Abstract The effects of the difference (DIF) between day/night temperature (DT/NT) and end-of-day (EOD) light quality on growth, morphology, dry matter (DM) content and carbohydrate status in cucumber (Cucumis sativus L.) were examined. Plants were grown under a 12 h high light intensity period and DT/NT of 25/19 °C (positive DIF) or 19/25 °C (negative DIF) in combination with an exposure of 30 min EOD-red (EOD-R) or far-red (EOD-FR) light. A significant interaction between DIF and EOD light quality was found on morphology, DM and carbohydrate content in axial plant organs like stems and petioles, but not in leaf blades and roots. Positive DIF induced taller stems, and higher DM and carbohydrate content than negative DIF when the plants were grown under EOD-R. The stems developed under EOD-FR were tall and accumulated the highest content of DM and carbohydrates and only small differences were found between positive DIF and negative DIF. Under EOD-R a higher content of glucose and fructose was found under positive than negative DIF, while EOD-FR light resulted in a high glucose and fructose content under both positive and negative DIF and thereby equalized the effect of the two temperature treatments. The results show that positive DIF can induce similar responses in elongation growth, DM and carbohydrate accumulation as EOD-FR, and further that phytochrome status interact with the responses to alternating DT and NT.