The objective of this study was to assess the genetic diversity and genetic relationships among 59 Turkish barley (Hordeum vulgare L.) germplasm lines (maintained for scald disease resistance breeding) using sequence-related amplified polymorphism (SRAP) markers. Seventeen SRAP primer combinations produced 83 polymorphic markers, with a mean polymorphism of 73.5%. The dendrogram created based on Nei's unweighted-pair group method using arithmetic average (UPGMA) indicated that there were 4 main clusters, which was supported by principle component analysis (PCA). Cluster I primarily included only scald-resistant germplasm lines, while clusters III and IV consisted of only scald-sensitive lines, but cluster 11 had both scald-resistant and scald-sensitive barley germplasm lines. The coefficients of genetic similarity among the genotypes ranged from 0.58 to 0.96, with a cophenetic correlation (r = 0.71) suggesting that the cluster analysis moderately represented the similarity matrix. The results indicate that a large amount of the genetic diversity present could be of great use in the development of future scald-resistant barley lines.