Radishes with black hearts will lose edible value and cause food safety problems, so it is important to detect and remove the defective ones before processing and consumption. A hyperspectral transmittance imaging system with 420 wavelengths was developed to capture images from white radishes. A successive-projections algorithm (SPA) was applied with 10 wavelengths selected to distinguish defective radishes with black hearts from normal samples. Pearson linear correlation coefficients were calculated to further refine the set of wavelengths with 4 wavelengths determined. Four chemometric classifiers were developed for classification of normal and defective radishes, using 420, 10 and 4 wavelengths as input variables. The overall classifying accuracy based on the four classifiers were 95.6%-100%. The highest classification with 100% was obtained with a back propagation artificial neural network (BPANN) for both calibration and prediction using 420 and 10 wavelengths. Overall accuracies of 98.4% and 97.8% were obtained for calibration and prediction, respectively, with Fisher's linear discriminant analysis (FLDA) based on 4 wavelengths, and was better than the other three classifiers. This indicated that the developed hyperspectral transmittance imaging was suitable for black heart detection in white radishes with the optimal wavelengths, which has potential for fast on-line discrimination before food processing or reaching storage shelves.