Perennial crops harvested in short rotations provide substantial amounts of biomass. This study determined the survival rate, biometric features and yield of fresh and dry biomass of 15 willow genotypes (including seven varieties and eight clones), cultivated at two different sites in two consecutive three-year harvest rotations. The study revealed the very high impact of the genotype (81% of the total variance) on the willow yield. The harvest rotation, along with the genotype, had a significant impact on the plant survival rate and the number of shoots per stool. Willow biomass was mainly affected by the plant height, its survival rate and shoot diameter. The significantly highest fresh (106 Mg ha−1) and dry biomass yield (54.0 Mg ha−1) was obtained from the Żubr variety of S. viminalis, which distinguished this variety from the other genotypes. The mean yield for the best three and five genotypes was 13% and 17% lower, respectively, and the mean yield for the whole experiment was 37% lower compared to the mean yield of the best variety (Żubr). Therefore, the choice of a willow genotype is of key importance for successful willow production.