Based On Molecular Phylogeography To Analysis Cyathula Officinalis Kuan Varieties And Quality

Cyathula officinalis Kuan to produce authentic Sichuan commonly used in Chinese medicine.It is the dry root of the plant Cyathula officinalis Kuan, which is a perennial herb belong to family Amaranthaceae and genus Cyathula. Is currently facing major decline in quality, hybrid varieties, origin and other changes of the status quo. The topic by collecting each main production areas Cyathula officinalis Kuan and its hybrid varieties and wild C. officinalis Kuan samples for quality analysis in terms of resources, medicinal properties, composition, etc; by chloroplast matK gene sequence, ITS sequences and SSR molecular markers, genetic diversity C. officinalis Kuan analysis, application software, and phylogenetic analysis phylogeography analysis method C. officinalis Kuan mesh evolutionary lineage and geography for analysis. Combined with the quality of the geographical division of the geography and genealogy C. officinalis Kuan quality differences in depth analysis to explore the causes and mechanisms of formation of quality differences.1. C. officinalis Kuan quality geographic variation pattern analysisC. officinalis Kuan each main production areas of environmental factors comparison, the major producing areas currently Sichuan Jinkouhe. Yaan Baoxing, Tianquan County compared with Chongqing, Hunan, Hubei. the areas gradually formed the scale, the study of environmental factors of climate, soil, elevation etc. closer, preliminary speculated that the ecological environment caused by differences in the quality not the dominant factor.From C. officinalis Kuan medicinal traits analysis, we can see the different origins of herbs showed cylindrical taproot obvious, few branches, sweet, the main difference in the diameter. C. officinalis Kuan difference with adulterant main root of the number of branches, taste and color. In terms of taste, C. officinalis Kuan sweet taste, Cyathula officinalis* Cyathula capitata and Cyathula capitata Mop.no sweet, slightly bitter taste hemp, Achyranthes agpera L the taste lighter. Smart electronic tongue sensory systems of different origin, different age, commercially C. officinalis Kuan and adulterants "flavor" analysis, through its response value differences can determine the origin, fixed number of year, and whether there is a confusion of the sample. Through the analysis of the electronic tongue, fixed number of year of the medicinal materials of collecting large influence on its "taste" difference. Medicinal traits verified by analysis of a " thick, branches few, toughening, section pale yellow, sweet" better to determine the traditional experience of authentic C. officinalis Kuan.C. officinalis Kuan has special secondary structure, different origin C. qfficinalis Kuan in the cork layer of cell on the number of columns, the secondary structure of the central vascular system more obvious differences. C. officinalis Kuan difference with hybrid varieties mainly in cork and juniors vascular bundles. Sansei rounds of vascular bundle C. officinalis Kuan different years of observation, shows as the growth of the fixed number of year of the root growth, the number of rounds of vascular Sansei and the number gradually increased.By C. officinalis Kuan cyasterone content analysis shows that there are some differences of origin, while C. officinalis Kuan cyasterone content is higherthan its adulterants. By comparing different fixed number of year samples of C.officinalis Kuan found cyasterone content is increasing along with the increaseof growth period, reached after three years reduced its growth, becoming steady. Hunan. Hubei. Chongqing and other places C. officinalis Kuan collecting fixed number of year is generally 1 to 2 years, which is cyasterone content, quite different; Sichuan collecting fixed number of year is more than three years and three years, quality stabilized. Therefore, collecting fixed number of year is an important factor affecting C. officinalis Kuan ingredients differences. The measurement results show a commercially available medicinal materials purchased six shipments of commercial C. officinalis Kuan only two batches of medicinal materials content up to standard, indicating that medicine currently marketed C. officinalis Kuan poor quality, market supervision needs to be improved. The UPLC was used to construct C. officinalis Kuan fingerprint, found different origin C. officinalis Kuan its fingerprint similarity in 0.889～0.974. its high similarity, cluster analysis showed that C. officinalis Kuan fixed number of year of fingerprint similarity effect more obvious than the effect of different producing areas.2. Based on SSR analysis of genetic diversity C. officinalis KuanSSR is a simple sequence repeat, because of the different number of repeating sequences or repeated degree exhibit polymorphism. This experiment adopts SSR analysis of genetic diversity of C. officinalis Kuan. By Amaranthaceae plant 39197 EST sequences were screened out 286 SSR