The extracted proteins from the hardy kiwifruit, which stored at room temperature (25°C) for up to 7 days, were used for proteomic analysis in order to understand the changes of post-harvest hardy kiwifruit at proteomic level. After two dimensional gels electrophoresis (2-DE) was conducted, more than 1300 protein spots were detected. Among them, 39 differently expressed spots (P<0.05) were selected to be excised and analyzed using MALDI-TOF/MS, and finally 5 protein spots were confidently identified according to NCBI database. The results demonstrated that among the 39 protein spots expressed particularly induced in fruits softening.