Cup plant (Silphium perfoliatum L.) represents a promising alternative to silage maize as an energy crop for biogas production. This non-food plant possesses a highly ecological value due to its long blooming period, ability to grow in low-input agriculture and positive influence on soil structure. So far, there have been almost no breeding attempts for the cup plant, and all field experiments showing its high biomass yield were conducted by using only a few cultivated populations of unclear ancestry.Comprehensive assessment of five such populations for their biomass and methane yield parameters revealed substantial genetic variations indicating the possibility of improving these traits through selection and breeding. Higher biomass yield is likely to be achieved by breeding for secondary traits such as plant height, shoot diameter and internode number as well as photoperiod response. For increasing the methane production, reduced lignin or fibre content in the biomass seems to be important.Genetic relationships among the populations were estimated using tunable genotyping by sequencing (tGBS) technology. Genetic structure and phylogeny analyses revealed that all the plants belong to the same gene pool and share a common ancestry. Four out of five populations demonstrate a low genetic differentiation, whereas the fifth one represents a clear example of population stratification. To achieve a successful domestication and breeding of this new high-yielding perennial crop, a broader base of genetic diversity needs to be ensured and complemented by innovative breeding strategies driven by molecular genetic and modern genomics approaches.