Biodiversity is important in maintaining ecosystem viability, and the availability of adequate biodiversity data is a prerequisite for the sustainable management of natural resources. As such, there is a clear need to map biodiversity at high spatial resolutions across large areas. Airborne and spaceborne optical remote sensing is a potential tool to provide such biodiversity data. The spectral variation hypothesis (SVH) predicts a positive correlation between spectral variability (SV) of a remotely sensed image and biodiversity. The SVH has only been tested on a few terrestrial plant communities. Our study is the first attempt to apply the SVH in the marine environment using hyperspectral imagery recorded by Compact Airborne Spectrographic Imager (CASI). All coverage-based diversity measures of benthic macrophytes and invertebrates showed low but statistically significant positive correlations with SV whereas the relationship between biomass-based diversity measures and SV were weak or lacking. The observed relationships did not vary with spatial scale. SV had the highest independent effect among predictor variables in the statistical models of coverage-derived total benthic species richness and Shannon index. Thus, the relevance of SVH in marine benthic habitats was proved and this forms a prerequisite for the future use of SV in benthic biodiversity assessments.