This study describes the effects of environmental factors: the average and highest daily temperature, average relative air humidity, and the duration of the daily solar radiation, on the glucosinolate content in Brassicaceae. The results of our study indicate that the content of indole glucosinolate, glucobrassicin, is influenced by the average daily and the highest air temperature. Indole glucosinolates were much more susceptible to environmental factors than aliphatic or aromatic glucosinolates. Although the impact of the environment on the groups of aliphatic and aromatic glucosinolates was variable, there was a significant impact of the environment on specific aliphatic or aromatic glucosinolates. We conclude that climatic conditions cannot be neglected in the future planning of cropping systems, as the results of our investigation showed the important effect of environmental factors on the glucosinolate content in Brassicaceae.