In breeding programs, classification of genotypes and genetic information within or between species of crop plants were useful for any selection and hybridization perspectives. In the present study 17 barley lines and cultivars were used for studing seed storage protein by electrophoresis technique and some physiological traits. Based on ANOVA results, studding genotypes had significant differences at level of P ≤0.01. The resulting dendrogram using WARD method indicated that the studding lines could be divided into three groups. First group lines had a high Fv/Fm value. Second group contained lines numbered 10, 11, 12 and 8, had a low chlorophyll content values. The results of electrophoresis profile showed 11 protein bands with maximum in number 2 and 17 lines (10 bands) and minimum in number 6 and 16 lines (1 band). Based on jaccard’s similarity coefficient, There was the most similarity (or the least difference) among number 8 and 10 lines (1), 2 and 17 (1), 11 and 17 (0.9), 11 and 2 (0.9) and the least similarity (or highest difference) between number 1 and 3 lines (0), 3 and 6 (0), 3 and 8 (0), 3 and 10 (0), 3 and 16 (0), 4 and 6 (0), 6 and 8 (0), 6 and 10 (0) and 6 and 16 (0) respectively. Also principal coord inate analysis confirmed that 80.9 percent of total variance of barley storage proteins was explained by the first three components. Finally it is clear that genetic variance was existed between studding lines