The altitude gradient limits the growth and distribution of alpine plants.Alpine plants have developed strategies to survive the extremely cold conditions prevailing at high altitudes; however, the mechanism underlying the evolution of these strategies remains unknown. The alpine plant Potentilla saundersiana is widespread in the Northwestern Tibetan Plateau. In this study, we conducted a comparative proteomics analysis to investigate the dynamic patterns of protein expression of P. saundersiana located at five different altitudes. We detected and functionally characterized 118 differentially expressed proteins. Our study confirmed that increasing levels of antioxidant proteins, and their respective activities, and accumulation of primary metabolites, such as proline and sugar, confer tolerance to the alpine environment in P. saundersiana. Proteins species associated with the epigenetic regulation of DNA stability and post-translational protein degradation were also involved in this process. Furthermore, our results showed that P. saundersiana modulated the root architecture and leaf phenotype to enhance adaptation to alpine environmental stress through mechanisms that involved hormone synthesis and signal transduction, particularly the cross-talk between auxin and strictosidine. Based on these findings, we conclude that P. saundersiana uses multiple strategies to adapt to the high-altitude environment of the Northwestern Tibetan Plateau.