A two-year field screening analysis was carried out in of Nubaria Agricultural Research Station during the two growing seasons 2015/ 2016 and 2016/2017 to determine the genetic diversity and relationships among 19 Egyptian barley cultivars using multivariable analysis for some physiological, quality, agronomic characters, grain yield and Sequence-Related Amplified Polymorphism (SRAP) marker analysis. Variance analysis of the traits showed that there are significant differences among the genotypes under study with respect to all traits. Principal component analysis was carried out for all traits under study clarifying about 53.6% of total variance. Cluster analysis revealed four distinct groups which were clustered according to their morphological traits Similarity levels among the pairs of cultivars ranged between 39.77 and 98.36% .Seven SRAP combination primers were used to assess the genetic variation among all cultivars. The primers showed that the average percentage of polymorphic loci was 65.2% and the average band number amplified from each pair of primers was 5.14 bands, of which included 6.2 polymorphic bands. Highest PIC was related to primer me6+em5 was (0.94) indicating that this primer is highly informative. The average percentage of polymorphic loci for all cultivars was 79.3%. The average of both genetic diversity indicters such as effective number of alleles and Shannon’s diversity index were (1.205 and 3.802) respectively. The dendrogram of SRAP markers had clustered all the Egyptian cultivars into four groups each group include the most closed cultivars together with genetic similarity coefficients (GSC) ranging from (0.66) to (1.00). The results of the present study showed that there were high genetic differences among Egyptian barley cultivars which provide new information about the relationships among Egyptian barley cultivars which are useful for cultivar identification and for their utilization in further barley breeding programs.