Distinct co-tolerance responses to combined salinity and cadmium exposure in metallicolous and non-metallicolous ecotypes of Silene vulgaris

This study compared co-tolerance to salinity and cadmium and investigated its mechanisms in a facultative metallophyte Silene vulgaris originating from distinct habitats. Shoots of calamine (Cal) and non-metallicolous (N-Cal) ecotypes grown in vitro were exposed to 10 and 100 mM NaCl, 5 μM CdCl₂ and their combinations. Stress effects were evaluated based on growth, oxidative stress parameters, and DNA content and damage. Tolerance mechanisms were assessed by analyzing non-enzymatic antioxidants, osmolytes and ion accumulation. Irrespective of the ecotype, Cd stimulated shoot proliferation (micropropagation coefficients MC = 15.2 and 12.1 for Cal and N-Cal, respectively, growth tolerance index GTI = 148.1 and 156.7%). In Cal ecotype this was attributed to an increase in glutathione content and reorganization of cell membrane structures under Cd exposure, whereas in N-Cal to enhanced synthesis of other non-enzymatic antioxidants, mainly carotenoids and ascorbate. Low salinity stimulated growth of Cal ecotype due to optimizing Cl^- content. High salinity inhibited growth, especially in Cal ecotype, where it enhanced DNA damage and disturbed ionic homeostasis. Species-specific reaction to combined salinity and Cd involved a mutual inhibition of Na⁺, Cl^- and Cd²⁺ uptake. N-Cal ecotype responded to combined stresses by enhancing its antioxidant defense, presumably induced by Cd, whereas the metallicolous ecotype triggered osmotic adjustment. The study revealed that in S. vulgaris Cd application ameliorated metabolic responses to simultaneous salinity exposure. It also shed a light on distinct strategies of coping with combined abiotic stresses in two ecotypes of the species showing high plasticity in environmental conditions.

Keywords: Calamine ecotype; Cross-tolerance; Heavy metal; Multi-stress; Salinity.