This paper evaluates, in the greenhouse and under natural conditions, barley yield losses due to defoliation treatments of the upper three leaves either healthy or infected at the boot stage by Pyrenophora teres f. teres. Defoliation was assumed as a loss of a similar leaf area caused by net blotch disease severity of 100%. Contribution to grain yield was defined herein as a difference between defoliation treatments and a treatment where plants lost all their upper three leaves. In contrast, yield losses referred to differences in yield between defoliations and the control. In the greenhouse, removal of the antepenultimate leaf did not affect any yield component. For main stems, defoliating upper three leaves reduced grain yield by 30% and this was mainly due to flag leaf removal. These losses were similar to those induced by net blotch disease under natural conditions, but were of 42% for all tillers. Grain yield losses due to disease severity were not equivalent to the defoliation effect of a similar healthy leaf area. On the other hand and for a significant contribution to grain yield, flag leaf was dependent on the presence of the other two leaves. Inoculation and defoliation of 21 cultivars induced similar grain yield losses of 32%. However, biotic stress reduced by 40% the contribution of their upper three leaves. Under field conditions, yield losses were not significant until barley plants lost more than one upper leaf and flag leaf contribution was equivalent to that of the remaining leaves. Characteristic roots, defined as leaf coefficients for plant performance, were 0.13, 0.06 and 0.01 for the flag, penultimate and antepenultimate leaves, respectively. Because antepenultimate leaves become trivial at the boot stage, we propose that coefficients of the remaining leaves should be used when modelling yield losses due to barley foliar diseases.