Hydrogen sulfide (H2S) has been proven to be a multifunctional signaling molecule in plants. In this study, we attempted to explore the effects of H2S on the climacteric fruit tomato during postharvest storage. H2S fumigation for 1 d was found to delay the peel color transition from green to red and decreased fruit firmness induced by ethylene. Further investigation showed that H2S fumigation downregulated the activities and gene expressions of cell wall–degrading enzymes pectin lyase (PL), polygalacturonase (PG), and cellulase. Furthermore, H2S fumigation downregulated the expression of ethylene biosynthesis genes SlACS2 and SlACS3. Ethylene treatment for 1 d was found to induce the expression of SlACO1, SlACO3, and SlACO4 genes, whereas the increase was significantly inhibited by H2S combined with ethylene. Furthermore, H2S decreased the transcript accumulation of ethylene receptor genes SlETR5 and SlETR6 and ethylene transcription factors SlCRF2 and SlERF2. The correlation analysis suggested that the fruit firmness was negatively correlated with ethylene biosynthesis and signaling pathway. The current study showed that exogenous H2S could inhibit the synthesis of endogenous ethylene and regulate ethylene signal transduction, thereby delaying fruit softening and the ripening process of tomato fruit during postharvest storage.