MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)

Low-temperature stress is a major environmental factor that limits the yield, quality, and geographical distribution of forage crops and restricts the development of the forage industry. As a core component of plant circadian clocks, Late Elongated Hypocotyl (LHY) plays a crucial role in regulating...

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Main Authors: Jikai Li, Lu Chai, Mei Yang, Hailing Zhang, Chen Shang, Yuxuan Liu, Kailin Qian, Jiuding Sun, Weibo Han, Pan Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2025.1559988/full
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author Jikai Li
Lu Chai
Mei Yang
Hailing Zhang
Chen Shang
Yuxuan Liu
Kailin Qian
Jiuding Sun
Weibo Han
Pan Zhang
author_facet Jikai Li
Lu Chai
Mei Yang
Hailing Zhang
Chen Shang
Yuxuan Liu
Kailin Qian
Jiuding Sun
Weibo Han
Pan Zhang
author_sort Jikai Li
collection DOAJ
description Low-temperature stress is a major environmental factor that limits the yield, quality, and geographical distribution of forage crops and restricts the development of the forage industry. As a core component of plant circadian clocks, Late Elongated Hypocotyl (LHY) plays a crucial role in regulating plant rhythms and responses to abiotic stress. However, the molecular mechanism by which LHY regulates the cold tolerance of alfalfa has not been reported. In this study, MsLHY, which is 2,235 bp in length and encodes 744 amino acids, was isolated from alfalfa. MsLHY was highly expressed in roots and stems and was significantly induced by low temperature. Transgenic MsLHY-overexpressing (OE) and RNAi alfalfa plants were obtained via Agrobacterium-mediated transformation. Under low-temperature stress, OE plants presented reduced reactive oxygen species accumulation and more osmotic regulatory substances, as well as greater antioxidant enzyme activity, to combat cold stress. Conversely, the RNAi plants presented trends opposite those of the OE plants. Furthermore, under cold stress, the overexpression of MsLHY upregulated the expression of the cold-responsive genes MsICE1, MsCBF1, MsCOR15A, and MsCML10, as well as the expression of the antioxidant-synthesizing genes MsSOD1 and MsCAT1, thereby increasing the cold tolerance of transgenic alfalfa. These results suggest that MsLHY plays an important role in increasing the cold tolerance of alfalfa.
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spelling doaj-art-041660e4fcc94d9f968df4ee1ac978102025-08-20T02:55:39ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-05-011610.3389/fpls.2025.15599881559988MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)Jikai Li0Lu Chai1Mei Yang2Hailing Zhang3Chen Shang4Yuxuan Liu5Kailin Qian6Jiuding Sun7Weibo Han8Pan Zhang9Institute of Grass Research, Heilongjiang Academy of Agricultural Sciences, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaInstitute of Grass Research, Heilongjiang Academy of Agricultural Sciences, Harbin, ChinaInstitute of Grass Research, Heilongjiang Academy of Agricultural Sciences, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaInstitute of Grass Research, Heilongjiang Academy of Agricultural Sciences, Harbin, ChinaDepartment of Grassland Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, ChinaLow-temperature stress is a major environmental factor that limits the yield, quality, and geographical distribution of forage crops and restricts the development of the forage industry. As a core component of plant circadian clocks, Late Elongated Hypocotyl (LHY) plays a crucial role in regulating plant rhythms and responses to abiotic stress. However, the molecular mechanism by which LHY regulates the cold tolerance of alfalfa has not been reported. In this study, MsLHY, which is 2,235 bp in length and encodes 744 amino acids, was isolated from alfalfa. MsLHY was highly expressed in roots and stems and was significantly induced by low temperature. Transgenic MsLHY-overexpressing (OE) and RNAi alfalfa plants were obtained via Agrobacterium-mediated transformation. Under low-temperature stress, OE plants presented reduced reactive oxygen species accumulation and more osmotic regulatory substances, as well as greater antioxidant enzyme activity, to combat cold stress. Conversely, the RNAi plants presented trends opposite those of the OE plants. Furthermore, under cold stress, the overexpression of MsLHY upregulated the expression of the cold-responsive genes MsICE1, MsCBF1, MsCOR15A, and MsCML10, as well as the expression of the antioxidant-synthesizing genes MsSOD1 and MsCAT1, thereby increasing the cold tolerance of transgenic alfalfa. These results suggest that MsLHY plays an important role in increasing the cold tolerance of alfalfa.https://www.frontiersin.org/articles/10.3389/fpls.2025.1559988/fullMedicago sativa L.late-elongated hypocotyllow temperatureinteracting proteintransgenic alfalfa
spellingShingle Jikai Li
Lu Chai
Mei Yang
Hailing Zhang
Chen Shang
Yuxuan Liu
Kailin Qian
Jiuding Sun
Weibo Han
Pan Zhang
MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
Frontiers in Plant Science
Medicago sativa L.
late-elongated hypocotyl
low temperature
interacting protein
transgenic alfalfa
title MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
title_full MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
title_fullStr MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
title_full_unstemmed MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
title_short MsLHY is an active regulator of cold resistance in alfalfa (Medicago sativa L.)
title_sort mslhy is an active regulator of cold resistance in alfalfa medicago sativa l
topic Medicago sativa L.
late-elongated hypocotyl
low temperature
interacting protein
transgenic alfalfa
url https://www.frontiersin.org/articles/10.3389/fpls.2025.1559988/full
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