| Ginkgo biloba L. is also called white fruitage tree, and it is one of the higher plants, which have the longest life. The leaves from Ginkgo biloba L. mainly included some effective constituents, such as flavonoids and ginkgolides, which can aggrandize the blood vessel, increase the outflow of blood, improve the kinetic of blood-circulating, decrease the resisting force of cerebrovascular, suppress thrombisis and clean free radical of human body. Ginkgolide B especially is the most effective agonist of plaque activation factor (PAF) and now become a hot research issue in the medicine field of the world.Now, Flavonoids and ginkgolides were mainly extracted from the leaves of Ginkgo biloba., but the leaves often contain high content of corious and cereous, and the effective constituent is very different under the various conditions. In addition, the constituent of ginkgolides is very low in the leaves, and thus the quality of extraction is difficult to reach the international standards. The large area plantation of Ginkgo biloba occupied the rare cropland. In order to solve the problem above, in this study, the callus and cell suspension culture clone were constructed using plant cell engineering technology. After several generation selection and domestication, the cell suspension clone, which can vigorous grow and contained higher content of flavonoides and ginkgolides, were constructed.The results showed the contents of flavone and ginkgolides in the callus culture substances were affected by the factors, such as genotype, expalnt category, season and so on. Thus, the trees with higher content of flavone and ginkgolides should be selected at first time.After a ten days in April, the germination leaves, which grow on 3-5 years old tree, was selected as explant inducting callus. After research on the kinds of culture media .various content of hermone, ultraviolet irridation, temperature and intensity of light in the culture, the callus clone with higher content of flavonoids and ginkgolides of Ginkgo biloba spiraster screened using photometer and HPLC combined.As the growth rate of callus is slow in the solid medium in the process of culture, callus is not fit for industrialization of secondary metabolite. The cell suspension culture line was established using the callus in the solid medium. The results of physiology and biochemical modulation on cell suspension culture line show that the primary medium was B5, pH value of culture was 5.8-6.2, and then NAA 0.5-1 .Omg/L. 6-BA 0.2~0.5mg/L, sucrose 30g/L and 15g/L glucose were added. In the experiment, thecontent of Cu + was increased ten times; the content of Fe + was increased half of one time; the content of ammoniacal nitrogen was increased one time and the content of nitrate nitrogen was decreased half of one time. The inoculation weight was 30-40 FW/L, and inoculate the 150ml flask which contained 50ml culture liquid. At 25? 癈, SOOOlux continuous light treatment, conducted suspension cell culture, at Hie 15th day, add 0.1~0.2g/L phenylalanine, after 20 days, the product will be harvest, which is the optimium condition of producing flavone and ginkgolides in the suspension cell culture solution. In the production, the content of flavonoides can reach 3.35%, the content of ginkgolides can reach 0.0758%.In this experiment, active constituents in the leaves, tissue and cell culture substration of Ginkgo biloba were extracted, purified and detected. The leaves of Ginkgo biloba were pulverized in 40-60 orders, and were extracted in 70% ethanol solution at the rate of material and solution 1:15, pretreated 20s by microwave, and extracted 5min by ultrasonic, and repeated 2 times above. The extraction rate of active constituent can reach 98.5%. After the extraction by liquid-liquid method diluted the concentration of flavone to 1.0~1.2mg/ml and using 10-times of volume colume at the flow velocity of 4 times of colume volume per hour absorbed by HPD 100, and then washed by distilled water and 20% ethanol, and using 3-times of volume colume at the f...
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