Producing quantitative and reliable measures of crop disease is essential for resistance breeding, but is challenging and time consuming using traditional phenotyping methods. Hyperspectral remote sensing has shown potential for the detection of plant diseases,but its utility for phenotyping large and diverse populations of plants under field conditions requires further evaluation. In this study, we collected canopy hyperspectral data from 335 wheat varieties using a spectroradiometer, and we investigated the use of canopy reflectance for detecting the Septoria tritici blotch (STB) isease and for quantifying the severity of infection. Canopy- and leaf-level infection metrics of STB based on traditional visual assessments and automated nalyses of leaf images were used as ground truth data. Results showed (i) that canopy reflectance and the selected spectral indices show promise for quantifying STB infections, and (ii) that the normalized difference water index (NDWI) showed the best performance in detecting STB compared to other spectral indices. Moreover, artial least squares (PLS) regression models allowed for an improvement in the prediction of STB metrics. The PLS discriminant analysis (PLSDA) model calibrated based on the spectral data of four reference varieties was able to discriminate between the diseased and healthy canopies among the 335 varieties with an accuracy of 93%(Kappa = 0.60). Finally, the PLSDAmodel predictions allowed for the identification of wheat genotypes that are potentiallymore susceptible to STB, which was confirmed by the STB visual assessment. This study demonstrates the great potential of using canopy hyperspectral remote sensing to improve foliar disease ssessment and to facilitate plant breeding for disease resistance.