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Seed germination is a complex physiological process that allows the seed embryo to grow and develop into a photosynthetic organism. The two major constituents of rice seed include the embryo and endosperm, with embryo being of much significance despite its small size. In this study, we conducted a systematic proteomic analysis of the embryos dissected from rice seed at different stages of germination through a combination of gel-based and gel-free strategies. In total, 343 differentially expressed proteins were identified. Among them, 191 were decreased and 152 were increased in terms of expression levels. All these proteins could be sorted into 11 functional groups based on MapMan analysis. Some starch biosynthesis-related enzymes such as starch branching enzyme, granule-bound starch synthase 1 and starch synthase increased during the early stage of germination and then decreased at the late stage, which was similar to the expressional patterns of glycolysis-related enzymes. However, tricarboxylic acid cycle-related enzymes only increased at the later stage. It was also found that sucrose might be an important intermediate for the biosynthesis of starch in embryos. Furthermore, gel-based proteomic analysis of the dissected endosperm showed that the biological processes in the endosperm were heavily regulated by the embryo. This study could provide some new insights into the distinct roles of the embryo and endosperm in rice seed germination. [ABSTRACT FROM AUTHOR]

Seed germination is a well regulated process, which incorporates many events including signal transduction, mobilization of reserves, reactive oxygen species scavenging and cell division. Although many transcriptomic and proteomic studies have been conducted on this process, regulation of protein modification has not been studied. To better understand the mechanism, a gel-based comparative phosphoproteomic study was performed on rice embryo during the germination process. In total, 168 protein spots exhibited significantly changed Pro-Q staining intensity during germination. Using matrix-assisted laser deionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis, 193 proteins were identified. By combining Pro-Q and Coomassie brilliant blue stain intensity analyses, 109 proteins were verified to be phosphorylation regulation proteins. Functional analyses indicated that phosphorylation of proteins involved in stress response and storage was gradually enhanced. Phosphorylation of signal transduction proteins was mainly activated during the early stage of germination, while stress response and storage protein phosphorylation were enhanced at the late stage. Enzyme assays proved that the phosphorylation of fructokinase, pyruvate kinase, malate dehydrogenase, GDP-mannose 3,5-epimerase1, ascorbate peroxidase and glutathione S-transferase could consistently enhance their activity. This study showed the dynamic changes of protein phosphorylation status in rice embryo during germination and provided new insight into understanding the mechanism underlying this process. [ABSTRACT FROM PUBLISHER]