Application of a low-relative-humidity treatment (LHT) to seedlings can reduce water stress on cuttings harvested from the seedlings, after the cuttings are planted. Effects of illumination during LHT and LHT duration on leaf water potential and leaf conductance in cucumber (Cucumis sativus L.) seedlings used as the model plant material and on growth of harvested cuttings were investigated to determine optimal LHT conditions. The seedlings received LHT for 12 or 24 h in a lighted or dark growth chamber at air temperatures of 28 to 31 °C and relative humidity of 12% to 25%. Cuttings including a foliage leaf and two cotyledons were harvested by cutting the hypocotyl of the seedlings immediately after the treatment, and then the cuttings were planted in vermiculite medium. Four days after planting, the total fresh weight of the cuttings from seedlings that had received LHT in the lighted chamber was 2.2 times that of cuttings from seedlings that had not received LHT, whereas the total fresh weight of those that had received LHT in the dark increased by 1.3 to 1.8 times. Significant effects of illumination during LHT were also observed in the transpiration rate and growth of the cuttings, harvested following the treatment, after they were planted. By varying LHT duration, it was also found that leaf water potential and leaf conductance of the seedlings decreased as LHT duration increased up to 18 h. Thus, illumination during LHT increased the growth of cuttings taken following the treatment, and optimal treatment duration of around 18 h was estimated from the seedlings' leaf conductance and leaf water potential.