Abstract: Horses’ fear reactions are hazardous to both horses and human beings, but it is not clear whether fear is influenced more by training or by other factors such as genetics. The following study was designed to detect differences between young, untrained (U) and older, well-trained (T) horses of dressage (D), show-jumping (J), and mixed (M) genetic lines with regard to intensity of reaction and ease of habituation to a frightening stimulus. In five consecutive trials, 90 horses were exposed to a standardized fear-eliciting stimulus where intensity and duration of the reactions were recorded. Repeated measures analysis showed that flight reactions by J were less intense (p < 0.05) than those by D or M regardless of training status or age. Habituation to the stimulus over time was not significantly (p > 0.1) different between the disciplines, as indicated by similar slopes for all measurements, but reaction vigour declined faster for T than for U. These findings indicate that there may be a genetic basis for less strong, though not shorter-lasting, fear reactions in J compared to D or M lines of horses. Research including the estimation of genetic correlations between traits related to fearfulness and to performance would be required to verify this assumption.