| Owing of the variation with in the speeies, easily natuer eross between speeies and wide generation, dieffrent planting enviromnent in the world, many speeies populations and Variety groups of penoy have been forming in the long history of penoy evolution and culturing, which make taxonomy and identification of penoy difficult and naming confusing. The study was undertaken to identify penoy material of several grapes of pengzhou area in sichuan provice. Though molecular markers technology of RAPD to studies the genetic diversity of 39 series penoys. The aim of the studies on the amplification patterns of RAPD got by NTSYS pc2.10e is to identificate and taxonomy penoy,study linkage distance.The main results as follows:(1)The methods-CTAB method were used to extract genomic DNA from leaves of grape. The DNA samples of the three methods were tested by spectrophotometer,agarose gelelectrophoresis and RAPD. The result was that the DNA integrality extract with those methods was good,the DNA trap was distinct, the amplified band of the RAPD is clear, the number of A260/A280 lied between 1.8and 2.0,which is in accordance with reaction requirements of RAPD.(2)The different PCR program and the ingredients in RAPD reation system such as DNA and primer,Taq DNA polymerase, dNTP,Mg2+, were sceened by different concentrations. Suitable reaction system and program of the RAPD for penoy genomic DNA was settled. The volumn of amplified reation was DNA 1μL (20ng/μL), 10×PCR buffer 2μL, Random primer 0.8μL(2.5μmol/L),dNTPs0.8μL(2.5mmol/μL),MgCl21.6μL(25mmol/L), Taq-DNA Polymerase 0.25μL(5U/μL), Double distilled water 13.55μL,total reaction 20μL. The reaction program was devised for 45 cycles,each with 94℃Pre-denaturation for 5 min,94℃denaturation for 1 min,36℃annealing for 1 min, 72℃extension for 2 min, at last 72℃elongation for 10 min,4℃for preservation.(3) 20 primers which are the better of 20 primers in the experiment are used to analyze the genetic diversity of 39 series penoy using RAPD markers.139 sites were found. Among which there are 118 polymorphic loci. The percentage of polymorphic loci is 84.89%. The average number DNA bands produced by each primer was 17.38, and theat of polymorphic loci was 14.75. The percentage of polymorphic loci among different penoy varies from 79.36% to 88.19%. The percentage of polymorphic loci of primers including S30 is the highest as the same time the lowest one is S85.Those above results showed that the genetic diversity of 39 accessions was high.(4) 39 kinds of Peony are researched in classification by RAPD technology, according to the result of cluster analysis, estimate the genetic distance, set up tree chart, when the dissimilarity coefficient is 0.46,39 kinds of Peony are obviously gathered into 2 kinds. Local Peony belongs to the first kind, while Japanese Peony belongs to the second kind. The first kind includs 35 varieties, which are divided into 2 subgroups, the first subgroup includs 31 varieties that are zhongyuan Peony. The second subgroup includs 4 varieties that are local Peony of Pengzhou. The conclusion based on the result bofore are that the variety of Peony coming from same place can show a close relatives relationship. |
PDF Full Text Request