Sulfur (S) metabolism plays a vital role in Cd detoxification, but the collaboration between melatonin biosynthesis and S metabolism under Cd stress remains unaddressed. Using exogenous melatonin, melatonin-deficient tomato plants with a silenced caffeic acid O-methyltransferase (COMT) gene, and COMT-overexpressing plants with cosuppression of sulfate transporter (SUT)1and SUT2genes, we found that melatonin deficiency decreased S accumulation and aggravated Cd phytotoxicity, whereas exogenous melatonin or overexpression of COMTincreased S uptake and assimilation, resulting in an improved plant growth and Cd tolerance. Melatonin deficiency promoted Cd translocation from root to shoot, but COMToverexpression caused the opposite effect. COMToverexpression failed to compensate the functional hierarchy of S when its uptake was inhibited by cosilencing of transporter SUT1and SUT2.Our study provides genetic evidence that melatonin-mediated tolerance to Cd is closely associated with the efficient regulation of S metabolism, redox homeostasis, and Cd translocation in tomato plants.