Abstract Biodiversity relates to ecosystem functioning by modulating biogeochemical cycles of carbon, water, energy, and nutrients within and between multiple biotic and abiotic components of the ecosystems. However, large-scale, systematic measurements of plant biodiversity are still lacking, and the effects of biodiversity on measured biogeochemical processes are understudied. Here, we combined alpha (α) and beta (β) taxonomic measurements, spectral diversity from satellite observations, structural properties of the vegetation, and climatic drivers to assess the effect of biodiversity on ecosystem functional properties. Ecosystem functional properties were computed from eddy-covariance fluxes at 44 sites of the National Ecological Observatory Network. Based on the spectral variation hypothesis, we used the near-infrared reflectance of vegetation (NIRv) derived from Sentinel-2 satellite imagery to compute Rao’s quadratic entropy (Rao Q), a distance metric related to spatial heterogeneity. Using an automatic model averaging technique, we found that biodiversity proxies hold substantial explanatory power when predicting several ecosystem functions related to carbon and water exchange. In particular, NIRv-based Rao Q (RaoQNIRv) reflected positive biodiversity effects on productivity, as expected from the literature. In contrast, traditional taxonomic α-diversity indices were generally not selected as relevant predictors of the ecosystem functional properties. Yet, β-diversity strongly contributed to the prediction of carbon use efficiency, surface conductance, and water use efficiency. We also found that the RaoQNIRv is less affected by issues of saturation and bare soil contribution compared to RaoQNDVI. We show that spectral heterogeneity based on remotely sensed NIRv holds the potential for globally characterizing the biodiversity-ecosystem functioning relationship (BEF). While systematic measurements of taxonomic diversity co-located at biogeochemical measurement stations could reduce the uncertainty surrounding the BEF relationship at whole-ecosystem scale, remotely- sensed metrics characterizing important functional and structural diversity aspects of the landscape will be crucial for continuous spatiotemporal monitoring of biodiversity with relevant implications for ecosystem services to humankind.