Supporting materials for rooting have a considerable influence on the growth and quality of in vitro plantlets. Various supporting materials (rockwool, perlite, vermiculite, and polyurethane) and nutrient supply cycles (12, 24, 36, and 48 hours) were examined to find the optimum conditions for photoautotrophic micropropagation of potato plantlets in the nutrient-circulated micropropagation (NCM) system. In the NCM system, nutrient solution was circulated between the culture vessel and the nutrient reservoir. A plug cell tray with 70 plantlets was placed inside. The number of air exchanges was 10 hours under forced ventilation. Nodal leafy cuttings of plantlets were cultured at CO2 concentrations (mol·mol-1)/PPF s (mol·m-2·s-1) of 350/80, 700/120, and 1500/250 on day 5-11, 12-18, and 19-28, respectively, for all treatments. All growth factors of in vitro plantlets grown for 28 days using rockwool, perlite and vermiculite were greater than those grown using polyurethane. Dry weight of plantlets grown using rockwool was eight times greater than those grown using polyurethane. The same results were obtained in the growth and survival percentages 14 days after transplanting to ex vitro conditions. Optimum nutrient supply cycles were 12, 24, and 48 hours when perlite, rockwool, and vermiculite were used as supporting materials, respectively. It was considered that the range of optimum nutrient supply cycle was affected by water retention characteristics of supporting materials. This study proved that the supporting material and the nutrient supply cycle were very important environmental factors in photoautotrophic mass propagation.