分子生物学技术在污水处理微生物检测中的应用

,.[J].,2005,25(9):1143-1147PENGYongzhen,GAOShouyou.Theapplicationsofmolecularbiologicaltoolstomicrobialecosystemanalysesinwastewatertreatmentprocess[J].ActaScientiaeCircumstantiae,2005,25(9):1143-1147[]:1,2,3,111,15009021,100022:2005204216:2005207204:2005207204:,PCRFISH.,,,,.,DNA,,.:;;:025322468(2005)0921143205:X832:ATheapplicationsofmolecularbiologicaltoolstomicrobialecosystemanalysesinwastewatertreatmentprocessPENGYongzhen1,2,3,GAOShouyou111SchoolofMunicipal&EnvironmentalEngineering,HarbinInstituteofTechnology,Harbin15009021KeyLaboratoryofBeijingforWaterQualityScienceandWaterEnvironmentRecoveryEngineering,BeijingPolytechnicUniversity,Beijing100022Received16April2005;receivedinrevisedform4July2005;accepted4July2005Abstract:Theuseofmolecularbiologicaltoolstomicroorganismdetectionsandcommunityanalysesinthebiologicalwastewatertreatmentprocesshadbeendiscussed,andthemajorpointswereputonthemorefrequentlyusedtechniquessuchaspolymerasechainreaction(PCR)andfluorescenceinsituhybridization(FISH).Theadvantagesanddisadvantagesofeachtechniquewereanalyzed.Throughthedetectionsandanalyses,theinformationoftheamounts,thecommunitystructuresandtheactivitiesofthemicroorganisminthesystemcouldbeabtained,sotheseinformationscanbeusedtobuildenhancedecosystemandtostrengthentheefficiencyoftheexistedsystem.Itisbelievedthat,themolecularbiologicaltoolsbasedontheDNAsequencesandtherelevantstructuregeneshavebeensuccessfullyusedincommunityanalysesinthebiologicalwastewatertreatmentprocess,andwillsurelyhelptogetadeeperinsightintoandoptimizethebiologicaltreatmentprocess.Furthermore,itwillpromotethedevelopmentsandapplicationsofnoveltechniques,andplayanevenmoreimportantroleintheprocesscontrollingofthebiologicalwastewatertreatments.Keywords:biologicalwastewatertreatment;moleculartools;analysesofmicroorganismcommunity:(No.50521140075);863(No.2003AA601010):(1949),,(),E2mail:[e-mail];3()Foundationitem:NationalNaturalScienceFoundationofChina(No.50521140075);NationalBasicResearchProgramofChina(No.2003AA601010)Biography:PENGYongzhen(1949),male,professor(Ph.D.),E2mail:[e-mail];3Correspondingauthor.,,:,;;,;,(Schramm25920059ActaScientiaeCircumstantiaeVol.25,No.9Sep.,2005etal.,1996)..2090,,.16rDNA,.,,.,DNA,..1(PCR)(PolymeraseChainReaction)2080K.MullisDNA,PCR,DNA.(2530),..PCR,DNA(Metcalfetal.,2003).PCR,.PCR,RT2PCRPCRPCR2DGGEPCR2SSCPPCR.RT2PCRRNA,RNADNA.Selvaratnam(1997)dmpNPCR(Pseudomonas).,RT2PCR,dmpN,.,.PCR(competitivePCR)PCR,PCR,.PCR22,32dmpB(Trperetal.,1992).PCRdmpB,40bp,PCR.,PCRPCR.PCR,,dmpB.,,.,,,.PCR2DGGE(PolymeraseChainReactionDenaturingGradientGelElectrphoreses),DNA,16SrDNA.,.(2004)PCR2DGGE,DNA;Marsh(1998)PCR2DGGE;Nicolaisen(2002)PCR2DGGENitrosomonas2like.,PCR2DGGE.(Single2StrandConformationPolymorphism,SSCP).,SSCP.PCRSSCPPCRrRNA.Zumstein(2001)PCR2SSCP,457;,,.Delbes(2001)PCR2SSCP441125,,(Clostridiumsp),,.PCRDNA,,.PCR,,,.PCR(Norwalkvirus),PCR,(Trperetal.,1992):;;,;(,PCR).,PCR2kb;PCR2DGGEDNA;PCR;TaqDNA(Zumsteinetal.,2000).,PCR.PCR.2(RFLP)(RestrictiveFragmentLengthPolymorphism,RFLP),DNA,.DNA;DNA;DNA,.RELP.Liu(1998)RFLP2SBR,,Glycogen2acumulatingorganisms(GAOs)Polyphosphate2acumulatingorganisms(PAOs);Jane(1998)PCRRFLPPoliovirus.3,rRNADNA.rRNA.DNA,1518,100.RNA.,DNARNA.RNA,,RNA;rRNA.,,RNA.RNA.(FluorescenceInSituHybridization,FISH),,.FISH,.Nielsen(2001),FISH.Konuma(2001)FISH(ammonia2oxidizingbacteria),,FISH,.1FISH.1FISHTable1TheapplicationsofFISHinthemicroorganismdetectionsinwastewaterMicrothrixparvicella(Eberletal.,1997)EBPR,(PAOs)(Minoetal.,1998)(Silyn2Robertsetal.,2001)UASB(Sekiguchietal.,2002;Syutsuboetal.,2001)(Coskuneretal.,2002)SBR,,(DNPAOs)(Johuanetal.,2002)54119:,.,FISH,,.,,,.,FISH.,.FISH.,,.,,FISH.ZhangDan(2004)PCR2DGGEFISH2,,,,Nitrosomonas7215%.FISH;FISH.4DNA.(Mascini,2001).,DNA.DNA,DNADNA,DNA,.Chiti(2001)DNAaromaticamines;Wang(1996)DNA,CryptosporidiumEscherichiacoli..DNA.,,,.5DNA2070,DNA.DNA,DNADNA.:DNA;;DNA;,DNA;,DNA..,;,;.,..,.,,.,:;;.HwangFarrand(Hwangetal.,1994)(Pseudomonas)641125,,,,PCR,.6DNA2080,,,.,.,,.(References):CoskunerG,CurtisTP.2002.Insitucharacterizationofnitrifiersinanactivatedsludgeplant:DetectionofNitrobacterSpp[J].JournalofAppliedMicrobiology,93:431437DelbesC,MolettaR,GodonJJ.2001.Bacterialandarcheal16SrDNAand16SrRNAdynamicsduringanacetatecrisisinananaerobicdigestorecosystem[J].FemsMicrobiolEcol,35:1628GiacomoC,GiovannaM,MarcoM.2001.ElectrochemicalDNAbiosensorforenvironmentalmonitoring[J].AnalyticaChimicaActa,427:155164HwangI,FarrandSK.1994.Molecularbiologyofbacterialbioluminescence[J].ApplEnvironMicrobiol,60:913920JohwanA,TomotakaD,SatoshiT,etal.2002.Characterizationofdenitrifyingphosphate2accumulatingorganismscultivatedunderdifferentelectronacceptorconditionsusingpolymerasechainreaction2denaturinggradientgelelectrophoresisassay[J].WatRes,36:403412KonumaS,satohH,MinoT.2001.Comparisonofenumerationmethodsforammonia2oxidizingbacteria[J].WatSciTech,43(1):107114LeoE,RenateS,AldoA,etal.1997.Useofgreenfluorescentproteinasamarkerforecologicalstudiesofactivatedsludgecommunities[J].FemsMicrobiologyLetters,149(1):20:7783Liu