柑桔dna指纹图谱库的构建与分析

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1、西南大学硕士学位论文cultiVars，usingthetechn0109ysystema_bOVe．Itwilllayafoundationfordetectionofauthenticityandpurityamongcitrusseedings．ThemainresultswereaSf01lows：EstablishmentOfDNAfiⅡgerprintingdetectionsystemFourmolecularmarI∞rsystemshaVebeenestablishedbyPCRreactionfktorSOptimization．RAPDmarkersystem：tem

2、plateDNA50ng，lo×PcRBufrer2．5pL，25mMM∥+1．5”L，2．5mMdNTP1．5肛L，勋gDNApolymerase1．OU，20n∥岫primer1pLin25“Lsystem．．SSRmarkerSystem：templateDNA50ng，10×PCRBufrer2．5pL，25mMMgp2．O皿，2．5mMdNTP2．0¨L，砀gI)NApolymerase0．8U，20ng／pLprimerslpLin25pLsystem．．1ssRmarkersystem：templateDNA50ng，1o×PcRBufrer2．5灿，25mMM旷+1．6m一

3、，2．5mMdNTP2．0此，聊DNApolymerase1．0U，20n∥此primerl此in25儿syStem．．sRAPma暾erSyStem：templateDNA50ng，1o×PcRBuffer2．5IJL，25mMM∥+．1．2此，2．5mMdNTP1．2止，购DNApolymerase1．OU，20n∥此primersl止in25此system—Prime璐screeningByscreeningprimerpairs，34outofl05SSRprimerpairs，6outof40RAPDprimers，4outof20SRAPpimerpairsand3outof4

4、0ISSRprimerSwereobtained，、vhichwithhighpolymorphisma11ds诎ili吼Polymo叩hicprimersaccountedfor32．38％，l5％，5％andl7．5％respectiVely-It’scontaining14SSRprimersdesignedbyourselVes．SSRmarkerwaSconsideredasthekeymethod锄ongcitrusDNA6nge叩rinting，锄dISSRmarkeraStheaccessorialmethod．CitrusDNAnngerprintingl26citrus

5、cultiVarswereanalyzeduSingSSRmarker锄dISSRmarker．241emoncultiVarS，14pomelocultiVarS，22hybridcultiVarsand6clementinacultivarscouldbeidentifiedbySSRmarker．1SSRmarkercouldidentified5unshuicultiVars，18sweetorangecultiVarsaIld4ponkancultiVarswhichcouldnotbeidemifiedbySSRm矾∞r．93outof126citmscultivarswbre

6、identifiedwitllSSRandISSRtOgether．COnstructionOfDNAfingerprintingdatabaseincitruscuKVarsl2SSRpimerpairsand2ISSRprimerswithhighpolymorphismandnolinkagerelationshipwereconsideredaLsthecoreprimersforconstmctionofcitrusDNAfingerprintingdatabase，includingSS2，SS15，SSl6，SSl7，SS18，Cssr015，Cssr036，Cssr037，

7、Cssr038，Cssr050，Cssr05l，Cssr052，IS09andIS22．Amodefigerprintd砒abaSecontained93citmscultiVarshaSbeen