Hydrogen sulfide is required for salicylic acid-induced chilling tolerance of cucumber seedlings

Salicylic acid (SA) and hydrogen sulfide (H₂S) have been proved to be multifunctional signal molecules to participate in the response of plants to abiotic stresses. However, it is still unclear whether there is interaction between SA and H₂S in response to chilling intensity of cucumber seedlings. Here, we found SA was sensitive to chilling intensity. Under normal condition, NaHS (H₂S donor) or removing endogenous H₂S with hypotaurine (HT, a specific scavenger of H₂S) and DL-propargylglycine (PAG, a specific inhibitor of H₂S) has no effect on endogenous SA level; however, SA induced endogenous H₂S content and activated the activities and mRNA level of L-/D-cysteine desulfhydrase (L-/D-CD), and inhibiting endogenous SA with paclobutrazol (PAC) or 2-aminoindan-2-phosphonic acid (AIP) blocked this effect, implying H₂S may play a role after SA signal. Further studies showed that both SA and NaHS notably alleviated chilling injury, which was evidenced by lower electrolyte leakage (EL), MDA content, and ROS accumulation, compared with H₂O treatment. Of note, SA and H₂S improved the activities and mRNA level of antioxidant enzymes (SOD, POD, CAT, APX, and GR) as well as the contents of AsA and GSH. Additionally, the chilling-response genes (ICE, CBF1, and COR) were obviously upregulated by exogenous SA and NaHS. However, the positive effect of SA on chilling tolerance was inhibited by HT, whereas PAC or AIP did not affect NaHS-induced chilling tolerance. Taken together, the data reveals that H₂S acts as a downstream signal of SA-induced chilling tolerance of cucumber via modulating antioxidant system and chilling-response genes.