Cloning and Expression Analysis of Salt-tolerant Gene NHX1 in Vicia villosa Roth.

doi:10.11733/j.issn.1007-0435.2024.05.010

Acta Agrestia Sinica ›› 2024, Vol. 32 ›› Issue (5): 1401-1409.DOI: 10.11733/j.issn.1007-0435.2024.05.010

Cloning and Expression Analysis of Salt-tolerant Gene NHX1 in Vicia villosa Roth.

YAO Na¹, YUN Lan^1,2, AI Qian¹, REN Xiao-min¹, ZHOU Cheng-mei¹, LI Bo-hua¹

1. College of Grassland Resource and Environment, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia 010010, China；;
2. Key Laboratory of Grassland Resources of Education Ministry, Hohhot, Inner Mongolia 010010, China

Received:2024-03-04 Revised:2024-03-28 Published:2024-06-01

冬箭筈豌豆耐盐基因NHX1克隆及表达分析

姚娜¹, 云岚^1,2, 艾芊¹, 任晓敏¹, 周成美¹, 李博华¹

1. 内蒙古农业大学草原与资源环境学院, 内蒙古呼和浩特 010010;
2. 草地资源教育部重点实验室, 内蒙古呼和浩特 010010

通讯作者: 云岚，E-mail:yunlan@imau.edu.cn
作者简介:姚娜(1998-)，女，汉族，内蒙古乌海人，硕士研究生，主要从事草种质资源与育种研究，E-mail:m18847319047_1@163.com
基金资助:
内蒙古自治区自然科学基金重点项目(2023ZD07)；内蒙古自治区种业创新重大示范工程项目(2022 JBGS00400303)资助

Abstract

Abstract: In order to analyze and excavate more salt-tolerant genes of legume forage and breed salt-tolerant varieties,the NHX1 gene was cloned from the cDNA of Vicia villosa Roth. by homologous cloning PCR method,and its biological information was analyzed. The CDS sequence length of the gene NHX1 of Vicia villosa Roth. is 1 641 bp,encoding 546 amino acids,and it is an alkaline hydrophobic protein with the highest homology similarity to pea. The protein structure is dominated by alpha helix and contains 10 transmembrane domains. Relative quantitative analysis showed that NHX1 was expressed in root,stem and leaf tissues. The highest expression was found in leaves and the maximum expression was found at 48 hours after salt stress treatment. Subcellular localization of Agrobacterium 1300-cYFP-NHX1 transformed tobacco showed that it was localized to the cell membrane,and the protein expression was verified by western blot. The NHX1 gene is involved in the salt stress response and is relatively highly expressed in salt-tolerant materials. This study laid a foundation for the improvement of salt tolerance of legume forage.

Key words: Vicia villosa Roth., NHX1, Gene cloning, Expression analysis

摘要： 为了分析和挖掘更多豆科牧草耐盐基因，选育耐盐品种，本研究通过同源克隆PCR方法从冬箭筈豌豆(Vicia villosa Roth.)cDNA中克隆NHX1基因，并对其进行生物信息分析。结果表明：冬箭筈豌豆NHX1基因CDS序列长度为1 641 bp，编码546个氨基酸，为碱性疏水蛋白，与豌豆同源相似度最高；蛋白结构以α螺旋为主，包含10个跨膜结构域；相对定量分析表明NHX1在根、茎、叶组织中均有表达，盐胁迫下叶中表达量最高并在盐胁迫后48 h达到最大值；1300-cYFP-NHX1农杆菌转化烟草亚细胞定位显示其定位于细胞膜，并通过蛋白印迹法验证蛋白表达。冬箭筈豌豆NHX1基因参与盐胁迫响应，在耐盐性强的材料中相对高表达。本研究为豆科牧草耐盐性改良奠定基础。

关键词: 冬箭筈豌豆, NHX1, 基因克隆, 表达分析

CLC Number:

S542

YAO Na, YUN Lan, AI Qian, REN Xiao-min, ZHOU Cheng-mei, LI Bo-hua. Cloning and Expression Analysis of Salt-tolerant Gene NHX1 in Vicia villosa Roth.[J]. Acta Agrestia Sinica, 2024, 32(5): 1401-1409.

姚娜, 云岚, 艾芊, 任晓敏, 周成美, 李博华. 冬箭筈豌豆耐盐基因NHX1克隆及表达分析[J]. 草地学报, 2024, 32(5): 1401-1409.

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Cloning and Expression Analysis of Salt-tolerant Gene NHX1 in Vicia villosa Roth.

冬箭筈豌豆耐盐基因NHX1克隆及表达分析

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