Effects of explant type （leaf and stem segment） and hormone concentration （0.1， 0.2 and 0.3 mg·L^-1 6-BA； 1.0， 1.5 and 2.0 mg·L^-1 2，4-D） in medium on callus induction of Isodon amethystoides （Benth.） H. Hara were studied by using single-factor and L₉（3²） orthogonal experiments. The results show that compared with callus induction from leaves， calli can be induced from stem segments within a short time， which are loose and large in quantity. The results of orthogonal experiment show that the callus induction rate of stem segments of I. amethystoides varies from 72％ to 93％ in different media， in which， mass concentration of 2，4-D has a significant （P＜0.05） effect on callus induction rate， while that of 6-BA has no significant effect. According to the results， MS medium （containing 30 g·L^-1 sucrose and 7 g·L^-1 agar， pH 5.8） with 0.1 mg·L^-1 6-BA and 1.5 mg·L^-1 2，4-D is considered as an optimum medium for callus induction from stem segments of I. amethystoides.