To evaluate the effects of defoliation on green forage performance and grain yield (GY) variation in barley, five barley genotypes were imposed on three levels of defoliation treatments over two consecutive growing seasons in this study. The results indicated that green forage yields were significantly improved by repeated defoliation. The traits of green forage quality, including the ratio of dry weight to fresh weight, crude ash and calcium content were improved, while crude protein and crude fat were reduced, and crude fiber and phosphorus contents appeared not to be influenced by repeated defoliation. Plant height (PH), GY and other yield components, grain number per spike and thousand kernel weight, were significantly reduced by defoliation over the two growing seasons, while internode length below spike was less affected. Reduction in spike length and the number of spikes per plant were identified in only one year. Correlation analysis revealed that only PH exhibited a positive correlation with GY. Effects of genotype, interaction between genotype and defoliation, and environments on changes of forage yield and quality and GY were also discussed. Our current work provides a feasible approach to select elite barley cultivars with optimal defoliation treatments for both forage and grain uses in barley breeding programme.