Background: Salinity is one of the most challenging abiotic stresses restricting the growth of plants. In vitro screening will increase the efficiency and speed of salinity tolerant genotypes identifications. The response of four tomato cultivars under salinity was analyzed in vitro to evaluate the seedlings growth, biochemical, and gene expression responses as well as the effect of nano zinc and iron on callus induction and plant regeneration. Results: The results showed that an increase in salinity stress in the medium decreased the germination percentage, fresh and dry weight of shoot, root length, chlorophyll a, b and carotenoids content, K and Ca content, and on the other hand, Na content was increased. MDA content (‘Nora’, ‘PS-10’, ‘Peto’ and ‘Roma’: 1.71, 1.78, 1.66 and 2.16 folds, respectively), electrolyte leakage (‘PS-10’: 33.33%; ‘Roma’: 56.33%), were increased with salinity of 100 mM compared to control. Proline content was increased in 50 mM NaCl (10.8 fold). The most activity of antioxidant enzymes including CAT, SOD, APX, GPX, and GR was observed in the ‘PS-10’ cultivar, and the lowest activity of these enzymes was observed in ‘Roma’ under salinity stress. The AsA and GSH were decreased and DHA and GSSG were increased with the increased intensity of salinity. The relative expression of SOD, APX, and GR genes varied in different cultivars at different salinity concentrations. The most percentage of callus induction was observed with applying iron oxide nanoparticles, and the most regeneration rate was recorded using zinc oxide nanoparticles. Conclusion: The results showed that salt-tolerant cultivars such as ‘PS-10’ with better osmotic adjustment, are suitable candidates for the future production and breeding programs.
