Land plants are continuously exposed to multiple abiotic stress factors like drought, heat, and salinity. Nitric oxide (NO) and hydrogen sulfide (H₂S) are two well-examined signaling molecules that act as priming agents, regulating the response of plants to stressful conditions. Several chemical donors exist that provide plants with NO and H₂S separately. NOSH is a remarkable novel donor as it can donate NO and H₂S simultaneously to plants, while NOSH-aspirin additionally provides the pharmaceutical molecule acetylsalicylic acid. The current study aimed to investigate the potential synergistic effect of these molecules in drought-stressed Medicago sativa L. plants by following a pharmacological approach. Plants were initially pre-treated with both donors (NOSH and NOSH-aspirin) via foliar spraying, and were then subsequently exposed to a moderate water deficit while NO and H₂S inhibitors (cPTIO and HA, respectively) were also employed. Phenotypic and physiological data showed that pre-treatment with NOSH synthetic compounds induced acclimation to subsequent drought stress and improved the recovery following rewatering. This was accompanied by modified reactive-oxygen and nitrogen-species signaling and metabolism, as well as attenuation of cellular damage, as evidenced by altered lipid peroxidation and proline accumulation levels. Furthermore, real-time RT-qPCR analysis revealed the differential regulation of multiple defense-related transcripts, including antioxidant enzymes. Overall, the present study proposed a novel role for NOSH compounds as efficient plant priming agents against environmental constraints through the coordinated regulation of multiple defense components, thus opening new horizons in the field of chemical priming research toward the use of target-selected compounds for stress tolerance enhancement.

Under the era of climate change, plants are forced to survive under increasingly adverse conditions. Application of biostimulants in plants is shown to mitigate the deleterious effects of abiotic stresses including salinity, enhancing plant tolerance and performance. The present study focuses on the effects of five biostimulants based on biocompost and biofertilizer compounds that have been applied to tomato plants grown in the presence (salt-stressed plants) or absence of salt stress (control plants). To study the beneficial effects of the biostimulants in tomato plants, a series of analyses were performed, including phenotypic and agronomic observations, physiological, biochemical and enzymatic activity measurements, as well as gene expression analysis (RT-qPCR) including genes involved in antioxidant defense (SlCu/ZnSOD, SlFeSOD, SlCAT1, SlcAPX), nitrogen (SlNR, SlNiR, SlGTS1) and proline metabolism (p5CS), potassium transporters (HKT1.1, HKT1.2), and stress-inducible TFs (SlWRKY8, SlWRKY31). Among all the biostimulant solutions applied to the plants, the composition of 70% biofertilizer and 30% biocompost (Bf70/Bc30) as well as 70% biocompost and 30% biofertilizer (Bc70/Bf30) formulations garnered interest, since the former showed growth promoting features while the latter displayed better defense responses at the time of harvesting compared with the other treatments and controls. Taken together, current findings provide new insight into the beneficial effects of biostimulants, encouraging future field studies to further evaluate the biostimulant effects in plants under a real environment which is compromised by a combination of abiotic and biotic stresses.

Salinity constitutes one of the most important causes leading to severe reduction in plant yield. Several reports correlate the accumulation of polyamines in plants with tolerance to abiotic stress cues. The present study examined three Medicago truncatula genotypes with differing sensitivities to salinity (TN1.11, tolerant; Jemalong A17, moderately sensitive; TN6.18, sensitive), with the aim of examining the genotype-specific involvement of the polyamine metabolic pathway in plant response to salinity. The study was carried out with leaves harvested 48 h after watering plants with 200 mM NaCl. A comprehensive profile of free polyamines was determined using high performance liquid chromatography. All genotypes showed spermidine and spermine as the most abundant polyamines under control conditions. In salinity conditions, spermine levels increased at the expense of putrescine and spermidine, indicating a drift of polyamine metabolism towards the synthesis of increasing polycationic forms as a stress response. The increasing balance between high and low polycationic forms was clearly diminished in the salt-sensitive genotype TN6.18, showing a clear correlation with its sensitive phenotype. The polyamine metabolic profile was then supported by molecular evidence through the examination of polyamine metabolism transcript levels by RT-qPCR. General suppression of genes that are involved upstream in the PA biosynthetic pathway was determined. Contrarily, an induction in the expression of genes involved in the biosynthesis of spermine and spermidine was observed, in agreement with the metabolic analysis. A significant induction in diamino oxidase expression, involved in the catabolism of putrescine, was specifically found in the sensitive genotype ΤΝ6.18, indicating a distinct metabolic response to stress. Present findings highlight the involvement of polyamines in the defense response of Medicago genotypes showing sensitivity to salt stress.