Nitric oxide (NO) and hydrogen sulfide (H2 S) are signalling molecules that regulate adventitious rooting in plants. However, little is known about the cross-talk between NO and H2 S during adventitious rooting. Tomato (Solanum lycopersicum L.) explants were used to investigate the roles of and relationships between NO and H2 S during rooting. Effects of the NO donor sodium nitroprusside (SNP) and the H2 S donor sodium hydrosulfide (NaHS) on adventitious rooting were dose-dependent, and the greatest biological responses were observed under 25μM SNP and 50μM NaHS. The positive effect of NaHS was reversed by the NO scavenger 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), indicating that the H2 S-induced response was partially NO-dependent. Peroxidase (POD), polyphenol oxidase (PPO), and superoxide dismutase (SOD) activities significantly increased by SNP and NaHS treatment, and indoleacetic acid oxidase (IAAO) activity and the O2 - and H2 O2 content significantly decreased by SNP and NaHS treatment. SNP and NaHS treatment also increased the content of soluble sugar and protein and indole-3-acetic acid (IAA). cPTIO significantly mitigated the increases in POD, PPO and SOD activity and soluble sugar, protein and IAA content induced by NaHS. SNP and NaHS upregulated the expression of auxin-related genes (ARF4 and ARF16 ), cell cycle-related genes (CYCD3 , CYCA3 and CDKA1 ), and antioxidant-related genes (TPX2 , SOD and POD ); whereas cPTIO significantly inhibited the increase in the expression of these genes induced by NaHS. Overall, these results show that NO may be involved in H2 S-induced adventitious rooting by regulating the activity of rooting-related enzymes, the expression of related genes, and the content of various nutrients.