The objectives of this study were: (1) to monitoring the CO2 consumption dynamics and the critical CO2
concentration level at which CO2 has to be added before photosynthesis of plantlets is maintained during
the initial stage of tissue-cultured plantlets; (2) to quantify the influences of the enriched CO2 concentration
and the media moisture content on sugar-free tissue-cultured seedlings in a large-scaled open growth chamber;
and (3) to demonstrate the possibility of tissue culture with sugar-free media plantlets grown in an open
growth chamber under CO2 enriched environments.