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Lebanese barley landrace Baladi is not sufficiently analyzed morphologically and genetically. This investigation was carried out to evaluate Baladi in its center of diversification by using morphological traits and microsatellite markers (SSR). The genetic diversity of 237 accessions of Lebanese barley landrace Baladi was evaluated by using 19 morphological traits. Principal components analysis showed that plant height, awn color, number of rows, length of rachilla hairs, number of spikelets per spike, number of seeds per spike and shape of lemma tip for sterile spikelets were the most significant traits. Regarding the relationship between plants, dendrogram constructed according to Jaccard distance, clearly separated the studied spikelets into 19 distinguished morphological profiles. A total of 55 genomic DNA extracted from selected accessions were analyzed using seven microsatellite markers (SSRs). A high level of polymorphism information content (PIC; average = 0.66) was observed. The polymorphic primers revealed 16 alleles with an average of 5.3 alleles per polymorphic primer. The highest number of polymorphic bands was developed by the primer pairs MGB402 (7 alleles). In the dendrogram constructed based on the SSR data, the evaluated lines were classified into two major clusters corresponding to the number of rows per spike (two-rows or six-rows). Based on this study, it can be concluded that there is a high level of genetic diversity within Baladi barley landrace in Lebanon. These results will be useful for barley germplasm management in terms of biodiversity protection and as a valuable source of gene pool for new useful alleles for crop improvement.