| Tripterygium Wilfordii Hook is the root, foliage and flower of celastraceae plants Tripterygium Wilfordii. Its flavor is bitter and property is warm cool. It can enter people’s liver, spleen, and can pass through 12 meridians and collaterals. It has the ability of cleaning heat and disintoxicating, expelling wind, removing damness, promoting blood circulation, freeing channel, bruise trimming, and hemagogue and so on.The traditional research indicates that Tripterygium Wilfordii mainly used in Rheumatoid Arthritis, Lupus Erythematosus and some other autoimmune diseases in clinical. At present, there aren’t other medicines to treat those kinds of diseases both in Traditional Chinese medicine and Western medicine. The modern reseach indicates that Tripterygium Wilfordii can be used as the anti-organ rejects medicine and the effect is extremely good. Therefore, this paper does the research of genetic diversity and the main active composition of Tripterygium Wilfordii germ plasm. The main content is as follows:1. Establish the genetic resources center. This program starts on the headstream of medicinal plants Tripterygium Wilfordii, which integrates resource protecting, developing and using in a systematic way. This program collects Tripterygium Wilfordii which distributing in 7 provinces,30 countys(towns) and 227 seed sources in Jiangxi, Hunan, Guangxi, Guizhou, Fujian, Zhejiang and Yunnan. It constructs the most representative Tripterygium Wilfordii idioplasm gene in China.2. Through the comparison on the main photosynthesis target of Tripterygium Wilfordii from different geography seedling source, the result indicates that photosynthetic rate (Pn), transpiration rate (Vpd), stomatal conductance (Gs) and intercellular CO2 consistency (Ci) of southeast areas are higher than which of southwest areas. However intercellular CO2 of southwest areas is higher than which of southeast areas. The analysis of physiological factors indicates that it has remarkably significant relationship between photosynthetic rate and stomatal conductance and transpiration rate. It also has remarkably significant relationship between transpiration rate and stomatal conductance.3. Taking Tripterygium Wilfordii from different areas as material of study, using Bradford method, Giannoplitis NBT actinic reduction method and TBA method, we comprehensively analyse the active oxygen metabolism of seedling sources, which shows differences on different degree. The difference of POD activity is the biggest, the difference of MDA content is the smallest. The difference of soluble protein content, SOD activity and MDA content of seedling source in the southwest area is obviously bigger than which in the southeast area. Tongxin, Rongshui, Yizhou and Xiangyun with the highest SOD activity are all in southwest area; Luliang and Jinzhou with the lowest SOD activity are also in the southwest area. These targets of the seedling source of Tongxin show out obvious outness. The soluble protein content, SOD activity is 80.15/g fresh weight/min, MDA content is 30.766μmol/g fresh weight. This is the highest, but the POD activity is on the low level. While the four targets of in Fujian Taining, Liancheng, and Dehua are on the average level, the differences are not obvious.4. Taking Tripterygium Wilfordii from different areas as material of study, by using CTAB to extract the general DNA from Tripterygium Wilfordii materials and then utilizing molecular marker technology,153 pairs of primer will be got, from which 8 pairs, which is more polymorphic and stable, will be choosed. By amplifying the DNA from different Tripterygium Wilfordii materials,88 belts will be produced, each pair of primer composing and producing 11 belts on average, which can reflect good polymorphism. By analyzing genetic diversity, most Tripterygium Wilfordii samples can be classified in the same group for the similar geographical origin and genetic background. The 36 Tripterygium Wilfordii samples can be divided into 5 categories at the dissimilarity coefficient of 0.68. Samples with completely same genetic background haven’t appeared. The closest relationship in two samples is from Dabu, Taining and Xin qiao, Taining, whose dissimilarity coefficient is 0.111. And the farthest relationship is from Tongzi, Guizhou and Xiangxiang, Hunan, whose dissimilarity coefficient is 0.9714.5. Set up HPLC test system for Triptolide, and adopt high performance liquid chromatograph of Model Agilent 1100 and diode-array detector of