| Objective:In order to clarify the origin of Pueraria Lobata Radix,Pueraria Thomsonii Radix and Pueraria Flos.the three varieties,Pueraria montana var.lobata(Willd.)Sanjappa & Pradeep,P.montana var.thomsonii(Benth.)Wiersema ex D.B.Ward and P.montana var.montana(Lour)Van der Maesen,were comprehensively analyzed from the aspects of resource distribution,morphology and phenology.At the same time,on the basis of germplasm evaluation,the medicinal components of root and flower from 3 varieties of P.montana(Lour.)Merr.were analyzed to provide reference for the resource development and protection of P.montana(Lour.)Merr.Methods1.By consulting the samples of three varieties of P.montana(Lour.)Merr.in China and combining field investigations in several provinces,we sorted out the distribution of varieties of P.montana(Lour.)Merr.in China,investigated and collected the germplasm resources of P.montana(Lour.)Merr.,and conducted a preliminary assessment on the germplasm diversity of P.montana(Lour.)Merr.2.From February 2020 to January 2021,11 germplasm of P.montana(Lour.)Merr.planted in Jiangxi University of Traditional Chinese Medicine Germplasm Resources Garden were counted,and the morphological traits of stem,leaf,flower and fruit were measured.The phenology and morphological differences of 3 varieties of P.montana(Lour.)Merr.were compared and analyzed.3.UPLC-Q-TOF-MS was used to analyze and identify the chemical components of root from25 germplasms of P.montana(Lour.)Merr.which from several provinces and cities,and OPLSDA model was established to analyze the differences in the MS data obtained by detection,and the main differential compounds in the root of 3 varieties of P.montana(Lour.)Merr.were obtained through VIP value screening.At the same time,a HPLC method was established to detect the contents of 10 isoflavones in the roots to evaluate the quality of root medicinal materials of different germplasm and different varieties.4.UPLC-Q-TOF-MS was used to detect,analyze and identify the chemical components of flower of 28 germplasms from 3 varieties of P.montana(Lour.)Merr.,and OPLS-DA model was established to analyze the differences of the detected MS data,and VIP value was used to screen the main compounds in flower of the 3 varieties of P.montana(Lour.)Merr..At the same time,the contents of 8 isoflavones in flowers were detected by HPLC to evaluate the quality difference of different varieties of medicinal materials from flower parts.Results:1.The distribution of the three varieties of P.montana(Lour.)Merr.showed obvious differences from north to south.P.montana var.lobata was mainly found in northern mountains and forests,while P.montana var.thomsonii and P.montana var.montana had a broad common distribution area,mainly in the south of the Yangtze River.At the same time,the three varieties of P.montana(Lour.)Merr.formed a common mixed distribution in southwest China.It was found that there were abundant differences in root,leaf,flower and other morphological traits among the varieties of P.montana(Lour.)Merr.in the wild state.2.There were some differences in the phenological periods of different varieties.The germination of P.montana var.lobata began in mid-February and began to blossom in late June.Germinates of P.montana var.thomsonii in early March and enters the flowering period in early September.Among the P.montana var.montana,JXLB* had the phenomenon of early germination and late defoliation,while other germination of P.montana var.montana had similar phenology to germination of P.montana var.thomsonii.There were significant differences in inflorescence axial length,corolla size and fruit shape among the three varieties,which could be used as the basis for distinguishing the three varieties at different stages.3.A total of 84 compounds were identified from the roots of the three varieties P.montana(Lour.)Merr.,46 of which are isoflavones,7 kinds of flavones,17 kinds of triterpenoid saponins,8 kinds of carotenol glycosides,2 kinds of coumarins and 4 kinds of sesquiterpenoids.The root of P.montana var.lobata,P.montana var.thomsonii and P.montana var.montana have 84,81,70,respectively.18 different compounds were screened under the positive and negative ion modes,including Puerarin,Puerarin-6?-O-xyloside,3?-methoxy puerarin,3?-hydroxy puerarin、Daidzin、6?-O-malonyl genistin,6?-O-malonyldaidzin,Genistein,Daidzein-4?,7-di-glucoside,Puerarin-7-O-glucoside,5,6,7-trihydroxy-4?-methoxy isoflavone-6,7-di-O-β-D-glucopyranoside,kudzusaponin A5,Genistein-8-C-apiofuranosyl(1→6)glucoside,sophoraside A,Glycitin,Genistin,daidzein-4?-β-D-glucopyranoside,3?-hydroxy puerarin-4?-O-deoxy Hexoside,Apigenin7-glucoside,Daidzein,Soyasaponin I,Isoorientin,Sissotrin,Ononin,most of which are isoflavones.4.A total of 35 compounds were identified by composition analysis,22 of which were isoflavones,6 kinds of flavonoids and 7 kinds of saponins.The flowers of P.montana var.lobata,P.montana var.thomsonii and P.montana var.montana have 32,35,33,respectively.18 different compounds were screened under the positive and negative ion modes,including Kakkalide,Tectoridin,6″-O-xylosyl-tectoridin,Glycitin,4’-methyltectorigenin 7-glucoside,6″-O-xylosylglycitin,Irisolidone,Kaikasaponin III,6″-O-malonylglycitin,Kakkalidone,Tectorigenin,Rutin,Soyasaponin BB,Vitexin,Biochanin A,Genistin,Kakkatin,Azukisaponin II.Conclusion:The three varieties of P.montana(Lour.)Merr.are widely distributed in China,and the regional distribution of different varieties shows a north-south difference,and there are mixed distribution in the mountainous areas of southwest China.Different varieties of P.montana(Lour.)Merr.have different growth habits and different phenology,so we should pay attention to the difference management in the process of germplasm acclimation and cultivation.At the same time,the inflorescence axial length,corolla size and fruit inflorescence characteristics of the three varieties are different,which has certain reference significance for the identification of wild plants.At the same time,24 compounds with root differences and 18 compounds with flower differences were screened by comparing the components of root and flower parts,which had certain reference value in the identification of root and flower of P.montana(Lour.)Merr..In the application of medicinal materials,although Pueraria lobata Radix and Pueraria thomsonii Radix are used separately as two traditional Chinese medicines,there is no difference in efficacy,and there is a great difference in component content between the two.Therefore,it is suggested to further distinguish the efficacy of medicinal materials in clinical use.At present,the flowers of P.montana var.lobata and P.montana var.thomsonii are both included as the source of medicinal materials,but there are significant differences in the contents of main functional components such as kakkalide and tectoridin,suggesting that they should be used separately.At the same time,the quality of flower from P.montana var.thomsonii,which is the main source of the market,is greatly affected by the difference of germplasm,so it is urgent to establish a standard quality evaluation system for the control of Pueraria Flos.This study is the first time to systematically study the P.montana var.montana,and evaluate the application value of this variety through the study of chemical constituents.Although the content of puerarin and other components in root is low,it has higher content of genistein and genistein,which has certain development value.Although the Flowers of it is a fake product,the contents of tectoridin and 6″-O-xylosyl-tectoridin are high,which has a potential for development. |
PDF Full Text Request