Summary: Hydrogen peroxide (H₂O₂) and nitric oxide (˙NO) are key reactive species in signal transduction pathways leading to activation of plant defense against biotic or abiotic stress. Here, we investigated the effect of pre‐treating citrus plants (Citrus aurantium L.) with either of these two molecules on plant acclimation to salinity and show that both pre‐treatments strongly reduced the detrimental phenotypical and physiological effects accompanying this stress. A proteomic analysis disclosed 85 leaf proteins that underwent significant quantitative variations in plants directly exposed to salt stress. A large part of these changes was not observed with salt‐stressed plants pre‐treated with either H₂O₂ or sodium nitroprusside (SNP; a ˙NO‐releasing chemical). We also identified several proteins undergoing changes either in their oxidation (carbonylation; 40 proteins) and/or S‐nitrosylation (49 proteins) status in response to salinity stress. Both H₂O₂ and SNP pre‐treatments before salinity stress alleviated salinity‐induced protein carbonylation and shifted the accumulation levels of leaf S‐nitrosylated proteins to those of unstressed control plants. Altogether, the results indicate an overlap between H₂O₂‐ and ˙NO‐signaling pathways in acclimation to salinity and suggest that the oxidation and S‐nitrosylation patterns of leaf proteins are specific molecular signatures of citrus plant vigour under stressful conditions.

Summary: Reactive oxygen and nitrogen species are involved in a plethora of cellular responses in plants; however, our knowledge on the outcomes of oxidative and nitrosative signaling is still unclear. To better understand how oxidative and nitrosative signals are integrated to regulate cellular adjustments to external conditions, local and systemic responses were investigated in the roots and leaves of sour orange plants (Citrus aurantium L.) after root treatment with hydrogen peroxide (H₂O₂) or sodium nitroprusside (a nitric oxide donor), followed by NaCl stress for 8 days. Phenotypic and physiological data showed that pre‐exposure to these treatments induced an acclimation to subsequent salinity stress that was accompanied by both local and systemic H₂O₂ and nitric oxide (NO) accumulation. Combined histochemical and fluorescent probe approaches showed the existence of a vascular‐driven long‐distance reactive oxygen species and NO signaling pathway. Transcriptional analysis of genes diagnostic for H₂O₂ and NO signaling just after treatments or after 8 days of salt stress revealed tissue‐ and time‐specific mechanisms controlling internal H₂O₂ and NO homeostasis. Furthermore, evidence is presented showing that protein carbonylation, nitration and S‐nitrosylation are involved in acclimation to salinity stress. In addition, this work enabled characterization of potential carbonylated, nitrated and nitrosylated proteins with distinct or overlapping signatures. This work provides a framework to better understand the oxidative and nitrosative priming network in citrus plants subjected to salinity conditions.