Samples of 30 plants were used for the molecular analyses: eight from SWE, eight from SOB, eight from PRA and six from NOR. Genomic DNA was isolated from dry leaves using about 20-50 mg of plant material, following the protocol of the genomic DNA purification kit (Thermo Scientific™, Waltham, MA, USA). We amplified three noncoding plastid DNA regions: psbD-trnT spacer, rpL32-trnL spacer and trnL-trnF spacer previously used for testing the variability of these regions in three different angiosperm lineages [Atropa vs. Nicotiana,Lotus vs. Medicago and Saccharum vs. Oryza; Shaw et al. (2007)] and for testing the genetic diversity ofSilene acaulis (Gussarova et al. 2015). The primers were specifically designated for V. alpina from the 10 most similar sequences in GenBank [including Lychnis alpina (synonymV. alpina), Atocion rupestre Silene acaulis,S. aprica, S. capitata and other Silene species; Table 1]. PCR was performed in 25-µL volume with 2 µL template DNA, 7.5 µL distilled H2O, 2 µL 25 mM MgCl2, 12.5 µL VWR Red Taq DNA Polymerase Master Mix containing the Taq polymerase (VWR Life Science, Milan, Italy) and 0.5 µL (10 pmol/µL) of each primer. The PCR reaction was performed using a BioRad T100 Thermal Cycler as follows: 94°C, 3 min; 40 cycles (94°C, 30 s; annealing temperature, 1 min; 72°C, 1 min); 72°C, 5 min. The annealing temperatures were 55°C for psbD-trnT, 51°C for rpL32-trnL and 56°C for trnL+ trnL-F.