| Rhodiola kirilowii(Tibetan name"Gaduer Manba")is customarily used for Tibetan.R.kirilowii contains gallic acid,salidroside,tyrosine and other components,with anti anoxia,anti aging effect,main treatment of plague,lung heat,poisoning and limb swelling and so on.R.kirilowii was recorded in the Tibetan medicine classics,such as Yue Wang Yao Zhen,Si Bu Yi Dian,and Jing Zhu Ben Cao.Dictionary of Chinese Ethnic Medicine recorded the use of Rhodiola sachalinensis in Kazakh,Qiang,Uygur,and Tibetan.It was also collected in the standard of Tibetan medicinal materials in Sichuan(2014 Edition).The content determination index component is salidroside.In recent years,due to excessive mining,in addition to the restriction of the growth environment and its own conditions,the wild resources of Rhodiola have been reduced dramatically.R.kirilowii,as an alternative variety of Rhodiola crenulata,is very necessary for its resource status,drug safety and cultivation technology.In order to provide scientific basis for the protection of wild Rhodiola of Tibetan medicine and cultivation of artificial cultivation of Rhodiola,the study of resources,ecological suitability,the correlation of Chemical Components Accumulation and ecological factors,and ~1H-NMR metabolomics research are carried out.Objective:1.To investigate the distribution of wild resources and ecological suitability of R.kirilowii in order to provide evidence for the utilization of R.kirilowii.2.To determine the effective components of R.kirilowii from different habitats,and to analyze the correlation with ecological factors,so as to provide basis for breeding of R.kirilowii varieties.3.The metabolic components of R.kirilowii in different habitats and different growing years were evaluated and analyzed by ~1H-NMR technology,which provided a basis for the artificial cultivation of R.kirilowii.Method:1.The investigation of R.kirilowii germplasm resources was conducted by means of field investigation and visiting hospital.2.The ecological factors were extracted from the global climate database,and the ROC evaluation curve was set up based on the Maxent model.At the same time,the suitability of the R.kirilowii was studied by using the ArcGIS software.3.The total flavonoids and polysaccharides in R.kirilowii were determined by colorimetric method and sulfuric acid anthrone method,and the content of gallic acid,glucoside,salidroside and tyrosalin in R.kirilowii was determined by HPLC-UV-ELSD method.4.The data of average pressure,annual precipitation,average wind speed,sunshine hours,relative humidity,annual mean temperature,annual maximum temperature,minimum annual temperature and other 9 ecological factors are obtained by using China Meteorological scientific data sharing service network platform.The data are divided by PLS and RDA.The correlation between chemical composition and ecological factors was analyzed.5.~1H-NMR analysis technology was used to establish fingerprints,and the similarity coefficient of Rhodiola sachalinensis was calculated by correlation coefficient method.A field test was used to determine the growth index of Rhodiola sachalensis with different growth years,and ~1H-NMR technique was used to analyze the metabolic components of Rhodiola sachalidensis with different growth years.Result:1.Through field investigation,it was found that the density of wild leaved R.kirilowii populations in 4 provinces was very different,and the reserves of wild resources decreased gradually in many areas.According to the results of the survey,the wild resources of R.kirilowii in 16 counties of 4 provinces are about 1100 t,and the resources of R.kirilowii in Sichuan province are the largest.2.Combined with Maxent model and ArcGIS analysis,the results show that Rhodiola sachalensis in the Qinghai Tibet Plateau,part of Xinjiang,Shaanxi,Shanxi,Hebei and other regions,mainly influenced by the altitude,the warmest quarter precipitation,the coldest quarter precipitation,and the soil sediment ratio.In China,the suitable area is 717 thousand square kilometers,and the suitable area is 1 million 882thousand square kilometers.3.The method of determining the content of gallic acid,salidroside,tyrosine and glucoside in R.kirilowii was established.The method was short of time and good separation effect.The method of study and sample recovery were all in accordance with the requirement of content determination,and the content of 4 components in 13batches of different habitats was determined by this method.PLS and RDA analysis showed that salidroside had a strong positive correlation with altitude and sunshine hours.There is a strong negative correlation with relative humidity.4.The 23 primary and secondary metabolites were identified from the extract of R.kirilowii,including acids,flavonoids,amino acids,carbohydrates and fatty acids.In addition,the average similarity of 13 batches of fingerprints was 0.979.Integrating ~1H-NMR data with Simca-P,PCA analysis and PLS-DA analysis can clearly distinguish the different years of R.kirilowii.Conclusion:1.Based on the Maxent model and ArcGIS technology,the suitability division of R.kirilowii was studied,and the classification of the suitability grade was carried out to provide scientific basis for the selection of the wild R.kirilowii and the artificial normalized planting base of the R.kirilowii.2.There are obvious differences in the climate of different habitats.The chemical constituents of R.kirilowii from different habitats are obviously different.PLS and RDA were applied to analyze the influence of ecological factors on content and correlation.3.The ~1H-NMR analysis method has the characteristics of the whole,and the sample preparation is simple,the analysis is fast and the reproducibility is good.The established fingerprint provides the scientific basis for the quality control and evaluation of R.kirilowii.The combination of relative content method and principal component analysis method can be used to evaluate and analyze the active components of R.kirilowii at different growth stages,which can be used as a method to study the metabolomics of plant medicinal materials. |
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