North China Blue Potted Tissue Culture And The Flavonoids Composition Studies

Huabei Scabiosa, Scabiosa tschiliensis Grunning, is a plant in the family of Dipsacaceae. The flower, which has an effect of anti-inflammatory, antipyretic and anti-oxidation, has been used as an herb medicine in history and had widely been used in Mongolian and China.In this paper, Scabiosa tschiliensis Grunning leaves and petiole were used as explants, and we discussed the inductive effect of callus, adventitious bud and root in different plant growth regulators and different concentration combination culture. The system of rapid regeneration and propagation of tissue culture of Scabiosa tschiliensis Grunning was established. The chromosome number of the wild plant, callus and the regeneration plants has been observed. The content of flavonoids from wild plant, callus and regeneration plant were analysised by ultraviolet spectrophotometry, and free DPPH radicals ability were determined by ELIASA. All the results set up the foundation for future exploitation and research of Scabiosa tschiliensis Grunning.The main experimental results are as follows:1. Scabiosa tschiliensis Grunning leaves and petiole were used as explants, the best medium of callus induction was MS medium with2.0mg/L TDZ and0.5mg/L2,4-D by leaves,the induction rate was94.05%and the callus were faint yellow and loose in good condition.2. Scabiosa tschiliensis Grunning leaves and petiole were used as explants, the best medium of adventitious bud induction was MS medium with4.0mg/L6-BA by leaves which has84.3%of induction rate, and MS medium with2.0mg/L6-BA by petiole which has83.33% of induction rate.3. The adventitious buds were used in rootage cultures, the best medium of rooting induction was MS medium with2.0mg/L IBA, the rooting rate was73.85%. The regeneration plant has been transplanted after rooting, survival rate was70%.4. The callus and root tip of seedings and regeneration plant were used by chromosome section, and the result suggest that chromosome number of Scabiosa tschiliensis Grunning was14.5. The content of flavonoids in Scabiosa tschiliensis Grunning determined by ultraviolet spectrophotometry, using rutin as the standards. The result shows that the content of flavonoids in wild plant was9.21mg/g. The highest content of flavonoids in adventitious buds was those cultured in MS medium with1.0mg/L6-BA, content of flavonoids was7.53mg/g. The content of flavonoids in callus which were cultured in MS medium with TDZ was higher than wild plant, the highest one was15.03mg/g, which was cultured in MS medium with1.0mg/L TDZ.The content of flavonoids in callus with TDZ and2,4-D concentration combination were lower than wild plant, the effect of concentration combination with high TDZ and Low2,4-D was better. The content of flavonoids in callus was8.58mg/g in the condition of MS medium with2.0mg/L TDZ and0.5mg/L2,4-D.6. There were obvious antioxidant activity in wild plant, callus and regeneration plant. In the18samples, the most efficient of free DPPH radicals ability was found in the alcohol extracts of the adventitious buds with MS medium with0.5mg/L6-BA, the callus with MS medium with0.5mg/L TDZ, the callus with MS medium with1.0mg/L TDZ and the wild plant. Their IC50were31.11μg/ml,23.94μg/ml,27.95μg/ml and26.5μg/ml.