The article presents the results of biochemical and QTL (Quantitative Trait Loci) analysis of dry matter content, nutrient and biologically active compounds: sugars, ascorbic acid, chlorophylls a and b, anthocyanins and carotenoids in populations of doubled haploid lines of leaf, root crops, and oilseeds of the Brassica rapa L. species grown in optimal light culture conditions, but with different photoperiod durations. The purpose of this study was to evaluate the effect of the photoperiod on the transition to bolting and the accumulation of biologically active substances, as well as how the localization and identification of chromosomal loci determined the content of certain phytochemicals. The influence of the length of daylight hours on the content of components of the biochemical composition was assessed. It was shown that growing under conditions of a 16 h photoperiod increased the content of dry matter, sugars, vitamin C, and anthocyanins. On the contrary, the content of photosynthetic pigments was higher under the conditions of a 12 h photoperiod. Valuable lines that can be sources of biologically active compounds were revealed. Based on the results of the obtained data, 102 QTLs were mapped, which determine the manifestation of the studied biochemical quality traits in the B. rapa doubled haploid lines under conditions of short and long daylight hours. Molecular markers genetically linked to the selected QTLs were determined. It was revealed that the identified loci controlling all the studied biochemical traits were mainly in the fifth, sixth, seventh, and ninth linkage groups, which correlated with the data obtained in the field and greenhouse. Most of the identified loci controlled several studied traits simultaneously. The identified QTLs and identified molecular markers are of interest for further study of the genetic control of the economically valuable traits determined by them and for the implementation of marker-assisted selection in B. rapa. The data obtained can be used in genetic and breeding work, including for the obtaining of new genotypes, lines and cultivars with a valuable biochemical composition, adapted for cultivation under specific photoperiodic conditions.