Dry grassland sites are amongst the most species-rich habitats of central Europe and it is necessary to design effective management schemes for monitoring of their biomass production. This study explored the potential of hyperspectral remote sensing for mapping aboveground biomass in grassland habitats along a dry-mesic gradient, independent of a specific type or phenological period. Statistical models were developed between biomass samples and spectral reflectance collected with a field spectroradiometer, and it was further investigated to what degree the calibrated biomass models could be scaled to Hyperion data. Furthermore, biomass prediction was used as a surrogate for productivity for grassland habitats and the relationship between biomass and plant species richness was explored. Grassland samples were collected at four time steps during the growing season to capture normally occurring variation due to canopy growth stage and management factors. The relationships were investigated between biomass and (1) existing broad- and narrowband vegetation indices, (2) narrowband normalized difference vegetation index (NDVI) type indices, and (3) multiple linear regression (MLR) with individual spectral bands. Best models were obtained from the MLR and narrowband NDVI-type indices. Spectral regions related to plant water content were identified as the best estimators of biomass. Models calibrated with narrowband NDVI indices were best for up-scaling the field-developed models to the Hyperion scene. Furthermore, promising results were obtained from linking biomass estimations from the Hyperion scene with plant species richness of grassland habitats. Overall, it is concluded that ratio-based NDVI-type indices are less prone to scaling errors and thus offer higher potential for mapping grassland biomass using hyperspectral data from space-borne sensors.