Trees of Scots pine (Pinus sylvestris L.) are known for their effective phytoextraction capabilities. The results obtained in this study point to the significant role of substrate composition and chemical characteristics in the phytoextraction potential of this species. A multi-elemental (53 elements) analysis of pines from unpolluted (soil) and polluted (post-flotation tailings) sites was performed using inductively coupled plasma optical emission spectrometry. The analyzed flotation tailings were characterized by alkaline pH (7.19 ± 0.06) and significantly higher conductivity (277.7 ± 2.9 µS cm-1) than the soil (pH = 5.11 ± 0.09; 81.3 ± 4.9 µS cm-1). The two substrates also differed with respect to the contribution of the clay fraction (0% in the unpolluted and 8% in the polluted substrate). The specimens of P. sylvestris growing on flotation tailings had significantly smaller height (381 ± 58 cm) and total aboveground biomass (4.78 ± 0.66 kg) than the trees growing in soil (699 ± 80 cm and 10.24 ± 2.10 kg). The biomass of the trunk, twigs and branches, and needles of the trees from polluted sites was between 40.0% and 48.7% of the biomass of the same organs of the control trees. Generally, the organs (trunk, twigs and branches, needles) of the P. sylvestris specimens from polluted sites had significantly higher concentrations of Au, Al, Ba, Cd, Co, La, Lu, Ni, Pd, Sc, Zn, and lower concentrations of B, Bi, Ca, Ce, Er, In, K, Mg, Na, Nd, P, Pr, Re, Se, Sr, Te than in the control plants, these metals being accumulated effectively in the whole of the aboveground biomass (BCF>1). Although the concentration of the majority of elements was significantly higher in the flotation tailings, significantly higher concentrations of these elements were observed in the tree organs from unpolluted sites, which points to the important role of substrate characteristics in the phytoextraction efficiency of P. sylvestris.