In this experiment, we investigated the effects of various carbon sources and concentrations on the microtuber induction and acclimatization of the yam (Dioscorea cayenensis). First, the effects of the in vitro carbon sources and concentrations on the microtuber induction were examined. The highest efficiency of the microtuber induction was obtained in the 7% sucrose treatment, whereas the glucose treatment shows no effect on the microtuber formation. Secondly, the effects of the survival rate and the microtuber formation rate after the acclimatization were examined. The diameter (6.1 mm) and fresh weight (0.5g) of the tuberous root are the highest in the pretreatment of the 7% sucrose. Although the survival rate of the pretreatment of the low concentration sucrose (3% sucrose) is 100 %, the growth and development were inhibited. These results suggest the 7% sucrose treatment is appropriate for the yam microtuber formation and acclimatization. In addition, this protocol could be used for the propagation of virus-or disease-free clones and the multiplication of elite yam cultivars. which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.