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The synergistic roles of MsRCI2B and MsRCI2E in the regulation of ion balance and ROS homeostasis in alfalfa under salt stress

文献类型： 外文期刊

作者： Zhang, Depeng 1 ; Shen, Zhongbao 2 ; He, Pin 1 ; Wang, Jianli 2 ; Li, Donghuan 1 ; Meng, Jing 1 ; Zhang, Dongmei 2 ; You, Jia 2 ; Luo, Yaqin 1 ; Wang, Xinsheng 1 ; Zhuang, Xu 2 ; Mu, Linlin 2 ; Zhang, Shichao 1 ; Han, Weibo 2 ; Cai, Hua 1 ;

作者机构： 1.Northeast Agr Univ, Coll Life Sci, Harbin 150030, Peoples R China

2.Heilongjiang Acad Agr Sci, Inst Forage & Grassland Sci, Harbin 150086, Peoples R China

关键词： Medicago sativa; RCI2; Salt stress

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 300 卷

页码：

收录情况： SCI

摘要： Under salt stress, plasma membrane proteins regulate ion homeostasis and the balance between reactive oxygen species (ROS). In this study, we investigated the functions of two small membrane proteins-MsRCI2B (tailless) and MsRCI2E (tailed)-encoded by the RCI2 (Rare Cold Inducible 2) gene family in Medicago sativa (alfalfa). We identified the distinct subcellular localization and expression patterns of these proteins under salt stress. Using yeast two-hybrid (Y2H), GST pull-down, and bimolecular fluorescence complementation (BiFC) assays, we confirmed the physical interactions between MsRCI2B and MsRCI2E. Transgenic alfalfa lines overexpressing MsRCI2(OE#RCI2) and co-expressing both MsRCI2B and MsRCI2E (OE#RCI2E-2B) were developed to explore their roles in salt tolerance. Interestingly, the C-terminal tail of MsRCI2E negatively affects salt tolerance; however, its interaction with MsRCI2B mitigates this adverse effect. To further understand the regulatory mechanisms, we screened for plasma membrane proteins (PMPs) that interact with MsRCI2B or MsRCI2E using a DUALmembrane yeast two-hybrid system. MsCaM1 interacts with MsRCI2B, whereas MsPIP1;4 and MsHVP1 specifically interact with MsRCI2E. Notably, the MsRCI2E-PIP1;4 interaction influenced the intracellular trafficking of PIP1;4, reducing its presence on the plasma membrane and thereby limiting the export of H2O2, which helps maintain ROS homeostasis. Additionally, the interaction between MsRCI2E and HVP1 stabilized ion homeostasis by decreasing Na+ concentration in the cytoplasm under salt stress. Overall, our study provides new insights into the molecular mechanisms through which MsRCI2B and MsRCI2E coordinate the ion and ROS balance under salt stress and offering promising strategies for enhancing crop tolerance to salinity.

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