| Paclitaxel (Taxol) is an anti-cancer drugs extracted from the yew, it have a significant effects for treatment of ovarian cancer, breast cancer and lung cancer and other cancer. At present, more of the extraction of taxol in yew bark as raw material, not only destroying the environment, but endanger the yew resources, lack of resources and yew plants, and it also far from being able to meet the needs of the market, so the source is very short of drugs, which is very expensive; chemical synthetic method has successful, but its production cost is extremely expensive for the actual production are subject to certain limitations; plant fermentation mass production of taxol was able to get the fungi to be in-depth study, and more difficult to obtain high-yield strains, this production did not put large production-scale applications. Therefore, we must find alternative ways of production, the Taxus plant cell culture technology to the production of paclitaxel is to open up a new source of paclitaxel good way. The current study of Taxus cell cultures are in a laboratory shake flask stage, we cultured cells of Taxus environment, conditions and the key to control technology to further improve the large-scale production will provide an important source of information.We carried out the production of plant cell culture of Taxus paclitaxel research, choose a high taxol content in the Northeast outside the plastid dwarf yew, Taxus callus induction on media and culture conditions to optimize the choice; through the optimization of experimental suspension of Taxus cuspidata elicitors of culture and control the level of the best types.We mainly research the following aspects:(1) Taxus callus induction training, the establishment of the production of callus growth factor system and its analysis.(2) Taxus callus induction training to optimize the foreign implant research, to find the best explants.(3) Search for laboratory operations (factoryt) method of detection of paclitaxel.(4) In suspension cultures of Taxus cuspidata cells control the production of Taxol and technological research and find the best medium for the production of paclitaxel and control program.Taxus in a large number of distribution in Northeast China, the main product of Taxol is an important cancer drugs. The study of the production Taxus of paclitaxel at home and abroad still in development stage, so the production of Taxus cuspidata cell culture study of paclitaxel with a very good geographical advantage and economic value. The current study of Taxus cell cultures are in a laboratory phase, on cell growth regulation and control environmental conditions and the key technology to further improve the large-scale production will provide a theoretical basis.In this paper, we main researched the effects of different elicitors on the induction of callus culture, as well as the impact of cell suspension culture,we get the impact of degrees through the orthogonal experiment of qualitative and quantitative analysis of various elicitors on Taxus cuspidata cell culture, with a view to further establish and improve the Taxus cuspidata cell culture conditions and the production of paclitaxel, in order to further advance the industrialization of research results and provide an important reference.To take advantage of callus tissue of Taxus paclitaxel to production culture, we studied TPL, orthogonal experiment, callus diameter measurement, weighing and other methods of Taxus tender stem, young leaves, the top three kinds of meristem explants body of callus induction training, medium optimization, the growth of callus and liquid suspension culture in taxol biosynthesis, such as the volume control to a more optimized system. The results showed that:The highest rate of callus induction medium to add programs for the NAA hormone concentration is 1.0mg/L,6-BA concentration is 0.4mg/L,2,4-D concentration is1.5mg/L, 3 factors on the number of days out of the more affected relationship between primary and secondary: NAA>BA>2,4-D, was the fastest out of the program for a few more days,NAA concentration 1.5mg/L,6-BA concentration of 0.3mg/L,2,4-D concentration of 2.00mg/L.Available through the analysis of the highest induction rate and the number of days the fastest of the more hormones are basically the same.At the same time we come through the study also Taxus tender stem, young leaves, the top three kinds of meristem explants of the more time and rate of callus induction there is a big difference, the more time, the callus induction are the highest rates of tender stem, followed by the apical meristem, leaves the worst.We also made use of spectrophotometer were determined to Taxus callus rough material and content of taxol was 0.66‰and 0.19‰.Finally, the experiments in cell culture of Taxus cuspidata were added 6-BA, NAA,2,4-D,salicylic acid, sodium acetate, and their cell culture species are on average taxol content of0.75‰,0.77‰,0.93‰,0.80‰,0.80‰, an average of 0.81‰, than the calli of taxol content (0.66‰) increased significantly; in the mixed orthogonal experiment, the cell cultures for the content of paclitaxel was 1.35‰, is the raw material(0.19‰)of 7.10 times ,and of callus 0.24(0.66‰)times.In this study, Taxus cuspidata cell culture for production of paclitaxel to provide key technologies. |
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