Establishment Of Tissue Culture System For Dictamnus Dasycarpus

In this experiment,wild Dictamnus dasycarpus seeds in Yanbian area were used as explants,by means of plant tissue culture,studies were carried out on seed germination,callus induction and differentiation,adventitious bud proliferation and hair roots,and regenerated plants transplanting matrix,HPLC method was used to determine the contents of the medicinal components,such as dictamnine,obacunone and fraxinellone in the callus induced by roots,stems and leaves of the regenerated plants.In addition,the contents and production of medicinal components of the callus induced by seeds with different concentrations of plant growth regulators were studied.The suitable medium for seed germination and growth was selected as MS basic medium;The medium for callus induction was MS+2.0 mg/L NAA＋2.0 mg/L 6-BA,2.0 mg/L 2,4-D+2.0 mg/L 6-BA;The medium for callus differentiation was MS+1.5mg/L NAA+1.0mg/L KT;The culture medium of adventitious bud proliferation was MS+2.0 mg/L NAA+1.0 mg/L 6-BA;The culture medium of adventitious bud roots was 1/2 MS+1.0 mg/L NAA+1.0mg/L IAA;The transplanting medium for the regenerated plants to survive was humus soil:turfy soil:vermiculite=2:2:l(V:V:V).The results of content determination of medicinal components showed that there were significant differences among the contents of dictamninein,obacunone and fraxinellone callus induced by roots,stems and leaves.The content of dictamninein in stem was significantly higher than that in root and leaf.The contents of obacunone and fraxinellone in root were significantly higher than those in stem and leaf,the content of root was significantly higher than that of stem and leaf.Among the callus induced by seeds,the content of dictamnine in the medium of MS+1.5 mg/L NAA+2.0 mg/L 6-BA was the highest.The highest content of obacunone was found in the medium of MS+2.0 mg/L 2,4-D+2.0 mg/L 6-BA.The concentration of fraxinellone in MS+1.5 mg/L NAA+2.0 mg/L 6-BA medium was the highest.Further analysis on the production amount showed that the production amount of dictamnine was the highest in the medium of MS+2.0 mg/L 2,4-D+1.0 mg/L 6-BA.In the medium of MS+2.0 mg/L 2,4-d+2.0 mg/L 6-BA,the production of obacunone was the highest.In MS+2.0 mg/L 2,4-D+1.0 mg/L 6-BA medium,the production of fraxinellone was the highest.