Hydrogen sulfide regulates the activity of antioxidant enzymes through persulfidation and improves the resistance of tomato seedling to Copper Oxide nanoparticles (CuO NPs)-induced oxidative stress

Hydrogen sulfide (H2S), a small gaseous signaling molecule, regulates antioxidase activity and improves plant tolerance to oxidative stress. The phytotoxic effect of Copper Oxide (II) nanoparticles (CuO NPs) is due to oxidative stress. Here, we show that H2S-mediated persulfidation of antioxidase is essential for an effective stress response of tomato exposed to CuO NPs. The CuO NP-induced increase in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was significantly reduced by treatment with the H2S donor NaHS. In vivo, NaHS increased superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POD) activities under CuO NP stress. In vitro, NaHS increased APX and POD activities but decreased catalase (CAT) activity. Persulfidation existed in recombinant SlCAT1, SlcAPX1 and SlPOD5 proteins. The persulfidatied cysteine (Cys) residues were verified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), revealing their position on the protein surface. Cys234 of SlCAT1 is located in the immune-responsive domain and close to the enzyme activity domain. Cys234 of SlcAPX1 and Cys 61 SlPOD5 are located in the enzyme activity domain. Persulfidation increased SlcAPX1 and SlPOD5 activities but decreased SlCAT1 activity. These data indicate that H2S-mediated persulfidation posttranslationally regulates the activities of CAT, APX and POD, thereby enhancing the plant's response to oxidative stress. Additionally, this work provides an experimental approach for the study of persulfidation in plants.