Imaging spectroscopy has the potential to map closely related plant taxa at landscape scales. Although spectral investigations at the leaf and canopy levels have revealed relationships between phylogeny and reflectance, understanding how spectra differ across, and are inherited from, genotypes of a single species has received less attention. We used a common-garden population of four varieties of the keystone canopy tree,Metrosideros polymorpha, from Hawaii Island and four F1-hybrid genotypes derived from controlled crosses to determine if reflectance spectra discriminate sympatric, conspecific varieties of this species and their hybrids. With a single exception, pairwise comparisons of leaf reflectance patterns successfully distinguished varieties ofM. polymorphaon Hawaii Island as well as populations of the same variety from different islands. Further, spectral variability within a single variety from Hawaii Island and the older island of Oahu was greater than that observed among the four varieties on Hawaii Island. F1 hybrids most frequently displayed leaf spectral patterns intermediate to those of their parent taxa. Spectral reflectance patterns distinguished each of two of the hybrid genotypes from one of their parent varieties, indicating that classifying hybrids may be possible, particularly if sample sizes are increased. This work quantifies a baseline in spectral variability for an endemic Hawaiian tree species and advances the use of imaging spectroscopy in biodiversity studies at the genetic level.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00442-023-05374-1.