Dracocephalum rupestre is an important medicinal plant.However,its leaf explant regeneration system has yet to be established.Here we investigated the impact of plant growth regulators on callus induction,differentiation,ad-ventitious bud proliferation,and rooting using field planted leaves and tissue-cultured seedling leaves of D.rupestre as explants.We found that the optimal medium for inducing callus from field planted leaves was MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 2,4-D+1.0 mg·L-1 IAA,achieving an induction rate of 84.51％.For adventitious bud differentiation,the pre-ferred medium comprised MS+3.0 mg·L-1 6-BA+0.5 mg·L-1 TDZ+0.5 mg·L-1 IAA,resulting in a differentiation rate of 66.37％.Similarly,for inducing callus from tissue-cultured seedling leaves,the optimal medium included MS+2.0 mg·L-16-BA+0.1 mg·L-1 2,4-D+0.5 mg·L-1 IAA,with an induction rate of 86.73％.The medium conductive to adventitious bud differentiation consisted of MS+2.0 mg·L-1 6-BA+2.0 mg·L-1 TDZ+0.05 mg·L-1 IAA,yielding a differentiation rate of 53.48％.Furthermore,the appropriate medium for adventitious bud proliferation was MS+2.0 mg·L-1 6-BA+0.05 mg·L-1 NAA,achieving a proliferation rate of 83.57％.The rooting medium was 1/2MS+0.1 mg·L-1 NAA+0.1 mg·L-1 IBA,resulting in a rooting rate of 86.97％.Tissue-cultured seedlings exhibited optimal growth in a mixed substrate of peat and vermicu-lite at a ratio of 1∶1(v/v).This study successfully established a leaf explant regeneration system for D.rupestre,providing valuable technical support for the conservation and rapid propagation of its germplasm resources.
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