参薯金属硫蛋白基因DaMT2a的克隆与表达分析
Molecular Cloning and Expressional Analysis of Metallothionein Gene DaMT2a from Dioscorea alata
黄亚成 1任东立 1鲁明秋 1曾林 1黄东益 2刘林娅1
作者信息
- 1. 六盘水师范学院生物科学与技术学院,六盘水,553000
- 2. 海南大学热带作物学院,海口,570100
- 折叠
摘要
金属硫蛋白属于一类富含半胱氨酸,能够与重金属离子结合的低分子量蛋白质,在植物抗逆过程中起着重要作用.本研究从参薯中克隆了一个金属硫蛋白基因,命名为DaMT2a(与金属硫蛋白MT2a同源),序列分析显示克隆获得的金属硫蛋白基因的cDNA长度为540 bp,包含了 3个外显子和2个内含子,编码79个氨基酸,具有金属硫蛋白基因家族的保守结构域.进化树分析表明DaMT2a属于金属硫蛋白Type2类成员.利用实时荧光定量PCR技术研究了其在正常组织及胁迫条件下的表达特性.结果表明DaMT2a主要在参薯的茎和雄花中大量表达,机械伤害可显著上调DaMT2a基因表达,高温处理则明显下调DaMT2a基因表达,乙烯利和脱落酸处理上调DaWT2a基因的表达.本研究为参薯抗逆相关金属硫蛋白基因的筛选和功能鉴定提供了参考依据.
Abstract
Metallothioneins(MTs),a class of small,cysteine(Cys)-rich and metal-binding proteins,play important roles in stress response.In this study,the full-length cDNAs encoding a metallothionein gene was obtained by RT-PCR in Dioscorea alata,which is homologous to MT2a and named as DaMT2a.Sequence analysis revealed that DaMT2a was 540 bp long,contained 3 exons and 2 introns,predicted to encode proteins of 79 amino acids,and had the conserved domains of the metallothionein.Phylogenetic analysis showed that DaMT2a was clustered into Type2 of the metallothionein family.The DaMT2a gene was explored by RT-qPCR for its expressional patterns in different tissues and in tissue-cultured seedling responding to stresses.The results indicated that DaMT2a showed a predominance expression in the stems and male flowers.The treatments of mechanical wounding,ethylene and ab-scisic acid markedly up-regulated the expression of DaMT2a,whereas the high-temperature treatment down-regu-lated its expression.This study provides a reference for the screening and functional identification of stress-related MT genes in Dioscorea alata.
关键词
参薯/金属硫蛋白/基因克隆/表达分析Key words
Dioscorea alata L./Metallothioneins/Gene cloning/Expressional analysis引用本文复制引用
基金项目
贵州省科学技术基金项目(黔科合基础[2020]1Y115)
贵州省普通高等学校青年科技人才成长项目(黔教合KY字[2019]134)
六盘水师范学院高层次人才科研启动基金项目(LPSSYKYJJ201705)
六盘水师范学院高层次人才科研启动基金项目(LPSSYKYJJ201706)
出版年
2024
NETL
NSTL
万方数据