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为了从蛋白质分子生物学方面研究白菜型冬油菜不同抗寒品种低温逆境及恢复条件下相关蛋白的响应机制,揭示大田生产中冬油菜抵御突发骤变低温伤害的抗寒机理,以超强抗寒冬油菜品种陇油7号和弱抗寒品种天油2号为材料,在培养箱培养至幼苗五叶期时,模拟北方寒旱区冬油菜返青期倒春寒环境,运用双向电泳结合质谱技术,分析其幼苗在低温(4℃)-恢复(20℃)-低温(4℃)-恢复(20℃)过程中存在的蛋白及其差异表达量。研究表明低温胁迫与恢复过程中,有2个蛋白点发生了显著差异的丰度变化,它们分别是半胱氨酸型氧化还蛋白过氧化物酶(定义为1号蛋白)和一种未知蛋白(2号蛋白)。且不同抗寒性品种存在2种不同蛋白的差异表达机制,1号蛋白在抗寒性较弱的天油2号低温胁迫恢复过程中表现出上调-下调-上调的变化趋势,抗寒性较强的陇油7号中此蛋白在第一次低温胁迫后消失,而出现了另外一种未知蛋白(蛋白2),且在低温胁迫恢复过程中也表现出上调-下调-上调的变化趋势,并且表达量高于天油2号中的蛋白1,因此,这种未知蛋白2可能是陇油7号抗寒性强于天油2号的原因之一。
In order to study the response mechanism of related proteins under cold stress and recovery conditions of different cold-tolerant varieties of cabbage-type rapeseed from the aspects of protein molecular biology, the cold-resistant mechanism of winter rapeseed against sudden sudden cold damage in field production was revealed, Winter oilseed rape variety Longyou No.7 and weak tolerant variety Tianyou No.2 were used as materials to simulate the late spring cold rapeseed winter greening environment in the cold and dry areas of North China when the incubator was cultured to the seedling five-leaf stage. Two-dimensional electrophoresis combined with mass spectrometry The proteins present in the seedlings during low temperature (4 ℃) -recovery (20 ℃) -low temperature (4 ℃) -recovery (20 ℃) and their differential expression levels. The results showed that there were two distinct abundance changes in the process of low temperature stress and recovery. They were cysteine-type oxidoreductase peroxidase (defined as protein No.1) and an unknown protein (Protein number 2). There were two different expression mechanisms of different proteins in different cold resistance varieties. The protein No.1 showed a trend of up-down-up-regulation in the process of cold-stress-resistant Tianyou 2, In Longyou 7, the protein disappeared after the first low temperature stress, and another unknown protein (protein 2) appeared, and showed a tendency of up-down-up-regulation during the process of low temperature stress recovery. And the expression level was higher than that of Tianyou 2. Therefore, this unknown protein 2 may be one of the reasons why the cold resistance of Longyou 7 is stronger than Tianyou2.