Melatonin and nitric oxide: Dual players inhibiting hazardous metal toxicity in soybean plants via molecular and antioxidant signaling cascades

Melatonin (MT), a ubiquitous signaling molecule, is known to improve plant growth. Its regulatory function alongside nitric oxide (NO) is known to induce heavy metal (Cd and Pb) stress tolerance, although the underlying mechanisms remain unknown. Here, we observed that the combined application of MT and NO remarkably enhanced plant biomass by reducing oxidative stress. Both MT and NO minimized metal toxicity by significantly lowering the levels of endogenous abscisic acid and jasmonic acid via downregulating NCED3 and upregulating catabolic genes (CYP707A1 and CYP707A2). MT/NO-induced mitigation of Cd and Pb stress was associated with increased endo-melatonin and variable endo-S-nitrosothiol levels caused by enhanced expression of gmNR and gmGSNOR mRNAs. Remarkably, the combined application of MT/NO reduced soil Cd and Pb mobilization by increasing the uptake of Ca²⁺ and K⁺ and increasing the exudation of organic acids into the rhizosphere. These results correlated with the upregulation of MTF-1 and WARKY27 during metal translocation. MT/NO regulates the MAPK and CDPK cascades to promote plant cell survival and Ca²⁺ signaling, thereby imparting resistance to heavy metal toxicity. In conclusion, MT/NO modulates the stress-resistance machinery to mitigate Cd and Pb toxicity by regulating the activation of antioxidant and molecular transcription factors.