| Plants in the genus Tripterygium are important medicinal herb. Pharmacological studies have shown the anti-inflammatory, anti-tumor and immunosuppressive effects of these plants, and they are widely used for the treatment of rheumatoid arthritis,nephrotic syndrome and systemic lupus erythematosus in clinical. The source of traditional Chinese medicine Leigongteng mainly comes from Tripterygium wilfordii Hook. f. and Tripterygium hypoglaucum (Levl.) Hutch. However, there are several controversial issues such as the taxonomy of the two species and how should we deal with the substantial changes for proper pharmaceutical management.In this paper we focused on the relationship between Tripterygium Wilfordii and Tripterygium hypoglaucum. We collected samples from 16 sample sites of Tripterygium wilfordii and Tripterygium hypoglaucum (Levl.). Analysis methods such as plant metabolomics and molecular genetic analysis were used to apart them. The main research contents four parts:1 Compared the plants morphological characteristics and growth habit; and investigated their wild resources conditions, planting information, easy-confused varieties and different commodity features.2 The whole metabolic profiles of roots of Tripterygium wilfordii and Tripterygium hypoglaucum (Levl.) Hutch from 9 sample sites were analyzed based on UPLC-Q-TOF-MS. Five populations which three with typical characteristics of T. wilfordii blade and two with the typical characteristics of T. hypoglaucum (Levl.) Hutch blades were also analyzed based on UPLC-Q-TOF-MS, and the potential markers to distinguish two species were picked out.3 By phylogeography methods, we analysis the genetic relationship of 14 populations of two species with space linear distance over 50 kilometers between 14 populations, and furthermore we research genetic diversity and genetic differentiation of the populations by SSR methods.Results:1(1) T. wilfordii and T. hypoglaucum were mainly collected under woods or on the edge of woodland. Light and moisture attribute to their distribution to some extent. (2) Wild resources of T. wilfordii and T. hypoglaucum are shrinking, and both of their cultivation history are relatively short and their cultivation technique are still in low level. (3) Due to lack of harvesting and processing standards, decoction pieces varing from roots, rhizomes, to stems of plants, are all sold as commercial medicines.2 The cluster analysis and PCA analysis results showed that Tripterygium Wilfordii and Tripterygium hypoglaucum (Levl.) Hutch metabolic components exist obvious differences, they can overly be divided into two categories. But whole metabolic profiles between the two do not exist obvious boundaries while present a sort of regional distribution.Fujian Province, Taining (FTN) is considered the real estate areas of Tripterygium wilfordii. Systematic cluster analysis and PCA analysis results both showed that the metabolic characteristics of Tripterygium wilfordii of the region is different from the other regions, while have similar metabolic features to some parts of the origin.23 compounds were screening as an important biological marker to distinguish the Tripterygium Wilfordii and Tripterygium hypoglaucum (Levl.) Hutch leaves based on UPLC-Q-TOFMS and these compounds were alkaloids, oligomeric flavanols, quercetin and kaempferol phenolic compounds.314 populations were divided into 8 haplotype. Haplotype network tree and Nei’s phylogenetic tree show the similar structure, two kinds of calculation results can both divided the 14 populations into 4 groups and 8 haplotypes. Considering of geographical distribution, the four groups showed regional and have different ecological characteristics, which indicates that environmental factors are related to the genetic differentiation between Tripterygium Wilfordii and Tripterygium hypoglaucum (Levl.) Hutch. The population ZLX, XYY (Tripterygium Wilfordii) and population XLH (Tripterygium hypoglaucum (Levl.) Hutch, shared haplotypes Hap4, wich support that Tripterygium Wilfordii and Tripterygium hypoglaucum combine into a single species. The result of SSR showed that Tripterygium wilfordii populations had abundant genetic diversity (Ho=0.46255, He=0.4016, A=3.0500, Ne=2.0796, 1=0.6958)。The results show that 42% genetic differentiation of Tripterygium wilfordii among the populations (Gst=0.420, Fst=0.436),which is close to molecular ANOVA (41% of the variation among populations).and the reslut indicated the populations have already exist significant genetic differentiation.Gene flow Nm=0.324 (Nra< 1) further indicated that due to genetic drift, the populations have occur significant genetic differentiation. Based on Nei’s (1972) method, the genetic similarity data was used to carry on Mantel test with the geographical distance, altitude respectively. The results showed that the genetic similarity was very significantly correlated to altitude (r=0.609, P< 0.01, y= 5.719x+16.812, R2= 0.2148), while significant to geographical distance (r=-1.1191, P< 0.05, y= 0.0038x+22.801, R2=0.0761)Conclusion:1 The human factor contributed to the rapidly shrinking of resource from Tripterygium Wilfordii and Tripterygium Hypoglaucum in some areas. Conducting in-depth artificial cultivation of Tripterygium Wilfordii and Tripterygium Hypoglaucum will help alleviate the rapid decline of wild resources. Currently commodities of Tripterygium Wilfordii have no uniform standard, which roots, rhizomes and stems are used as the commodities. It is necessary to strengthen the oversight and monitoring of Tripterygium Wilfordii in order to ensure safety and rational drug use of different medicative parts from this plant.2 The metabolites of roots from Tripterygium Wilfordii and Tripterygium Hypoglaucum present certain regional characteristic. The whole metabolic profiles showed no obvious boundary (no significant difference). Altitude, soil and all kinds of ecological factors are likely to lead to the metabolic differences in two species of plants and plant from different regions. The chemical composition of Tripterygium wilfordii and Tripterygium Hypoglaucum has a certain similarity. The metabolites such as flavanol, quercetin and kaempferol, which are important photoprotection material (such as against the strong light radiation, filter ultraviolet radiation, remove free radicals, etc.), and the significant differences of relative content proves that the environmental factors (such as altitude, lighting, etc.) are important effectors contribute to the differences of the metabolite in the leaves from Tripterygium wilfordii and Tripterygium Hypoglaucum.3 The Tripterygium Wilfordii and Tripterygium Hypoglaucum sharing the one haplotypes support that they should be combined into one species. The genetic diversity of Tripterygium Wilfordii is rich, that means high ecological adaptability and the larger evolution potential and huge potential for genetic improvement. Populations from Tripterygium Wilfordii have obvious genetic differentiation. Geographical and ecological environment factors, the population inbreeding or selfing way of breeding may be the important factors of genetic diversity and genetic differentiation. |
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