In water-limited areas of Tunisia, more than 30% of subsurface water used for irrigation is saline water, leading to a long-term salinization and degradation processes. To prevent the problems and to minimize the negative impact of using saline water, selection of proper germplasm as well as integrated soil and water management are essential. Understanding the diversity for salt tolerance in barley (Hordeum vulgare L.) genotypes will facilitate their use in genetic improvement. Our objectives were to evaluate, in green, house-salt tolerance for 14 barley cultivars under three different salinity levels: (tap water with an Ec = 0.73 dSm(-1), tap water with 102 mM of NaCl, Ec = 10.76 dSm(-1) and tap water with 151 mM of NaCl, Ec = 15.38 dSm(-1)), and quantify genetic variation based on salt tolerance index, morphological traits, molecular and factorial analysis of correspondence (FAC). The study was conducted in a randomized complete block design arranged as a split plot. The results indicate a great genetic variability to salt tolerance among used barley genotypes. Consequently, it is possible to identify superior cultivars, and evaluate the genotypic performance under salinities conditions. In addition, results obtained were confirmed by clustering made by SSR tool on the base of DNA analysis, which is compatible with the arrangement obtained with statistical method.