Heavy metals increase in nature due to anthropogenic activities and negatively impact the growth, progress, and efficiency of plants. Among the toxic metal pollutants that can cause dangerous effects when accumulated by plants, mercury (Hg) and cadmium (Cd) were investigated in this study. These metals typically inhibit important enzymes and halt their functioning, thereby adversely affecting the capability of plants to achieve photosynthesis, respiration, and produce quality crops. Acetylcholine (ACh) serves as a potent neurotransmitter present in both primitive and advanced plant species. Its significant involvement in diverse metabolic processes, particularly in regulating growth and adaptation to stress, needs to be further elucidated. For this aim, effects of acetylcholine (ACh1, 10 μM; ACh2, 100 μM) were survey in Triticum aestivum under Hg and/or Cd stress (Hg, 50 μM; Cd, 100 μM). Wheat seedlings exhibited a growth retardation of about 24% under Hg or Cd stress. Combined stress conditions (Cd + Hg) resulted in a decrease in RWC by approximately 16%. Two different doses of ACh treatment to stressed plants positively affected growth parameters and regulated the water relations. Gas exchange was limited in stress groups, and the photochemical quantum competency of PSII (Fv/Fm) was suppressed. Cd + ACh1 and Cd + ACh2 treatments resulted in approximately 2-fold and 1.5-fold improvement in stomatal conductance and carbon assimilation rate, respectively. Similarly, improvement was observed with ACh treatments in wheat seedlings under Hg stress. Under Cd and/or Hg stress, high levels of H2O2 accumulated and lipid peroxidation occurred. According to our results, ACh treatment upon Cd and Hg stresses improved the activities of SOD, POX, and APX, thereby reducing oxidative damage. In conclusion, ACh treatment was found to ensure stress tolerance and limit the adverse effects caused by heavy metals.