甜菜铵转运蛋白BvAMT1-3基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Beta vulgaris Ammonium Transporter 1-3(BvAMT1-3)Gene

作　　者：魏多 刘铭曦 高卓[1,4] 李佳佳 刘大丽 马龙彪[1,2] WEI Duo;LIU Mingxi;GAO Zhuo;LI Jiajia;LIU Dali;MA Longbiao(Key Laboratory of Sugar Beet Genetics and Breeding,Heilongjiang Province Common College/College of Advanced Agriculture and Ecological Environment,Heilongjiang University,Harbin 150080;Key Laboratory of Northern Sugar Crop Resources and Utilization of Chinese Academy of Agricultural Sciences/Sugar Beet Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150080;College of Life Science and Technology,Harbin Normal University,Harbin 150025;College of Life Sciences,Heilongjiang University,Harbin 150080)

机构地区：[1]黑龙江省普通高等学校甜菜遗传育种重点实验室/黑龙江大学现代农业与生态环境学院,哈尔滨150080 [2]中国农业科学院北方糖料作物资源与利用重点实验室/中国农业科学院甜菜研究所,哈尔滨150080 [3]哈尔滨师范大学生命科学与技术学院,哈尔滨150025 [4]黑龙江大学生命科学学院,哈尔滨150080

出　　处：《中国糖料》2021年第4期1-8,共8页Sugar Crops of China

基　　金：财政部和农业农村部国家现代农业产业技术体系(糖料)项目“甜菜养分管理与土壤肥料”(CARS-170204);黑龙江省高校基本科研业务费黑龙江大学专项资金项目“能源甜菜BvMPT11基因的Cd污染生物修复应答机制研究”(KJCX201920);黑龙江省自然基金“BvHIPP24基因在能源甜菜重金属镉污染生物修复中的分子机制研究”(LH2019C057);国家作物种质资源库“甜菜分库运行服务”(NCGRC-2021-017);农业农村部“甜菜种质资源的收集、鉴定、编目、繁种与入库(圃)保存”(2130135);黑龙江省普通本科高等学校青年创新人才培养计划“甜菜抗旱遗传资源评价及优异基因挖掘”(UNPYSCT-2020014)资助。

摘　　要：在植物体内,由铵转运蛋白(Ammonium transporters,AMTs)介导植株对铵态氮的吸收和转运。氮素在甜菜糖代谢的过程中起着重要作用,对甜菜AMTs蛋白的功能特性及调控机制进行生物信息学分析对于解析甜菜AMT基因家族的分子功能具有重要意义。本研究通过RT-PCR的方法,在甜菜体内克隆获得了Beta vulgaris ammonium transporter 1-3(BvAMT1-3)基因的全长序列。生物信息学分析结果显示该基因大小为1388 bp,编码462个氨基酸,分子量约为49.729 kDa,亚细胞定位分析其位于内质网膜结构上的可能性较大。系统进化分析显示5个甜菜AMT蛋白序列可分成2个亚族,AMT3亚族仅有1个成员,其余为AMT1亚族成员,同时甜菜BvAMT1-3蛋白与菠菜SoAMT1-3蛋白的亲缘关系最近。5个AMT基因家族成员均由α-螺旋、β-折叠、扩展链以及无规则卷曲4种形式组成。BvAMTs基因家族间的等电点、亲水性、相对分子质量等理化性质存在差异。此外,BvAMT1-3基因的启动子区检测到与植物逆境胁迫、光反应和生长发育相关的顺式作用元件。研究结果可为后续甜菜BvAMT1-3基因在氮胁迫逆境下分子机制的研究提供一定的科学依据。In plants,ammonium transporter(AMTs)mediates ammonium uptake and transport.Nitrogen plays an important role in sugar metabolism of sugar beet.Bioinformatics analysis of the functional characteristics and regulatory mechanism of Beta vulgaris AMT is of great significance to analyze the molecular function of Beta vulgarisAMT gene family.In this study,the full-length sequence of Beta vulgaris ammonium transporter 1-3(BvAMT1-3)gene was cloned in sugar beet by RT-PCR.The results of bioinformatics analysis showed that the gene was 1388 bp,which encoded 462 amino acids with molecular weight of 49.729 kDa.Subcellular localization showed that it was more likely to be located on the endoplasmic reticulum.Phylogenetic analysis showed that the five Beta vulgaris AMT protein sequences could be divided into two subfamilies.There was only one member of AMT3 subfamily,and the rest were members of AMT1 subfamily.At the same time,the genetic relationship between Beta vulgaris BvAMT1-3 protein and SoAMT1-3 protein in spinach was the closest.The five members of AMT gene family were composed ofα-helix,β-fold,extended chain and random coil.There were differences in physical and chemical properties such as isoelectric point,hydrophilicity and relative molecular weight among BvAMTs gene families.In addition,cis-acting elements related to stress,light response,and growth were detected in the promoter region of BvAMT1-3 gene.The results provide a scientific basis for the further study on molecular mechanism of BvAMT1-3 gene under nitrogen stress.

关 键 词：甜菜 铵转运蛋白 基因 克隆 生物信息学 

分 类 号：S566.3[农业科学—作物学]