Accumulating evidence has shown that hydrogen sulfide (H₂S) acts as a signaling regulator in plants. Here we show that H₂S delays the postharvest senescence of broccoli in a dose-dependent manner. H₂S maintains higher levels of metabolites, such as carotenoids, anthocyanin, and ascorbate, and reduces the accumulation of malondialdehyde, H₂O₂, and the superoxide anion. Further investigations showed that H₂S sustained higher activities of guaiacol peroxidase, ascorbate peroxidase, catalase, and glutathione reductase and lower activities of lipoxygenase, polyphenol oxidase, phenylalanine ammonia lyase, and protease than those of water control. Moreover, the expression of the chlorophyll degradation related genes BoSGR, BoCLH2, BoPaO, BoRCCR, as well as cysteine protease BoCP1 and lipoxygenase gene BoLOX1, was down-regulated in postharvest broccoli treated with H₂S. The functions of H₂S on the senescence of other vegetables and fruits suggest its universal role acting as a senescence regulator.