Beneficial behavior of nitric oxide in copper-treated medicinal plants

Highlights: • Endogenous NO and ROS accumulation were inversely related. • Selected amino acids in the roots were increased by SNP. • NO induced regulation of phenolic metabolism for protection against Cu toxicity. • SNP improved the vincristine, vinblastine and total alkaloid contents in Cu-treated plants. - Abstract: Despite numerous reports implicating nitric oxide (NO) in the environmental-stress responses of plants, the specific metabolic and ionic mechanisms of NO-mediated adaptation to metal stress remain unclear. Here, the impacts of copper (Cu) and NO donor (SNP, 50 μM) alone or in combination on the well-known medicinal plant Catharanthus roseus L. were investigated. Our results showed that Cu markedly increased Cu²⁺ accumulation, decreased NO production, and disrupted mineral equilibrium and proton pumps, thereby stimulating a burst of ROS; in addition, SNP ameliorates the negative toxicity of Cu, and cPTIO reverses this action. Furthermore, the accumulations of ROS and NO resulted in reciprocal changes. Interestingly, nearly all of the investigated amino acids and the total phenolic content in the roots were promoted by the SNP treatment but were depleted by the Cu + SNP treatment, which is consistent with the self-evident increases in phenylalanine ammonia-lyase activity and total soluble phenol content induced by SNP. Unexpectedly, leaf vincristine and vinblastine as well as the total alkaloid content (ca. 1.5-fold) were decreased by Cu but markedly increased by SNP (+38% and +49% of the control levels). This study provides the first evidence of the beneficial behavior of NO, rather than other compounds, in depleting Cu toxicity by regulating mineral absorption, reestablishing ATPase activities, and stimulating secondary metabolites.