Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L
IntroductionAlfalfa is the most widely cultivated high-quality perennial leguminous forage crop in the world. In China, saline-alkali land represents an important yet underutilized land resource. Cultivating salt-tolerant alfalfa varieties is crucial for the effective development and utilization of...
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Frontiers Media S.A.
2025-04-01
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| author | Zhiguang Li Qianqian Yu Yue Ma Fuhong Miao Lichao Ma Lichao Ma Shuo Li Huajie Zhang Huajie Zhang Zeng-Yu Wang Guofeng Yang Guofeng Yang Kunlong Su |
| author_facet | Zhiguang Li Qianqian Yu Yue Ma Fuhong Miao Lichao Ma Lichao Ma Shuo Li Huajie Zhang Huajie Zhang Zeng-Yu Wang Guofeng Yang Guofeng Yang Kunlong Su |
| author_sort | Zhiguang Li |
| collection | DOAJ |
| description | IntroductionAlfalfa is the most widely cultivated high-quality perennial leguminous forage crop in the world. In China, saline-alkali land represents an important yet underutilized land resource. Cultivating salt-tolerant alfalfa varieties is crucial for the effective development and utilization of saline-alkali soils and for promoting the sustainable growth of grassland-livestock farming in these regions. The NAC (NAM, ATAF, and CUC) family of transcription factors plays a key role in regulating gene expression in response to various abiotic stresses, such as drought, salinity and extreme temperatures, thereby enhancing plant stress tolerance.MethodsThis study evaluated the structure and evolutionary relationship of the members of the NAC-like transcription factor family in alfalfa using bioinformatics. We identified 114 members of the NAC gene family in the Zhongmu No.1 genome and classified them into 13 subclasses ranging from I to XIII. The bioinformatics analysis showed that subfamily V might be related to the response to salt stress. Gene expression analysis was conducted using RNA-seq and qRT-PCR, and MsNAC40 from subfamily V was chosen for further investigation into salt tolerance.ResultsMsNAC40 gene had an open reading frame of 990 bp and encoded a protein containing 329 amino acids, with a molecular weight of 3.70 KDa and a conserved NAM structural domain. The protein was hydrophilic with no transmembrane structure.After treating both the MsNAC40 overexpressing plants and the control group with 150 mmol/L NaCl for 15 days, physiological and biochemical measurements revealed that these plants had significantly greater height, net photosynthetic rate, stomatal conductance, and transpiration rate compared to the control group, while their conductivity was significantly lower. Additionally, the levels of abscisic acid in the roots and leaves, along with the activities of peroxidase, superoxide dismutase, and catalase in the leaves, were significantly higher in the overexpressing plants, whereas the malondialdehyde content was significantly lower. Moreover, the Na+ content in the overexpressing plants was significantly reduced, while the K+/Na+ ratio was significantly increased compared to the control group.DiscussionThese results indicated that the MsNAC40 gene improved the salt tolerance of Pioneer Alfalfa SY4D, but its potential mechanism of action still needs to be further explored. |
| format | Article |
| id | doaj-art-6cfed866ce97464c986f3177daf1ca86 |
| institution | DOAJ |
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| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj-art-6cfed866ce97464c986f3177daf1ca862025-08-20T02:53:44ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-04-011610.3389/fpls.2025.14617351461735Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa LZhiguang Li0Qianqian Yu1Yue Ma2Fuhong Miao3Lichao Ma4Lichao Ma5Shuo Li6Huajie Zhang7Huajie Zhang8Zeng-Yu Wang9Guofeng Yang10Guofeng Yang11Kunlong Su12Key Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaAcademy of Dongying Efficient Agricultural Technology and Industry on Saline and Alkaline Land in Collaboration with Qingdao Agricultural University, Dongying, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaWeihai Animal Epidemic Disease Prevention and Control Center, Weihai, ChinaWeihai Academy of Agricultural Sciences, Weihai, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaWeihai Animal Epidemic Disease Prevention and Control Center, Weihai, ChinaKey Laboratory of National Forestry and Grassland Administration on Grassland Resources and Ecology in the Yellow River Delta, College of Grassland Science, Qingdao Agricultural University, Qingdao, ChinaIntroductionAlfalfa is the most widely cultivated high-quality perennial leguminous forage crop in the world. In China, saline-alkali land represents an important yet underutilized land resource. Cultivating salt-tolerant alfalfa varieties is crucial for the effective development and utilization of saline-alkali soils and for promoting the sustainable growth of grassland-livestock farming in these regions. The NAC (NAM, ATAF, and CUC) family of transcription factors plays a key role in regulating gene expression in response to various abiotic stresses, such as drought, salinity and extreme temperatures, thereby enhancing plant stress tolerance.MethodsThis study evaluated the structure and evolutionary relationship of the members of the NAC-like transcription factor family in alfalfa using bioinformatics. We identified 114 members of the NAC gene family in the Zhongmu No.1 genome and classified them into 13 subclasses ranging from I to XIII. The bioinformatics analysis showed that subfamily V might be related to the response to salt stress. Gene expression analysis was conducted using RNA-seq and qRT-PCR, and MsNAC40 from subfamily V was chosen for further investigation into salt tolerance.ResultsMsNAC40 gene had an open reading frame of 990 bp and encoded a protein containing 329 amino acids, with a molecular weight of 3.70 KDa and a conserved NAM structural domain. The protein was hydrophilic with no transmembrane structure.After treating both the MsNAC40 overexpressing plants and the control group with 150 mmol/L NaCl for 15 days, physiological and biochemical measurements revealed that these plants had significantly greater height, net photosynthetic rate, stomatal conductance, and transpiration rate compared to the control group, while their conductivity was significantly lower. Additionally, the levels of abscisic acid in the roots and leaves, along with the activities of peroxidase, superoxide dismutase, and catalase in the leaves, were significantly higher in the overexpressing plants, whereas the malondialdehyde content was significantly lower. Moreover, the Na+ content in the overexpressing plants was significantly reduced, while the K+/Na+ ratio was significantly increased compared to the control group.DiscussionThese results indicated that the MsNAC40 gene improved the salt tolerance of Pioneer Alfalfa SY4D, but its potential mechanism of action still needs to be further explored.https://www.frontiersin.org/articles/10.3389/fpls.2025.1461735/fullalfalfaNAcsalt tolerancephylogenetic analysisMsNAC40 |
| spellingShingle | Zhiguang Li Qianqian Yu Yue Ma Fuhong Miao Lichao Ma Lichao Ma Shuo Li Huajie Zhang Huajie Zhang Zeng-Yu Wang Guofeng Yang Guofeng Yang Kunlong Su Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L Frontiers in Plant Science alfalfa NAc salt tolerance phylogenetic analysis MsNAC40 |
| title | Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L |
| title_full | Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L |
| title_fullStr | Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L |
| title_full_unstemmed | Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L |
| title_short | Screening and functional characterization of salt-tolerant NAC gene family members in Medicago sativa L |
| title_sort | screening and functional characterization of salt tolerant nac gene family members in medicago sativa l |
| topic | alfalfa NAc salt tolerance phylogenetic analysis MsNAC40 |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2025.1461735/full |
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