| The dry roots of Follopia multiflora( Thunb.) Harald. are used as Radix Polygoni Multifiori,which is a common ingredient in Chinese prescriptions. It is used for dizziness with tinnitus, premature greying of the hair, soreness and weakness of the knee and lower back, and for limb numbness. The crude extracts of F. multiflora containing phospholipids, anthraquinones, and bianthraquinonyl glucosides exhibit prominent actions in many clinical therapies, particularly on cardiovascular diseases. Radix Polygoni Multifiori that distributing in different places had significant differences in content of bioactive components and in the effect of lowering anti-decrepitude. So the quality of Radix Polygoni Multifiori was not stable. So far, there was no report about the genetic variation and the evaluation of idioplasmatic resources of Radix Polygoni Multifiori. The quality standards of Radix Polygoni Multifiori were too simple. In the present research, the botanically taxonomic and anatomic approaches, chemical analytical methods and the molecular marking techniques were used to find out the variations of F. multiflora in the tissue structures, chemical components and genetic characters, to search for the good resources and to establish the specific methods for identification of Radix Polygoni Multifiori. Data of this research will be useful for the quality control of Radix Polygoni Multifiori, and assure that the safety and efficiency of crude drugs in clinical practice.1.Base on the cluster analysis method, 10 places (Jinyunshan, Jinggangshan, Deqing, Enshi, Shibing, Emeishan, Jingxi, Jiyuan, Dengfeng and Tianyang) of F. multiflora were divided into three groups by leaf characters. Those growing in north China were the firsr group, such as Jiyuan's and Dengfeng's. Those in south China were the second group, and Tianyang's was the third group. The distribution of three groups showed latitude zonation.2.The morphological features of leaf abaxial epidermis and micromorphological features of stem were observed with scanning electron microscope and light microscope. Cuticular membrane near stomata apparatuses was serried striate, while the way of arraying, thickness of stripe, grade of curve had no significant difference. Especially, F. multiflora from Tianyang had papilla on the spot of nervation in leaf abaxial epidermis and stem. The results indicated that characters of foliar surface under SEM are very useful evidence in identifying F. multiflora from different places.The epidermal characters of mature leaves of F. multiflora from different places were observed with light-microscope. Multi-regression analysis was used to determine the relationship between the leaves morphological characters of F. multiflora distributing in different locations and climate factors of different locations. The characters of leaf surface including the stomata density, stomata index, stomata width and stomata length were determined and compared. As a result, stomata were rarely observed at the upper epidermis. The stomata apparatuses on the lower epidermis were non-typical anisocytic type and anomocytic type. Stomata length was 31.1(20 - 44)μm, stomata width 23.1(16-38)μm, stomata index 17.1(7.5-26.5)%, stomata density 241.7(64– 573)个/mm2, respectively. The variations of anatomical characters for the lower of leaves of F. multiflora from different districts were definitely. The results of multi-regression analysis showed that the stomata length, width and stomata density were significant correlated to the latitude. With the latitude increasing, the length and width of stomata consistent decreased, while stomata density consistent increased.3.The medicinal materials from 10 places can be classified into two categories according to microstructure features of root. The root of F. multiflora from Tianyang had special characters, more fiber in the transverse section of root. The characters of transverse section of others were the same with the F. multiflora recorded in Chinese Pharmacopoeia.4.The contents of 15 inorganic elements (Al, B, Ba, Ca, Cu, Fe, K, Mg, Mn, Ni, P, Sr, Ti, Zn ) in soil and F. multiflora collected from 10 places were detected with ICP. The contents of inorganic element and their accumulation in F. multiflora between different places were compared. The results showed the contents of these elements in soil or plant had significant difference between 10 places. The content of Mn in plants from Deqing, Jinggangshan, and Jingxi is higher than that from other places. As to the accumulation coefficient of F. multiflora to these inorganic elements, there is no significant difference between 10 places. The accumulation coefficient of. F. multiflora to various inorganic elements had significant difference. Ca, K, Mg and Sr especially accumulated in F. multiflora.5.The effective components extracted by MeOH were separated by two kinds of mobile phase on TLC. Then the TLC fingerprints were scanned by Camag TLC scanner, and the results were analyzed by cluster analysis. The TLC chromatograms were constituted 14 characteristic common peaks include anthraquinone and stilbene glucoside.6.The bioactive components of F. multiflora determined its quality. Through measuring 17 environmental factors and 5 bioactive components (measured by HPLC), the influence of environmental factors on the bioactive components of F. multiflora (collected from ten places) was studied. Correlation, stepwise regression and principal component analysis were used to analyze the influence. Results showed that average temperature of Jan.,accumulated temperature, available potassium and Zn significant correlated with emodin. Average temperature of Jan.,accumulated temperature and organic matter significant negatively correlated with physcion. Stepwise regression showed that accumulated temperature was the main factor influencing the contents of emodin and physcion. But the environmental ecological factors did not significant correlated with 2,3,5,4′-Terahydroxystilbene-2-O-β-D-glucoside , emodin-8-O-β-D-glucoside and physcion-8-O-β-D-glucoside. Results indicated that the difference of bioactive components is mainly influenced by local specialized genotype. The HPLC data had been analyzed by principal components analysis, and results showed that F. multiflora from ten different places could be classified into four categories. The quality of F. multiflora from Deqing and Emeishan were the best.7. PCR products were sequenced directly and the sequences were analyzed with PAUP4.0. In order to find the patterns of the rDNA ITS sequence variation of F. multiflora, and to search of the method for distinguishing the Chinese crude drug Radix Polygoni Multifiori by molecular biological technique. The total length of ITS sequence is 648bp in the different samples. The lengths of three fragments, ITS1, 5.8S, ITS2, are 195bp, 164bp, 189bp respectively. There are twenty-one variable sites. The rDNA internal transcribed spacer is a good marker to distinguish the F. multiflora.. |
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