Total and individual glucosinolate contents of broccoli cv Marathon were assessed at ambient CO2 (430-480 ppm) and elevated atmospheric CO2 (685-820 ppm) to determine the ecological relationship between changing atmospheric CO2 concentrations and phytochemicals. Elevated atmospheric CO2 concentration had a differing effect on individual glucosinolates and glucosinolate groups. Total glucosinolate content increased at elevated atmospheric CO2 concentration as a result of a strong increase in both methylsulfinylalkyl glucosinolates glucoraphanin and glucoiberin. In contrast, indole glucosinolates simultaneously decreased, predominantly because of a reduction of glucobrassicin and 4-methoxy-glucobrassicin contents. We conclude that changes in N content and N/S ratios as well as alterations in photochemical processes at elevated atmospheric CO2 concentration can influence total and individual glucosinolates contents of Brassicaceae, as demonstrated in the greenhouse, for broccoli.