Acta Agrestia Sinica

Acta Agrestia Sinica ›› 2025, Vol. 33 ›› Issue (8): 2456-2466.DOI: 10.11733/j.issn.1007-0435.2025.08.005

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Cloning and Salt Tolerance Analysis of MsNF-YB8 Gene in Alfalfa

AN Wen1,2,3, ZHANG Xin-yu1,2,3, MA Dong-mei1,2,3, MA Qiao-li4, LANG Si-rui5,6, LIU Xiao-xia1,2,3   

  1. 1. Cultivation Base of State Key Laboratory of Northwest Land Degradation and Ecosystem Restoration, Ningxia University, Yinchuan, Ningxia 750021, China;
    2. Key Laboratory of Restoration and Reconstruction of Degraded Ecosystems in Northwest China, Ministry of Education, Yinchuan, Ningxia 750021, China;
    3. College of Ecology and Environment, Ningxia University, Yinchuan, Ningxia 750021, China;
    4. College of Forestry and Grassland, Ningxia University, Yinchuan, Ningxia 750021, China;
    5. School of Life Sciences, Ningxia University, Yinchuan, Ningxia 750021, China;
    6. Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, Ningxia University, Yinchuan, Ningxia 750021, China
  • Received:2024-09-02 Revised:2024-12-01 Online:2025-08-15 Published:2025-08-27

紫花苜蓿MsNF-YB8基因的克隆及耐盐性分析

安雯1,2,3, 张新宇1,2,3, 麻冬梅1,2,3, 马巧利4, 郎思睿5,6, 刘晓霞1,2,3   

  1. 1. 宁夏大学西北土地退化与生态系统恢复国家重点实验室培养基地, 宁夏 银川 750021;
    2. 西北退化生态系统恢复与重建教育部重点实验室, 宁夏 银川 750021;
    3. 宁夏大学生态环境学院, 宁夏 银川 750021;
    4. 宁夏大学林业与草业学院, 宁夏 银川 750021;
    5. 宁夏大学生命科学学院, 宁夏 银川 750021;
    6. 宁夏大学西部特色生物资源保护与利用教育部重点实验室, 宁夏 银川 750021
  • 通讯作者: 刘晓霞, E-mail:liuxiaoxia0856@163.com
  • 作者简介:安雯(1997-),女,汉族,宁夏石嘴山人,硕士研究生,主要从事作物逆境生理与分子生态学研究,E-mail:xinan9080@163.com
  • 基金资助:
    国家自然科学基金青年基金项目（32401473）；宁夏回族自治区重点研发计划（引才专项）项目（2021BEB04075）资助

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Abstract: To explore the function of MsNF-YB8 gene in alfalfa in response to salt stress， the MsNF-YB8 gene of alfalfa ‘Zhongmu No. 1’ was cloned in this study， and its molecular properties structural characteristics and evolutionary relationship were analyzed by using bioinformatics tools. Real-time quantitative PCR （qRT-PCR） was used to analyze its expression pattern under different salt stress conditions. Finally， the MsNF-YB8 gene was transformed into Arabidopsis thaliana by using Agrobacterium-mediated genetic transformation method. The tolerance of transgenic Arabidopsis thaliana lines to salt stress were analyzed. The results showed that the full length of coding region of MsNF-YB8 gene was 411 bp （GenBank accession number： PQ448238.1）， encoding 136 amino acids， the protein had a molecular weight of 15.3 kDa and an isoelectric point of 5.31， which was high sequence homology with the MtNF-YB8 gene of Medicago truncatula. qRT-PCR showed that the expression of MsNF-YB8 gene was significantly induced by salt stress. Transgenic Arabidopsis plants showed enhanced tolerance to NaCl stress. Compared to wild type line， the transgenic plants overexpressing MsNF-YB8 showed enhanced growth of seedling roots. As the same time， physiological parameter measurements revealed that the transgenic Arabidopsis plants exhibited a lower degree of membrane lipid peroxidation， relatively higher reactive oxygen species （ROS） scavenging capacity， photosynthetic ability， and osmotic adjustment capability. Therefore， the MsNF-YB8 gene is a potential candidate gene for molecular breeding of alfalfa for salt tolerance.

Key words: Alfalfa, MsNF-YB8, Salt tolerance, Gene expression

摘要： 为探究紫花苜蓿（Medicago sativa L.）MsNF-YB8基因在盐胁迫响应时的功能，本研究克隆‘中苜1号’紫花苜蓿MsNF-YB8基因，并利用生物信息学工具分析其分子特性、结构特征和进化关系等信息；进一步采用实时荧光定量PCR技术（qRT-PCR）监测该基因在不同盐胁迫条件下的表达模式；最后利用农杆菌介导的转化方法，对拟南芥转化株系的耐盐性进行分析。结果表明，MsNF-YB8基因编码区全长411 bp（GenBank登录号： PQ448238.1），编码136个氨基酸，蛋白分子量为15.3 kDa，等电点为5.31，与蒺藜苜蓿MtNF-YB8基因具有较高的序列同源性。qRT-PCR检测发现，MsNF-YB8基因表达量受到盐胁迫的显著诱导。在拟南芥中过表达MsNF-YB8基因能够增强盐胁迫时幼根的生长，同时生理指标测定发现转基因拟南芥中的膜脂过氧化反应程度较低，活性氧清除能力、光合作用能力及渗透调节能力相对较高。因此，MsNF-YB8基因是一个有潜力的候选基因，可用于紫花苜蓿的耐盐性分子育种。

关键词: 紫花苜蓿, MsNF-YB8, 耐盐性, 基因表达

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