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大豆GmARA6a的克隆及响应盐胁迫的功能分析

张晴1，杨昱2，郭茜2，岳霈尧2，殷丛丛1，牛景萍3，赵晋忠1，杜维俊2，岳爱琴2,*   

  1. 1 山西农业大学基础部, 山西太谷030801; 2 山西农业大学农学院, 山西太谷030801; 3 山西农业大学生命科学学院, 山西太谷030801
  • 收稿日期:2025-06-24 修回日期:2025-10-30 接受日期:2025-10-30 网络出版日期:2025-11-10
  • 通讯作者: 岳爱琴, E-mail: yueaiqinnd@126.com
  • 基金资助:
    本研究由山西省科技重大专项计划揭榜挂帅项目子课题(202201140601025-3-06), 山西省农业关键核心技术攻关子课题(NYGG27-04-01)、山西农业大学科技创新提升工程项目(CXGC2023004), 山西省现代农业产业技术体系建设项目(2025CYJSTX05-07), 山西农业大学育种工程项目(YZGC096)和山西“1331工程”作物科学一级学科建设项目资助。

Cloning and functional analysis of the soybean GmARA6a gene in response to salt stress

Zhang Qing1,Yang Yu2,Guo Qian2,Yue Pei-Yao2,Yin Cong-Cong1,Niu Jing-Ping3,Zhao Jin-Zhong1,Du Wei-Jun2,Yue Ai-Qin2,*   

  1. 1 College of Basic Sciences, Shanxi Agricultural University, Taigu 030801, Shanxi, China; 2 College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi, China; 3 College of Life Sciences, Shanxi Agricultural University, Taigu 030801, Shanxi, China
  • Received:2025-06-24 Revised:2025-10-30 Accepted:2025-10-30 Published online:2025-11-10
  • Supported by:
    This study was supported by the Sub-project of the Shanxi Province Science and Technology Major Special Plan Unveiling Project (202201140601025-3-06), Sub-project on Key Core Technology Breakthroughs in Agriculture of Shanxi Province (NYGG27-04-01), Shanxi Agricultural University Scientific and Technological Innovation Enhancement Program (CXGC2023004), the Construction Project of Modern Agricultural Industrial Technology System of Shanxi Province (2025CYJSTX05-07), Breeding Project of Shanxi Agricultural University (YZGC096), and the Shanxi “1331 Project” Crop Science First-Class Discipline Construction Project.

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摘要： 囊泡运输在植物响应盐胁迫过程中起关键作用，ARA6 (RabF1)通过促进内体与质膜间囊泡介导的物质转运，在调节植物的耐盐能力上发挥积极影响。本研究对大豆GmARA6a基因进行克隆及生物信息学分析；通过BFA、WM处理以及与AtARA6共定位探究GmARA6a的亚细胞定位；探究该基因在不同组织及盐胁迫下的表达模式；利用拟南芥ara6突变株、GmARA6a回补株系和过表达株系研究GmARA6a基因在调控植株耐盐性方面的作用。结果表明，大豆中克隆的GmARA6a基因长603 bp，编码200个氨基酸，具有ARA6家族的4段共有序列、1段效应结合区以及N端独特的“MGCXSS”结构域。GmARA6a定位于细胞膜以及多囊泡体(MVB)上。在大豆根中GmARA6a被盐胁迫诱导上调表达。盐胁迫下，拟南芥ara6突变株出现盐敏表型；而GmARA6a过表达株系生长良好，抗氧化能力增强，膜损伤、脂质过氧化程度降低，H2O2、O2?积累量下降；通过分析相关基因表达水平，表明GmARA6a可能通过SOS信号通路、囊泡运输途径调控植株耐盐性。本研究结果为进一步探究GmARA6a的功能提供参考。

关键词: 大豆, GmARA6a, 亚细胞定位, 盐胁迫, 功能分析

Abstract:

Vesicle trafficking plays a crucial role in plant responses to salt stress. ARA6 (RabF1) positively regulates salt tolerance by mediating vesicle transport between endosomes and the plasma membrane. In this study, GmARA6a was cloned from soybean and analyzed using bioinformatics approaches. Subcellular localization was examined through BFA and WM treatments, as well as colocalization with AtARA6. The expression pattern of GmARA6a was assessed across various tissues and under salt stress conditions. To evaluate its functional role in salt tolerance, we used Arabidopsis ara6 mutants, complemented lines (Com-1, Com-2), and overexpression lines (OE-1, OE-2). The GmARA6a gene, with a full-length coding sequence of 603 bp encoding a 200-amino-acid protein, was successfully cloned. The protein contains four conserved ARA6-family domains, an effector-binding region, and a unique N-terminal “MGCXSS” motif. Subcellular localization analysis revealed that GmARA6a localizes to the plasma membrane and multivesicular bodies (MVBs). Its expression was induced by salt stress, particularly in soybean roots. Under salt stress conditions, Arabidopsis ara6 mutants exhibited a salt-sensitive phenotype compared to wild-type plants, while GmARA6a overexpression lines showed improved growth, enhanced antioxidant enzyme activity, reduced membrane damage and lipid peroxidation, and lower levels of H2O2 and O2? accumulation. Expression analysis of related genes suggested that GmARA6a may contribute to salt tolerance by modulating the SOS signaling pathway and vesicle trafficking. These findings provide new insights into the functional role of GmARA6a in plant salt stress responses.

Key words: soybean, GmARA6a, subcellular localization, salt stress, functional analysis

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