| Astragalus is dried root of Astragalus membranaceus legumes（Astragalus membranaceus(Fisch.)Bge.Var. mongholicus (Bge)） or Astragalus membranaceus legumes （Astragalusmembranaceus(Fisch.) Bge）. Hedysarum is dried root of multi-sequence Hedysarum（Hedysarumpolybotrys Hand Mazz.）. In Gansu Province, the Astragalus and Radix Hedysari were verywidespread with wild species and cultivatedvarieties. Astragalus and Radix Hedysari were in thesame family and in different genera, but Astragalus and Radix Hedysari were used mix in Chineseclinical medicine. What’s the material basis of Astragalus and Radix Hedysari? What’s theGenetic kinship between the Astragalus and Radix Hedysari? Therefore, this study comparativeanalysis Astragalus and Radix Hedysari by the morphological comparison, ISSR DNA molecularmarkers, the chloroplast genome of psbA-trnH sequence comparison and combination of infraredspectroscopy detection methods. In order to provide the necessary biological basis on Gansugeoherbs Astragalus and Radix Hedysari, and theoretical basis on its further research anddevelopment and widely used. The results were as following:1．Comparative Study on morphology of Astragalus and Radix Hedysari27different origin Astragalus and Radix Hedysari leaf sample were collected from Weiyuan,Longxi, Dangchang, Minxian of Gansu Province,20Astragalus,7Radix Hedysari. Thedifferences of morphology According to the five main characters:leaves, flowers, stems, podsand roots of Astragalus and Radix Hedysari, Astragalus with oval, obovate, elliptic-ovate oroval-shaped, obovate leaves and more smaller leaves than Radix Hedysari; Radix Hedysari withmore lobular, mostly ovate-lanceolate or ovate-oblong, rounded or obtuse apex, usually with thetip. The different colors of petals can visually distinguish between two forms, Astragalus petalsare yellow and Radix Hedysari was pale yellow. The pod coat of Astragalus glabrous was smoothpod and the pod coat of Radix Hedysari have pubescent with obvious reticulate.2. The genetic differences on ISSR analysisThree ISSR primers were screened from eight ISSR primers by ISSR molecular markertechnology, then DNA were amplified. Amplified DNA detectied by polyacrylamide gelelectrophoresis and got clear and stable fragments. Statistical analysis showed that23loci,16polymorphic loci, the average polymorm rate was71.3%, and amplified17loci,12polymorphicloci, in20Astragalus with polymorphism rate of70.5%; and13loci,8polymorphic sites, in7 Radix Hedysari with polymorphism rate of61.5%. The genetic diversity was0.49-0.88of27Astragalus and Radix Hedysari, an average of0.53. Construct the cluster analysis by UPGMA(unweighted pair arithmetic average) found that the genetic distance of0.49. According to this,27Astragalus and Radix Hedysari samples were divided into two classes: The first, geneticrelationship between Radix Hedysari H1(Dangchang magistrate goose ditch) and H7(WeiyuanCounty Chuanzhen) was nearest, H6(children Longxi County Beach Township) was a separategenetic with distance of0.58; the second, Astragalus h1(Minxian Qi Chuang) and h20(WeiyuanCounty, Qingyuan town), h3(MINXIAN meichuan town) and h12(13Longxi County, whereshop), h15(Weiyuan CountyMr Yeung Ka Zhai) and h18(Weiyuan County Chuanzhen) with theclosest genetic relationship and the genetic distance of0.64. The results showed that there were awealthy genetic diversity between Astragalus and Radix Hedysari, the genetic relationshipbetween them was further than Astragalus cohabitation group, genetic relationship of ISSR caneffectively analyse genetic diversity of Astragalus and Radix Hedysari.3. Astragalus and Radix Hedysari genetic differences based on psbA-trnH sequence ofchloroplastTo investigate the different populations of Gansu Astragalus and Radix Hedysari, Chloroplastgenome of psbA-trnH intergenic region sequences were amplified and the sequence identity wasanalysed by the molecular identification methods. The result showed that psbA-trnH of Astragalusand Radix Hedysari was331bp, with eight variants and many information variable sites.Dendrogram was constructed by the UPGMA, the results showed that there was big variationbetween them and the difference of psbA-trnH sequence could be used to identify Astragalus andRadix Hedysari.4. Infrared Spectroscopy analysis Astragalus and Radix HedysariInfrared spectroscopy was used to obtain infrared spectral fingerprint of the Gansu differentorigin Astragalus and Radix Hedysari,11common characteristics of peak absorbance wasselected and by second-order derivative spectrum, principal component analysis and neuralnetworks used to evaluat Astragalus and Radix Hedysari of classification model. The results ofprincipal component analysis show that selected the absorption peak1635.34cm-1and1035.58cm-1can be used to identify Astragalus and Radix Hedysari. Using the filter node ofrandomly selected65%of the sample as training set of them and difference analysed, the results show that the classification rate up to100%, indicating that the model accurate. Randomlyselected35%of the samples as a validation set, the Astragalus and Radix Hedysari discriminantanalysis, the results show the correct rate of90%. The conclusion indicate that the principalcomponent analysis and neural network analysis can be a accurated and reliable identification andvalidation of them to provide a scientific basis for quality evaluation of Astragalus and RadixHedysari.This experiment evaluated the different populations of Astragalus and Radix Hedysari, thekinship of them by morphology, ISSR molecular marker technology，chloroplast of psbA-trnHgene sequence analysis, and infrared spectroscopy detection methods. The difference betweenAstragalus and Radix Hedysari morphology can intuitively distinguish them; ISSR analysis of thenuclear genome level can clustering distinguish and explains the kinship distant between them;chloroplast genome of psbA-trnH sequence analysis showed that the chloroplast genome ofpsbA-trnH gene as molecular identification of Gansu Astragalus and Radix Hedysari; infraredspectroscopy combined with chemometrics methods can identify Astragalus and Radix Hedysari.This study revealed that the differences between Astragalus and Radix Hedysari on morphological,genetic and material basis can provide foundation for future clinical use and research. |
PDF Full Text Request