Separation and identification of Brassica rapa BrAFP and its gene cloning and expression under freezing stress
Separation and identification of Brassica rapa BrAFP and its gene cloning and expression under freezing stress
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白菜BrAFP的分离鉴定及其基因克隆与冷冻胁迫下的表达
DOI：
10.1111/pbr.12677
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发表时间：
2019-04-01
期刊：
影响因子：
2
通讯作者：
Li, Xuecai
中科院分区：
文献类型：
--
作者：
Liu, Zigang;Dong, Xiaoyun;Li, Xuecai
抗冻蛋白（AFPs）是一类通过与冰非依数性结合来抑制冰核形成、抑制冰生长并改变冰晶形态的蛋白质。在本研究中，以耐寒的‘陇油6号’（芜菁）为模型。在电泳图谱上，从低温处理的植物叶片中提取的一种38kDa的蛋白质条带增强。这种38kDa的蛋白质能够显著抑制冰晶生长并改变其形态，因此表明这种蛋白质在冬油菜中具有抗冻蛋白的特性。通过质谱鉴定该蛋白质为β - 1,3 - 葡聚糖酶，命名为BrAFP1。其相应基因包含1032个碱基，编码343个氨基酸，分子量为38102kDa。BrAFP1是一种预测的具有信号肽的疏水蛋白。在冷胁迫下，植物表现出耐寒的形态特征，并且与对照（室温）相比，在 - 4℃下从0.5天到4天，BrAFP的表达显著上调，这表明BrAFP基因在芜菁的冷驯化中起重要作用。
Antifreeze proteins (AFPs) are proteins that inhibit ice nucleation by non-colligative combined with ice, to inhibit its growth and to modify ice crystal morphology. In this study, 'Longyou 6' (Brassica rapa) with the cold resistance was used as a model. A 38kDa band of protein extracted from leaves of plants exposed to low temperature was enhanced on the electropherogram. The 38kDa protein can significantly inhibit growth of ice crystals and modify its morphology, thus suggesting this protein has an AFP in winter turnip rape. The protein was identified as beta-1,3-glucanase by mass spectrometry, named BrAFP1. Its corresponding gene contains 1,032bp bases, coding 343 amino acids with a molecular weight of 38,102kDa. The BrAFP1 is a predicted hydrophobic protein with a signal peptide. Under cold stress, the plants showed morphological characteristics of cold resistance, and BrAFP expression was significantly up-regulated at -4 degrees C from 0.5 to 4days compared with the control (at room temperature), suggesting that the BrAFP gene played an important role in cold acclimation in B.rapa.