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X1,2,2(1.,510275;2.,510275):,,,:;:Q78:A:100828873(2001)01,220115206,(geneticallyengineeredmicroorgan2isms,GEMs),,,,:,,,[13],,[4,5],,,DNA,,1DNARNA,DNA(NaOH),RNARNADNADNADNARNAX:2001202227;:(1971-),,;:.201,220016EcologicScienceVol120No11,2Jun12001,,DNARNAcDNA,PCR()Southern(slotblots)DNA,,-,;,,,()[6],(),,,,,,,2,()(),[7][8],DNA[9]CotRotDNACOTDNA,()[10],DNA,DNA[11],Torsvik[12],200,ROT,DNARNA[13]DNADNA,RNARNA(),DNA61120RNARNA[14]DNARNADNA,,(,homologousprobing),(,heterol2ogousprobing),,,(DNA),[15],,()DNA,DNA,DNArRNArRNArDNA,rRNA,,,,16SrRNA1500,(1000),,,,DNArRNA,rDNArRNA(cDNA)Southern,[1],,[2,3,16,17]3:;,,,,,;,;,(),7111,2:RNA,RNA(RNA)DNA,,DNARNA,rRNA,,rRNA[1],(),,,,,4DNARNA,DNA[15](most-proba2ble-number,MPN)DNA[18][19],DNArRNA,16S/23SrRNA,(),[20],PCR5,,,(),,:;;,,81120,,PCR,:[1]AMANNRI,LUDWIGW,SCHLEIFERKH.Phylogeneticidentificationandinsitudetectionofindividualmicro2bialcellswithoutcultivation[J].Microbiol.Rev.1995.59(1):143-169.[2]GIOVANNONISJ,BRISCHGITB,MOYERCL,etal.GeneticdiversityinSargassoSeabacteraplankton[J].Nature(London),1990,345:60-63.[3]WARDDM,WELLERR,BATESONMM.16SrRNAsequencesrevealnumerousunculturedmicroorganismsinanaturalcommunity[J].Nature,1990,345:63-65.[4]STALEYJT,KONOPKAA.Measurementofinsituactivitiesofnonphotosyntheticmicroorganismsinaquaticandterrestrialhabitats[J].NnnuRevMicrobiol,1985,39:321-346.[5]ROSZAKDB,COLWELLRR.Survivalstrategiesofbacteriainthenaturalenvironment[J].MicrobiolRev,1987,51:365-379.[6]SAMBROOKJ,FRITSCHEF,MANIATIST.MolecularCloning:Alaboratorymanual[M].NewYork:ColdSpringHarborLab,1989.[7]TILGHMANSM,TIENEIERDC,SEIDMANJG,etal.InterveningsequenceofDNAidentifiedinthestructuralportionofamouseB-globingene[J].ProceedingsoftheNationalAcademyofSciences(USA),197875:725-729.[8]SHAPIROJA.Mobilegeneticelements[M].NewYork:AcademicPress,1983.[9]WILLIAMSONVM,YOUNGET,GRIACYM.Transposableelementsassociatedwithconstitutiveexpressionofyeastalcoholdehydrogenase[J].Cell,1981,23:605-614.[10]CHELMBK.DNA-DNArenaturationkinetics[A].In:EDELMANM,etaleds.Methodsinchloroplastmolecularbiology[M].NewYork:Elsevierbiomedical,1982.301-313.[11]BRITTENRJ,GRAHAMDE,NEUFELDBR.AnalysisofrepeationDNAsequencesbyreassociation[J].MethodsinEnzymology,1974,29:363-406.[12]TORSVIKV,GOKSOYRJ,DAAEFL.HighdiversityinDNAofsoilbacteria[J].ApplEnvironMicrobiol,1990,56:782-787.[13]CHELMBK.AnalysisofchloroplastDNAtranscriptionbyRNAdrivenhybridizationkinetics[A].In:EDEL2MANM,etaleds.Methodsinchloroplastmolecularbiology[M].NewYork:ElsevierBiomedical,1982.551-557.[14]SHARROCKRA,GOURSERL,NOMURAM.Inhibitoryeffectofhigh-leveltranscriptionofthebacteriophagelambdanutLregionoftranscriptionofrRNAinEscherichiacoli[J].JBacteriol,1985,163:704-708.[15]HOLBENWE,JANSSONJK,CHELMBK,etal.DNAprobemethodforthedetectionofspecificmicroorganismsinthesoilbacterialcommunity[J].ApplEnvironMicrobiol,1988,54:703-711.[16]SCHMIDTTM,DELONGEF,PACENR.Analysisofamirinepicoplanktoncommunityby16SrRNAgenecloningandsequencing[J].JBacteriol,1991,173:4371-4378.[17]LLOYD-JONESG,LAUPCK.AmolecularviewofmicrobialdiversityinadynamiclandfillinQuebec[J].9111,2:FEMSMicrobiolLett,1998,162:219-226.[18]DRAKETA,HINDLERJA,BERLINGW,etal.RapididentificationofMycobacteriumaviumcomplexincultureusingDNAprobes[J].JClinMicrobiol,1987,25:1442-1448.[19]RATNERM.DNAprobes:Themarketversusthemagic[J].Bio/Technology,1988,6:1369-1370.[20]LEES,MALONEC,KEMPPF.Useofmultiple16SrRNA-targetedfluorescentprobestoincreasesignalstrengthandmeasurecellularRNAfromnaturalplanktonicbacteria[J].MarEcolProgSer,1993,101:193-201.UseofNucleicAcidHybridizationinMicrobialEcologyHUANGLi-nan1,NIEXiang-ping2,LANChong-yu2(1.DepartmentofEnvirenmentalSciences,ZhongshanUniversity,Guangzhou510275,China;2.SchoolofLifeSciences,ZhongshanUniversity,Guangzhou510275,China)Abstract:Duetothefactthatmostofthemicrobialspeciesinnaturecannotbecultured,thetradi2tionaltechniquesbaseoncultivationandpure-cultureisolationhadbecomethemajorlimitationtothestudyofmicrobialecology.Advancesinmolecularbiologyprovideresolutionstothequestionsthathadhinderedthemicrobiologicalstudyforalongtime.Nucleicacidhybridizationhasgreatpotentialitiesandhasbeenwidelyappliedinthefieldofsystematictaxonomyandecologyofmicrobiology.Herewereviewedthebasicprinciplesofnucleicacidhybridizationanditswideapplication,majoradvantagesandlimitations,andhowtoimproveitsdetectingsensitivityandspecificity.Theapplicativeprospectofthistechniquewasalsodiscussed.Keywords:nucleicacidhybridization;microbialecology02120