Purpose <p>Salinity is a hazardous abiotic stress that arrests crop productivity by damaging its physiological and molecular mechanisms. However, it is unclear how the osmolytes and proteomic indicators of antioxidant proteins regulate Na⁺ partitioning and detoxify reactive oxygen species (ROS) to increase the salt tolerance of wheat seedlings.</p> Methods <p>We examined the proteomic profile, osmoprotectants, salinity-responsive genes, ion markers, and essential plant hormones in wheat seedlings subjected to 200 mM NaCl stress.</p> Results <p>Our findings showed that the high expression status of Phosphatase 2&#xa0;C protein (PP2C) and ABA-responsive element binding factor (ABF) proteins markedly increased ABA hormone accumulation in response to salinity, resulting in ABA-mediated downstream actions that stimulate ion homeostasis in wheat roots. Moreover, it was demonstrated that the up-regulation of 14-3-3 proteins and vacuolar Na⁺/H⁺antiporters was crucial for salt stress tolerance via important mechanisms of osmotic adjustments and Na⁺ compartmentalization within the vacuole under salt treatments. Salt stress also reduced the oxidative damage of wheat roots, improving the levels of multiple enzymes that detoxify ROS, particularly glutathione-S-transferase, ascorbate peroxidase, and thioredoxins.</p> Conclusions <p>According to KEGG enrichment analyses, salt stress positively increased protein abundance in numerous protein pathways, including metabolic processes, ADP metabolic process, ATP metabolic process, and glutathione metabolism, which helps protect cells from oxidative stress. This research provides new insights into how wheat roots mitigate the hazardous effects of salt stress through protein synthesis and biochemical responsive mechanisms, which may be helpful in the development of salinity-tolerant species.</p>