Establishment Of Saposhnikovia Divaricate(turcz.) Schisck.Plant Regeneration System And Determination Of Active Constituents

Saposhnikovia divaricate?Turcz.?Schisck.is Perennial herb of Umbelliferae Saposhnikovia,whose root was seleted as medicine.Most Saposhnikovia divaricate are distributed in North-east China?North China etc..Prim-O-glucosylcimifugin and 5-O-methylvisammioside in Saposhnikovia divaricate?Turcz.?Schisck have an obvious efficacy on antipyretic?analgesic?anti-inflammatory?Anti-tumor and so on.Several physiological indexes like the weight of one thousand seeds,pure rate,content of moisture,the rate of water absorption,the vitality of seeds were measured.Different density of6-BA?GA₃?KNO₃?NAA and different environmental culture temperature were selected to find out the effect on Saposhnikovia divaricate?Turcz.?Schisck.seed.According to result:the pure rate of Saposhnikovia divaricate?Turcz.?Schisck.seeds was 71%;the weight of one thousand seeds was?3.975±0.12?g;the amount of aqueous was 8.4%;the highest rate of water absorption was 140.67%,which was about 2.4 times more than seed dry weight;the vitality of seeds was77.83%.Soak seeds in 3mg/L KNO₃ and constant temperature culture at 25°C had promoted effect on seeds germination and seedling growth.In the state of natural,mature and full-dry windproof seeds of Saposhnikovia divaricate?Turcz.?Schisck.were selected in this study.Aseptic seedlings were successfully obtained through investigating types of disinfectant,duration of action and the methods of treating seeds.In order to select healthy aseptic seedlings,it should investigate the influence of different explants,explants age,culture mediums,culture temperature,light conditions and hormone ratios on callus induction to filter out the best condition of Saposhnikovia divaricate?Turcz.?Schisck.Callus induction:lay the aseptic seedling blade of 41d on the culture medium surface of the MS+0.90 mg/L 6-BA+1.05 mg/L 2,4-D+0.96 mg/L KT;25°C culture temperature;12h/d light culture.Callus of better color and texture were selected from the above experiment,to optimize the conditions of Saposhnikovia divaricate?Turcz.?Schisck.callus proliferation.The best conditions were obtained after investigating the effect of light time,carbon source and its content,subculture cycle and hormone ratio on the growth of callus:MS+0.56 mg/L 6-BA+0.48 mg/L 2,4-D+0.40 mg/L KT;12h/d light culture;30g/L sucrose as a carbon source;30d sub-generation cycle.Good-grown callus of Saposhnikovia divaricate?Turcz.?Schisck.were transfered torooting medium with different hormone ratios.The cluster bud was given different times of light,put into rooting medium with different hormone ratios and obtained complete plants.The rooting medium suitable for growth of Saposhnikovia divaricate?Turcz.?Schisck.callus was:MS+0.6 mg/L NAA+1.0 mg/L 6-BA.The rooting medium suitable for growth of Saposhnikovia divaricate cluster bud was:MS+0.4 mg/L NAA+0.1 mg/L 6-BA;12h/d light culture.According to Pharmacopoeia,use HPLC chromatographic conditions the content ofprim-O-glucosylcimifugin and 5-O-methylvisammioside was measured.Saposhnikovia divaricate?Turcz.?Schisck.dried roots and callus were determined and compared.The total prim-O-glucosylcimifugin content of the whole Saposhnikovia divaricat callus was 0.89 times more than that of its dried roots;The total 5-O-methylvisammioside content of the whole Saposhnikovia divaricate?Turcz.?Schisck.callus was 0.74 times more than that of its dried roots.According to this study,the efficient and stable of Saposhnikovia divaricate?Turcz.?Schisck.callus culture system,laid the foundation for industrial production of Saposhnikovia divaricate?Turcz.?Schisck.secondary metabolites.