Cloning and Functional Analysis of MsHSP18.2 Gene in Medicago sativa

doi:10.11733/j.issn.1007-0435.2026.04.003

Acta Agrestia Sinica ›› 2026, Vol. 34 ›› Issue (4): 1158-1167.DOI: 10.11733/j.issn.1007-0435.2026.04.003

Cloning and Functional Analysis of MsHSP18.2 Gene in Medicago sativa

LIU Jin-nan¹, ZHANG Chi-hao¹, DONG Jia-le², AI Ye¹, CHAO Yue-hui¹

1. Beijing Forestry University School of Grassland Science, Beijing 100083, China;
2. Sichuan Jinjiang Electronic Medical Device Technology Co, Ltd, Chengdu, Sichuan Province 610045, China

Received:2025-06-16 Revised:2025-08-10 Published:2026-04-15

紫花苜蓿MsHSP18.2基因的克隆与功能分析

刘晋囡¹, 张驰昊¹, 董家乐², 艾晔¹, 晁跃辉¹

1. 北京林业大学草业与草原学院, 北京 100083;
2. 四川锦江电子医疗器械科技股份有限公司, 四川成都 610045

通讯作者: 晁跃辉，E-mail：chaoyuehui@bjfu.edu.cn
作者简介:刘晋囡（1998-），女，汉族，山西朔州人，硕士研究生，主要从事草地植物育种研究，E-mail：ljn19981002@163.com；
基金资助:
内蒙古自治区科技重大专项项目专题“抗寒高产苜蓿新品种培育”（2022JBGS00160302）资助

Abstract

Abstract: Small heat shock proteins （Small heat shock proteins， sHSPs） are a class of stress-induced proteins widely present in plants， play crucial roles in plant growth， development， and response to drought and salt stresses. To investigate the function of the MsHSP18.2 gene in Medicago sativa， bioinformatics tools were employed to predict and analyze its theoretical properties， structure， and function. Plants were treated with methyl jasmonate （MeJA）， salicylic acid （SA）， abscisic acid （ABA）， indole-3-acetic acid （IAA）， gibberellin （GA）， and cytokinin （6-BA）， as well as subjected to drought and salt stress. Gene expression levels were analyzed by qRT-PCR at 0， 0.5， 1， 3， 6， 9， and 12 hours post-treatment. Transcriptional autoactivation was examined by using a yeast expression system. The stress resistance function was verified using a heterologous yeast system. The results showed that this protein has a molecular weight of 18.18 KDa and a theoretical isoelectric point of 5.55， exhibiting relatively high instability （instability index： 54.67）. Its secondary structure was predominantly composed of random coils. Expression analysis demonstrated that MsHSP18.2 was upregulated under drought and salt stress treatments and displayed differential responses to various plant hormones. Yeast autoactivation assays confirmed its autoactivation activity. Heterologous overexpression of MsHSP18.2 gene significantly enhanced the salt tolerance （NaCl） and osmotic stress （PEG） tolerance of yeast cells.

Key words: MsHSP18.2, Medicago sativa, Bioinformatics analysis, Expression analysis, Self-activation detection

摘要： 小热休克蛋白（Small heat shock proteins，sHSPs），作为植物体内重要的应激诱导因子，在植物生长发育及响应各类胁迫中起重要调控作用。为探究紫花苜蓿MsHSP18.2基因功能，利用生物信息学工具进行蛋白理论性质、结构和功能预测分析；对植物进行茉莉酸甲酯（MeJA）、水杨酸（SA）、脱落酸（ABA）、吲哚乙酸（IAA）、赤霉素（GA）、细胞分裂素（6-BA）等激素，以及干旱胁迫和盐胁迫的处理，利用RT-qPCR方法分别对处理0，0.5，1，3，6，9，12 h时进行基因表达水平分析；利用酵母表达方法检测其转录自激活特性；利用异源酵母系统初步验证其耐逆功能。生物信息学分析结果表明，该蛋白分子量为18.18 kDa，理论等电点为5.55，具有较高的不稳定性（不稳定指数54.67），其二级结构以无规卷曲为主。表达分析结果显示，MsHSP18.2在干旱和盐胁迫处理下表达增强，并对不同植物激素呈现差异响应。酵母自激活活性分析检测显示该蛋白具有自激活特性。异源过量表达MsHSP18.2基因显著增强了酵母细胞的耐盐（NaCl）和耐渗透胁迫（PEG）能力。

关键词: MsHSP18.2, 紫花苜蓿, 生物学信息分析, 表达分析, 自激活检测

CLC Number:

S541.9

LIU Jin-nan, ZHANG Chi-hao, DONG Jia-le, AI Ye, CHAO Yue-hui. Cloning and Functional Analysis of MsHSP18.2 Gene in Medicago sativa[J]. Acta Agrestia Sinica, 2026, 34(4): 1158-1167.

刘晋囡, 张驰昊, 董家乐, 艾晔, 晁跃辉. 紫花苜蓿MsHSP18.2基因的克隆与功能分析[J]. 草地学报, 2026, 34(4): 1158-1167.

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Cloning and Functional Analysis of MsHSP18.2 Gene in Medicago sativa

紫花苜蓿MsHSP18.2基因的克隆与功能分析

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