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采用双向聚丙烯酰胺凝胶电泳(2D-PAGE)及质谱技术对橡胶种子萌发后蛋白质表达谱的变化进行了检测分析。结果显示,种子萌发前后的2个蛋白质组在蛋白质总数上并无显著差异,但各自均检测到特异性蛋白质的存在。成熟干种子蛋白质组含有2个分子量约为36.5kDa和23kDa的蛋白质簇(pH4-7)以及一组碱性蛋白质(大约pH10),而已萌发种子蛋白质组中23kDa蛋白质的表达量出现明显下降。已萌发种子蛋白质组中大约有60%的蛋白质与成熟干种子一致,其余40%则不相同或为其所独有。此外,本研究通过质谱分析从已检测到的蛋白点中鉴定出了3种在萌发种子蛋白质组中丰度有所下降的蛋白质,同时还发现了2种仅存在于成熟干种子蛋白质组中的蛋白质。前3种蛋白质包括1种推定的β-葡萄糖苷酶,淀粉分枝酶,以及1种MutT/nudix家族蛋白质,后2种蛋白质则分别为1种酸性植物凝集素和赤霉素20-氧化酶。本研究目的旨在探索2D-PAGE及质谱分析技术在巴西橡胶树蛋白质鉴定中的应用潜力,并根据蛋白质的变化动态藉以了解橡胶种子萌发的内在机制。
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry were used to detect the changes of protein expression profile after germination of rubber seeds. The results showed that there was no significant difference in the total number of proteins between the two proteomes before and after germination, but the presence of specific proteins was detected in each of the two proteomes. The mature dry seed proteome contains two protein clusters (pH 4-7) with molecular weights of about 36.5 kDa and 23 kDa and a set of basic proteins (about pH 10), whereas the 23 kDa protein expression in the germinating seed proteome is significantly reduced. Approximately 60% of the proteins in the germinating seed proteome are identical to the mature ones and the remaining 40% are not the same or unique to them. In addition, three kinds of proteins with decreased abundance in the germinating seed proteome were identified from the detected protein spots by mass spectrometry. In addition, two kinds of proteins that existed only in the mature dry seed proteome protein. The first three proteins include one putative beta-glucosidase, amylin, and one MutT / nudix family of proteins, the latter two proteins being one acidic phytohemagglutinin and gibberellin 20-oxidase . The aim of this study was to explore the potential of 2D-PAGE and mass spectrometry in the identification of the proteins of Hevea brasiliensis and to understand the underlying mechanisms underlying the germination of rubber seeds based on the dynamics of the proteins.