Interactive effect of silicon and nitric oxide effectively contracts copper toxicity in Salvia officinalis L.

Excess copper (Cu) causes the toxic effects in plants and health hazards to humans. Therefore, in this study, the effect of sodium silicate (1 mM Si) and sodium nitroprusside (200 µM SNP as a releasing NO), was assessed on Cu tolerance in Salvia officinalis L. plants exposed to 400 µM CuSO4. Results revealed that the combined supplementation with Si and SNP rather than the single application of these chemicals lowered Cu concentrations and translocation factor and increased Mg, Zn, and Fe concentrations in roots and shoots. Furthermore, combined treatment more efficiently decreased electrolyte leakage enhanced the activities of POD and APX in the leaves and roots, and improved relative water content and the content of Chl. a and Chl. b in leaves and consequently further increased tolerance index. Silicon supply enhanced NO content and applying Si + SNP more than the treatment of Si alone increased Si concentrations in the roots and shoots under Cu stress. Therefore, the reciprocal interaction of Si and NO might enhance Cu tolerance in plants, and the combined application of Si and SNP might be a promising strategy to decrease heavy metal accumulation in medicinal plants grown in polluted lands.

In most studies, co-precipitation of silicon and heavy metals in medium has been suggested as a reason for reducing heavy metal uptake in plants. In this study, the impact of Si on NO generation and the role of NO signaling in regulating Cu uptake and translocation and defensive responses were assessed to clarify another mechanism of Si in inducing Cu tolerance in sage. Furthermore, the combined application of Si and SNP has been indicated as an innovative strategy to enhance Cu tolerance and decrease heavy metal accumulation in medicinal plants grown in polluted lands.

Silicon and nitric oxide synergistically modulate the production of essential oil and rosmarinic acid in Salvia officinalis under Cu stress.

The individual impact of silicon (Si) and nitric oxide (NO) on secondary metabolism in several plant species has been reported, but their combined effect has never been evaluated yet. Therefore, in this study, single and combined impacts of NO and Si on the biosynthesis of rosmarinic acid (RA) and essential oil (EO) content in leaves of Salvia officinalis were investigated under both non-stress and Cu stress conditions. The results indicated that high Cu concentration decreased biomass and the content of polyphenols, but elevated electrolyte leakage, while lower Cu concentrations, especially 200 µM Cu, increased the content of polyphenols, EO, and antioxidant capacity in leaves of S. officinalis. The foliar application of sodium silicate (1 mM Si) and sodium nitroprusside (200 µM SNP as a NO donor) alone and particularly in combination improved shoot dry biomass, restored chlorophyll and carotenoids, increased EO content, the amounts of flavonoids, and phenolic compounds especially RA, and enhanced antioxidant capacity in the leaves of S. officinalis under both non-stress and Cu stress conditions. Copper treatment increased NO content, upregulated expression of PAL, TAT, and RAS genes, and enhanced phenylalanine ammonia-lyase activity in the leaves, which were responsible for improving the production of phenolic compounds, particularly rosmarinic acid. Foliar spraying with Si and SNP intensified these attributes. All responses were more pronounced when NO and Si were simultaneously applied under Cu stress. These findings suggest that NO and Si synergistically modulate secondary metabolism through upregulation of related gene expression and enzyme activities under both non-stress and Cu stress conditions.

Silicon- induced nitric oxide burst modulates systemic defensive responses of Salvia officinalis under copper toxicity.

In this study, the role of nitric oxide (NO) burst in modulating Si-induced defensive responses in leaves and roots of Salvia officinalis under copper (Cu) stress were investigated. The result showed that 400 µM Cu markedly reduced shoot dry weight, but increased electrolyte leakage (EL) in leaves and both Si and sodium nitroprusside (SNP as the NO donor) improved these attributes in a dose-dependent manner. Interestingly, Cu toxicity systemically boosted a NO burst in both roots and shoots and applying Si and SNP markedly intensified it. The application of Si and SNP alone as well as their combination improved growth parameters and systemically alleviated Cu-induced lipid peroxidation and H2O2 accumulation through lowering Cu accumulation, increasing proline content, enhancing the activities of catalase (CAT) and superoxide dismutase (SOD) in both roots and leaves and up-regulating expression of SOD gene in leaves of S. officinalis. NO generation was substantially arrested and the responses induced by Si were significantly suppressed by pretreatment with 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxy l-3-oxide (cPTIO) as a NO scavenger, Nx-Nitro- L-arginine methyl ester hydrochloride (L-NAME) as a nitric oxide synthase inhibitor, and tungstate as a nitrate reductase inhibitor. These novel results indicate that Si can induce Cu tolerance through triggering NO generation which systemically modulates defensive reactions in both roots and leaves of Salvia officinalis.