羊草锌转运蛋白LcZNE1功能的鉴定  

Functional Identification of Zinc Transporter LcZNE1 in Leymus chinensis

作　　者：亢燕 王耀辉 李俊 祁智[1] KANG Yan;WANG Yao-hui;LI Jun;QI Zhi(Key Laboratory of Herbage and Endemic Crop Biology,Ministry of Education,Inner Mongolia University,Hohhot 010070,China)

机构地区：[1]内蒙古大学牧草与特色作物生物学教育部重点实验室,内蒙古呼和浩特010070

出　　处：《中国草地学报》2021年第11期1-9,共9页Chinese Journal of Grassland

基　　金：内蒙古自治区科技创新引导项目(21400-5173208);国家自然科学基金(32060063);内蒙古自然科学基金(2018MS03075)共同资助。

摘　　要：从羊草转录组数据库中筛选出基因Lc240210,发现其编码蛋白与拟南芥Zn2+转运蛋白AtZNE1同源,因此将其命名为LcZNE1。生物信息学分析表明,LcZNE1是膜蛋白,有15个跨膜域,与二穗短柄草亲缘关系最近,具有Zn2+配基。通过瞬时转染烟草叶表皮细胞和羊草叶原生质体发现,LcZNE1定位于内质网。实时荧光定量PCR分析发现,在高锌和低铁条件下LcZNE1基因表达上调,LcZNE1参与Zn和Fe营养调控。酵母功能互补试验证明,LcZNE1能够增强高锌敏感酵母突变体(△zrc1/△cot1)对高锌条件的抗性,具有Zn2+转运功能。等温滴定量热法(ITC)发现,LcZNE1有8条非跨膜域短肽与Zn2+结合,包括胞质面和非胞质面各4条短肽,其中Non-CytoⅡ与Zn2+结合能力强。In this study,via screening the gene Lc240210 was screened from the Leymus chinensis tran-scriptome database and found its coding protein wan homologous with zinc transporter ZNE1(Zinc Nutrition Essential 1)of Arabidopsis thaliana,so it was renamed Lc240210 as LcZNE1.Bioinformatics analysis indicated that LcZNE1 encoded a membrane protein with 15 transmembrane domains,and had the closest genetic relationship with Brachypodium distachyon and Zn^(2+) ligands.Transient expression of the LcZNE1 protein in tobacco leaf epidermal cells and Leymus chinensis leaf protoplast indicated that LcZNE1 was located in the endoplasmic reticulum.Real-time RT-PCR analysis showed that the expression of gene LcZNE1 was up-regulated under high zinc and low iron concentrations,and LcZNE1 was involved in the regulation of Zn and Fe nutrition.Yeast functional complementarity test proved that LcZNE1 could enhance the resistance of hyper-zinc-sensitive yeast mutant strain(△zrc1/△cot1)to excessive Zn conditions,and had the function of Zn^(2+) transport.Isothermal titration calorimetry(ITC)analysis showed that LcZNE1 had 8 non-transmembrane domains short peptides that bind to Zn^(2+),including 4 short peptides on the non-cytoplasmic surface and the cytoplasmic surface,respectively.Among them,Non-cytoⅡhad the most strongest binding ability to Zn^(2+).

关 键 词：锌 LcZNE1 亚细胞定位 基因表达 

分 类 号：S543.9[农业科学—作物学]