Silene vulgaris is a pseudometallophyte that spontaneously occurs in various ecological niches. Therefore, three ecotypes of this species representing calamine (CAL), serpentine (SER), and non-metallicolous (NM) populations were investigated in this study. Owing to the presence of Pb or Ni ions in natural habitats from metallicolous populations originated, we used these metals as model stressors to determine the survival strategy of tested ecotypes and analyze metal distribution at various levels of organism organization. We focused on growth tolerance, non-enzymatic antioxidants, and photosynthetic apparatus efficiency as well as anatomical and ultrastructural changes occurred in contrasting ecotypes exposed in vitro to excess amounts of Pb2+ and Ni2+. Although Ni application contributed to shoot culture death, the study revealed that the mechanisms of Pb detoxification differed between ecotypes. The unspecific reaction of both metallicolous specimens relied on the formation of effective mechanical barrier against toxic ion penetration, while the Pb appearance in the protoplasts led to the activation of ecotype-specific intracellular defense mechanisms. Hence, the response of CAL and SER ecotypes was almost unchanged under Pb treatment, whereas the reaction of NM one resulted in growth disturbances and physiological alternations. Moreover, both metallicolous ecotypes exhibited increase generation of reactive oxygen species (ROS) in leaves, even before the harmful ions got into these parts of plants. It may implicate the potential role of ROS in CAL and SER adaptation to heavy metals and, for the first time, indicate on integral function of ROS as signaling molecules in metal-tolerant species.
Keywords: Heavy metal toxicity; Microscopic imaging; Non-enzymatic antioxidants; Photosynthetic apparatus; ROS metabolisms; Shoot culture.