油用向日葵铵转运基因HaAMT1.1的克隆及功能分析  

Cloning and Functional Analysis of an Ammonium Transporter Gene HaAMT1.1 in Helianthus annuus L.

作　　者：张爽 李一 谭定权 王淑冉 张胜 Zhang Shuang;Li Yi;Tan Dingquan;Wang Shuran;Zhang Sheng(College of Life Science and Technology,Xinjiang University,Urumqi,830046;College of Forestry,Northwest A&F University,Yangling,712100)

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]西北农林科技大学林学院,杨陵712100

出　　处：《分子植物育种》2022年第13期4305-4313,共9页Molecular Plant Breeding

基　　金：新疆维吾尔自治区自然科学基金项目(2019D01C017);自治区高层次天池百人计划项目(042419003);新疆大学自然科学基金项目(62031224612)共同资助。

摘　　要：AMT1.1是铵转运家族的基因之一,在植物吸收转运NH_(4)^(+)中起到重要作用。为了明确氮素在植物体的转运吸收机制,本研究以油用向日葵‘早矮大头’为研究材料,成功从中克隆向日葵AMT1.1的同源基因HaAMT1.1。生物信息学分析表明该基因CDS区长1497 bp,编码491个氨基酸;正电荷残基28个,占氨基酸总数的5.7%,负电荷残基30个,占氨基酸总数的6.1%;其分子式为C_(2417)H_(3625)N_(597)O_(668)S_(22),预测分子量为52439.29 Da,理论等电点为6.58;HaAMT1.1为疏水性蛋白,无跨膜结构域,具有Ammonium Transp超家族保守结构域,与黄花蒿AMT1.1亲缘关系最近。亚细胞定位结果表明HaAMT1.1位于细胞质膜和细胞核上。实时荧光定量PCR分析表明,不同组织中Ha AMT1.1基因在根系中表达量最高;不同NH_(4)^(+)浓度处理下,该基因在经低浓度NH_(4)^(+)处理的表达量显著高于经高浓度NH_(4)^(+)处理的组织。本研究结果为进一步分析HaAMT1.1基因的生物学功能提供参考。AMT1.1 is one of the genes of Ammonium transports family,which plays an significant role in absorbing and transporting NH_(4)^(+)in plants.In this study,in order to clarify the transport and absorption mechanism of nitrogen in plants,we cloned the homologous gene HaAMT1.1 of sunflower AMT1.1 from Helianthus annuus L’Zaoaidatou’.Bioinformatics analysis showed that the CDS region of the gene was 1497 bp encoding 491 amino acids.Positively charged residues was 28,accounting for 5.7%of the total amino acids;negatively charged residues was 30,accounting for 6.1%of the total amino acids.Its molecular formula was C_(2417)H_(3625)N_(597)O_(668)S_(22),and its molecular weight was 52439.29 Da,theoretical isoelectric point was 6.58.HaAMT1.1 was a hydrophobic protein with no trans-membrane domain and has the conserved domain of ammonium transp superfamily.HaAMT1.1was highly conserved with Artemisia annua AMT1.1.The result of subcellular localization showed that HaAMT1.1was located in the plasma membrane and nucleus.Quantitative Real-time PCR analysis showed that the expression level of HaAMT1.1 gene in roots was the highest in different tissues;under different NH_(4)^(+)concentrations,the expression level of HaAMT1.1 gene in low concentration NH_(4)^(+)treatment was significantly higher than that in high concentration NH_(4)^(+)treatment.This study provides a reference for further analysis of the biological function of HaAMT1.1 gene.

关 键 词：油用向日葵 铵转运家族 HaAMT1.1 亚细胞定位 表达分析 

分 类 号：S565.5[农业科学—作物学]