| Paris polyphylla var.yunnanensis(Franch.)Hand.Mazz is a herbaceous perennid which belongs to Liliaceae family,Paris genus.It is recorded as Rhizoma Paridis with Paris polyphylla Smith var.chinensis(Franch).Hara in Pharmacopoeia of the People’s Republic of China.Paris plants have low rate of natural reproduction,low rate and long cycle of the roots’ natural growth.In recent years,with the growth demand of pharmaceutical industry for Rhizoma Paridis,the excavation speed is far exceed than the natural growth rate of Paris plants,which caused the scarcity of wild resources and the flooding phenomenon of closely related species used as Rhizoma Paridis in the marke.In the past ten years,the prices soaring of Rhizoma Paridis also stimulated the Paris plantation began to rise.Although some enterprises grow Paris,but most are individual farmers.They lack experience in planting and cultivation technology standardization,resulting in uneven variety of herbs and quality.However the character of Paris closely related species are highly converging,and the conventional character and microscopic identification are indistinguishable.The index chemical composition is also widely distributed and overlapped in multiple Paris species,which results in the lack of specificity for its conventional identification methods,and caused confusion in true and false identification and quality control.In response to the above problems,this study conducted a survey of resources in Sichuan and Yunnan to clarify the species composition of P.polyphylla var.yunnanensis and its common adulterant species,and established accurate,intuitive and reliable systems to authenticate P.polyphylla var.yunnanensis and Rhizoma Paridis’ legal origins based on DNA molecular marker technology.The results are as follows:1.Conducted resource surveys on artificial cultivation bases in various parts of Sichuan and Yunnan,and clarified that the species composition of medicinal plants of Paris in Sichuan and Yunnan Province were mainly nine species and varieties: P.polyphylla var.yunnanensis,P.polyphylla var.chinensis,P.polyphylla var.polyphylla,P.polyphylla var.stenophylla,P.axialis var.axialis,P.vaniotii,P.fargesii,P.thibetica var.thibetica and P.forrestii.2.Based on ITS barcoding,we have established a fluorescent visual identification system.It is proposed for the first time that P.polyphylla var.yunnanensis should be divided into two types of genotypes: YN-I and YN-II.In order to take into account two types of genotypes and avoid false-negative results,we designed two pairs of specific primers respectively,and established specific visual fluorescent identification systems which can be observed directly with the naked eye of fluorescent color.It has realized the rapid and directly visual identification of P.polyphylla var.yunnanensis and its common adulterant products,at the same time,it laid the foundation for the establishment of the identification system of Rhizoma Paridis’ legal origins.3.Established a multiple PCR identification system of Rhizoma Paridis’ legal origins for the first time.Compared to the conventional PCR method in which only 1pair of primers and 3 PCRs are required to complete a sample identification,the established multiplex PCR system has high efficiency and low cost,and can be accurately,quickly,and reliably identify the two legal origins of P.polyphylla var.yunnanensis and P.polyphylla var.chinensis only by one PCR amplification.4.Established a method for simultaneous determination of 7 kind of saponins content in Paris polyphylla var.yunnanensis based on HPLC.The determination of the two genotypes of P.polyphylla var.yunnanensis showed that 81.08% of the samples conformed to the Chinese Pharmacopoeia standards.Cluster analysis and principal component analysis were applied to the determination of samples and the results showed that the genotype differentiation of Paris polyphylla var.yunnanensis did not lead to differentiation of chemical components and the quality of medicinal materials. |
PDF Full Text Request