Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa
IntroductionThe cold tolerance of winter rapeseed cultivars is critically important for winter survival and yield formation in northern area. BrAFP1, an antifreeze protein in Brassica rapa, is hypothesized to stabilize membranes and inhibit ice crystal formation.Methodswe cloned the BrAFP1 promoter...
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Frontiers Media S.A.
2025-08-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2025.1527754/full |
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| author | Xiaoyun Dong Jinxiong Wang Jiaping Wei Guoqiang Zheng Zefeng Wu Junmei Cui Xuezhen Yang Baojin Li Shujun Zhu Ermei Sa Fengpeng Yang Zigang Liu |
| author_facet | Xiaoyun Dong Jinxiong Wang Jiaping Wei Guoqiang Zheng Zefeng Wu Junmei Cui Xuezhen Yang Baojin Li Shujun Zhu Ermei Sa Fengpeng Yang Zigang Liu |
| author_sort | Xiaoyun Dong |
| collection | DOAJ |
| description | IntroductionThe cold tolerance of winter rapeseed cultivars is critically important for winter survival and yield formation in northern area. BrAFP1, an antifreeze protein in Brassica rapa, is hypothesized to stabilize membranes and inhibit ice crystal formation.Methodswe cloned the BrAFP1 promoter from the cold-tolerant cultivar Longyou 7 (L7) and constructed the proBrAFP1::GUS expression vector to investigate the impact of membrane state changes on BrAFP1 expression and the cold tolerance in winter rapeseed. Ten independent transgenic T3 lines were generated, among which T3-5 and T3-7 were selected for subsequent analysis.ResultsThe dimethyl sulfoxide (DMSO) treatment in the absence of cold exposure activated the transcriptional activity of proBrAFP1, a cold-inducible promoter; in contrast, benzyl alcohol (BA) treatment eliminated its cold-induced activation. The expression levels of cold-responsive genes, including cyclic nucleotide-gated channel 1 (CNGC1), open stomata 1 (OST1), and inducer of CBF expression 1 (ICE1), as well as membrane fluidity-related genes, such as acyl-lipid desaturase 2 (ADS2), fatty acid desaturase 2 (FAD2), and sensitive to freezing 2 (SFR2), were significantly increased following DMSO pretreatment, while BA treatment significantly inhibited the expression of these genes. Furthermore, ABA and SA levels are closely linked to alterations in the membrane state, compared to untreated plants, the levels of ABA and SA in the leaves markedly increased at 4°C after DMSO and BA treatment but decreased at -4°C.ConclusionCollectively, DMSO pretreatment enhanced cold tolerance, while BA pretreatment improved cell survival under cold stress, which is important for practise of keeping the rapeseed yields. |
| format | Article |
| id | doaj-art-710b4f13841843378d274f1107fc23d9 |
| institution | Kabale University |
| issn | 1664-462X |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj-art-710b4f13841843378d274f1107fc23d92025-08-20T04:00:54ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-08-011610.3389/fpls.2025.15277541527754Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapaXiaoyun Dong0Jinxiong Wang1Jiaping Wei2Guoqiang Zheng3Zefeng Wu4Junmei Cui5Xuezhen Yang6Baojin Li7Shujun Zhu8Ermei Sa9Fengpeng Yang10Zigang Liu11State Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaAgricultural Research Institute, Tibet Academy of Agriculture and Animal Husbandry Sciences, Lasa, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaSeed Industry Development Section, Pingliang Seed Station, Pingliang, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaResearch and Development Center, Gansu Seed Industry Company Limited, Lanzhou, ChinaState Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, ChinaIntroductionThe cold tolerance of winter rapeseed cultivars is critically important for winter survival and yield formation in northern area. BrAFP1, an antifreeze protein in Brassica rapa, is hypothesized to stabilize membranes and inhibit ice crystal formation.Methodswe cloned the BrAFP1 promoter from the cold-tolerant cultivar Longyou 7 (L7) and constructed the proBrAFP1::GUS expression vector to investigate the impact of membrane state changes on BrAFP1 expression and the cold tolerance in winter rapeseed. Ten independent transgenic T3 lines were generated, among which T3-5 and T3-7 were selected for subsequent analysis.ResultsThe dimethyl sulfoxide (DMSO) treatment in the absence of cold exposure activated the transcriptional activity of proBrAFP1, a cold-inducible promoter; in contrast, benzyl alcohol (BA) treatment eliminated its cold-induced activation. The expression levels of cold-responsive genes, including cyclic nucleotide-gated channel 1 (CNGC1), open stomata 1 (OST1), and inducer of CBF expression 1 (ICE1), as well as membrane fluidity-related genes, such as acyl-lipid desaturase 2 (ADS2), fatty acid desaturase 2 (FAD2), and sensitive to freezing 2 (SFR2), were significantly increased following DMSO pretreatment, while BA treatment significantly inhibited the expression of these genes. Furthermore, ABA and SA levels are closely linked to alterations in the membrane state, compared to untreated plants, the levels of ABA and SA in the leaves markedly increased at 4°C after DMSO and BA treatment but decreased at -4°C.ConclusionCollectively, DMSO pretreatment enhanced cold tolerance, while BA pretreatment improved cell survival under cold stress, which is important for practise of keeping the rapeseed yields.https://www.frontiersin.org/articles/10.3389/fpls.2025.1527754/fullmembrane fluiditycold tolerancedimethyl sulfoxidebenzyl alcoholproBrAFP1 activity |
| spellingShingle | Xiaoyun Dong Jinxiong Wang Jiaping Wei Guoqiang Zheng Zefeng Wu Junmei Cui Xuezhen Yang Baojin Li Shujun Zhu Ermei Sa Fengpeng Yang Zigang Liu Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa Frontiers in Plant Science membrane fluidity cold tolerance dimethyl sulfoxide benzyl alcohol proBrAFP1 activity |
| title | Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa |
| title_full | Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa |
| title_fullStr | Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa |
| title_full_unstemmed | Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa |
| title_short | Effect of membrane rigidification on the BrAFP1 expression and cold-tolerance in Brassica rapa |
| title_sort | effect of membrane rigidification on the brafp1 expression and cold tolerance in brassica rapa |
| topic | membrane fluidity cold tolerance dimethyl sulfoxide benzyl alcohol proBrAFP1 activity |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2025.1527754/full |
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