Salinity is one of the most important abiotic stresses because it causes zinc to precipitate in an unusable form for plants and is influenced by saline-calcareous soils. This experiment was carried out in a strip split block design with three replications at the Esfahan Rodasht Drainage and Salinity Research Station to investigate the effects of agrophysiological responses of barley genotypes to zinc fertilization and water saline irrigation. As vertical factors, water irrigation quality at three levels, 2, 10, and 18, dS/m, were used. Fertilizer application included Nano zinc-oxide, zinc-chelate, a mixture of Nano zinc-oxide and zinc-chelate, and water as a control. Within vertical factors, three different barley genotypes are arranged, including Morocco (moderate semi-sensitive), Nosrat (moderate tolerant), and Khatam (tolerant). The results showed that the application of Zn-chelate fertilizer resulted in the highest grain yield, K⁺ concentration, and K⁺/Na⁺ ratio in shoots. In Khatam, stomatal conductance (gs), the maximum quantum efficiency of PSII (Fv/Fm), K⁺ and Zn²⁺ concentrations, and the K⁺/Na⁺ ratio were all higher than in Morocco. In comparison to Morocco, Khatam had lower Na⁺ and Ca²⁺+Na⁺ contents. Furthermore, as salinity stress increased, all barley genotypes showed a decreasing trend in K⁺ content and the K⁺/Na⁺ ratio in shoots.