Putrescine regulates stomatal opening of cucumber leaves under salt stress via the HO-mediated signaling pathway.

The stomatal aperture is imperative for photosynthesis in higher plants. The function of polyamines (PAs) in stomatal regulation under a stressful environment has not been fully determined. In this study, we demonstrated the mechanism by which putrescine (Put) regulates stomatal changes in cucumber leaves under salt stress. The results showed that foliar application of Put alleviated the decrease of stomatal aperture and photosynthesis caused by salt stress and promoted plant growth. Exogenous Put caused a significant increase in endogenous PAs and hydrogen peroxide (HO) levels by 105.43% and 27.97%, respectively, while decreased abscisic acid (ABA) content by 67.68% under salt stress. However, application of inhibitors of aminoguanidine hydrochloride (AG), 1, 8-diaminooctane (1, 8-DO), diphenyleneiodonium chloride (DPI) and salicylhydroxamic acid (SHAM) upregulated the 9-cis-cyclocarotenoid dioxygenase (NCED) gene and downregulated the reduced glutathione synthetase (GSHS) gene. These inhibitors also decreased the stomatal aperture, levels of HO and reduced glutathione (GSH), but increased the ABA content under salt stress and Put treatment conditions. The order of influence is AG > 1, 8-DO > DPI > SHAM. However, Put-induced downregulation of ABA content and upregulation of GSH content under salt stress were effectively blocked by N, N'-dimethylthiourea (DMTU, HO scavenger) and 1-chloro-2,4-dinitrobenzene (CDNB, GSH scavenger) treatments. Taken together, these results suggest that Put induced the formation of HO signaling mediates the degradation of PAs by diamine oxidase (DAO), increasing GSH content and inhibiting the accumulation of ABA in leaves, thereby promoting stomatal opening in salt-stressed cucumber leaves.