Studies Of Germplasm And Preliminary Construction Of Core Collection Of Fallopia Multiflora (Thunb.) Harald

Fallopia multiflora is a perennial herbaceous plant,which is distributed in most parts of China.The study shows that Fallopia multiflora is in an active evolutionary period with a fast evolutionary speed,and it forms different geographical provenance when it adapts to the complex and changeable living environment.Therefore,in the natural environment,Fallopia multiflora contains rich and diverse germplasm resources and contains excellent genes to resist adversity.The preparation of Fallopia multiflora which has been recorded in the book of materia medica for thousands of years has the functions of enriching blood,strengthening kidney and black hear.As the mainstream of this product,wild Fallopia multiflora is decreasing at a rate of about 15% per year while the demand for Fallopia multiflora is increasing.The depletion of Fallopia multiflora resources,on the one hand,will reduce species diversity,viability,but also lead to the loss of good genes,seriously affect the sustainable development and utilization of Fallopia multiflora.Therefore,the core task that needs to be completed urgently is tostrengthen the research of Fallopia multiflora germplasm,and then build a core germplasm bank to protect the core germplasm in a targeted manner,laying a foundation for subsequent research and excavation of excellent germplasm.This experimental study is mainly divided into four parts.The methods and results are as follows:(?)Genetic diversity and population structure analysis of Fallopia multiflora.In this experiment,SRAP molecular marker method was used to explore the germplasm diversity and population structure of Fallopia multiflora germplasm by 327 Fallopia multiflora samples from 44 populations in 12 provinces in China,and the analysis results are as follows :(1)The genetic diversity of wild population of Fallopia multiflora is greater than that of cultivated population,and the diversity mainly comes from the interpopulation.In the wild population,the genetic diversity was high in Sichuan population and Chongqing population,and the genetic distance between samples of Fallopia multiflora had no obvious correlation with geographical distance.(2)The NJ clustering results of Fallopia multiflora samples were consistent with geographical distribution,and samples from the same collection points could be clustered together.In addition,the results also showed that the cultivars of peasant households had close relationship,indicating that there was a situation of mutual introduction.(3)Population structureanalysis showed that most Fallopia multiflora samples had a single consanguineous composition,and when K=16,the sample classification effect was the best,and the classification results were basically consistent with NJ clustering.(4)SRAP molecular markers are an effective tool for estimating genetic diversity and population relationships of Fallopia multiflora.Terrain complexity and less gene communication are important reasons for the degree of genetic diversity of Fallopia multiflora.(5)Polygonum multiflorum Thunb var angulatum S.Y.Liu from Guangxi shows specificity in Fallopia multiflora population and has a large genetic distance from other provenances,therefore,it is suggested to classify it as a variety of Fallopia multiflora.(?)Quality study on root of Fallopia multiflora from different sources.The contents of 2,3,5,4'-tetrahydroxystibene-2-O-D-glucopyranoside,free anthraquinone and conjugated anthraquinone in root of Fallopia multiflora from different sources were determined by HPLC.The analysis results included :(1)In the content determination,the variation coefficient of the contents of 2,3,5,4'-tetrahydroxystibene-2-O-D-glucopyranoside,free anthraquinone and conjugated anthraquinone of Fallopia multiflora from different sources was very high,and the three components of Fallopia multiflora from different years of cultivation were significantly different,but the wild and cultivated Fallopia multiflora growing inGuangdong province met the requirements of pharmacopoeia regardless of the growth years.(2)When roots were planted for 7 months to 2 years,the content of 2,3,5,4'-tetrahydroxystibene-2-O-D-glucopyranoside and conjugated anthraquinone had a very significant and high positive correlation,while free anthraquinone showed a non-significant negative correlation with the 2,3,5,4'-tetrahydroxystibene-2-O-D-glucopyranoside and conjugated anthraquinone.However,when the roots were planted for3-4 years,there is no significant correlation among the three.The relationship between free anthraquinone and the other two components changes from negative correlation to positive correlation.(3)Polygonum multiflorum Thunb var angulatum S.Y.Liu does not contain anthraquinones,and the content of2,3,5,4'-tetrahydroxystibene-2-O-D-glucopyranoside does not meet the requirements of the Pharmacopoeia.(4)The root of Fallopia multiflora from different sources show obvious differences in color,texture,and structure.The same or similar provenances also show certain differences when they are cultivated or grown in different places.(?)Analysis on the difference of root metabolites of Fallopia multiflora from different sources.The UPLC-Q-Orbitrap technique was used to study the differences of root metabolites of Fallopia multiflora from different sources,and to select the main different components of Fallopia multiflora root fromdifferent habitats,so as to provide a reference for the formation mechanism of Fallopia multiflora in authentic production areas.The results are as follows:(1)Thirty-five Fallopia multiflora samples were clustered into two categories.The first type was Deqing provenance.In the second major group,the metabolites of Polygonum multiflorum Thunb var angulatum S.Y.Liu(LZ)showed significant differences from other samples and formed a small branch.In addition,The cultivated and wild products in Guizhou are close to each other and their metabolites are clustered in Hetai 1-2,Mocun 7,and Maoming samples.The samples from Shantangjiao Village,Xintiantang Town,Guangdong,and the samples from Guigang,Guangxi are similar.The samples were gathered into a small group in Yuncheng,Shanxi and Chongqing Medicinal Botanical Gardens.The clustering results showed that the roots of Fallopia multiflora from the same provenance or the same cultivation place had similar metabolites.(2)The classification results of unsupervised principal component analysis were consistent with the results of cluster analysis.On this basis,the differential metabolites of Fallopia multiflora grown in Deqing provenance or guangdong province and other producing areas or provenances were investigated.Thus,7differential metabolites,mainly catechins,were selected.Gallocatechin-(4??8)-epicatechin?Epigallocatechin-(4??8)-catechin?5,7,3',4',5'-Pentahydroxyflavanone ? Epigallocatechin-(4? ?8)-epicatechin-3-O-gallate ester?(+)-Gallocatechin(-)-Epigallocatechin(?)Preliminary construction of core germplasm of Fallopia multiflora.The core germplasm was constructed based on the results of genetic diversity and population structure of Fallopia multiflora germplasm,using three sampling strategies that include "NJ(adjacent method)clustering-proportional sampling","STRUCTURE classification-proportional sampling" and "minimum distance stepwise cluster sampling" and using a total of 6 proportion sampling modes of 5%-30% to construct the core germplasm.At last,the t-test is used to compare the four genetic parameters(Na,Ne,H,and I)of the initial germplasm with the original germplasm,A good core germplasm construction method was screened out,and a representative and reasonable core germplasm sample was selected based on the chemical composition research results.The results of core germplasm construction are as follows:(1)The germplasm library constructed by "STRUCTURE taxation-proportional sampling" and 10% sampling proportion has high retention rate of genetic diversity,relatively small sample quantity and strong representativeness.(2)The core germplasm bank consists of 37 samples,including 9 cultivated products and 28 wild products.The cultivated products include Polygonum multiflorum Thunb var angulatumS.Y.Liu and two farmer planted varieties.The wild products are mainly composed of samples from Sichuan,Chongqing and Guizhou.In addition,Hetai No.1,Hetai No.2 and Sankeshu samples are special and excellent samples in the study of composition and also included in the core germplasm bank,which can provide materials for future selection of excellent germplasm.