A photoautotrophic or sugar-free medium micropropagation system (PAM) using five large culture vessels (volume = 120 L each) with a forced ventilation unit for supplying CO2-enriched air was developed and applied to commercial production of calla lily (Zantedeschia elliottiana) and china fir (Cunninghamia lanceolata) plantlets. The culture period of calla lily plantlets in the PAM was reduced by 50%, compared with that in a conventional, photomixotrophic micropropagation system (PMM) using small vessels each containing a sugar-containing medium. Percent survival ex vitro of calla lily plantlets from the PAM was 95%, while that from the PMM was 60%. The production cost of calla lily in the PAM was reduced by about 40%, compared with that in the PMM, and the initial investment per plantlet for the PAM was ≈10% lower than that for the PMM. The sales price of ex vitro acclimatized calla lily plantlet was increased by 25% due to its higher quality, compared with plantlets produced in the PMM.