| Ginseng(Panax ginseng C. A. Mey) is a famous and precious Chinese traditional medicinal material. Ginsenoside is the main effective composition of ginseng, and the pharmacology activation that ginseng displays is mainly the fruiting of the ginsenoside function. Ginsenoside is a secondary supersession result of ginseng. Ginsenoside Rb1 is a kind of ginsenosides among them, having much pharmacology function, for example, ginsenoside Rb1 not only has function of treating the disease in the nervous system of centre, but also hurts to have the result of treatment to the memory that senile dementia and medicine cause. Meanwhile, ginsenoside Rb1 is still forebody thing and raw materials of anticancer drug Rg3, Rh2, Compound K, etc by biochemistry transform.The ginseng hairy root was induced from the root explants of ginseng by Agrobacterium rhizogenes A4 with directly inoculating, non- natural plant. Compared with the cell development, its growth speed is quick, the live material content is high and the hereditary form is stability. This experiment added Phytohormones and Elicitors to culture medium of MS, which is to improve the total ginsenoside content and ginsenoside Rb1 of panax ginseng hairy root. This thesis studied as the main material with the panax ginseng hairy root. First, we added Phytohormones (2,4-D, IBA and NAA)and Elicitors (SA and MJ)to culture medium of MS. Then, we used Soxhlet Extraction to draw the total ginsenoside. And macroreticular resin is used to divide sugar, at the same time, total ginsenoside is getted by vanillic aldehyde colorimetry. Through HPLC, the final strength of NAA is 0.050mg/L in culture medium of MS, which is advantageous to accumulation of valid substance. The main contents and conclusions are as follows:1) This thesis studied culture of the hairy root. The hairy root grew vigorous. Through incessant generation and extension culture, the ginseng hairy root materials that follow-up's testing needed were obtained.2) Got the method of drawing total ginsenoside: panax ginseng hairy root→rubbing ( filtration of 40 )→weighing up 0.2g→wrapping by filter paper→putting it in the drawing vessel of Soxhlet Extraction→adding a lot of chloroform→heating and recycling for 1 hour→throw away chloroform→drugresidue volatilizing chloroform→putting a lot of methanol in flask→heating and recycling for 3 hour→making drawing water dry through decompressing and distilling→making it dissolve through adding 10ml water→adding petroleum ether(30～60℃) to draw for twice→throw away petroleum ether→water flowing into column of macroreticular resin of AB-8→adding a lot of water to wash and throwing away→adding a lot of 20% ethanol to wash and throwing away→last washing through 50% ethanol and getting→making menstruum dry through decompressing and distilling→total ginsenoside.3) Choosing method of HPLC about ginsenoside Rb1 of panax ginseng hairy root: column temperature is 25℃, cheching wavelength is 203nm, volume of adding sample is 10ul, flowing partcomprise acetonitrile and 0.05% Phosphoric acid, flowing velocity is 1.0ml/min, washing program of grads is 0～35min, percent of acetonitrile is 81%; 35～55min, percent of acetonitrile is 81%～71%; 55～70min, percent of acetonitrile is 71%; 70～100min, percent of acetonitrile is 71%～60%, computing by area of peak.4) Through detecting total biomass, total ginsenoside content and ginsenoside Rb1 of panax ginseng hairy root without phytohormones and elicitors, we found biomass, yield of total ginsenoside and ginsenoside Rb1 of ginseng hairy root simultaneously increase with time, but total ginsenoside content and ginsenoside Rb1 content simultaneously decrease with time. In addition, biomass, yield of total ginsenoside and ginsenoside Rb1 of ginseng hairy root which culture for four weeks got maximum which is 34.52g/L, 0.0846g/L and 0.00842g/L. For that, we choosed four weeks as time of sampling and checking.5) After detecting content and yield of total ginsenoside and ginsenoside Rb1 of panax ginseng hairy root which cultured for 4 weeks, we found that total ginsenoside content is 1.93%, yield is 0.0846g/L; ginsenoside Rb1 content is 0.192%, yield is 0.00842g/L.6) Through