Researching On The Effect Of Precursors On Growth And Metabolism In Callus Of Panax Quinquefolium L.

This experiment takes 3 years root and 10 days young stem and leaf ofpanax quinquefolium L.as explant to induce the formation of callus. The result indicated that at each of explant is able to induce callus under the same induction condition. The inductivity of root is lower than stem and leaf.We optimize the cultural condition of panax quinquefolium L. callus from the sucrose density and the growth regulator. The best medium is improved MS medium add 3.0mg/L2, 4-D,0.5mg/LBA and 3% sucrose. Through the examination of culture's yield and the saponin content, we have determined the most suitable growth cycle ofpanax quinquefolium L.callus is 35d. At this time, saponin yield reach the highest level is 1.5%.We use the method of precursor feeding to study on the regulation of four kinds of precursors the sodium pyruvate, pyruvic acid, alpha - pinene and the squalene about the growth and saponin synthesis of callus of panax quinquefolium L.. And at the same time we study the effect of the four precursors on enzyme activity of peroxydase enzyme, catalase and polyphenoloxidase in panax quinquefolium L.culture. The result indicated that besides pyruvic acid, other three kinds of precursors can promote the saponin synthesis and the growth ofpanax quinquefolium L. callus culture.We add sodium pyruvate to the medium in different concentration on the initial period of culture, and reap one cultivated cycle later. Detected the yield, saponin content and enzyme activity of the callus, and determine the optimum adding density of sodium pyruvate is 5mmol/L. Under this density, the growth yield, saponin content and peroxydase, catalase activity all reach supreme value. It is 1.38 times, 2.2 times, 4.8 times, 2.2 times of control group. Polyphenol oxidase activity reaches the minimum value, is 57% of the contrast. We carry on the analysis of the single saponin to the culture resulting saponin sample which has added 5mmol/L sodium pyruvate through the method of HPLC. Finally we found saponin Rg1 and Rg2of ginseng increased notably in the sample after adding sodium pyruvate, and the content of Rb2 obviously drops. It is proved that sodium pyruvate is favorable to the formation of Rg1 and Rg2.After 14 days culture, we add pyruvate in different concentration in the medium, and study dynamics characteristic of the growth of culture, saponin synthesis and variation of peroxydase, catalase,Polyphenol oxidase activity. The result reveals that pyruvate has strong inhibitory activity in the growth of culture. No matter the size of the adding concentration and the length of action time, the growth amount, saponin content and peroxydase, catalase activity of the culture are all less than the control group except polyphenol oxidase activity, thus confirm the regulation of pyruvate on metabolism of panax quinquefolium L. callus is negative.We have studied the regulation ofα-pinene in different concentration, added time and action time on the panax quinquefolium L. callus. Determine the optimal concentration is 0.2mmol/L, the optimal adding time is 14 days after culture, and the best action time is 15d. The saponin content in culture treated by this way is 88.8% higher than the control. At the same time we detect the peroxydase, catalase and polyphenol oxidase activity in the culture treated byα-pinene. The result is further verified that the optimal concentration, optimal adding time and best action time we determined are correct.When callus culture for 14d, we add squqlene in different concentration in medium, culture one cycle time later, measure the growth amount and saponin content of culture, and at the same time detect peroxydase, catalase and polyphenol oxidase activity in the callus treated by squqlene. Selected out the best adding concentration is 0.2mmol/L, at this time the saponin content, peroxydase and catalase activity all reach the peak. They are 123%, 24% and 2% higher than control. Polyphenol oxidase activity reach the lowest level.Study sodium pyruvate,α-pinene and squqlene through orthogonal experimental design. The analytic result shows that the result of sodium pyruvate to the culture is most notable,α-pinene takes the second place, the effective result of the squqlene is the worst but also reach the notable lexel. The best association of three kinds of precursor is 5mmol/L sodium pyruvate, 0.2mmol/Lα- pinene, 0.2mmol/L squqlene.