Plant small molecules, such as nitric oxide (NO) and melatonin (MN) as natural and human health-friendly compounds, play important roles in the mitigation of abiotic stresses in plants. Heavy metals such as chromium (Cr) are hazardous for the survival of ornamentals, especially edible flowers. This study evaluated the effects of NO (50 µM; sourced as sodium nitroprusside) and MN (50 µM) applied two times through foliar spraying at 1-week intervals on alleviating Cr (120 µM; K2Cr2O7)-induced oxidative stress in edible flowers of Calendula officinalis cv. Orange King. Cr stress decreased plant dry mass, leaf SPAD values, net photosynthetic rates, and the maximum photochemical quantum yield (Fv/Fm), and increased the oxidative stress markers. The individual application of NO or MN significantly mitigated the adverse effects, and the combined application of NO and MN synergistically enhanced plant tolerance to Cr stress, including increased activities of antioxidant enzymes in plants and concentrations of carbohydrate, ascorbic acid, sugar, total protein, as well as ash contents of edible flowers. The co-application also significantly elevated the concentrations of total phenolics, flavonoids, free reducing power, antioxidant capacity DPPH, and total carotenoids in Cr-treated plants compared with those in Cr-stressed plants. Additionally, the essential oil contents in flowers increased in response to the signaling molecule treatment under Cr stress. Compared with individual applications, the co-application of NO and MN had more significant effects. Our results indicate that the combination of signaling molecules, such as MN and NO, can not only increase the biomass of edible calendula plants but also improve flower quality for use as a novel food.
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