朱淑俊,王小霞,魏家萍,崔俊美,武泽峰,王晋雄,方彦,董小云,郑国强,撒二梅,李宝晶,魏鸿燕,罗倩,张欣怡,秦将源,梁欢,杨峰鹏,刘自刚.BrCTL1基因增强白菜型冬油菜抗寒性[J].干旱地区农业研究,2025,(5):59~68
BrCTL1基因增强白菜型冬油菜抗寒性
BrCTL1 gene enhances the cold tolerance of Winter Brassica rapa L.
  
DOI：10.7606/j.issn.1000-7601.2025.05.06
中文关键词:  BrCTL1基因  白菜型冬油菜  抗冻蛋白  几丁质酶  低温胁迫  抗寒性
英文关键词:BrCTL1 gene  Winter Brassica rarp L.  antifreeze protein  chitinase  low\|temperature stress  cold resistance
基金项目:国家自然科学基金(32360520)；甘肃省自然科学基金重点项目(23JRRA1408)；甘肃省科技重大专项(22ZD6NA009)；甘肃省重点人才项目(2023RCXM33)；甘肃省科技计划项目（23CXJA0001）；林芝市科技计划项目（LZZX-04）
作者单位
朱淑俊 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
王小霞 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
魏家萍 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
崔俊美 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
武泽峰 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
王晋雄 西藏自治区农牧科学院农业研究所，西藏 拉萨 850030 
方彦 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
董小云 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
郑国强 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
撒二梅 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
李宝晶 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
魏鸿燕 甘肃农业大学农学院，兰州 甘肃 730070；省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
罗倩 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
张欣怡 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
秦将源 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
梁欢 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
杨峰鹏 甘肃种业有限公司，兰州 甘肃 730070 
刘自刚 省部共建干旱生境作物学国家重点实验室, 兰州 甘肃 730070 
摘要点击次数: 271
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中文摘要:
      通过显微镜观察强抗寒冬油菜叶片几丁质酶蛋白冰晶形态，分析其编码基因家族冷胁迫下的表达模式。克隆几丁质酶基因（BrCTL1），进行亚细胞定位与过表达载体构建，测定过表达株系低温胁迫下生理变化及基因低温表达模式。结果表明，在冷胁迫环境下，从强抗寒白菜型冬油菜叶片中提取的蛋白液，具备显著的重结晶抑制活性，有效抑制冰晶生长，同时含有高活性的抗冻蛋白。SDS-PAGE电泳将该蛋白鉴定为几丁质酶（chitinase，CTL1）。同源克隆油菜BrCTL1（Brapa08T003472.1）基因，其ORF全长963 bp，编码321个氨基酸，BrCTL1蛋白被定位在叶绿体上。低温胁迫下，BrCTL1基因过表达植株形态无明显变化，突变体与野生型植株出现明显冻害症状，过表达拟南芥株系SOD、POD、CAT活性较野生型分别提高20%、17%、48%，CTL1突变体则分别降低65%、32%、67%。低温胁迫下，BrCTL1过表达株系相对电导率和MDA含量较野生型分别降低19%和45%，CTL1突变体分别提高9.4%和5.7%。综上，BrCTL1基因能够显著增强转基因拟南芥植株的抗寒性。
英文摘要:
      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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