Agilent G1315A; use water and carbinol as mobile phase (water:carbinol=72:28(v/v)) for gradient elution and test samples in 220mm wavelength. The average recovery of this method is 93%, RSD=4.37%. Sample size of Triptolide shows a good linear relationship in the range of 6.34ng-317ng. The analysis result indicates that the highest Triptolide is 6.337μg/g in Wuyi Mountain of Fujian, the lowest is 0.119μg/g in Dali of Yunnan. The average is 1.900μg/g. Xiangdong in Jiangxi, Wuyi Mountain in Fujian, Liancheng, Youxi, Changxing in Taining, Supper seeding and supper seedling in Taining, Xiangxiang in Hunan, Hechi in Guangxi, Songyang in Zhejiang are over the average. Purity of Tripterygium Wilfordii also differs in different position in the same production area. The contents of Triptolid in different parts of Tripterygium Wilfordii from the same place are also different. Triptolid in the root core is higher than in root velamen or root cauline. The proportion among root core, root velamen and root cauline is 6:3:1.6. Set up HPLC test system for wilforlide A, use water and carbinol as mobile phase (water:carbinol=72:28(v/v)) for gradient elution and test samples in 220mm wavelength; the average recovery of this method is 96.01%, RSD=4.49%. Sample size of wilforlide A shows a good linear relationship in the range of 10.74ng-537ng. Analysis result indicates that the highest content is 17.355μg/g of the material from Datian of Fujian and the lowest one is 0.471 μg/g from the supper seeding in Taining of Fujian. The average is 3.157μg/g. Among which,10 materials from Rongshui of Guangxi, Leigong of Guizhou, Tissue culture plant in Tongzhi and Guizhou are over the average. Purity of wilforlide A also differs in different position in the same production area. The contents of wilforlide A of different parts of Tripterygium Wilfordii from the same place are also different. It mainly distributed in root and husk, about 65.7%. And it is also affected in some way by the factors of longitude and latitude, and elevation.7. Set up HPLC test system for Tripterine in Tripterygium Wilfordii, use the method of gradient elution in flowing water and methanol (water:carbinol=13:87 (v: v) and in the place of wavelength 425nm; in this method, the rate of average recovery is 95.12%, RSD=4.71%, the sample size of Tripterine in Tripterygium Wilfordii is good in Linear relationship ranging from 65.1ng-2170ng. Analysis result indicates that the highest content is 1.89mg/g of the material from Liancheng of Fujian and the lowest one is 0.28mg/g from Longhui of Hunan. And there are 13 materials, that is, Lead mountain in Jiangxi, Jinzhou in Guangxi, Leigong in Guizhou and its seedling cultivation, wuyi mountain and some other areas in Fujian, such as liancheng, yong’an, mingxi, qingliu, youxi, Taining’s super seedling and super cutting seedlings, and Songyang in Zhejiang, are all above the average content which is 0.83mg/g. The contents of Tripterine from the same origin but different parts of Tripterygium Wilfordii are different, and this difference is significant. In its velamen, there is about 94.2%. The proportion of Tripterine between its velamen and root is around 20:1.8. Use EXCEL to do the relativity analyzing, the result shows the relationship of distributing of different components from the same part. The relativity of triptolide and wilforlide A in the caudex is the biggest, the coefficient is 0.514. The relativity coefficient of triptolide and wilforlide A in the root and husk is 0.409, the relativity coefficient of triptolide and Tripterine is 0.347, the relativity coefficient of wilforlide A and Tripterine is 0.355. The relativity coefficient of different components in other parts is not existed.9. According to the ecological benefit and social efficiency of Tripterygium wilfordii idioplasm resources research, using Net Present Value (NPV) analysis, and Marginal Cost and Marginal Revenue comparison analysis the different plant models economic efficiency of the Tripterygium wilfordii field cultivation, the mountainous region pure crop cultivation, the mountainous region interplanting, obtained NPVFC NPVMRPCC, NPVMRIC are positive, and the marginal cost is smaller than the marginal revenue, namely, three kind of plant models were feasible, Tripterygium wilfordii field cultivation models was most superior, mountainous region pure crop cultivation and mountainous region interplanting cultivation models next best. |
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