A study was conducted to determine the implication of nitrate reductase (NR) and glutamine synthetase (GS) during the transition of micropropagated plantlets from heterotrophy to photoautotrophy to document how nitrogen metabolism interfaces with photosynthetic and anaplerotic CO2 fixation. The activity of the two enzymes was determined in different tissues at different organogenic stages during the development of plantlets transferred onto rooting media containing varying quantities of sucrose. Under 3% sucrose, NR activity was much higher in leaves than in crown tissues. When roots are initiating, there is a shift in the proportion of nitrate reduction from leaves to crown. As roots mature, the proportion of nitrate reduction increases in roots. Similar trends were observed under 5% sucrose. In contrast, under 1% sucrose, a higher proportion of the nitrate is reduced in the leaf tissues throughout the culture period. This suggests that nitrate is reduced mainly in leaves in photoautotrophic plantlets, while it is reduced in crowns and root tissues for mixotrophic plantlets. In general, the GS activity follows the pattern of NR, but is always in excess, to enable rapid assimilation of ammonium derived from metabolism and medium absorption.