Rosa damascena is one of the most important medicinal and ornamental plants in Iran which is tolerant of salinity to some extent. However, the selection of genotypes that are more tolerant to salinity will influence on Damask cultivation in salt stress-affected regions. For this purpose, a factorial experiment in a completely randomized design with three replicates was performed under in vitro conditions on four Damask rose genotypes (Atashi, Bi-Khar, Chahar-Fasl and Kashan) at 5 concentrations of NaCl (0, 25, 50, 75, and 100 mM), and the physico-chemical traits were measured 14 and 28 days after treatment.The results showed that Atashi genotype with high levels of Chl a, Chl b, total Chl content, carotenoids, relative leaf water content, proline, total soluble protein, TPC, TFC, TAA, and the highest increase in the activity of antioxidant enzymes such as GPX, APX, CAT, SOD, and POD as well as the lowest amount of hydrogen peroxide showed a better protection mechanism against oxidative damage than the other three genotypes (Bi-Khar, Chahar-Fasl and Kashan) in the 14th and 28th days by maintaining the constructive and induced activities of antioxidant enzymes, it was shown that Bi-Khar genotype had moderate tolerance and Kashan and Chahar-Fasl genotypes had low tolerance to salinity stress. In vitro selection methods can be used effectively for salt tolerant screening of Damask rose genotypes, although the same experiment should be conducted in open filed cultures to verify the in vitro experimental results.