The callus induction of Glechoma longituba was preliminary explored in this paper. Bud, leaf and leafstalk were cultured on different cultural conditions. The results showed that the callus of G. longituba could be induced on MS or LS medium supplemented with different kinds of hormones: 2，4-D, NAA, KT, BA. The frequency of callus induction was different when different explants were cultivated in medium supplemented with different types and different concentrations of hormones. The callus was induced by using leafstalk and leaves as explants; 2，4-D could promote to dedifferentiation, but NAA could restraint the callus to form. KT or BA mixed with 2，4-D both promoted the induction. MS+2，4-D, darkness and light in turn or LS+2，4-D,darkness were both good for callus induction, and the optimized condition for induction and multiplication was MS+2，4-D(1.5 mg·L-1)+BA(1.0 mg·L-1) in darkness and light in turn (14 hours illumination every day). After 30 days,the inducing rates of leaf could reach 91.38% and The inducing rates of leafstalk could reach 100%.After being subcultured for 14 days,the average multiplication rates could be up to 202.2%.