The feasibility of rapid and non-destructive classification of five different tomato seed cultivars was investigated by using visible and short-wave near infrared (Vis-NIR) spectra combined with chemometric approaches. Vis-NIR spectra containing 19 different wavelengths ranging from 375 nm to 970 nm were extracted from multispectral images of tomato seeds. Principal component analysis (PCA) was used for data exploration, while partial least squares discriminant analysis (PLS-DA) and support vector machine discriminant analysis (SVM-DA) were used to classify the five different tomato cultivars. The results showed very good classification accuracy for two independent test sets ranging from 94% to 100% for all tomato cultivars irrespective of chemometric methods. The overall classification error rates were 3.2% and 0.4% for the PLS-DA and SVM-DA calibration models, respectively. The results indicate that Vis-NIR spectra have the potential to be used for non-destructive discrimination of tomato seed cultivars with an opportunity to integrate them into plant genetic resource management, plant variety protection or registration programmes.