This study was conducted to establish mass propagation system from the axillary bud culture of chrysanthemum zawadskii H. which was used as material of medicinal plants. Shoot egeneration was better on MS medium with NAA and BA. The optimum concentraions of growth regulator for shoot regeneration differed depending on accessionsof C. Zawadskii. Shoot regeneration in Keungucheolcho was better on MS Medium with NAA 0.01mg/1 and BA 0.1mg/1 while Hyangrobonggucheocho was better with NAA 0.1mg/1and BA 0.3mg/1. Addition of NAA into medium was effective for induction of root from shoots regenerated. Shoot multiplcation was more effective when 10mg/1 spermine was added into medium than when other polyamines were treated ino medium . Randomly and specifically amplified polymorphic DAC banding patterns based on polymerase chain reaction (PCR) analysis were used to assess the genetic variation of plants regenerated from in vitro culture.