Study On Genetic Diversity Of The Pinellia Ternata (Thunb.) Breit Based On Molecular Markers

Pinellia ternate（Thunb.）Breit, belonging to Araceae, is a kind of traditionalChinese medicinal materials, which has been used to modify cough, dispel phlegm,calm vomiting, anti-ulceration, anti-tumour, and terminate early-pregnancy etc.Special for East Asia, P. ternata distributes in South Korea, Japan and China, wheremainly in the East China and South China. Differentiation in populations andphenotype formed because of geographical distance and time, which resulted inconfusion in cultivars later. The quantity of wilding resources declined sharply dueto extensive commercial exploitation and lack of protecting rules. It is time for P.ternata germplasm to be arranged and evaluated, so as to establish P. ternata coregermplasm collection.Germplasm of Pinellia ternata distributed in main areas of China were collected.We exploited Sequence Related Amplified Polymorphism(SRAP), Target RegionAmplified Polymorphism（TRAP）and Restriction Site Amplified Polymorphism（RSAP） markers for P. ternata genetic diversity study. In addition, DNAbarcodes technology was an introduction. Two cpDNA non-coding sequences werescreened to test genetic variance within and among populations in P. ternata. Thestudy above laid the foundation of the phylogeny of the plants, as well as theformulation of protection rules of P. ternata. The main contents and results are asfollows:1. Analysis of the genetic diversity of P. ternata germplasm resources in China bySRAP+TRAP Molecular marker technology43germplasm resources were utilized from16provinces of P. ternata maindistribution.13primers screened from208SRAP primer combinations and3primers selected from116TRAP primer combinations were used to analyze genomeDNA. Total286bands are produced through detection, polymorphic sites accountsfor65.31%. The average value of Shannon index was0.4896, Nei’s gene diversityindex （He） is0.3191, effective number of alleles and observed numbers of alleleswas1.5269,2.000, respectively. Groups of genetic differentiation factor （GST） is0.4112, population gene flow（Nm=0.715）. Data above showed that much differentiation existed among populations. Geneflow may become the main course leading to high differentiation level. The sameresult come from analysis of molecular variance （AMOVA） also displayed that32.82%genetic variation existing among populations. Cluster analysis revealed noagreement with geographic distribution, which kept accordance with the outcome ofprincipal component analysis （PCA）.2. Analysis of the genetic diversity of P. ternata germplasm resources in China byRSAP Molecular marker technology13primers combinations were elected from45RSAP primers to demonstrategenetic diversity of Pinellia ternata.212polymorphic sites were generated, whichtook account for94.62%. Shannon’s information index, Nei’s gene diversity,effective number of alleles and observed numbers of alleles were0.4554,0.2971,1.4903, and1.9731, respectively. Based on POPGENE3.2, Gst=0.4232，Nm=0.6814.Cluster analysis using unweighted pair-group method with arithmetic means(UPGMA) divided all the populations into9groups, which showed no correlationwith geographic distribution, agreeing with the principal component analysis.3. chloroplast DNA non-coding sequence analyzing of Pinellia ternata andrelated speciesTo provide evidence for identification and genetic diversity of Pinellia,sequencing of cpDNA non-coding sequences from Pinellia ternata and relatedspecies were performed.43Pinellia ternata and one Pinellia Cordate one Pinelliapedatisecta were collected from main producing regions of China. psbK-psbI andatpF-atpH from leaf genome DNA were amplified by PCR. Comparative analysiswas carried out by bioinformatics software. The length of atpF-atpH region fromPinellia ternata varied from337to342bp. The number of variable sites andparsimony information sites was only7and1respectively, showing conservatives.The length of the other sequence of Pinellia ternata, psbK-psbI, was432to435bp,with37variable sites,17parsimony information sites. The genetic distance rangedfrom0~0.069. Cluster analysis did not agree with the phenotype or the geographicaldistributions. psbK-psbI was more discriminating than atpF-atpH, and could becomethe candidate sequence of DNA barcodes for identification of Pinellia.