This investigation was applied to find out the seven bread wheat lines that are more tolerant to saline stress (6195 ppm soil and 8934 ppm irrigated water) at Ras Sudr, South Sinai, Egypt. Data indicated that bread wheat lines greatly varied in their ability to grow and subsequently assimilate and translocate biochemical components. Line1 (L1) surpassed the other lines, it was recorded the maximum values of growth, yield, and proline as well as the lowest malondialdehyde content followed by L3 and L7. ISSR-PCR analysis was used for discriminating between lines. A total of 102 DNA bands are detected across the six ISSR primers, out of which only 72 bands (51 non-unique bands and 21 unique bands) are polymorphic with polymorphism percentages (70.58%). In this regard, primer ISSR2 gave the highest polymorphism (92%) followed by ISSR-13 (86%) and ISSR-11 (78%). Also, patterns of proteins (SDS-PAGE) and superoxide dismutase (SOD) have been successfully used in discriminating between salt-tolerant and salt-sensitive lines. Concerning SDS-PAGE, bands number in 7 wheat lines ranged from 13 to 14 with molecular weights ranging between 6 to 130 kDa, and the more intensive bands are presented at 25, 29, and 34 kDa. Moreover, SOD patterns indicated the presence of 5 bands; the major is presented at bands (No. 4 and 5) in all wheat lines. Based on the results obtained, L1 was the best wheat line followed by L3 and L7 which more tolerant to saline stress, and this was associated with increasing growth, yield, and proline as well as decreasing MDA content. In conclusion, SDS-PAGE, SOD, and ISSR analyses are useful biochemical and molecular tools to discriminate different bread wheat lines under saline conditions.