To eliminate the damages of metal toxicity by reducing metal uptake by plants, organic amendments are useful. The use of carbon-rich materials known as biochar (BC) is a strong candidate to enhance the plant tolerance against stress conditions. The current study examined the effects of BC in wheat hydroponically grown treated with BC (1 and 3 g L−1) alone or in combination with cobalt (Co, 150 and 300 μM). Stress reduced the relative growth rate (RGR), relative water content (RWC), osmotic potential (ΨΠ), and increased proline content (Pro). Besides, endogenous contents of Ca2+, K+, and Mn2+ in leaves decreased under stress. In response to Co stress, a decline in the activities of peroxidase (POX), ascorbate peroxidase (APX), and glutathione reductase (GR) resulted in the induction of hydrogen peroxide (H2O2) content. BC applied with stress decreased endogenous Co2+ content and increased RGR, RWC, chlorophyll fluorescence and Pro content. Also, the activities of superoxide dismutase (SOD), catalase (CAT), APX and GR were induced and the ascorbate (AsA) and glutathione (GSH) pool and their redox state were maintained by BC application under stress condition. While, with the addition of BC, H2O2 content and lipid peroxidation displayed remarkable decreased, the scavenging activity of hydroxyl radical (OH·) increased as compared to Co stress-treated wheat plants. Besides, in wheat leaves, BC application triggered AsA-GSH pathway including activities of monodehydroascorbate reductase, dehydroascorbate reductase, and the contents of dehydroascorbate, GSH, and GSH/GSSG ratio. The presented results supported the view that biochar under stress could minimize the Co-induced oxidative damages through modulation of the growth, water status, photosynthetic apparatus, and antioxidant enzyme activity found in cellular compartments and ascorbate-glutathione cycle in wheat leaves.