Drought stress significantly retards the plant production. Melatonin is a vital hormone, signaling molecule, and bio-regulator of diverse physiological growth and development processes. Its role in boosting agronomic traits under diverse stress conditions has received considerable attention. However, the underlying molecular mechanism of action and how they increase drought stress tolerance has not been fully interpreted. The current study aimed to ascertain the protective role of melatonin in fortifying the antioxidant defense system, modulating the phytohormone profile, and improving agronomic traits of tomato seedlings under drought stress. After the V1 stage (1st leaf fully emerged), tomato seedlings were exposed to PEG-6000 to mimic drought-induced stress (DR 10% and DR 20%), followed by exogenous application of 100 µM soil drench. Drought-induced stress negatively impacted tomato seedlings by reducing growth and development and biomass accumulation, diminishing salicylic acid (SA) and chlorophyll levels, and dramatically lowering the antioxidant defense ability. However, melatonin protected them by activating the defense system, which decreased the oxidative burst and increased the activities of SOD, CAT, and APX. Administration of 100 µM melatonin by soil drench most remarkably downregulated the transcription factors of SlDREB3 and SlNCED3. This study has validated the moderating potential of melatonin against drought-induced stress by maintaining plant growth and development, enhancing hormone levels, elevating antioxidant enzyme activities, and suppressing the relative expression of drought-responsive genes. These findings also provide a basis for the potential use of MT in agricultural research and other relevant fields of study.
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