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Nitric oxide (NO), an endogenous signaling molecule in plants and animals, mediates responses to abiotic and biotic stresses. This study was conducted in a nutrient solution to investigate 1) the effects of exogenous sodium nitroprusside (SNP), an NO donor, on free proline (Pro) and protein content; and 2) the enzymes involved in Pro metabolism [pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (PDH)] in cucumber (Cucumis sativus) seedling leaves and roots under NaCl stress. The results showed that the increases in free Pro and protein were significantly higher in the 50 mm NaCl solution but highly significant with the addition of 100 μM SNP to the 50 mm NaCl solution for the entire treatment period. Moreover, leaves maintained higher levels of free Pro and protein content than roots throughout the experiments. The P5CS activity increased in the saline treatment compared with the control, and this increase was greater in the 50 mm NaCl + 100 μM SNP solution than in the other treatments. On the other hand, the PDH activity was inhibited under NaCl stress but the reduction in activity was greater in the 50 mm NaCl + 100 μM SNP solution than in the others. These findings suggest that Pro metabolism was significantly altered during the exogenously applied NO under salt stress and that this alteration prompted the accumulation of higher levels of free Pro, which, in turn, maintained the turgor in the cucumber seedlings and helped protect them from salt stress. Moreover, the toxic effects generated by 50 mm NaCl were partially overcome by the application of NO, which could be used as a potential growth regulator to improve plant salinity tolerance. Therefore, it was concluded that NO could alleviate salinity damage in cucumber seedlings by regulating Pro metabolism. Overall, the adverse effects of salt stress could be lessened by the exogenous application of NO to cucumber seedlings.