茶树叶片类脱水素蛋白的Western-blot分析

为了解茶树脱水素种类与功能,采用Western-blot技术,研究了不同季节及越冬过程中茶树叶片脱水素蛋白家族的表达模式。结果显示:(1)茶树叶片总蛋白提取采用酚-甲醇/醋酸铵沉淀法,用时短、蛋白浓度高、SDSPAGE电泳条带清晰,背景干净,满足茶树Western-blot技术要求。(2)在不同季节及越冬期中发现14~95kD共9种不同分子量的茶树类脱水素蛋白,其中95、65、48、37、34和14kD等6种蛋白表达量较为稳定,季节与越冬期变化不明显;58kD脱水素仅在冬季表达,越冬期不断上升,2月份增加到最高,表达丰度高;28kD脱水素蛋白在冬季表达量高,越冬期与茶树抗寒力变化规律一致;21kD脱水素在夏季和越冬期后期有较高的表达。研究表明,这3种脱水素可能在茶树抗逆中起着重要作用。

Expression Patterns of Dehydrin-like Proteins in Tea Plant (Camellia sinensis) by Western-blot Analysis

To investigate the types and functions of dehydrin in tea plant Camellia sinensis (L.) O.Kuntze],we studied the expression profiling of tea dehydrin protein family in different seasons and wintering period by Western-blot.The results showed that:(1)the phenol-methanol/ammonium acetate precipitation method was suitable for total protein extraction from tea leaves,and met the requirements for Western-blot analysis,together with short time,high concentration,clear electrophoretic bands,and no contaminated background in SDS-PAGE.Nine kinds of dehydrin-like prteins in tea plant with the molecular weights ranging from 14-95 kD,were found in different seasons and during overwintering.The expressions levels were relatively stable in the 6 kinds of dehydrin-like proteins (molecular weights 95,65,48,37,34 and 14 kD).The 58 kD dehydrin occurred only in winter,and showed a continuous expression in overwintering period,which achieved the highest expression in February.The highest expression of 28 kD dehydrin was found in winter,whose variation pattern was in accordance with the cold resistance for tea plant.The 21 kD dehydrin expression level was higher in summer and the later period of overwintering.It provided that the three dehydrins (58,28 and 21 kD) might play important roles in stress tolerance for tea plants.