This study investigates the impact of three cold plasma treatments on barley seed germination: direct treatment of dry seeds (DDS), direct treatment of water-soaked seeds (DWS), and indirect treatment of seeds using plasma-activated water (IPAW). Our findings reveal that while DDS maintained the seed's imbibition structures, DWS significantly reduced water uptake. Notably, DWS-treated seeds exhibited a substantial increase in moisture content, potentially leading to an oxygen-deficient environment within the seed. This condition appeared to hinder germination, contrasting with the beneficial effects observed in DDS-treated seeds. Analyses of surface hydrophilicity, phenolic compounds, carbohydrate composition, protein storage and seedling growth were conducted. Interestingly, cold plasma treatment enhanced seed surface hydrophilicity, as evidenced by a decrease in water contact angles, particularly for DWS. We observed specific reductions in flavonoids and total phenolics, alongside shifts in carbohydrate composition, including increased sugar content up to 15.1% (DDS) and 13.5% (DWS), accompanied by a decrease in starch content to 189.00 ± 7.86 mg/g FM for DWS. Protein storage assessments revealed declines in albumin, globulin and prolamin fractions post-CP exposure. Finally, DDS and IPAW treatments markedly improved seedling shoot growth (from two to three-fold), underscoring the potential of these treatments in enhancing barley germination.