Crassulacean acid metabolism (CAM) plants use surplus CO2 generated by cooling and heating at night when ventilation is not needed in a greenhouse. Schlumbergera truncata ‘Pink Dew’ is a multi–flowering cactus that needs more phylloclades for high–quality production. This study examined photosynthetic characteristics by the phylloclade levels of S. truncata in a growth chamber and a greenhouse for use of night CO2 enrichment. The CO2 uptake rate of the S. truncata’s top phylloclade in a growth chamber exhibited a C3 pattern, and the second phylloclade exhibited a C3 –CAM pattern. The CO2 uptake rate of the top phylloclade in a greenhouse showed a negative value both day and night, but those of the second phylloclade exhibited a CAM pattern. The stomatal conductance and water–use efficiency (WUE) of S. truncata at both the top and second phylloclades were higher in a growth chamber than in a greenhouse. The WUE of S. truncata in a growth chamber and a greenhouse was higher at the second phylloclade, which is a CAM pattern compared with those of the top phylloclade. The daily total net CO2 uptake of S. truncata was higher in a growth chamber than in a greenhouse. The daily total net CO2 uptake of S. truncata at the second phylloclade had the highest value of 155 mmol·m–2·d–1 in a growth chamber. The night total CO2 uptake of S. truncata at the second phylloclade was 3–fold higher in a growth chamber than in a greenhouse. S. truncata’s second phylloclade exhibited a CAM pattern that uptake CO2 at night, and the second phylloclade, was more mature than the top phylloclade. A multi–flowering cactus S. truncata ‘Pink Dew’ efficiently uptake night surplus CO2 in the proper environmental condition with matured phylloclade.