建立快速鉴定凤尾菇菌株的SCAR标记(江西农业大学学报)

©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(3):[e-mail],,,3(,350002):DNA,9RAPDSRAP,32741760bp356bp1113bp400bp4,,4SCAR,SCAR,1d9,,SCAR:;;RAPD;SRAP;;SCAR:Q939.9:A:1000-2286(2010)03-0601-07DevelopmentofSCARMarkersBasedonRAPDandSRAPforRapidIdentificationofPleurotussajor-cajuStrainXIONGFang,ZHENGMin2jiang,LIUXin2rui,XIEBao2gui3(MycologicalResearchCenter,FujianAgricultureandForestryUniversity,Fuzhou350002,China)Abstract:ThisstudywasdesignedforthepurposeofestablishinganeffectivewayforrapididentificationofPleurotussajor-cajustrainsbasedonDNAmolecularmarker.TheRAPD(randomamplifiedpolymorphicDNA)andSRAP(sequence-relatedamplifiedpolymorphism)analysiswereconductedon9cultivatedstrainsofPleurotussajor-caju.Fourspecificfragmentswithalengthof1760bp,356bp,1113bpand400bpcouldbegeneratedfromtheBrazilianPingPong,TemperaturePleurotus,327strain,Pleurotus.Subsequent2ly,theywereconvertedintomorestableSCAR(sequencecharacterizedamplifiedregion)markersbycloning,sequencingandprimerdesign.TheresultsshowedthattheseSCARmarkersacquiredbasedonthisstudycouldbeusedasaspecificDNAfingerprinttoidentifythehomonymandsynonyminPleurotussajor-cajuwhithinoneday.Thefeasibilityandreliabilityofadoptingstrain-specificSCARmarkersfortherapididentificationofPleurotussajor-cajustrainswereconfirmed.Keywords:Pleurotussajor-caju;molecularmarker;RAPD;SRAP;polymorphism;SCARmarker(Pleurotussajor-caju),1974,1980,,,,,[1]:2010-03-16:2010-05-12:(2008SZ0002):(1983-),,,,E-mail:[e-mail];3:,,E-mail:[e-mail]©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[2],RAPDSRAPSCAR9,RAPDSRAP,4,SCARSCAR,,DNA,,11.11.1.1:Escherichiacoli.DH-59()11Tab.1StrainsofPleurotussajor-cajuusedinthestudyStrainNo.StrainnameOriginPl.p00011FengJieNo.1FujianlightindustryresearchinstitutePl.p0002Pleurotussajor-cajuFujianagricultureandforestryuniversityPl.p0003831Feng831FujianagricultureandforestryuniversityPl.p0004PF1PleurotusPF1GuizhouXijiumushroominstitutePl.p00055PleurotusNo.5GuizhouXijiumushroominstitutePl.p0006BrazilianPingPongFujianlightindustryresearchinstitutePl.p0007TemperaturePleurotusGuizhouXijiumushroominstitutePl.p0008327FujianSanmingmycologicalinstitutePl.p0009Pleurotussajor-cajuFASI1.1.2,,22Tab.2TheprimersusedinthestudyNo.RAPD(5-3)ThebasesequenceofRAPDprimer(5-3)No.SRAP(5-3)ThebasesequenceofSRAPprimer(5-3)S10CTGCTGGGACme5TGAGTCCAAACCGGAAGS22TGCCGAGCTGme6TGAGTCCAAACCGGTAGS23AGTCAGCCACem8GACTGCGTACGAATTAGCS62GTGAGGCGTCem6GACTGCGTACGAATTGCA1.1.3UNIQ-10,Sangon;pMD18-TTaqdNTPsTaKaRa():1.21.2.1DNACTAB[3]1.2.22RAPDSRAP,206©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.:945min;941min,1min(SCAR),721.5min,30:7210min12g/L,()22.1RAPD-PCRSRAP-PCRS10S22S62me6-em69RAPD-PCRSRAP-PCR,S1061760bp(1),S229356bp(2)S6283271113bp(3),me3-em67400bp(4),DNA,UNIQ-101S109Fig.1Amplificationresultofthe9strainsofPleurotussajor-cajubyprimerS102S229Fig.2Amplificationresultofthe9strainsofPleurotussajor-cajubyprimerS223S629Fig.3Amplificationresultofthe9strainsofPleurotussajor-cajubyprimerS624me3-em69Fig.4Amplificationresultofthe9strainsofPleurotussajor-cajubyprimerme3-em62.2SCAR,DNAMAN(5-8),SCAR,PrimerPremier5.04SCAR(3)306©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.