The growing interest in using everbearing (EB) strawberry cultivars to extend the cultivation period has faced some challenges. These include poor runner production due to its perpetual flowering nature; irregular flowering behavior and extended periods of high temperature have caused floral inhibition and reduced yield. As flowering is an interplay between temperature and photoperiod, it is important to investigate the effects of this interaction on the cultivation. Therefore, this study used meristem dissection as a tool to study the effect of temperature and photoperiod on meristem development. Tray plants of two EB strawberry cultivars ‘Florentina’ and ‘Favori’ were grown at 20 °C, 25 °C, and 30 °C under short day (SD) conditions, and subsequently at 20 °C under long day (LD) conditions. The meristem development was analysed every 6 weeks for a 15-week period in SD and for 14 weeks in LD conditions using meristem dissection. The plants showed similar flowering patterns to previously studied everbearing cultivars, which was qualitative LD plants at high temperatures and quantitative LD plants at lower temperatures. Our results show that meristem dissection can be used to determine the temperature and photoperiodic effect on meristem development, and for the occurrence of cropping peaks, and can therefore be used to decide the environmental input and to evaluate yield potential.