Pepper is a widely cultivated and consumed vegetable in Türkiye and globally. However, salinity has negative effects on the development of plants and is considered an important problem in the cultivation of many plant species. This study investigated the effects of varying concentrations of sodium chloride (NaCl) (0, 50, 100, 150, 200, and 250 mM) added to Murashige and Skoog (MS) medium on the in vitro growth of pepper plants. Key physiological and morphological parameters were evaluated, including germination rate (CO), average germination time (OCS), salt tolerance index (TTI), real water content (RWC), fresh and dry weights of shoots and roots, shoot and root lengths, and visual damage scores (1–5). Results showed that increasing NaCl concentrations negatively impacted pepper growth. The highest CO (82.33%) and the shortest MGT (3.50 days) were recorded in the control-BA medium, while the lowest CO (58.00%) occurred in the 250 mM NaCl-BF medium. The greatest visual damage (score of 3.80) was observed in the BF medium, followed by the 200 mM NaCl-BE medium (3.60). As a result, pepper growth was significantly inhibited by increasing NaCl concentrations, demonstrating the plant’s sensitivity to salinity stress.