摘要
多药和有毒化合物外排转运蛋白MATE能转运金属、激素、次生代谢物等多种底物,因而在植物的生长发育中发挥重要作用。本研究基于紫花苜蓿(Medicago sativa L.)基因组数据和缺铁胁迫的转录组数据,克隆获得紫花苜蓿MsMATE1及pMsMATE1启动子(GenBank登录号分别为MN547958和MT505313)。MsMATE1基因长1470 bp,编码489个氨基酸。系统进化分析表明,紫花苜蓿MsMATE1蛋白属于mate家族,与蒺藜苜蓿MtMATE(XP013453190.1)亲缘关系最近。生物信息学分析表明,MsMATE1具有典型的MATE_like超家族结构域,属于H+势驱动的真核亚类;MsMATE1为跨膜蛋白,二级结构的主要构成元件是α螺旋。pMsMATE1启动子长1598 bp,序列分析显示内含多个植物激素和逆境胁迫响应元件。MsMATE1在紫花苜蓿幼苗的根茎叶中都有表达,茎中的表达量最高。高铁和缺铁胁迫下,MsMATE1基因在紫花苜蓿幼苗各部位的表达显著上调,茎中上调最明显。高铁和缺铁胁迫,转MsMATE1基因烟草的3种抗氧化酶活性、叶绿素和可溶性蛋白含量显著增加,丙二醛含量显著降低。以上结果表明,MsMATE1在烟草中的异源表达提高了植物抵御高铁和缺铁胁迫的能力。MsMATE1可以作为应用基因工程方法改良植物铁胁迫耐受的重要候选基因,本研究为深入了解MsMATE1蛋白在植物响应铁胁迫中的分子机制奠定基础,MsMATE1基因应对其他金属胁迫或环境胁迫的功能有待于进一步鉴定。
Multidrug and toxic compound extrusion transporters(MATE)are able to transport a wide variety of substrates such as metals,hormones and secondary metabolites,and they thus play an important role in growth and development in plant.In this study,the full-length coding sequence of MsMATE1(NCBI GeneBank accessions:MN547958)and its promoter pMsMATE1(MT505313)were isolated based on genome and transcriptome sequences of Medicago sativa under iron deficiency.The MsMATE1 is 1470 bp which encodes a putative protein of 489 amino acids.According to bioinformatics analysis,the MsMATE1 protein contains a typical MATE_like superfamily domain,which belongs to the eukaryotic subtype.MsMATE1 is a transmembrane protein,and the main component of its secondary structure isαhelix.Phylogenetic analysis showed that MsMATE1 protein is closely related to MtMATE1(XP013453190.1).The pMsMATE1 is 1598 bp which were detected with multiple elements responding to hormone and abiotic stress.MsMATE1 was expressed in roots,shoots and leaves of alfalfa seedlings with the highest abundance in shoots.Under iron-deficient or iron-excess stress,MsMATE1 was significantly up-regulated in different tissues of alfalfa seedlings,with the highest upregulation in shoots.In Ms MATE1 transgenic tobacco plants,the activities of SOD,POD,and CAT,chlorophyll content and soluble protein content were significantly increased,while the significant reduction on content of MDA was observed under iron-deficient or iron-excess conditions,compared with those of wild type plants.The results showed that heterologous expression of Ms MATE1 improved the tolerant against iron-deficient or iron-excess in plants.Ms MATE1 might be a candidate gene for improving tolerance to iron stress by genetic engineering.Collectively,this study laid a foundation for future deciphering the Ms MATE1-involved molecular mechanism upon iron stresses.
作者
高铭
周思莹
李钧儒
石卓
郭长虹
郭东林
GAO Ming;ZHOU Si-ying;LI Jun-ru;SHI Zhuo;GUO Chang-hong;GUO Dong-lin(College of Life Science and Technology,Harbin Normal University/Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province,Harbin 150025)
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2021年第2期512-520,共9页
Journal of Plant Genetic Resources
基金
国家转基因生物新品种培育重大专项(2016ZX08004-002)
林木遗传育种国家重点实验室(东北林业大学)开放基金
哈尔滨市科技局青年后备人才项目(2017RAQXJ048)。
