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以抗稻瘟病水稻近等基因系C101LAC及CO39为材料,应用差异蛋白组学技术对外源茉莉酸甲酯(MeJA)诱导8 h和12 h后的水稻叶片蛋白质差异表达变化进行了分析。采用PEG-4000预沉淀法提取水稻叶片总蛋白,经双向电泳、凝胶染色和图像分析,结果表明:与对照相比MeJA处理后的2个水稻品系中共有21个蛋白质表现为2倍以上的表达差异。其中,MeJA处理8 h后,CO39中的差异表达蛋白质点有5个,C101LAC中有8个,U5为2个品系中均新增的蛋白质点。Me-JA处理12 h后,CO39中有6个差异表达蛋白质点,C101LAC中则有5个。经胶内酶解、MALDI-TOF/TOF质谱分析及数据库检索,成功对21个差异表达蛋白质进行了鉴定,其中新增的蛋白质点U5被鉴定为内切β-1,3-1,4-葡聚糖酶。依据GO数据库和UniProtKB数据库提供的蛋白质注释信息,按照功能可将21个差异表达蛋白质分为8类,分别在植物抗性、氨基酸代谢、信号转导、光合作用、光呼吸、糖代谢、蛋白质合成与分解及细胞代谢等方面发挥作用。
The rice near-isogenic lines C101LAC and CO39 were used as materials to analyze the differentially expressed proteins in leaves of rice induced by Exogenous MeJA 8 h and 12 h by differential proteomics. The total protein of rice leaves was extracted by PEG-4000 pre-precipitation method. The results of two-dimensional electrophoresis, gel staining and image analysis showed that compared with the control, 21 proteins in two rice lines treated with MeJA showed more than twice Differences in expression. Among them, 8 days after MeJA treatment, there were 5 protein spots differentially expressed in CO39, 8 in C101 LAC, and U5 as new protein spots in 2 lines. After Me-JA treatment for 12 h, there were 6 differentially expressed protein spots in CO39 and 5 in C101 LAC. Twenty-one differentially expressed proteins were successfully identified by in-gel digestion, MALDI-TOF / TOF mass spectrometry and database search. Among them, the new protein U5 was identified as endo-β-1,3-1,4- Dextranase. According to the protein annotation information provided by the GO database and the UniProtKB database, 21 differentially expressed proteins were classified into 8 groups according to their functions, respectively in plant resistance, amino acid metabolism, signal transduction, photosynthesis, photorespiration, glucose metabolism, protein synthesis And decomposition and cell metabolism play a role.