In vitro amplification protocol was developed for multiplication of Valeriana officinalis L. var. latifolia Miq. by using leaves，petiols，rhizome and roots derived from wild plants grown in pot as explants. Effects of different plant growth regulators on callus induction，adventitious bud differentiation and bud anatomy were studied. The results showed that：1. it was optimal for the explants to be disinfected with 70% ethanol for 30 s and then soaked with 0.1% mercuric chloride for 6 min. 2.The optimum medium for callus induction was MS 2,4-D 2mg/L 6-BA 0.5 mg/L，and callus induction rate of leave could arrived at 80%. however，buds regenerated difficultly from callus， the differential rate was 25% on the medium of MS 6-BA 4 mg/L NAA 0.5 mg/L. 3. Buds regenerated exogenous from callus epidermal cells.