Soil salinity is one of the major constraints affecting agricultural production and crop yield. A detailed understanding of the underlying physiological and molecular mechanisms of the different genotypic salt tolerance response in crops under salinity is therefore a prerequisite for enhancing this tolerance. In this study, we explored the changes in physiological and proteome profiles of salt-sensitive (S210) and salt-tolerant (T510) sugar beet cultivars in response to salt stress. T510 showed better growth status, higher antioxidant enzymes activities and proline level, less Na accumulation, and lower P levels after salt-stress treatments. With iTRAQ-based comparative proteomics method, 47 and 56 differentially expressed proteins were identified in the roots and leaves of S210, respectively. In T510, 56 and 50 proteins changed significantly in the roots and leaves of T510, respectively. These proteins were found to be involved in multiple aspects of functions such as photosynthesis, metabolism, stress and defense, protein synthesis, and signal transduction. Our proteome results indicated that sensitive and tolerant sugar beet cultivars respond differently to salt stress. The proteins that were mapped to the protein modification, amino acid metabolism, tricarboxylic acid cycle, cell wall synthesis, and reactive oxygen species scavenging changed differently between the sensitive and tolerant cultivars, suggesting that these pathways may promote salt tolerance in the latter. This work leads to a better understanding of the salinity mechanism in sugar beet and provides a list of potential markers for the further engineering of salt tolerance in crops.