Phylogenetic relationships between ten Hordeum vulgare L. , seven cultivars (Giza 123, Giza 126, Giza 127, Giza 128, Giza 129, Giza 130 and Giza 2000) and three landraces from Sinai (El-Kheroba, El-Sheikh Zuwaid, and Wadi Sedr) were carried out using two molecular genetic markers (RAPD-PCR and ISSR-PCR). The genetic distance between Ten genotype was also estimated from banding patterns Twenty two random primers were used in RAPD revealed 316 bands while 159 bands were detected of ISSR analysis using 10 primers. RAPD analysis among ten genotypes showed 41.77% polymorphism, while ISSR analysis showed 62.02% polymorphism.
           It was found that, ISSR was a more viable marker than RAPD in the detection of the
 genetic variability among Hordeum vulgare (barley) varieties. The genetic distance tree was detected using UPGMA based on both molecular markers (RAPD and ISSR) and analysis of combined data.In addition, the Band Shearing index (BSI) factor was calculated shows a marked difference between the ten genotypes, seven cultivars and three landraces of studied  Hordeum vulgare where BSI average reach 1.42% with RAPD markers while reaching 0.93 % in ISSR. The obtained data indicated that both RAPD and ISSR markers are efficient in identification and differentiation between selected taxa, but the efficiency of ISSR was the best one. Also, the present results, enhancing the available knowledge of  Hordeum vulgare genetic resources in Egypt, which may contribute to their conservation and utilization in breeding programs.