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Genetic Diversity Of Six Populations In Chinese Pine (Pinus Tabulaeformis) By RAPD Analysis

Posted on:2009-07-09Degree:MasterType:Thesis
Country:ChinaCandidate:F Q GaoFull Text:PDF
GTID:2120360272463640Subject:Ecology
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The genetic diversity and genetic structure of six Pinus tabulaeformis Carr.populations in Shanxi,Shaanxi,Hebei and Liaoning provinces were assessed by using RAPD markers.The gradient tests on the concentrations of random primers,template DNA,dNTP,TaqDNA polymerase and Mg2+ were conducted.An optimal reaction system suitable for the RAPD analysis of Chinese pine was established with 25μL volume,containing 2.5μL10×Buffer,3.0μL 25 mmol/L MgCl2.0.5μL10 mmol/LdNTP,0.5 U Taq DNA polymerase,40 ng template DNA,2.0μL 10μmol/L random primers and 14.9μL ddH2O.At the same time,19 primers were chosen from 60 kinds of 10 bp random primers, which could produce polymorphic,clear and stable bands.From 102 individuals of 6 populations,19 primers yielded 146 clearly identifiable bands,and among them 122 polymorphic bands were polymorphic.As analyzed by POPGENE 1.31,the percentage of polymorphic loci(PPB) was 83.56%,the Nei's gene diversity(h) and Shannon diversity index(I) were 0.2587 and 0.3786,respectively,in the species.These indicated that genetic diversity of these populations was high. The populations of Lingkong Mountain(LK)(PPB =66.44%,h=0.2735,I =0.3965) and Wuling Mountain(WL)(PPB=65.75%,h=0.2639,I=0.3855) had higher genetic diversity than other 4 populations of P.tabulaeformis. Effective number of alleles per locus(ne) was 1.4667 and observed number of alleles(na) was 1.6518 in the species level.The total genetic diversity(Ht) was 0.3263,and the genetic diversity of within populations(Hpop) was 0.2587.The coefficient of genetic differentiation(Gst) was 0.2072,and gene flow(Nm) was 1.9145.It was indicated that 20.72%of total genetic variation was among populations,and 79.3%within populations.These showed that majority variation occurred within populations,while there existed certain degree of gene exchange among populations.This study also indicated that genetic distance among populations of P. tabulaeformis was from 0.0791 to 0.1380,with a mean value of 0.10482. Genetic distance between LK and PQ was biggest(0.1380) and genetic distance between LK and WL was smallest(0.0791).Based on the values of Nei's genetic distance,the result of UPGMA cluster analysis showed that these 6 populations could be classified into two groups.The first group consists of populations LK and WL,and the second includes populations QM, PQ,HL and LN;or the first one consists of populations LK,WL,QM and LN,and the second includes populations PQ and HL.The effect of geographical separation on genetic structure was examined by Mantel test based on matrices of pairwise geographical and genetic distances among six populations ofP.tabulaeformis.The result indicated there was no significant correlation between geographic and genetic distances among populations.Based on the above-mentioned results,some strategies for the protection and exploitation of genetic diversity of P.tabulaeformis were put forward in this thesis.
Keywords/Search Tags:Pinus tabulaeformis Carr., Genetic diversity, RAPD, Conservation
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