Behavioural and physiological changes during diapause, an important strategy of insects for surviving harsh seasonal conditions, have been intensively studied. The genetic and molecular mechanisms underpinning diapause development are less well known. We took a candidate gene approach to study prediapause gene expression patterns in the Colorado potato beetle (Leptinotarsa decemlineata), an invasive insect that has rapidly spread northwards to high seasonality environments. Newly eclosed beetles originating from southern (Italy) and northern (Russia) Europe were reared under short- [12 h light (L):12 h dark (D)] and long-day (18L:6D) photoperiods for 10 days. This time period includes the sensitive period for the photoperiodic induction and initiation of diapause. Gene expression trajectories of 12 diapause-related genes (regulatory, metabolic and stress-resistance) were analysed from 0-, 5- and 10-day-old beetles. Gene expression differences increased with age, deviating significantly between populations and photoperiods in 10-day-old beetles. The gene expression profiles, particularly those related to energy metabolism and stress-resistance, indicate that beetles originating from Russia also prepare for diapause under the long-day photoperiod and show qualitative differences in the diapausing phenotype. Our study shows that population-dependent differences seen in behavioural and physiological traits connected with diapause in L. decemlineata are also evident in the expression trajectories of diapause-related genes.