With the intensification of global warming, rice production is facing new challenges. Field evidence indicates increased temperature during rice grain-filling lead to a further deterioration of grain quality. Clarifying the potential regulation mechanism of elevated temperature on rice development and quality formation will be contributed to develop suitable cultivation measures to better cope with climate warming in the future. In this study, open field warming and DIA mass spectrometry were conducted to explore the regulatory effects of high temperature on pathways related to grain development and material accumulation during the formation of rice quality. 840 differentially expressed proteins (fold change > 2, p-value < 0.05) were identified when exposed to high temperature. Among these, prolamin PPROL 14E, PSB28, granule-bound starch synthase 1 and 26.7 kDa heat shock protein were the most significantly regulated, and that ultimately affected the main substances accumulation of starch and protein in the kernel, and further degraded rice quality under high temperature. In addition, the results provided novel targets involved in regulating the metabolism of storage compounds under warming environment, and that will help us to better understand the regulation mechanism of global warming on the formation of rice quality.