In this report, we demonstrate that sodium hydrosulfide (NaHS), a hydrogen sulfide (H(2)S) donor, promoted adventitious root formation mediated by auxin and nitric oxide (NO). Application of the H(2)S donor to seedling cuttings of sweet potato (Ipomoea batatas L.) promoted the number and length of adventitious roots in a dose-dependent manner. It was also verified that H(2)S or HS(-) rather than other sulfur-containing components derived from NaHS could be attributed to the stimulation of adventitious root formation. A rapid increase in endogenous H(2)S, indole acetic acid (IAA) and NO were sequentially observed in shoot tips of sweet potato seedlings treated with HaHS. Further investigation showed that H(2)S-mediated root formation was alleviated by N-1-naphthylphthalamic acid (NPA), an IAA transport inhibitor, and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), an NO scavenger. Similar phenomena in H(2)S donor-dependent root organogenesis were observed in both excised willow (Salix matsudana var. tortuosa Vilm) shoots and soybean (Glycine max L.) seedlings. These results indicated that the process of H(2)S-induced adventitious root formation was likely mediated by IAA and NO, and that H(2)S acts upstream of IAA and NO signal transduction pathways.