紫花苜蓿MsNF-YB5基因的克隆及表达特性分析

doi:10.11733/j.issn.1007-0435.2025.03.003

草地学报 ›› 2025, Vol. 33 ›› Issue (3): 686-695.DOI: 10.11733/j.issn.1007-0435.2025.03.003

• 研究论文 • 上一篇

紫花苜蓿MsNF-YB5基因的克隆及表达特性分析

王之杰, 张万军

中国农业大学草业科学与技术学院, 北京 100193

收稿日期:2024-06-06 修回日期:2024-08-22 发布日期:2025-04-07
通讯作者: 张万军,E-mail:wjzhang@cau.edu.cn
作者简介:王之杰（2002-）,男,汉族,山东潍坊人,本科生,主要从事苜蓿转基因育种研究,E-mail：wangzhijie@cau.edu.cn
基金资助:
国家自然科学青年基金项目（32201454）;国家草业技术创新中心（筹）项目（CCPTZX2023B01）资助

Cloning and Expression Analysis of MsNF-YB5 in Alfalfa

WANG Zhi-jie, ZHANG Wan-jun

College of Grassland Science and Technology, China Agricultural University, Beijing 100193, China

Received:2024-06-06 Revised:2024-08-22 Published:2025-04-07

摘要/Abstract

摘要： 研究发现异源表达拟南芥（Arabidopsis thaliana）QQS基因可显著提高紫花苜蓿的抗逆性,而MsNF-YB5是其发挥功能的关键内源基因。为探究MsNF-YB5在紫花苜蓿（Medicago sativa L.）胁迫响应过程中的功能,本研究克隆并完成了MsNF-YB5的生物信息学分析、亚细胞定位、转录自激活检测及胁迫下的表达模式分析。结果显示,克隆获得的MsNF-YB5 cDNA编码156个氨基酸,与蒺藜苜蓿（Medicago truncatula）MtNF-YB5相似度最高,该基因启动子区域含有参与茉莉酸甲酯、脱落酸、防御与胁迫反应的顺式作用元件；亚细胞定位结果显示其具有细胞核定位,截短N端后具有酵母转录自激活活性。MsNF-YB5受盐胁迫、渗透胁迫、低氮胁迫、冷胁迫及脱落酸处理诱导表达。本研究为进一步研究MsNF-YB5功能以提高紫花苜蓿抗逆性提供了基础。

关键词: 紫花苜蓿, MsNF-YB5, 亚细胞定位, 自激活活性, 胁迫表达模式

Abstract: Our previous research has demonstrated that the heterologous expression of the QQS gene from Arabidopsis thaliana significantly enhances the stress resistance of alfalfa （Medicago sativa L.）, and MsNF-YB5 is identified as a critical endogenous gene contributing to this effect. To elucidate the role of MsNF-YB5 in the stress response of alfalfa, we cloned the MsNF-YB5 gene and conducted a comprehensive bioinformatics analysis, subcellular localization studies, transcriptional self-activation assessments, and expression pattern analyses under various stress conditions. The findings revealed that the cloned MsNF-YB5 cDNA encoded a protein consisting of 156 amino acids, exhibiting the highest similarity to MtNF-YB5 from Medicago truncatula. The promoter region of this gene contained cis-acting elements associated with methyl jasmonate, abscisic acid, and the responses to defense and stress. Subcellular localization studies indicated that the protein was localized in the nucleus, and exhibited yeast transcriptional self-activation activity, which was only observed following truncation of the N-terminus. Furthermore, MsNF-YB5 expression was found to be upregulated under salt stress, PEG treatment, abscisic acid （ABA） treatment, low nitrogen availability and cold stress. This study established a foundation for further investigation into the role of MsNF-YB5 in enhancing the stress resistance of alfalfa.

Key words: Alfalfa, MsNF-YB5, Subcellular localization, Self-activation activity, Stress expression

中图分类号:

S541.9

王之杰, 张万军. 紫花苜蓿MsNF-YB5基因的克隆及表达特性分析[J]. 草地学报, 2025, 33(3): 686-695.

WANG Zhi-jie, ZHANG Wan-jun. Cloning and Expression Analysis of MsNF-YB5 in Alfalfa[J]. Acta Agrestia Sinica, 2025, 33(3): 686-695.

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紫花苜蓿MsNF-YB5基因的克隆及表达特性分析

Cloning and Expression Analysis of MsNF-YB5 in Alfalfa

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