The aim of this work was to explore the contributions of phosphorus in mitigating the salinity impacts on osmotic potential, protein content and proline accumulation of cucumber grown in vitro on proliferation medium. Microshoots were grown on Murashige and Skoog (MS) proliferation media containing 1 mg/l kinetin and supplemented with a combination of NaCl at 0.0, 25, 50, 75, 100 mM with 0.5, 1.0 or 2.0 mM phosphorus using KH2PO4. Leaf osmotic potential was reduced with increasing NaCl concentration and within each of the 50, 75 and 100 mM NaCl concentrations, the incorporation of 2 mM KH2PO4 was needed to significantly increase leaf osmotic potential. Protein content of cucumber microshoots was shown to decrease with NaCl concentration, this reduction was lessened with increase in KH2PO4 level applied. In vitro-induced salinity caused profound proline accumulation in cucumber microshoots and combinations having 2 mM KH2PO4 showed lower proline accumulation especially at the highest two concentrations of NaCl.