Updated FTP location. Seed dormancy is an indispensable trait for plants. It of barley is gradually decayed by a period of dry storage (after-ripening) or certain environmental condition. Freshly harvested (FH) and after-ripened (AR) grains have been differentially characterized by numerous studies, such as light, temperature and imbibition. Although FH and AR grains are known to change their gene expression during imbibition, phosphosignaling remain unclear. In this study we aim to investigate the effect of after-ripening on phosphosignaling in our cereal model species, barley. Phosphoproteomic analysis was carried out by using following process. Isolated barley seed embryo was used for protein extraction. Trypsin digestion and phosphopeptide enrichment by HAMMOC method were performed, then purified peptide sample was analyzed by TripleTOF5600 (AB-SCIEX).