Abstract <p>Salinity is a major global issue that significantly alters plant growth and productivity by disrupting the physiological and metabolic processes of plants, leading to reduced wheat (<i>Triticum aestivum</i> L<i>.</i>) yield. In this study, the parent variety ‘Berbet’, six selected salt-tolerant ‘Berbet’ Introgression Lines (L1, L6, L10, L45, L47, and L72), along with two salt-tolerant genotypes (KRL 210, PBW 803) and one salt-susceptible genotype (PBW725), were evaluated under saline conditions (EC = 5.8–6.2 dSm⁻¹). The objective was to investigate key physiological and biochemical traits associated with salt tolerance and to identify reliable screening markers that differentiate tolerant from non-tolerant genotypes. This approach is expected to accelerate the identification of salt-tolerant cultivars and support breeding programs aimed at developing resilient wheat varieties. We assessed stomatal characteristics (stomatal density and stomatal aperture size), leaf thickness, total chlorophyll content, chlorophyll fluorescence, relative water content, membrane stability index, malondialdehyde content, Na⁺ and K⁺ contents (including the Na⁺/K⁺ ratio in roots and shoots), osmolytes (proline, free amino acids, glycine betaine, and trehalose), and the activities of antioxidant enzymes (superoxide dismutase, catalase, and ascorbate peroxidase). The experiment was arranged in a completely randomized block design with three replicates. ANOVA revealed differential responses among ‘Berbet’ Introgression Lines, highlighting that salt tolerance arises from the integration of traits into a coordinated stress-response network. Genotypes L47 and L72 demonstrated superior stomatal regulation, antioxidant defense, and osmotic adjustment, marking them as elite candidates for saline environments.</p>