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以映山红(R.simsii)、马银花(R.ovatum)、满山红(R.mariesii)和云锦杜鹃(R.fortunei)4种杜鹃为实验材料,采用TCA-丙酮法、酚法和TCA-丙酮-酚法3种蛋白质提取方法提取杜鹃叶片蛋白质,并应用双向电泳技术(2-DE)比较分析不同方法的蛋白质提取率、溶解性、完整性等的差异,确立适用于2-DE分析的杜鹃叶片蛋白质提取方法。结果表明,TCA-丙酮法提取的蛋白溶解性差且粘稠,得到的蛋白质点少,不适合杜鹃叶片蛋白的提取;酚法提得的蛋白质溶解性好,蛋白质点多,但是杂质较多,且此法不适用于映山红叶片蛋白的提取;TCA-丙酮-酚法提取的杜鹃叶片蛋白质位点较多且清晰,重复性也好,且4种杜鹃叶片蛋白的提取效果都比较好。因此,TCA-丙酮-酚法被认为最适于杜鹃属植物叶片蛋白质的提取及分析。
Four species of rhododendron (Rhododendron fortunei, R.simsii, R.ovatum, R.mariesii, and R.fortunei) were used as experimental materials. The effects of TCA-acetone, phenol and TCA- - Phenolic method to extract three kinds of protein extraction Rhododendron leaf protein, and the use of two-dimensional electrophoresis (2-DE) comparative analysis of different methods of protein extraction rate, solubility, integrity, etc., to establish suitable for 2-DE analysis Rhododendron Leaf protein extraction method. The results showed that the protein extracted by TCA-acetone method was poorly soluble and viscous, and the obtained protein spots were not suitable for the extraction of leaf protein from Rhododendron fortunei. The protein extracted by phenol method had better solubility and more protein spots but more impurities, and This method does not apply to the extraction of azalea leaf protein; TCA-acetone-phenol extraction of rhododendron leaves more and clear protein sites, repeatability is also good, and the four kinds of rhododendron leaf protein extraction effect is better. Therefore, TCA-acetone-phenol method is considered as the most suitable for the extraction and analysis of Rhododendron leaf protein.