The Survey Of Rhubarb Resources And The Study On Its Quality Feature And Formation Mechanism In Authentic Regions

Purpose: Rhubarb is one of the most important traditional Chinese medicine, and also thetypical representative of TCMs which have various functions and geo-authentic habitats.Liang Zhou, Quan Shui, He Zhou, Xi Ning, Quan Shui, Min Xian and Qing Shui are allgeo-authentic habitats recorded in many herbal works. But there exist many problems. TheRhuharb resource reserves and the current situation in those habitats are not clear. The qualityof Rhubarb is unequivocal, and the genetic and environmental mechanism that have causedthe difference in the Rhubarb quality are also unclear. Aimed at those situation, this researchmade a systematic study in the investigation of Rhubarb resource in different habitats and thequality evalution of Rhubarb, and on the basis of the investigation, we made an intensivestudy on the genetic and environmental mechanism which caused quality difference. Themethods and results are as follows:Method:In this study, we take the method of ecology community survey and TraditionalChinese medicine resources simple investigation, and finished the investigation of Qing Hai,Gan Su, Ning Xia, Si Chuan and Shan Xi, including5provinces and33city districts. Amongthis, the visit investigation included governmental agencies such as medicine companies,forestry bureau, agricultural bureau, grassland station, and non-government institution such asplanting enterprises and marketing establishment. In the field investigation, we use ecologysurvey method to examine the community characteristics of different Rhubarb habitats andestimate the standing stock of Rhubarb resource. Furthermore, we use HPLC method todetermine the content of14components in146the roots and rhizomes of Rhubarb medicinesamples. The components include5anthraquinones (Aloe-emodin, Rhein, Emodin,Chrysophanol, Physcion) and their glycoside composition,2Double anthraquinone glycosides(Sennoside A, Sennoside B), Catechin and gallic acid. Through this work, we analyse thequalitative characteristics of Rhubarb medicine from different habitats from the aspect of thecontent difference of functional components. Then we use matK gene as a carrier, and analysethe haplotype of431Rhubarb samples with the aid of molecular phylogeography method.From this, we elucidate the genetic mechanism of the qualitative characteristics of eachhabitat from the aspect of molecular phylogeography. Then with the statistics of climatic andsoil data in different geo-authentic habitat, through correlation analysis and path analysis, weanalyse ecological mechanism of the difference of functional components from differenthabitat.Result:(1) From the Rhubarb medicine resources investigation, we can find there existsignificant differences in the Rhubarb resources in various geo-authentic habitats. Xi Ning Da Huang and Ya Huang are primarily wild resources, and all the habitats in Gan Su areprimarily cultural resources. Our invesitgation shows that the wild resource of Rhubarb is onthe edge of extinction, and the cultivation techniques in Rhubarb lack of standardization, andthe cultural Rhubarb’s quality need to be improved.(2) Based on quality analysis of Rhubarb from different geo-authentic habitats, we discoveredthat there exist significant differences in the content of functional components from variousgeo-authentic habitats, and different habitat has diverse advantage in functional components.Among these, Zhuang Lang Rhubarb has an advantage in "Xie Xia Gong Ji", the content ofcombined anthraquinone can reach5.133%in average. Liang Huang showed superior effectin "Huo Xue Hua Yu", the polyphenols’ content can reach1.45%. In addition, some habitatshave overall advantage in many effect, for example, Xi Ning Da Huang showed bothadvantage in "Xie Xia Gong Ji" and "Huo Xue Hua Yu",the content of combinedanthraquinone can reach5.016%in average and the content of double anthrone compounds is0.362%, the content of polyphenol can reach0.913%. Ya Huang showed both advantage in"Qing Re Jie Du" and "Xie Xia Gong Ji", the content of combined anthraquinone and doubleanthrone compound is4.439%and0.492%, respectively. All these results incidate that,Rhubarb from differernt hebitat possess different advantage in effect, and this will help usmade the choice of habitat.(3) Our research on the genetic mechanism of quality differences in different geo-authentichabitat shows that hebitats which are close in geographical position, i.e. the matK haplotypein Rhubarb from the same habitat are analogous, and they are always clustered as the samegroup. For example, the haplotype inQuan Shui Rhubarb are concordant, this fully reflect thatquality difference in different geo-authentic habitat maybe closedly result from theirdifference in genetic mechanism. However, when it comes to Ya Huang and Xi Ning habitat,whose distribution is relatively wider, significant differentiation of hereditary basis exists notin the same habitat, but in different habitats. Our results infer that this might be associatedwith the Rhubarb resources’ disperse distribution. The distance between different habitat is far,and the mountain barrier, altitude difference are also limiting factor of gene flow. Those aboveresults illustrated that, on the basis of geo-authentic habitats proposed by our ancestors, thereis a necessity in exploring Ya Huang and Xi Ning habitat meticulously and intensively, thusmay facilitate the impactful usage of resources.(4) Our research on the ecological mechanism for the formation of quality difference indifferent habitats shows that, geographic area that are appropriate for the formation andaccumulation of functional component in Rhubarb are not broad. In this geographic region,high quality Rhubarb medicine mainly come from Gan Su, south of Shan Xi, east of Qing Hai,north of Si Chuan and north of Yun Nan. There are These areas have moderate rainfall andhigh altitude, so the duration of sunshine is long and can avoid humidity effectively. Heightfactor has a significant effect on the quality of Rhubarb medicine, generally speaking, in those regions, the higher the altitude, the better the quality of Rhubarb medicine.(5) The analysis of soil factors which affect the quality difference in different geo-authentichabitat shows the following conclusion. Soils which possess the following characteristics mayfacilitate the formation and accumulation of functional components in Rhubarb. Thecharacteristics are higher organic content, higer nitrogen content and higher availablepotassium content, all the microelement and the content are relatively high, alkalescent. Butthe appropriate soil condition are not the same, total nitrogen can facilitate the formation andaccumulation of free anthraquinones, but is disadvantageous to combined anthraquinone, andmake no influence to phenolics and double anthrone compounds. Effective magnesium isdisadvantageous to combined anthraquinone, but can facilitate the formation andaccumulation of free anthraquinones and phenolics notablely. The available potassium in thesoil can facilitate the formation and accumulation of free anthraquinones significantly, andexist no influence on the combined anthraquinone and double anthrone compounds, but isdisadvantageous to phenolics.Conclusion:From those above analysis, we can find that there exist significant differences inthe quality of Rhubarb medicines from different geo-authentic habitats, and the differencemainly result from the total effect of hereditary constitution and habitat environment. Theconclusion we get from this research can provide guidence for the choice of Rhubarb’ssuitable growing district according to their various funcions. And this may help the realizationof the" directed " clutivation of Rhubarb medicine with certain function in actual production.It will aid the goal of TCM’s accurate medication, and will guarantee the curative effect andsafety of traditional Chinese medicine.