Analysis of differential protein expression of tea callus with different catechins contents | 茶树不同儿茶素含量愈伤组织的蛋白差异分析

Chinese. 【目的】建立适合于茶树愈伤组织的蛋白质双向电泳技术,并比较不同愈伤组织间的蛋白表达差异,为深入开展茶树儿茶素生物合成的蛋白质组学研究奠定基础。【方法】在优化蛋白质提取技术的基础上,分别对“云茎63X”(儿茶素含量低)、“云茎63Y”(儿茶素含量较高)和光照诱导的“云茎63Y”(儿茶素含量最高)三类愈伤组织的蛋白质进行双向电泳分析,并对表达差异的蛋白质点进行质谱(LTQ-ESI-MS/MS)分析。【结果】通过对蛋白得率和SDS-PAGE单向电泳图谱的比较,发现TCA-丙酮法最适合茶树愈伤组织中蛋白质的提取;从三类愈伤组织的蛋白双向电泳图谱中,分析检测出14个差异较大的蛋白质;经质谱分析和数据库检索,14个蛋白中包括有谷胱甘肽S-转移酶、WD40蛋白、咖啡酸-O-转甲基酶、S-腺苷甲硫氨酸合成酶、果胶甲酯酶等。【结论】这些蛋白可能参与了苯丙烷及类黄酮途径的合成及其调控、乙烯合成、细胞壁代谢、糖酵解和信号转导等生理作用。

English. 【Objective】 To establish a two-dimensional electrophoresis (2-DE) of proteome suitable for tea callus and analyze the differential protein expression patterns of tea callus with different catechins contents, which could provide a basis for the further study of proteomics of the catechins biosynthesis. 【Method】 Using the improved protein extraction, 2-DE and LTQ-ESI-MS/MS method, the differential protein expression profiling was investigated using the tea callus of “Yunjing63X” (with low catechins contents), “Yunjing63Y” (with higher contents), and “Yunjing63Y” cultured under light (with the highest contents). 【Result】 Protein quantitative and SDS-PAGE electrophoresis results showed that TCA-acetone method was the best for extracting tea callus proteins. And 14 differential protein spots were identified by 2-DE electrophoresis, LTQ-ESI-MS/MS and database searching. The glutathione S-transferase, WD40 family protein, catechol O-methyltransferase, S-adenosylmethionine synthetase, pectin methylesterase and so on were identified among those differential protein spots. 【Conclusion】 The predicted function of these picked proteins were involved in many physiological reactions, such as phenylpropanoid and flavonoids biosynthesis, transport and regulation, ethylene biosynthesis, cell wall metabolism , glycolysis pathway, and signal transduction.