Department of Botany, Institute of Biosciences, University of Sao Paulo, 05422-970 Sao Paulo, SP, Brazil.
Phytochemistry. 2011 Apr;72(4-5):302-11. doi: 10.1016/j.phytochem.2010.12.007. Epub 2011 Jan 27.
Araucaria angustifolia is an endangered Brazilian native conifer tree. The aim of the present work was to identify differentially expressed proteins between mature and germinated embryos of A. angustifolia, using one and two dimensional gel electrophoresis approaches followed by protein identification by tandem mass spectrometry. The identities of 32 differentially expressed protein spots from two dimensional gel maps were successfully determined, including proteins and enzymes involved in storage mobilization such as the vicilin-like storage protein and proteases. A label free approach, based on spectral counts, resulted in detection of 10 and 14 mature and germinated enriched proteins, respectively. Identified proteins were mainly related to energetic metabolism pathways, translational processes, oxidative stress regulation and cellular signaling. The integrated use of both strategies permitted a comprehensive protein expression overview of changes in germinated embryos in relation to matures, providing insights into the this process in a recalcitrant seed species. Applications of the data generated on the monitoring and control of in vitro somatic embryos were discussed.
南洋杉是一种濒危的巴西原生针叶树。本研究的目的是使用一维和二维凝胶电泳方法，结合串联质谱技术，鉴定南洋杉成熟和萌发胚胎之间差异表达的蛋白质。从二维凝胶图谱中成功鉴定出 32 个差异表达的蛋白质斑点，其中包括参与储存动员的蛋白质和酶，如豆球蛋白样储存蛋白和蛋白酶。基于光谱计数的无标记方法分别检测到 10 个和 14 个成熟和萌发富集蛋白。鉴定出的蛋白质主要与能量代谢途径、翻译过程、氧化应激调节和细胞信号转导有关。两种策略的综合应用可以全面概述萌发胚胎与成熟胚胎相比的蛋白质表达变化，深入了解这种顽固种子物种的萌发过程。还讨论了所产生的数据在体外体细胞胚胎监测和控制方面的应用。
