RBOH1-dependent H2O2 mediates spermine-induced antioxidant enzyme system to enhance tomato seedling tolerance to salinity-alkalinity stress.

Salinity-alkalinity stress is a limiting factor in tomato production in the world. Plants perceive salinity-alkalinity stress by activating signaling pathways to increase plant tolerance (Xu et al., 2020). Here, we investigated whether spermine (Spm) induces respiratory burst oxidase homolog 1 (RBOH1) and hydrogen peroxide (H2O2) signaling in response to salinity-alkalinity stress in tomato. The results showed that exogenous Spm induced the expression of RBOH1 and the accumulation of H2O2 under normal condition. Accordingly, we tested the function of H2O2 signal in tomato seedlings and found that exogenous H2O2 increased the expression levels of Cu/Zn-superoxide dismutase (Cu/Zn-SOD), catalase 1 (CAT1), cytosolic ascorbate peroxidase (cAPX), and glutathione reductase 1 (GR1) and the activities of SOD (EC 1.15.1.1), CAT (EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), and GR (EC 1.6.4.2) in tomato seedlings under salinity-alkalinity stress. DMTU increased the malondialdehyde (MDA) content and relative electrical conductivity, and the relative water content (RWC), and accelerated leaf yellowing in tomato seedlings under salinity-alkalinity stress, even though we sprayed Spm on tomato leaves. We also found that RBOH1 silencing decreased the expression levels of Cu/Zn-SOD, CAT1, cAPX, and GR1 and the activities of SOD, CAT, APX, and GR when tomato seedlings were under salinity-alkalinity stress. Exogenous Spm did not increase RWC and decrease MDA content in RBOH1 silencing tomato seedlings under salinity-alkalinity stress.

Exogenous spermine-induced expression of SlSPMS gene improves salinity-alkalinity stress tolerance by regulating the antioxidant enzyme system and ion homeostasis in tomato.

The study tested the function of exogenous spermine (Spm) in resisting salinity-alkalinity stress in tomato seedlings and found that tomato Spm synthase gene (SlSPMS) was involved in this regulation. The tomato seedlings cultivated in normal conditions or salinity-alkalinity conditions were irrigated with 100 ml one strength Hoagland nutrient solution 100 ml mixed solution (5 ml 300 mmol/L NaCl, 45 ml 300 mmol/L Na2SO4, 45 ml 300 mmol/L NaHCO3, and 5 ml 300 mmol/L Na2CO3 (pH = 8.90)) every 2 days, respectively. The 0.5 mM Spm pretreatment improved superoxide dismutase (SOD; EC 1.15.1.1) activity, catalase (CAT; EC 1.11.1.6) activity, ascorbate peroxidase (APX; EC 1.11.1.11) activity, and glutathione reductase (GR; EC 1.6.4.2) activity and decreased endogenous hydrogen peroxide (H2O2) content, malondialdehyde (MDA) content, and relative electrical conductivity (REC) in tomato leaves. Na+ content declined and K+ concentration rose in tomato seedlings when pre-treated with Spm. However the results showed that under salinity-alkalinity stress, silencing of SlSPMS with virus-induced gene silencing had lower antioxidant enzyme activities and higher Na+ content and lower K+ content than normal tomato seedlings, meaning that they had low salinity-alkalinity tolerance. Exogenous Spm could not reconstruct the tolerance to salinity-alkalinity stress in SlSPMS gene-silencing tomato seedlings. Taken together, exogenous Spm could induce the expression level of SlSPMS, which regulated the antioxidant enzyme system and ion homeostasis in tomato seedlings living in salinity-alkalinity environment, thereby improving the ability of tomato seedlings to resist salinity-alkalinity stress.