Plants have a high ability to adjust their metabolism, growth and development to changes in the light environment and to photoperiodic variation, but the current knowledge on how changes in metabolite contents are associated with growth and development is limited. We investigated the effect of three different photoperiodic treatments with similar daily light integral (DLI) on the growth responses and diurnal patterns in detected leaf metabolites in the short day plant Chrysanthemum×morifolium Ramat. Treatments were long day (LD, 18h light/6h dark), short day (SD, 12h light/12h dark) and short day with irregular night interruptions (NI-SD,12h light/12h dark, applied in a weekly pattern, shifting from day-to-day). Photoperiodic variation resulted in changes in the phenotypic development of the plants. The plants grown in the SD treatment started to initiate reproductive development of the meristems and a decrease in leaf expansion resulted in lower leaf area of expanding leaves. In contrast, plants in the NI-SD and LD treatments did not show reproductive development at any stage and final leaf area of the expanding leaves was intermediate for the NI-SD plants and largest for the LD plants. Photoperiodic variation also resulted in changes in the leaf metabolic profile for most of the analysed metabolites, but only carbohydrates, citrate and some amino acids displayed a shift in their diurnal pattern. Further, our results illustrated that short days (SD) increased the diurnal turnover of 1-kestose after 2 weeks, and decreased the overall contents of leaf hexoses after 3 weeks. In the two other treatments a diurnal turnover of 1-kestose was not stimulated before after 3 weeks, and hexoses together with the hexose:sucrose ratio steadily increased during the experiment. Our results enlighten the plasticity of leaf growth and metabolism to environmental changes, and demonstrate that diurnally regulated metabolites not always respond to photoperiodic variation.