Globally, drought and salinity stress critically constrain potato (Solanum tuberosum L.) production. Considering the impact of these stresses on crops and increasing food demand, insight into both tolerance and susceptibility is essential. The present study screens two potato cultivars, BARI-401 and Spunta, for their tolerance to simulated salinity and drought by in vitro LiCl and mannitol exposure. Plantlets treated with a range of LiCl (0, 10, 30, and 40 mM) and mannitol (0, 50, 100, 200, and 250 mM) concentrations were biochemically and physiologically characterized to assess their tolerance capacity. Shoot number, shoot length, root number, and root length were affected in both cultivars under higher LiCl and mannitol concentrations, even though Spunta was able to better maintain a higher shoot length under the 40 mM of LiCl and 250 mM of mannitol compared to BARI-401. The total phenol contents (TPC) in both cultivars were increased at the highest treatment concentration and the total flavonoids content (TFC) was decreased in BARI-401 as compared to Spunta. Higher free radical scavenging capacity (FRSC, low IC50 value) was recorded in Spunta as compared to BARI-401 with increasing treatment concentrations, which supports the high antioxidant capacity of Spunta. An inverse correlation between polyphenol oxidase (PPO) and TPC was noted in both cultivars. Peroxidase dismutase (POD) activity was increased significantly in both cultivars for all treatments, but activity was highest overall in Spunta. These physiological and biochemical analyses of both cultivars suggest that cultivar Spunta is more tolerant to salinity and drought stress. Further open-field experiments are required to confirm these results.