Biomass is currently the main renewable energy source (RES) in the EU, particularly in Poland. Solid biomass for energy purposes is primarily sourced from forests, the wood processing industry, and agriculture. A significant source of this energy feedstock could also be short-rotation woody crops (SRWCs), including black locust, poplar, and willow. Since numerous factors determine the SRWC biomass quality, the current study aimed at assessing biomass thermophysical properties and elemental composition depending on the plant species, soil enrichment procedure, and the plant harvest rotation over a consecutive 12-year period of cultivation. The characteristics under study, including the moisture content, ash content, volatile matter, fixed carbon, higher heating value (HHV), and the carbon, hydrogen, nitrogen, sulfur, and chlorine contents, were significantly differentiated by all the main factors, i.e., the SRWC species, the soil enrichment procedure, the harvest rotation, and the interactions between these factors. The SRWC species accounted for the highest percentage of the variation in the biomass moisture content, ash content, HHV, and nitrogen content, while the harvest rotation made the largest contribution to the variation in carbon, hydrogen, and chlorine contents. The black locust biomass was characterized by the significantly lowest moisture content (an average of 38.89%) and the highest sulfur content (an average of 0.033% DM), nitrogen content (an average of 0.91% DM), and chlorine content (an average of 0.032% DM). However, poplar was characterized by the highest HHV (an average of 19.84 GJ Mg–1 DM) and the highest moisture content (56.52% DM), carbon content (56.52% DM), and ash content (an average of 1.67% DM). Willow was characterized by the lowest ash content (an average of 1.67% DM), a medium moisture content, and the lowest nitrogen content (an average of 0.38% DM) and chlorine content (an average of 0.19% DM).