| BrCTL1 gene enhances the cold tolerance of Winter Brassica rapa L. |
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| DOI：10.7606/j.issn.1000-7601.2025.05.06 |
| Key Words: BrCTL1 gene Winter Brassica rarp L. antifreeze protein chitinase low\|temperature stress cold resistance |
| Author Name | Affiliation | | ZHU Shujun | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | WANG Xiaoxia | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | WEI Jiaping | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | CUI Junmei | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | WU Zefeng | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | WANG Jinxiong | Institute of Agricultural Research, Tibetan Academy of Agricultural and Animal Husbandry Sciences, Lasa, Xizang 850030, China | | FANG Yan | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | DONG Xiaoyun | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | ZHENG Guoqiang | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | SA Ermei | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | LI Baojing | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | WEI Hongyan | Agronomy College, Gansu Agricultural University, Gansu, Lanzhou 730070, China
State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | LUO Qian | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | ZHANG Xinyi | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | QIN Jiangyuan | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | LIANG Huan | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China | | YANG Fengpeng | Gansu seed Industry Co., Ltd., Gansu, Lanzhou 730070, China | | LIU Zigang | State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Gansu, Lanzhou 730070, China |
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| Abstract: |
| This study observed the ice crystal morphology of chitinase protein in the leaves of highly cold\|resistant winter rapeseed through a microscope and analyzed the expression patterns of its encoding gene family under cold stress. The chitinase gene (BrCTL1) was cloned, the subcellular localization was carried out, and an overexpression vector was constructed. The physiological changes and gene expression patterns of overexpressing lines under low\|temperature stress were determined. The results showed that under cold\|stress conditions, the protein solution extracted from the leaves of highly cold\|resistant winter rapeseed had significant recrystallization inhibition activity, effectively inhibits ice\|crystal growth, and contains highly active antifreeze proteins. The protein was identified as Chitinase (CTL1) by SDS-PAGE electrophoresis. The BrCTL1 (Brapa08T003472.1) gene of rapeseed was cloned by the homologous cloning method. Its ORF was 963 bp in full length and encoded 321 amino acids. The BrCTL1 protein was located in the chloroplast. Under low\|temperature stress, there were no obvious morphological changes in the BrCTL1 gene overexpression plants, while obvious freezing injury symptoms occurred in the mutant and wild\|type plants. The activities of SOD, POD, and CAT in the overexpression Arabidopsis lines increased by 20%, 17%, and 48%, respectively, compared with the wild\|type, while those in the CTL1 mutant decreased by 65%, 32%, and 67%, respectively. Under low\|temperature stress, the relative conductivity and MDA content in the BrCTL1 overexpression lines decreased by 19% and 45%, respectively, compared with the wild\|type, while those in the CTL1 mutant increased by 9.4% and 5.7%, respectively. In conclusion, the BrCTL1 gene can significantly enhance the cold resistance of transgenic Arabidopsis thaliana plants. |
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