Barley (Hordeum vulgare) is a widely cultivated cereal crops, and its production is increasingly threatened by environmental stresses such as drought and salinity. Hydrogen sulfide is established as a signaling molecule that promotes tolerance to plant stress throught persulfidation, a post-translational modification of cysteine residues in proteins. The purpose of this study is to explore the impact of NaHS (sulfide donor) pretreatment on barley plants in enhancing tolerance to drought and salinity stresses, and determine if persulfidation is involved. In pretreated-plants, phenotypical traits and pigment contents showed an improvement in the survival of the plants under stress conditions. Quantification of stress-markers such as anthocyanin, proline, and reactive oxygen species also showed significant decreased contents in pretreated compared to untreated plants. In addition, the accumulation of amino acids under drought stress was significantly reduced when plants were pretreated with NaHS. Similarly, the increase of ABA content as a typical drought response was reduced in the pretreated plants. When plants are exposed to salt stress, the Na