ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Plant Abiotic Stress
This article is part of the Research TopicExploring Cold Tolerance and Stress in PlantsView all 12 articles
The Alfalfa U-box E3 Ligase MsPUB210 Interacts with MsICE1 and Positively Regulates Cold Tolerance in Transgenic Arabidopsis
Provisionally accepted
- College of Life Science and Technology, Harbin Normal University, Harbin, China
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Cold stress severely restricts the productivity of Alfalfa (Medicago sativa L.), prompting the evolution of complex resistance mechanisms involving post-translational modifications. In this study, we characterized MsPUB210, a gene encoding a U-box E3 ubiquitin ligase, to elucidate its role in cold adaptation. Phylogenetic analysis confirmed that MsPUB210 is homologous to Arabidopsis U-box proteins, and gene regulatory network (GRN)-guided protein-protein interaction (PPI) network analysis, together with co-expression investigation, first predicted a potential association between MsPUB210 and the transcription factor MsICE1. This prediction was further corroborated by AlphaFold2 (AF2)-predicted 3D structures of their heteromeric complexes, supporting the biological plausibility of this interaction. We demonstrated via yeast two-hybrid (Y2H) assays that MsPUB210 physically interacts with the transcription factor MsICE1. Heterologous overexpression of MsPUB210 in Arabidopsis thaliana significantly enhanced cold tolerance, characterized by reduced reactive oxygen species (ROS) accumulation and mitigated lipid peroxidation compared to wild-type plants. Furthermore, MsPUB210 was found to upregulate downstream cold-responsive genes, suggesting it functions as a positive regulator within the ICE-CBF-COR signaling cascade. Collectively, these findings highlight the pivotal role of MsPUB210 in ubiquitin-mediated cold signaling and identify it as a promising genetic target for breeding cold-resilient forage crops.
Keywords: alfalfa, cold tolerance, GRN, Ubiquitination, U-box E3 ubiquitin ligases
Received: 22 Dec 2025; Accepted: 13 Feb 2026.
Copyright: © 2026 Wang, Li, Zhu, Guo, Guo and Shu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yongjun Shu
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