Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress

Medicago sativa is the most widely cultivated high-protein forage crop globally. However, its cultivation in high-latitude and cold regions of China is significantly hindered by low-temperature stress, particularly impacting the root system, the primary functional tissue crucial for winter survival....

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Main Authors: Xiaolong Wang, Hua Chai, Yanxia Xu, Shasha Li, Yue Wu, Ruoding Wang, Zhao Yang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2025.1597949/full
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author Xiaolong Wang
Hua Chai
Yanxia Xu
Shasha Li
Yue Wu
Ruoding Wang
Zhao Yang
author_facet Xiaolong Wang
Hua Chai
Yanxia Xu
Shasha Li
Yue Wu
Ruoding Wang
Zhao Yang
author_sort Xiaolong Wang
collection DOAJ
description Medicago sativa is the most widely cultivated high-protein forage crop globally. However, its cultivation in high-latitude and cold regions of China is significantly hindered by low-temperature stress, particularly impacting the root system, the primary functional tissue crucial for winter survival. The physiological and molecular mechanisms underlying the root system’s adaptation and tolerance to low temperatures remain poorly understood. To this end, this study utilized cold-tolerant Medicago sativa “Lomgmu801” and the cold-sensitive Medicago sativa “Sardi” genotypes as experimental materials to investigate root physiological responses during the overwintering period. Physiological indices, including soluble sugars (SS), proline (Pro), glutathione (GSH), jasmonic acid (JA), abscisic acid (ABA) contents, and peroxidase (POD) activity, were quantified. RNA-seq revealed 743 differentially expressed genes (DEGs) between the cold-tolerant and sensitive genotypes. Subsequently, correlation analysis between DEGs and physiological indices revealed that DEGs in the yellow, blue, and turquoise modules were significantly correlated with the levels of POD, GSH, SS, Pro, JA, and ABA. The core genes were predominantly included in the “MAPK signaling pathway”, “glutathione metabolism”, “plant hormone signal transduction”, “arginine and proline metabolism”, and “phenylpropanoid biosynthesis”. Ultimately, DEGs responsive to low-temperature stress were identified, including MsGST, MsRboh, MsPOD, MsJAZ, MsP5CR, and MsPP2C. By integrating physiological characteristics with cold-tolerance-associated genes, this study elucidates the physiological and molecular mechanisms underlying Medicago sativa root adaptation to low temperatures. The RNA-seq data and the core DEGs identified provide valuable theoretical insights and targets for future molecular breeding efforts aimed at enhancing cold tolerance in Medicago sativa.
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spelling doaj-art-e91a81f17e784ba1a87349d781d994b42025-08-25T05:26:27ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-08-011610.3389/fpls.2025.15979491597949Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stressXiaolong WangHua ChaiYanxia XuShasha LiYue WuRuoding WangZhao YangMedicago sativa is the most widely cultivated high-protein forage crop globally. However, its cultivation in high-latitude and cold regions of China is significantly hindered by low-temperature stress, particularly impacting the root system, the primary functional tissue crucial for winter survival. The physiological and molecular mechanisms underlying the root system’s adaptation and tolerance to low temperatures remain poorly understood. To this end, this study utilized cold-tolerant Medicago sativa “Lomgmu801” and the cold-sensitive Medicago sativa “Sardi” genotypes as experimental materials to investigate root physiological responses during the overwintering period. Physiological indices, including soluble sugars (SS), proline (Pro), glutathione (GSH), jasmonic acid (JA), abscisic acid (ABA) contents, and peroxidase (POD) activity, were quantified. RNA-seq revealed 743 differentially expressed genes (DEGs) between the cold-tolerant and sensitive genotypes. Subsequently, correlation analysis between DEGs and physiological indices revealed that DEGs in the yellow, blue, and turquoise modules were significantly correlated with the levels of POD, GSH, SS, Pro, JA, and ABA. The core genes were predominantly included in the “MAPK signaling pathway”, “glutathione metabolism”, “plant hormone signal transduction”, “arginine and proline metabolism”, and “phenylpropanoid biosynthesis”. Ultimately, DEGs responsive to low-temperature stress were identified, including MsGST, MsRboh, MsPOD, MsJAZ, MsP5CR, and MsPP2C. By integrating physiological characteristics with cold-tolerance-associated genes, this study elucidates the physiological and molecular mechanisms underlying Medicago sativa root adaptation to low temperatures. The RNA-seq data and the core DEGs identified provide valuable theoretical insights and targets for future molecular breeding efforts aimed at enhancing cold tolerance in Medicago sativa.https://www.frontiersin.org/articles/10.3389/fpls.2025.1597949/fullMedicago sativaphysiological characteristicsgenescold tolerancecorrelation analysis
spellingShingle Xiaolong Wang
Hua Chai
Yanxia Xu
Shasha Li
Yue Wu
Ruoding Wang
Zhao Yang
Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
Frontiers in Plant Science
Medicago sativa
physiological characteristics
genes
cold tolerance
correlation analysis
title Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
title_full Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
title_fullStr Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
title_full_unstemmed Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
title_short Analysis of physiological characteristics and gene co-expression networks in Medicago sativa roots under low-temperature stress
title_sort analysis of physiological characteristics and gene co expression networks in medicago sativa roots under low temperature stress
topic Medicago sativa
physiological characteristics
genes
cold tolerance
correlation analysis
url https://www.frontiersin.org/articles/10.3389/fpls.2025.1597949/full
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