Deoxynivalenol (DON) levels in harvested grain samples are used to evaluate the Fusarium head blight (FHB) resistance of wheat cultivars and breeding lines. Fourier transform near-infrared (FT-NIR) calibrations were developed to estimate the DON level and moisture content (MC) of bulk wheat grain samples harvested from FHB screening trials. Grains in a rotating glass petri dish were scanned in the 10,000-4,000 cm-1 (1,000-2,500 nm) spectral range using a Perkin Elmer Spectrum 400 FTIR/ FT-NIR spectrometer. The DON calibration predicted the DON levels in test samples with R2 = 0.62 and root mean square error of prediction (RMSEP) = 8.01 ppm. When 5-25 ppm of DON was used as the cut-off to classify samples into low-And high-DON groups, 60.8-82.3% of the low-DON samples were correctly classified, whereas the classification accuracy of the high-DON group was 82.3-94.0%. The MC calibration predicted the MC in grain samples with R2 = 0.98 and RMSEP = 0.19%. Therefore, these FT-NIR calibrations can be used to rapidly prescreen wheat lines to identify low-DON lines for further evaluation using standard laboratory methods, thereby reducing the time and costs of analyzing samples from FHB screening trials.