Tissue culture technique was applied to propagate for a new Dahlia variety named TDL-05. Two types of tissues, shoot tip and stem tissues, were used as initiation materials for culture. Sterilization by H2O2 at concentration of 20 percent for 5 min or 7 min (shoot tip and stem tissues respectively) were the best period time to result in good ratio of cleaned tissues as well as the regeneration capacity of the cleaned tissues. Both of experimented cytokinins, BAP and kinetin, could be used for in-vitro shoot multiplication of TDL-05 variety. However, BAP seemed to be more effective than kinetin. In MS medium containing the best concentration of BAP, at 0.5 mg/l, multiplication coefficient of in vitro shoot reached at 3.9/4 weeks
while it was 3.23/4 weeks in the best concentration, at 1 mg/l, of kinetin. All of in-vitro shoots could induce multi-root in the MS medium without phytohormon. In nursery, a substrate containing fired rice husk and soil at the 1:1 ratio was suitable for survival as well as growth of in-vitro plants of the Dahlia TDL-05 variety.