Barley (Hordeum vulgare L.) is an important crop grown throughout the world. Quality and quantity of barley is compromised by abiotic stresses, salinity in particular which is already widespread in many regions. Mitogen Activated Protein Kinase (MAPK) cascades are signal transduction pathways involved in biotic and abiotic plant defence mechanisms. Here, the function of the barley gene HvBWMK1 has been investigated. Transgenic barley plants that overexpress HvBWMK1, or in which HvBWMK1 has been down-regulated by antisens, were generated, as well as empty vector control plants. The transformation efficiency using Agrobacterium was 10.4% independent transgenic plant per embryo. Northern blot analysis of HvBWMK1 primary overexpression lines showed up-regulation of mRNA level, conversely the antisense lines of HvBWMK1 showed clear down-regulation. Real-time PCR analysis in the T1 generation revealed that two over-expresser lines showed higher expression as compared with wild type (by 79% and 35%) while the HvBWMK1 antisense construct reduced endogenous gene expression partially (by 14%, 23% and 39% compared with wild type). Barley seedlings were subjected to two weeks of salt stress (150 mM NaCl) then biochemical and physiological parameters were measured. The overexpression lines showed an increase in tolerance to salinity stress compared to antisense lines and controls. Tolerance was accompanied with increasing endogenous proline and chlorophyll levels and a reduction in hydrogen peroxide content after salinity exposure. Overall these results suggest that the barley MAPK HvBWMK1 acts as a positive regulator in barley tolerance to salinity stress.