The Production Of Ginsenoside By Endophytes G22 And Used It As Elicitors To Affect Ginseng Adventitious Roots

Ginsenosides is a major component of ginseng. It has an important value in the pharmaceutical industry. Due to the shortage of ginseng and large demand from market, ginsenosides are insufficient. To improve the ginsenosides production endophytic fungi G22 and methyl jasmonate ginsenosides (MeJA) were used. At the same time, G22 was used as elicitors to co-cultivate with ginseng for increasing ginseng saponin production. The results will provide theoretical and technical support for improving ginsenosides.Finally a HPLC system used for detect ginseng saponins was established.(1)Establishing a high-performance liquid chromatography (HPLC) systemTo measure the ginsenosides contents of ginseng cultivation fibrous and endophytes, HPLC was used to determine 10 kinds of ginseng saponin monomer in plant bioreactors and endophyte cultures. The chromatographic separation was performed on a water Symmetry C18 Synergi column (250mm×4.6mm,5μm). The column temperature was set to 30℃. Acetonitrile as the mobile phase A and ultrapure water as the mobile phase B. The detection wavelength to 203 nm and the injection volume 10μl. The results showed 10 kinds of saponins could be effectively seperated. And linear relationship is good. The average recovery range was between 95.07% and 108.72%. The method is simple and reproducible. It can be used to detected multi-indicators in ginseng root and ginseng cultivation endophytes.(2) The characteristic of G22 that synthesis of ginsenosidesIn order to determine the best medium and harvest time of ginseng saponin production in G22,we used four common fungal culture to explore the production capacity of saponins in G22, at the same time used the MeJA to affect the production saponins. The results showed that Gauserime synthetic medium is the best medium for synthesis saponins,but Martin medium for Rb1 Rd. Overall We could see that MeJA suppressioned the total saponins and the monomer saponin’s synthesis,but a concentration of MeJA would be targeted to improve Rg1, Rg3, production and F2, for example MeJA(100 μmol/L) could accumulation the largest amount of Rg1 so as MeJA(50 μmol/L) for Rg3, and F2 under the experimental conditions induce synthesis has not yet reached the maximum value, we should do more further exploration. (3) Co-cultivation of G22 co-cultured with with ginseng, and effects of G22 to affect as elicitors on saponin synthetic of ginseng cultivation fibrous synthetic saponinIn order to improve the ginsenosides production by low-cost mass,we focused on the G22 to improve the ability of ginseng saponin yield adventitious roots. In the experiment of G22 used to co-cultivate with ginseng for increasing ginseng saponin production, we found the co-cultured would inhibited the accumulation of biomass, but the maximum promotion of total saponins、Rbl and Rh2 was happened at 300 μL bacteria, they respectively production reached at 13.69mg/g、0.33mg/g、0.05mg/g.The results from elicitor experiment showed that the addition of five elicitors inhibited the growth of fibrous roots,but the synthesis of total saponins could be promoted. MeJA(100 μm/L) with sterile elicitors(+0.3 g) was the suitable elicitor to improve saponins to reduce costs.