One of the quality parameters of chicory (Cichorium intybus L.) is its slightly bitter taste which is caused by sesquiterpene lactones. As the quality of chicons is highly dependent on the quality of the roots, the effects of the cultivation conditions of the roots on the levels of sesquiterpene lactones in chicons were investigated. Roots from a collection of 11 commercially available and 2 experimental chicory cultivars were grown at five different locations with two nitrogen manuring levels to evaluate the differences in sesquiterpene lactone levels as measured with enzyme-linked immunosorbent assays (ELISAs). There were significant differences (P<0.001) between cultivars in both lactucopicrin and lactucin-like (lactucin, 8-deoxylactucin and their 11β,13-dihydro derivatives) sesquiterpene lactone levels. In addition, a significant (P=0.006 and P=0.019, respectively) effect of additional nitrogen manuring was observed on the levels of lactucopicrin and lactucin-like sesquiterpene lactones. The interaction of cultivar with nitrogen manuring had a significant (P=0.013) effect on the level of lactucin-like sesquiterpene lactones. Furthermore, the lactucopicrin level was significantly influenced by the interactions of the cultivar with the nitrogen manuring level (P<0.001), with the location (P=0.001) and with both the nitrogen manuring level and location (P<0.001). The results indicate that it may be possible to influence the level of the bitter sesquiterpene lactones and, consequently, to influence the taste of chicons by cultivar choice and/or cultivation location and conditions.