Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time

Abstract Background The C-repeat binding factor (CBF)/dehydration-responsive element binding (DREB1) belongs to a subfamily of the AP2/ERF (APETALA2/ethylene-responsive factor) superfamily, which can regulate many physiological and biochemical processes in plants, such as plant growth and developmen...

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Main Authors: Jing Cui, Yajing Li, Hao Liu, Xu Jiang, Lili Zhang, Hongbo Dai, Xue Wang, Fei He, Mingna Li, Junmei Kang
Format: Article
Language:English
Published: BMC 2025-01-01
Series:BMC Plant Biology
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Online Access:https://doi.org/10.1186/s12870-025-06081-0
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author Jing Cui
Yajing Li
Hao Liu
Xu Jiang
Lili Zhang
Hongbo Dai
Xue Wang
Fei He
Mingna Li
Junmei Kang
author_facet Jing Cui
Yajing Li
Hao Liu
Xu Jiang
Lili Zhang
Hongbo Dai
Xue Wang
Fei He
Mingna Li
Junmei Kang
author_sort Jing Cui
collection DOAJ
description Abstract Background The C-repeat binding factor (CBF)/dehydration-responsive element binding (DREB1) belongs to a subfamily of the AP2/ERF (APETALA2/ethylene-responsive factor) superfamily, which can regulate many physiological and biochemical processes in plants, such as plant growth and development, hormone signal transduction and response to abiotic stress. Although the CBF/DREB1 family has been identified in many plants, studies of the CBF/DREB1 family in alfalfa are insufficient. Results In this study, 25 MsCBF genes were identified in the genome of alfalfa (“Zhongmu No. 4”). These genes were distributed on chromosomes 1, 5, 6 and unassembled scaffolds. Phylogenetics divided the CBF members of Medicago sativa, Arabidopsis thaliana, and Medicago truncatula into six groups, of which group VI had the most MsCBFs members, reaching 52% (13/25). Gene duplication analysis showed that 64% (16/25) of MsCBFs formed tandem duplications, and 32% (8/25) formed segment duplications. The expression pattern of MsCBF9 under different hormone treatments was verified by RT-qPCR, and it was found that MsCBF9 responded to GA3, IAA, SA, and MeJA. Overexpression of MsCBF9 in Arabidopsis significantly delayed the flowering time of Arabidopsis. In contrast, the flowering time of the cbfs mutant was earlier, and overexpression of MsCBF9 also increased the number and size of Arabidopsis rosette leaves. Conclusion In this study, the CBF/DREB1 family of alfalfa was comprehensively identified and analyzed, and the function of MsCBF9 in regulating flowering time was studied. This study laid a foundation for further analysis of the function of the CBF family in alfalfa. Clinical trial number Not applicable.
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institution Kabale University
issn 1471-2229
language English
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series BMC Plant Biology
spelling doaj-art-d432201ebdeb4966beb10ff1a58cef282025-01-26T12:23:23ZengBMCBMC Plant Biology1471-22292025-01-0125111510.1186/s12870-025-06081-0Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering timeJing Cui0Yajing Li1Hao Liu2Xu Jiang3Lili Zhang4Hongbo Dai5Xue Wang6Fei He7Mingna Li8Junmei Kang9Institute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesInstitute of Animal Science, Chinese Academy of Agricultural SciencesAbstract Background The C-repeat binding factor (CBF)/dehydration-responsive element binding (DREB1) belongs to a subfamily of the AP2/ERF (APETALA2/ethylene-responsive factor) superfamily, which can regulate many physiological and biochemical processes in plants, such as plant growth and development, hormone signal transduction and response to abiotic stress. Although the CBF/DREB1 family has been identified in many plants, studies of the CBF/DREB1 family in alfalfa are insufficient. Results In this study, 25 MsCBF genes were identified in the genome of alfalfa (“Zhongmu No. 4”). These genes were distributed on chromosomes 1, 5, 6 and unassembled scaffolds. Phylogenetics divided the CBF members of Medicago sativa, Arabidopsis thaliana, and Medicago truncatula into six groups, of which group VI had the most MsCBFs members, reaching 52% (13/25). Gene duplication analysis showed that 64% (16/25) of MsCBFs formed tandem duplications, and 32% (8/25) formed segment duplications. The expression pattern of MsCBF9 under different hormone treatments was verified by RT-qPCR, and it was found that MsCBF9 responded to GA3, IAA, SA, and MeJA. Overexpression of MsCBF9 in Arabidopsis significantly delayed the flowering time of Arabidopsis. In contrast, the flowering time of the cbfs mutant was earlier, and overexpression of MsCBF9 also increased the number and size of Arabidopsis rosette leaves. Conclusion In this study, the CBF/DREB1 family of alfalfa was comprehensively identified and analyzed, and the function of MsCBF9 in regulating flowering time was studied. This study laid a foundation for further analysis of the function of the CBF family in alfalfa. Clinical trial number Not applicable.https://doi.org/10.1186/s12870-025-06081-0AlfalfaMsCBF;genome-wideFlowering regulation
spellingShingle Jing Cui
Yajing Li
Hao Liu
Xu Jiang
Lili Zhang
Hongbo Dai
Xue Wang
Fei He
Mingna Li
Junmei Kang
Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
BMC Plant Biology
Alfalfa
MsCBF;genome-wide
Flowering regulation
title Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
title_full Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
title_fullStr Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
title_full_unstemmed Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
title_short Genome-wide identification and expression analysis of CBF/DREB1 gene family in Medicago sativa L. and functional verification of MsCBF9 affecting flowering time
title_sort genome wide identification and expression analysis of cbf dreb1 gene family in medicago sativa l and functional verification of mscbf9 affecting flowering time
topic Alfalfa
MsCBF;genome-wide
Flowering regulation
url https://doi.org/10.1186/s12870-025-06081-0
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