SlMT3 promoter drove GUS gene to accumulate specifically in Arabidopsis roots, which was accordance with the results of quantitative RT-PCR. These results will provide a foundation for further functional research of SlMT protein.

关 键 词：番茄 金属硫蛋白 基因家族 根特异性启动子 GUS组织化学染色 

分 类 号：S641.2[农业科学—蔬菜学]