detecting total ginsenoside of panax ginseng hairy root with Phytohormones, we found that 2,4-D, NAA and IBA can all boost accumulation of total ginsenoside. When consistency of 2,4-D is 2.000mg/L, total ginsenoside content is 5.14%. Compared with CK (1.93%), increased twice. In contrast, 2,4-D decreased biomass of ginseng hairy root with increase of consistency. Hence, total ginsenoside yield only is 0.522g/L. When consistency of NAA is 0.050mg/L, total ginsenoside content is 3.21%. When consistency of NAA is 0.500mg/L, totalginsenoside yield show maximum is 1.429g/L. When consistency of NAA is 0.500mg/L, total ginsenoside content is 3.31%. In addition, total biomass of ginseng hairy root increased with consistency of IBA. When consistency of IBA is 2.000mg/L, total ginsenoside yield show maximum is 1.474g/L.7) Through detecting ginsenoside Rb1 of panax ginseng hairy root with Phytohormones, we found that different consistency of phytohormones will lead different ginsenoside Rb1 content. 2,4-D, NAA and IBA can all boost accumulation of ginsenoside Rb1 content in feasible consistency. Especially when consistency of NAA is 0.050mg/L, ginsenoside Rb1 content is 0.3270%, and its yield is 15.31mg/L. In addition, 2,4-D will lead a different effect in ginsenoside Rb1 content. 2,4-D can lead a accumulation of ginsenoside Rb1 content in high consistency. When its consistency is 2.000mg/L, ginsenoside Rb1 content is 0.2895%. Because the consistency of 2,4-D decrease biomass of ginseng hairy root,ginsenoside Rb1 yield is 4.38mg/L. When consistency of IBA is 0.050mg/L, ginsenoside Rb1 yield is 12.15mg/L.8) After comparing with effect of three kind of phytohormones in content of total ginsenoside and ginsenoside Rb1 of panax ginseng hairy root, we found that NAA is the most valid to accumulation of ginsenoside Rb1 content, and IBA is the most valid to accumulation of total ginsenoside content of ginseng hairy root. When consistency of NAA is 0.050mg/L, yield of ginsenoside Rb1 is 15.31mg/L. When consistency of IBA is 0.500mg/L, yield of ginsenoside Rb1 is 0.1753g/L.9) Through detecting total ginsenoside of panax ginseng hairy root with elicitors,we found SA have no clearly effect on accumulation of total ginsenoside and biomass of panax ginseng hairy root, and SA decreased total ginsenoside and biomass of panax ginseng hairy root with increase of consistency. When consistency of SA is 0.100mmol/L, total ginsenoside content is 0.1008g/L. Being in low consistency, MJ can clearly boost accumulation of total ginsenoside and biomass of panax ginseng hairy root. When consistency of MJ is 0.001mmol/L, total ginsenoside content is 0.1207g/L.10) Through detecting ginsenoside Rb1 of panax ginseng hairy root with elicitors, we found that appropriate consistency of SA and MJ will lead a accumulation on ginsenoside Rb1 content, but the effect is not clearly. When consistency of SA is 0.100mmol/L, ginsenoside Rb1 content is 0.2457%, ginsenoside Rb1 yield is 11.49 mg/L. SA decreased ginsenoside Rb1 content ofpanax ginseng hairy root with increase of consistency. When consistency of SA is 1.000mmol/L, ginsenoside Rb1 yield only is 4.17 mg/L. When consistency of MJ is 0.001mmol/L, ginsenoside Rb1 content only is 0.2487% and ginsenoside Rb1 yield is 11.41mg/L. When consistency of MJ is 1.000mmol/L, ginsenoside Rb1 yield only is 6.5mg/L.11) The more important thing is that we compared yield of total ginsenoside and ginsenoside Rb1 in this thesis. According to the experiment data, yield of total ginsenoside is 0.1503g/L when consistency of NAA is 0.050mg/L, and yield of Rb1 is 15.31mg/L. Because our aim is to increase yield ginsenoside Rb1 of panax ginseng hairy root, the culture medium of MS in which is added consistency of 0.050mg/L of NAA is more favorable to production of ginsenoside Rb1. The cost is reduced in an all-round way, and value is relatively high.This thesis studied mainly on effect of MJ on ginsenoside content of panax ginseng hairy root, have being not reported before this, which offered the reference on the theory and practice for research work of the present ginsenoside.
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