Study On The Accumulation Of Saponins And Suspension Culture Of Panax Notoginseng Hairy Root

Panax notoginseng has long been used as traditional Chinese herbal medicine for its unique and obvious medical effect,such as detumescence analgesic, anti-inflammatory,antioxidant, enhance of immunity and other medicinal value. Here the regenerated plantlets was infected with strains of Agrobacterium rhizogenes for induction of hairy roots of P.notoginseng. The liquid culture system for P. notonigseng hairy roots was also established,and then its growth and biomass in this system was investigated too. Furthermore, the rare element of germanium was introduced to test its effect on saponins accumulation of P.notoginseng hairy roots. The results will lay basis on the development and utilization of P.notoginseng hairy roots.(1) On the basis of single factor experiment, response surface method was employed to optimize the factors affecting P. notoginseng hairy roots induction. The results showed that the best induction rate was gained when the leaves of P. notoginseng were infected by the strains of ACCC10060 compared with other cases of experiments; According to the regression analysis, the optimal induction conditions were as follows: 41.62 h of pre-culture time, 23.10 min of infection time, addition of 15.76 mg/L acetosyringone, 46.22 h of co-culture time. The above conclusion was verified and an induction rate of 37.5% was achieved.(2) On the basis of single factor experiment, biomass as parameter, the liquid culture system for hairy roots was established with the method of orthogonal experimental. It’s found that the formula of 1/2 MS,1.425 g/LKNO3,1.65 g/L NH4NO3,0.085 g/LCaCl2,30 g/L sucrose,pH 5.4 was proved to be the optimal liquid culture system; The above conclusion was also verified, and it could be observed that the hairy roots under these conditions possess the properties of dense and long root hairs, and heavy branching of lateral root, and it got the maximal yield at the 35 th day of culture.(3) Through the single factor experiments, we studied the effects of culture temperature,illumination and elements of germanium concentration on the yield and content of saponins of P. notoginseng hairy roots. It’s found that the peak yield and content were both achieved from hairy roots cultured at 24 ℃.Likely, the maximal yield was gained at 1500 lx lightstrength, and in addition the dry weight in this case was slightly less than that of other treatments of light strength, however, the total yield of saponins was still the peak value among all of illumination treatments.To stimulate the accumulation of saponins, rare element of Ge was employed and examined in this experiment. And it’s found that the content of saponins increased significantly and the maximal yield of saponins was also achieved in the presence of 0.8 mmol?L-1 Ge in the media in spite of slight decline in dry weight compared with that without Ge. Taken all data together, it could be concluded that the optimized parameters suitable for saponins accumulation were at 24 ℃,1500 lx,and 0.8 mmol?L-1 Ge.