The study aimed to investigate the role of hydrogen sulfide (H2S) in regulating chromium stress (Cr–S) tolerance of tomato plants treated with citric acid (CA). Prior to the Cr treatment, tomato plants were foliar-fed with CA (100 μM) daily for 3 days. Subsequently, the plants were grown for another ten days in a hydroponic system in a 50 μM Cr (VI) solution. Chromium treatment reduced photosynthetic pigments and plant biomass, but boosted the levels of hydrogen peroxide (H2O2) malondialdehyde (MDA), H2S, phytochelatins (PCs), and glutathione (GSH), electrolyte leakage (EL), and antioxidant enzyme activity in tomato plants. However, the foliar spray of CA mitigated the levels of H2O2, MDA, and EL, promoted plant growth and chlorophyll content, enhanced antioxidant enzymes’ activities, and increased H2S production in Cr–S-tomato plants. CA also increased the levels of GSH and PCs, potentially reducing the toxicity of Cr through regulated sequestration. Additionally, the application of sodium hydrogen sulfide (NaHS), a donor of H2S, improved CA-induced Cr stress tolerance. The addition of CA promoted Cr accumulation in root cell wall and leaf vacuoles to suppress its toxicity. To assess the involvement of H2S in CA-mediated Cr–S tolerance, 0.1 mM hypotaurine (HT), an H2S scavenger, was provided to the control and Cr–S-plants along with CA and CA + NaHS. HT reduced the beneficial effects of CA by decreasing H2S production in tomato plants. However, the NaHS addition with CA + HT inverted the adverse impacts of HT, indicating that H2S is required for CA-induced Cr–S tolerance in tomato plants.
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