Here we investigated the effect of hydrogen sulphide (H2S) on postharvest quality of mulberry fruit and possible underlying mechanisms. Endogenous H2S content first increased after harvest and then decreased sharply with the process of ripening and senescence. A fumigation with H2S released from 0.8 mm NaHS solution could significantly enhance the endogenous H2S content by increasing the activities of D-cysteine desulfhydrase and L-cysteine desulfhydrase. NaHS could significantly slow down the ripening rate of mulberry fruit and reduce the respiratory intensity and anthocyanin content. Moreover, H2S fumigation was able to obviously delay or slow down the decreases in soluble protein, titratable acidity and ascorbate contents. Further results showed that activities of representative antioxidant enzymes in H2S-treated sample were higher than those of control samples during storage, resulting in a decrease in superoxide anion production. Together, these results clearly indicate that H2S fumigation has a potential role in the preservation of mulberries.