分子生物学技术在污水处理微生物检测中的应用彭永臻

,.[J].,2005,25(9):1143-1147PENGYongzhen,GAOShouyou.Theapplicationsofmolecularbiologicaltoolstomicrobialecosystemanalysesinwastewatertreatmentprocess[J].ActaScientiaeCircumstantiae,2005,25(9):1143-1147[]:彭永臻1,2,＊,高守有11.哈尔滨工业大学市政环境工程学院,哈尔滨　1500902.北京工业大学北京市水质科学与水环境恢复工程重点实验室,北京　100022:2005-04-16　　　:2005-07-04　　　:2005-07-04:,PCR、FISH.,,,,.,DNA,,.:;;:0253-2468(2005)09-1143-05　　:X832　　:ATheapplicationsofmolecularbiologicaltoolstomicrobialecosystemanalysesinwastewatertreatmentprocessPENGYongzhen1,2,＊,GAOShouyou11.SchoolofMunicipal&EnvironmentalEngineering,HarbinInstituteofTechnology,Harbin1500902.KeyLaboratoryofBeijingforWaterQualityScienceandWaterEnvironmentRecoveryEngineering,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—),,(),E-mail:[e-mail];＊()Foundationitem:NationalNaturalScienceFoundationofChina(No.50521140075);NationalBasicResearchProgramofChina(No.2003AA601010)Biography:PENGYongzhen(1949—),male,professor(Ph.D.),E-mail:[e-mail];＊Correspondingauthor　　.,,:,;、;,;,(Schramm25920059　　　　　ActaScientiaeCircumstantiaeVol.25,No.9Sep.,2005DOI:10.13671/j.hjkxxb.2005.09.001etal.,1996)..2090,,、.16rDNA,.,,.,DNA,..1　(PCR)　　(PolymeraseChainReaction)2080K.MullisDNA,PCR,DNA.(25～30),.、、.PCR,DNA(Metcalfetal.,2003).PCR,、.PCR,RT-PCR、PCR、PCR-DGGE、PCR-SSCPPCR.RT-PCRRNA,RNADNA.Selvaratnam(1997)dmpNPCR(Pseudomonas).,RT-PCR,dmpN,.,、、.PCR(competitivePCR)PCR,PCR、,.PCR-2,3-dmpB(Trperetal.,1992).PCRdmpB,40bp,PCR.,PCRPCR.PCR,,dmpB.,,.,,,.PCR-DGGE(PolymeraseChainReactionDenaturingGradientGelElectrphoreses),DNA,16SrDNA.,.(2004)PCR-DGGE,DNA;Marsh(1998)PCR-DGGE;Nicolaisen(2002)PCR-DGGENitrosomonas-like.,PCR-DGGE.(Single-StrandConformationPolymorphism,SSCP).,SSCP.PCRSSCPPCRrRNA.Zumstein(2001)PCR-SSCP,457;,,.Delbes(2001)PCR-SSCP1144　　　　　　　　　　25,,(Clostridiumsp),,.PCRDNA,,.PCR,,,、、.PCR(Norwalkvirus),PCR,(Trperetal.,1992):;;,;(,PCR).,PCR2kb;PCR-DGGEDNA;PCR;TaqDNA(Zumsteinetal.,2000).,PCR.PCR.2　(RFLP)　　(RestrictiveFragmentLengthPolymorphism,RFLP),DNA,.DNA;DNA;DNA,.RELP.Liu(1998)RFLP-SBR,,Glycogen-acumulatingorganisms(GAOs)Polyphosphate-acumulatingorganisms(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(ammonia-oxidizingbacteria),,FISH,.1FISH.1　FISHTable1　TheapplicationsofFISHinthemicroorganismdetectionsinwastewater　　Microthrixparvicella(Eberletal.,1997)EBPR,(PAOs)(Minoetal.,1998)(Silyn-Robertsetal.,2001)UASB(Sekiguchietal.,2002;Syutsuboetal.,2001)(Coskuneretal.,2002)SBR,,(DNPAOs)(Johuanetal.,2002)11459:　　,.,FISH,,.,,,.,FISH.,、、、.FISH.,,.,,FISH.ZhangDan(2004)PCR-DGGEFISH-,,,,Nitrosomonas72.5%.FISH;FISH.4　DNA　　.(Mascini,2001).,DNA.DNA,DNADNA、,DNA,.Chiti(2001)DNAaromaticamines;Wang(1996)DNA,Cryptosporidium、Escherichiacoli.、、.DNA.,,、、、,.5　DNA　　2070,DNA.DNA,DNADNA.:DNA;;DNA;,DNA;,DNA..,;,;.,..,、.,,.,:、;;.HwangFarrand(Hwangetal.,1994)(Pseudomonas)1146　　　　　　　　　　25,,,,PCR,.6　　　DNA2080,,、、,.,.、,、、,.(References):CoskunerG,CurtisTP.2002.Insitucharacterizationofnitrifiersinanactivatedsludgeplant:DetectionofNitrobacterSpp[J].JournalofAppliedMicrobiology,93:431—437DelbesC,MolettaR,GodonJJ.2001.Bacterialandarcheal16SrDNAand16SrRNAdynamicsduringanacetatecrisisinananaerobicdigestorecosystem[J].FemsMicrobiolEcol,35:16—28GiacomoC,GiovannaM,MarcoM.2001.ElectrochemicalDNAbiosensorforenvironmentalmonitoring[J].AnalyticaChimicaActa,427:155—164HwangI,FarrandSK.1994.Molecularbiologyofbacterialbioluminescence[J].ApplEnvironMicrobiol,60:913—920JohwanA,TomotakaD,SatoshiT,etal.2002.Characterizationofdenitrifyingphosphate-accumulatingorganismscultivatedunderdifferentelectronacceptorconditionsusingpolymerasechainreaction-denaturinggradientgelelectrophoresisassay[J].WatRes,36:403—412KonumaS,satohH,MinoT.2001.Comparisonofenumerationmethodsforammonia-oxidizingba