phylogenetic relationships of eleven accessions of (Hordeum vulgare L.) collected from different region of Egypt were assessed. Fifteen quantitative morphological traits were used, the measured data were evaluated statistically using ANOVA, phylogenetic tree were constructed using UPGMA. Also Inter simple sequence repeat (ISSR) molecular marker technique were used for DNA fingerprinting and assessing genetic diversity and phylogenetic relationships in barley germplasm. The results showed that ISSR primers produced 140 bands their size ranged between 110-1600 bp with 39.8% polymorphism percentage. Polymorphic information content PIC was 0.74 for ISSR. UPGMA dendrogram was divided into two clusters by morphological traits and ISSR analysis. Genetic similarity matrix was examined with Jacard's coefficient, maximum similarity was found between B8 and B7 (98%) with morphological analysis both from (North Sinai) and between B9 and B10 (96%) with ISSR analysis. Determination of genetic diversity between barley is of major importance for characterization of barley germplasm, breeding programs and conservation purposes. Morphological traits and ISSR analysis are effective tools for detecting genetic variations. The results showed that H. vulgare have high ratio of variation. This study may be considered as reference study for further studies on H. vulgare and may contribute to species concept and breeding programs.