Pyrenophora graminea, the causal agent of leaf stripe disease, is an economically important pathogen of barley found worldwide. It is critical to clearly define and standardize the leaf stripe assessment methods to avoid subjectivity and variability between assessors. Therefore, in this study, a comparison between the proportion of diseased plants (DP) and the proportion of diseased leaves (DL) per plant was investigated. Disease assessments were made visually at multiple sample sites in artificially and naturally inoculated research and production fields during four growing seasons. There were significant differences (P = 0.001) among cultivars in mean DP and DL averages, which are consistently higher in susceptible barley. However, DP values increased linearly as DL increased. The slopes and intercepts of the DP-DL relationship were consistent over the four growing seasons. This result might make a significant contribution for leaf stripe assessment in barley breeding programme.