Abstract:Using Hemerocallis middendorfii’s 6 varieties as experimental materials, we the investigated effects of hormone combination, types of explants, genotype and culture media on rooting of adventitious shoot. The results showed that, the best medium of Little bumble bee for callus and adventitious shoot differentiation was MS+1 mg·L-1 6-BA+0.1 mg·L-1 NAA, its callus induction rate, differentiation rate and mean shoot number per explant was 63.33%,91.11% and 5.86, respectively; however, the above indexes varied from genotype to genotype; scape was the best explant for inducing adventitious shoot in the three;1/2MS+0.2 mg·L-1 NAA was optimal for root formation of regenerated shoots, with 81.11% rooting rate and 6.08 roots per shoot; the conclusion was that we successfully established plant regeneration of three genotypes, mean shoot number per explant was all more than 4 and Stelladaorp was the most, which was 6.88.