This study aimed to determine the genetic diversity and relationships among barley varieties (Hordeum vulgare L.) growing at Wollo Highland areas by using hordein and agro-morphological traits. Twenty (20) varieties were laid down in randomized complete block design (RCBD) design with three replications; they were planted by irrigation at Wollo University, Dessie Campus from January to May 2014. The genetic analysis using hordein was done in the laboratory of Ethiopian Biodiversity Institute (EBI) in July 2014. Ten (10) competitive random plants from the rows of the experimental plots were taken for recording their agromorphological characters. Electrophoretic separation of barley storage proteins or hordeins was done using acid polyacrylamide gel electrophoresis (A-PAGE). The traits: day of heading, day of maturity, grain yield (kg/ha), plant height, spike length, number of spiklet per spike, kernel number per spike, weight of seed per spike and biomass yield (g/plot) were highly significant for the diversity of barely, whereas thousand seed weight was less significant. The results reveal positive correlation between spike length and number of spiklet per spike (the highest correlations from the agro morphological traits); the next highly correlated traits were kernel weight per spike and thousand seed weight. The A-PAGE analysis showed limited variation among the analysed accessions. The Nei’s genetic distance for all varieties of barely varied from 0.0000 to 1.6094. It is found that the 20 genotypes of barely investigated in this research were having a gene diversity (h) of overall populations (0.138) using hordein. The cluster analysis grouped the 20 barely genotypes into three different clusters using agro-morphological traits and into four clusters using hordein. This indicates the presence of wide diversity among the tested genotypes. From cluster mean values of agro-morphological traits, genotypes in cluster III deserve consideration for directly developing high yielding barely varieties. The result of the principal components analysis revealed that the first three principal components having greater than 1 eigenvalue contributed 84.22% of the total variation. From this study, it can be concluded that the presence of high morphological variation indicated the potential of Wollo Highland areas in contributing to barley improvement and conservation activities of land areas.Key words: Acid polyacrylamide gel electrophoresis, agro-morphological traits, hordein, genetic distance, hordeum vulgare, variability
