Genetic diversity among some wild close relatives of wheat was estimated using inter simple sequence repeats (ISSR) and SDS-PAGE markers. Four Triticum, Secale cereale and Triticale genotypes including seven Triticum, Secale and Triticale accessions belonging to one diploid (Secale cereale), three tetraploid (Triticum durum (4x)) and two hexaploid (Triticale (6x)) in addition to one hexaploid wheat (Triticum aestivum) species sampled from different ecogeographical regions of Egypt; a durum wheat cultivars 'Baniswef 1 and 3 and Sohag 3', a local wheat cultivars growing in upper Egypt 'Giza 168', a bread wheat cultivar and two triticale strains beside one rye strain were evaluated. Genetic diversity among wheat accessions was estimated using 11 ISSR primers. Of the approximately 153 detected ISSR markers, 65 (42%) were polymorphic with 4 bands per used primer pair. Cluster analysis of 7 accessions belonging to the four species by UPGMA cluster analysis based on Jaccard's similarity estimates for ISSR data divided all accessions into one major cluster and Secale cereale separated from the major cluster. two major sub-clusters reflecting almost their genome composition. The first one included wheat species having AB genomes ('Baniswef 1 and 3 and Sohag 3'), while second cluster included wheat species having ABD and ABR genomes. The genetic similarity coefficients ranged from 0.36 between Triticale 1 and rye accession and 0.84 between Triticale 1 (6x) and Triticale 2 (6x). Two Triticum durum species of were ranked as the second most related species (0.81). The results of SDS-PAGE revealed a total number of 145 bands with molecular weights (MW) ranging from about 17 kDa to 158 kDa, which were not necessarily present in the 7 genotypes. Data revealed seven common bands (monomorphic), six unique bands while the remaining 90 band were polymorphic with 62% polymorphism. The unique Bands at the molecular weight 158 and 41 kd were the distinguishable band of Baniswef 3. The unique Band at the molecular weight 64 kd was the distinguishable band detected of Sohag 3.  The unique Bands at the molecular weight 62 and 55 kd were detected only in Rye. The unique Band at the molecular weight 17 kd was detected only in Baniswef 1.