烟草黑胫病抗性基因-Bsl-(-t-)-的-RAPD-标记

Vol.30,No.5pp.516518May,2004ACTAAGRONOMICASINICA30520045516518Bsl(t)RAPD3X(,650201)ScreeningandStudyofRAPDMarkersLinkedtoTobaccoBlackShankResistanceGeneBs1(t)LIUXiao2Xia,WANGLi3,WENGuo2Song,YANGYan2Qiong(CollegeofAgronomyandBiotechnology,YunnanAgriculturalUniversity,Kunming650201,Yunnan,China)VanBredadeHaan[1]1896,,[2],,5%12%,75%,[3,4],76373hm2,2.869,1.23,[5],,,,,,F2,RAPD(BSA),Bs1(t)RAPDOPB023000OPM112500,20.5cM29.9cM,11.1,[6,7](60Co2,,,)2000,2001F1,2002F2(Phytophthoraparasiticavar.nicotianae)01.2(Phytophthoraparasiticavar.nicotianae)0,(26,57d),,23d1,[3]1.3DNADNACTAB[8,9]1.4PCROperon,10bp,400,PCR20L,1030ngDNA(F2),5mmolPLMgCl2,0.2mmolPLdNTPs,0.2pmolPL,1(10mmolPLTris2HCl,pH9.0,50mmolPLKCl),1UTaqDNA,GradientPCR:945min,941min,361min,722min,44,7210min1.5OPB02OPM11F2,MapMakerPEXP3.0,X:(2000C0051M):(1977-),,,3:Tel:[phone]Received():2003208204,Accepted():2003211205.22.1[7]F1,:1F1F2Table1ThereactionofF1andF2toPhytophthoraparasitiavia.nicotianaeMaterialTotalResistantplantsSusceptibleplants23R1SF14504500F2222154683.75P0.05(3.84)(CK)1200120(1),F1100%,,F2,,,231(1),2.2400,79,OPB02OPM11,3.0kb2.5kb2.3F2RAPDOPB023000OPM112500,F2RAPD,OPB02OPM11222F212232F2OPB02Table2TherelationshipbetweenresistancesegregationandRAPDamplificationresultsofF2generationResistancemarkerNo.ofplantsOPB023000WithOPB023000OPB023000WithoutOPB023000Resistant15431123Susceptible686083F2OPM11RAPDTable3TherelationshipbetweenresistancesegregationandRAPDamplificationresultsofF2generationResistancemarkerNo.ofplantOPM112500WithOPM112500OPM112500WithoutOPM112500Resistant15411242Susceptible6825431OPB02RAPDFig.1RAPDamplificationoftobaccowithrandomprimerOPB02M:1kbMarker;114:F2;1522:F27155:Bs1(t)RAPD2OPM11RAPDFig.2RAPDamplificationoftobaccowithrandomprimerOPM11M:1kbMarker;113F2;1523F23RAPDFig.3AlinkagemapofRAPDmarkersandtheresistantgeneMapMakerPEXP3.0,OPB02OPM11,OPB0220.5cM,OPM1129.9cM(3)3Bs1(t),400,79,19.75%Bs1(t),,,,References[1]VanBredadeHaan.DeBibitxiekteindeDeli2tabakveroorzaaktdoorPhytophthoranicotianae.Mded.SlandsPlentuim,1896,15:1-107[2]KongF2Y(),ZhuX2C(),ShiJ2K().DevelopingtendencycausesandcontrolmeasuresoftobaccoinfectiousdiseasesinChina.ChinaTobacco(),1995,(1):31-34[3]ChenR2T(),ZhuX2C(),WangZ2F().Areportofinvestigatingandstudyingtobaccoinfectiousdiseasesof16maintobac2coproducingprovinces(regions)inChina.ChineseTobaccoScience(),1997,1(4):1-7[4]SuD2C().TobaccoDiseasesandInsectPests().Beijing:ChinaAgriculturePress,1992.87-92[5]WangG(),ZhengX2B(),LuJ2Y().DetectionofresistanceofblackshanktometalaxylinYunnanprovince.JournalofNanjingAgriculturalUniversity(),1997,20(4):105-107[6]WangL(),LuoW2F(),YangY2Q().Usingphyto2toxintoselectmutantsresistanttotobaccoPhytophthoraparasitiavar.nicotinaeSelectionofcallusresistanttophytotoxin.JournalofYun2nanAgriculturalUniversity(),1999,14(1):16-21[7]WangL(),YangY2Q(),YangD().Selectionofmu2tantsresistancetotobaccoPhytophthoraparasitiavar.nicotianaeusingphytotoxin.Resistancetoblackshrankfungusinoculationofregenera2tionplantsandtheirprogeniesoftoxin2resistantcallus.JournalofYunnanAgriculturalUniversity(),1999,14(3):273-278[8]OsberF,KingstonRE.AGuideofMolecularBiology().Beijing:SciencePress,1998[9]CsinosAS,FortrumBA,PowellNT,etal.Identificationandseparat2ingofRaceiPhytophthoranicotianaeresistantgene.TobSci,1984,26:153-15581530