loci, the SSR repeat type dinucleotide repeat the most primitive types, accounting for 90.56%. Through the experiment discovered C. officinalis Kuan SSR polymorphic loci total of six, the average of the PIC was 0.2946, showed each loci of C. officinalis Kuan moderate polymorphic loci. Coefficient of genetic differentiation among populations Gst value of 0.8623. indicating that variation among populations accounted C. officinalis Kuan genetic variation overall 86.23%, variation within populations accounted for 13.77% of the overall variation, illustrate C. qfficinalis Kuan genetic differentiation among populations in more abundant than within populations.3. C. officinalis Kuan molecular phylogeography analysisMolecular phylogeography research core is the historical characteristics of the spatial distribution of genetic lineage. Chloroplast genes and ribosomal DNA internal transcribed spacer （ITS） because of restrictions in functionality less, evolutionary rate faster, is often used to study molecular phylogeography.This experiment based on the chloroplast matk gene and ITS sequence discussed C. officinalis Kuan current geographic distribution patterns and genetic diversity.There are four haplotypes based on C. officinalis Kuan chloroplast gene matk, by the results of genetic differentiation among populations of known Nst value is less than the value of Gst, illustrate C. officinalis Kuan the haplotypes no significant phylogeographic structure in the distribution. Speculated that the reason for this may be due to the initial sequence of choice when primer design caused the sample distribution mainly concentrates in haploid type Hap₁ and Hap₂. Evolved from the system tree and reticulate evolution diagram shows haplotype Hap₃ and Hap₄ separate clustered together, both corresponding to Sichuan Emei Cyathula capitata Mop and Hubei Enshi Cyathula capitata Mop. Cyathula capitata Mop of C. officinalis Kuan common confusion, by the above analysis, although Cyathula capitata Mop and C. officinalis Kuan belong Amaranthaceae Cyathula, but there is a certain distance in the genetic relationship.C. officinalis Kuan based on ITS sequence consists of a haplotype of 24 kinds. genetic differentiation among populations Nst coefficient value is greater than the Gst, indicates that a significant phylogeographic structure between C. officinalis Kuan haploid type. Evolved from the system tree diagrams and reticulate evolution in connection with the haplotype corresponding sample location geopattern analysis, it can be divided into three parts:（1） the first part, the top of the reticulate evolution part, mainly composed of hunan, hubei, chongqing, sichuan corresponding haploid type in some parts of the sample,from the geographical analysis shows that the distribution presented above in several provinces of sichuan basin and basin area, the geographical position close, genetic relationship is relatively close to its sample; （2） the second part, reticulate evolution intermediate portion, which is mainly composed of Sichuan Tianquan, Baoxing, Jinkouhe, Miyi region corresponding sample haplotype composition, show that Sichuan Tianquan. Baoxing, Jinkouhe sample close genetic relationship with Miyi sample, early in the query data and field survey found that different areas have had largescale introduction of Sichuan, in 1976 Tianquan county in order to expand the scale of cultivation, from Xichang swapped in a large number of seeds, and later Baoxing, Jinkouhe are from Tianquan introduction, so the results of this study confirmed the introduction events have occurred in these areas. （3） The third part, reticulate evolution of the lower left side of the branch portion. Hap₇ and Hap₂3 separate clustered together, both corresponding to Sichuan Emei Cyathula capitata Mop and Hubei Enshi Cyathula capitata Mop. Hap₁7 corresponding sample Sichuan Jinkouhe Cyathula officinalis* Cyathula capitata, from two kinds of evolution in the diagram as the haploid type between C. officinalis Kuan and Cyathula capitata Mop corresponding between haploid type, Cyathula officinalis* Cyathula capitata is Miscellaneous C. officinalis Kuan and Cyathula capitata Mop, its distance in relationships with both are similar.In C. officinalis Kuan genetic diversity analysis, matk chloroplast gene and ITS sequence analysis showed rich genetic diversity among populations than the populations of the interior. The formation of the genetic diversity of populations of many factors, reproduction, ecological geography and gene flow and other factors. C. qfficinalis Kuan for the cross-pollination, increased the gene flow between the populations and species; Geographic range widely, C. officinalis Kuan distribution around the ecological environment, planting cultivation mode and so on all differences; With C. officinalis Kuan planting, cultivation history of accumulation, the origin of the occurrence of certain C. officinalis Kuan migration, comprehensive the above several factors cause the genetic diversity of C. officinalis Kuan more and more rich.