Finding suitable models of canopy reflectance in forward simulation mode is a prerequisite for their use in inverse mode to characterize canopy variables of interest, such as leaf area index (LAI) or chlorophyll content. In this study, the accuracy of the three-dimensional reflectance model DART (Discrete Anisotropic Radiative Transfer) was assessed for canopies of different genotypes of Eucalyptus, having distinct biophysical and biochemical characteristics, to improve the knowledge on how these characteristics are influencing the reflectance signal as measured by passive orbital sensors. The first step was to test the model suitability to simulate reflectance images in the visible and near infrared. We parameterized DART model using extensive measurements from Eucalyptus plantations including 16 contrasted genotypes. Forest inventories were conducted and leaf, bark, and forest floor optical properties were measured. Simulation accuracy was evaluated by comparing the mean top of canopy (TOC) bidirectional reflectance of DART with TOC reflectance extracted from a Pleiades very high resolution satellite image. Results showed a good performance of DART with mean reflectance absolute error lower than 2%. Intergenotype reflectance variability was correctly simulated, but the model did not succeed at catching the slight spatial variation for a given genotype, excepted when large gaps appeared due to tree mortality. The second step consisted of sensitivity analysis to explore which biochemical or biophysical characteristics influenced more the canopy reflectance between genotypes. Perspectives for using DART model in inversion mode in these ecosystems were discussed.