This paper presents the effects of irrigating barley plants with different type of water solutions saturated with gaseous ozone generated from atmospheric air. The study investigated the effects of the applied types of water on the modulation of the biosynthesis of selected bioactive compounds (content of total polyphenols, small molecule antioxidants, vitamin C) in the produced plant material. A number of transformations of reactive oxygen species (ROS) and nitrogen compounds have also been postulated; these are observed during the saturation of water with gaseous O3and 30 min after the end of the process. It was shown that after the process of water saturation with gaseous O3, the gas later is converted to compounds with high oxidative potential and good stability; these, in turn, lead to the oxidation of oxidates generated from atmospheric nitrogen into nitrates, which exhibit fertilising properties. Thirty minutes after the process of H2O saturation with gaseous O3was completed, the tests showed the highest concentrations of nitrates and the relatively high oxidative potential of the solution originating from H2O2with a low concentration of the dissolved O3. This solution exhibited the highest activity modulating the biosynthesis of polyphenols, small molecule antioxidants and vitamin C in young barley plants. The resulting differences were significant, and they were reflected by 15% higher total polyphenol content, 35% higher antioxidative potential and 57% greater content of vitamin C compared to the control specimens (plants treated with fresh H2O).