Salt stress obstructs plant’s growth by affecting metabolic processes, ion homeostasis and over-production of reactive oxygen species. In this regard silicon (Si) has been known to augment a plant’s antioxidant defense system to combat adverse effects of salinity stress. In order to quantify the Si-mediated salinity tolerance, we studied the role of Si (200 ppm) applied through rooting media on antioxidant battery system of barley genotypes; B-10008 (salt-tolerant) and B-14011 (salt-sensitive) subjected to salt stress (200 mM NaCl). A significant decline in the accumulation of shoot (35–74%) and root (30–85%) biomass was observed under salinity stress, while Si application through rooting media enhancing biomass accumulation of shoots (33–49%) and root (32–37%) under salinity stress. The over-accumulation reactive oxygen species i.e., hydrogen peroxide (H₂O₂) is an inevitable process resulting into lipid peroxidation, which was evident by enhanced malondialdehyde levels (13–67%) under salinity stress. These events activated a defense system, which was marked by higher levels of total soluble proteins and uplifted activities of antioxidants enzymatic (SOD, POD, CAT, GR and APX) and non-enzymatic (α-tocopherol, total phenolics, AsA, total glutathione, GSH, GSSG and proline) in roots and leaves under salinity stress. The Si application through rooting media further strengthened the salt stressed barley plant’s defense system by up-regulating the activities of enzymatic and non-enzymatic antioxidant in order to mitigate excessive H₂O₂ efficiently. The results revealed that although salt-tolerant genotype (B-10008) was best adopted to tolerate salt stress, comparably the response of salt-sensitive genotype (B-14011) was more prominent (accumulation of antioxidant) after application of Si through rooting media under salinity stress.