Molecular Identification Of Medicinal Ligusticum And Exploitation Of Its Substitute-

In the field of traditional Chinese medicine and Chinese patent medicine,adulteration,unstable quality and other factors have severely restricted the sustainable development of related industries.For the safety and effectiveness of traditional Chinese medicine and Chinese patent medicine,DNA barcode technology was used to clarify the botanical identity of medicinal Ligusticum plants,and explored its possible substitutes;at the same time,the prescription components of Chinese patent medicine Renshen Jianpi Wan were analyzed.Therefore,DNA barcode technology provided the basis for the establishment of quality control of traditional Chinese medicine and Chinese patent medicine and for the ensuring the clinical safety:(1)Molecular Identification of Medicinal Ligusticum PlantsIn this study,the ITS2 region was used to identify 43 accessions representing Ligustici Rhizoma et Radix and its adulterants,and 25 commercial 'Gao-ben'products purchased from different medicinal markets in China.Firstly,the ITS2 secondary structure can be used for the identification of genuine 'Gao-ben' and to distinguish it from common substitutes and adulterants;Secondly,the twenty-five samples from medicine markets were identified as Ligusticum sinense&L.jeholense,Conioselinum vaginatum and Meeboldia yunnanensis,indicating that crude drugs available in the market have complex origins(76%of which are adulteration and substitution).(2)Exploration Substitutes of Medicinal LigusticumBased on the molecular phylogenetic framework of the Ligusticum and the analysis of chemical constituents,this study explored the substitutes for the genuine Ligusticum.Ligusticum pteridophyllum and Cnidium officinale(distributed in Japan)are closely related to the genuine Ligusticum,and contain similar chemical composition,so they can be substituted for true 'Gao-ben'.(3)Application of Metabarcoding Technology in Component Analysis of Ren shen Jianpi WanIn this study,a total of 23 batches of Renshen Jianpi Wan from 11 manufacturers were used to test the prescription components by the DNA metabarcding technology.High-throughput sequencing was carried out using ITS2 as barcode,and QIIME2 software for data analysis.The results showed that:1)DNA metabarcoding technology can detect all prescription components in Renshen Jianpi Wan except Atractylodes macrocephala,Aucklandia costus and Poria cocos;2)Substitutes or adulterants,such as Panax quinquefolius,Panax notoginseng,Ziziphus mauritiana,were detected in some samples;3)The species composition detected by different manufacturers varied greatly,and there is no significant difference for different batches of the same manufacturer.Therefore,DNA metabarcoding technology can be used as an important tool for the analysis of prescription component for Chinese patent medicine.