Abstract: Tree peony is known as china’s national flower which is very beautiful, graceful and looked as the best of all the flowers. Tree peony Var. spontanea Rehd was micropropagated by using lateral buds as explants, the effect of pruned and cut method on developing adventitious shoots was investigated, a regeneration system which is the first report was developed for dwarf peony from a lateral bud. A good disinfection protocol for lateral buds was as follows: Disinfecting with 75% alcohol for 30 sec and then soaking with 0.1% HgCl2 for 7 min adventitious shoots regenerated from lateral buds with different pruned and cut treatments and the control, the highest mean number of shoots (5.0) was obtained on pruned and cut wound. A high-frequency multiplication rate was achieved on woody plant medium supplemented with BA 3.0 mg L-1 and IAA 0.2 mg L-1 multiple shoots were formed within 5 weeks; adventious shoots were efficiently rooted (75.62%) on half-strength woody plant medium with NAA 1.0 and IBA 4.0 mg L-1 and rooted plantlets survived after acclimatization to the greenhouse.