樟樹居群遺傳多樣性及遺傳結構的RAPD分析.pdf

1、福建師范大學碩士學位論文樟樹居群遺傳多樣性及遺傳結構的RAPD分析姓名：潘曉華申請學位級別：碩士專業(yè)：植物學指導教師：朱錦懋黃儒珠20040401AbstractThegeneticdiversityandgeneticstructureofCinnanlonffllCamphora(L)PreslpopulationwerestudiedbyrandomamplifiedpolymorphicDNA(RAPD)Thesiswascom

2、posedof4parts：1BasedontheconventionaICTABmethodanefficientprocedurewasdevelopedThemajorimprovementincludedeliminationaIotofsecondarymetaboIitessuchaspolyphenolsetcandcollectionofnucleibeforefreeDNAThen3CTABbuf艷rwhichcons

3、istedof100mrnollLTris(PH8∞，25mmol／LEDTA15mol／LNaCI，30％C11ABwasaddedtoextracttotaIDNATheresultsfromratiosofA260andA2so，electrophoresis，digestionwithrestrictionendonueleasesandPCRamplificationsuggestedthatisolatedDNAwassui

4、tableforRAPDanalysis2ApreservativemediumwhichconsistedofsaturatedNaCl一C1AB2％(w：VPVP2％(V：V)8一sulthydrylethanol，50mmol／LEDTA(pH8o)，wasappliedtofieldpreservationofleavestissueforDNApreparationofCCamphoraItwasindicatedthatth

5、eisolatedDNAfromtheleavestissuepreservedinthemediumfor7d、14dwassimilartothatfromfreshleavestissue3EfrectsofthecontentofDNAtemplateprimerdNTPMg“andTaqDNApolymerase011experimentalresultweretestedbyuniformtestdesign。andtheo

6、ptimalreactionsystemofRAPDforCCamphoraWasdetemainedas協(xié)llows：8ng／uLDNAtemplate，02umol／Lprimer，02mmol／LdNTP，15mmol／LMgCIzandlU7bqDNA，polymeraseintotal20pLreactionvolumeEffectsot’thedenaturationtimeandannealingtemperatureon

7、experimentalresultweretestedaswellTheoptimalamplificationprocedureconditionswere：180sat94℃30sat94℃，90sat35℃，120sat72℃，cycling42times300sat72℃，thenstoredat4℃4111egeneticdiversityandgeneticstructureofCCamphorapopulationwer

8、estudiedbyRAPDneanalysisdataof168RAPDlOCiof2lrandomprimersindicatedthatleveIofgeneticvariabilityinthe68samplesfrom7populationsinFujianwasnothighThenumberofpolymorphicbandswas75Thepercentageofpolymorphicbands(PPB)was4464％

9、ThegeneticdiversityofShaowupopulation(SW)Wasthehighest，whichPPBwas3333％AndFujianAcadenryofForestrypopulation(vs)wasthelowest，whichPPBwas2262％Asanalyzedbypopulationgeneticanalysis(POPGENE)132，Shannon’SInformationindex，Nei