高浓度硝酸盐对城市污水活性污泥中微生态种群结构的影响

,,,.[J].,2005,25(3):372-378FUYigang,WANGFeng,ZHAOJianfu,etal.Influenceofhighconcentrationnitrateiononthemicrobialecosystemofactivatedsludgeofmunicipalsewage[J].ActaScientiaeCircumstantiae,2005,25(3):372-3783,,,,,200092:,.DNA,16SrDNAV3PCR,DGGE().16SrDNAV3,NCBI(),.,,.:16SrDNA;;:025322468(2005)0320372207:X703:AInfluenceofhighconcentrationnitrateiononthemicrobialecosystemofactivatedsludgeofmunicipalsewageFUYigang3,WANGFeng,ZHAOJianfu,XIASiqingStateKeyLaboratoryofPollutionControlandResourceReuse,CollegeofEnvironmentalScienceandEngineering,TongjiUniversity,Shanghai200092Abstract:Thecommunitystructureandthebehaviorcharacteristicoftheactivatedsludgeinmineralsubstrateandinbasicsubstrateconditionsarestudied.There2sultshowsnotabledifferencebetweenthetwoconditionsinregardtocommunitystructureandwholecharacteristicofthemicroorganism.Whenimposedtohighcon2centrationNO-3,themicroorganismrapidlychangesitscommunitystructuretoadaptthenewenvironmentandtriestokeepitsintrinsicmicrobialdiversity.Further2more,somebacteriaspeciesareconfirmedbycomparingthesequencesofseveralpiecesof16SrDNAV3fragmentswiththeDNAsequencesinGeneBank(NCBI).Keywords:16SrDNA;microbialecosystem;activatedsludge:2004203215;:2004211220:2004(0400241015);863(2002AA601320);:(1971),,(),[e-mail];3(),.,.,[1],,.,..,,1.11.1.200mL6000rmin-1-30.1.1.11g,100g.,80mL,1.015mL33,8025320053ACTASCIENTIAECIRCUMSTANTIAEVol.25,No.3Mar.,2005rmin-1.50mL7000rmin-1,DNAPCR2DGGE.1.1.2.,,33,80rmin-110d.50mL7000rmin-1,DNAPCR2DGGE.1.2DNADNA[2,3]:DNA:100mmolL-1Tris2HCl(pH=810);100mmolL-1(pH=810);115molL-1NaCl;1%CTAB();TE:10mmolL-1Tris2HCl,pH=716,1mmolL-1EDTA(),pH=810.:115mL,012013g(111)016mLDNA10LK(10mgmL-1).37,225rmin-130min.100L20%SDS(),652h,1520min1.255000rmin-110min,115mL.P(24P1),5000rmin-110min,;016,1h,9000rmin-12520min,500L(pH=716)TE[2].1NaNO3Table1TheconcentrationofnitrateionsadditivetoincubatedactivatedsludgegL-1A1A2A3A4A50810152025113PCR942min;94,45s;60,45s;72,90s.30,7210min.PCR:39L,10PCR5L,215mmolL-1dNTP1L,10mmolL-1P1P21L,Taq1,DNA2L[4,5].114DNAPCR1%(1g,015TBE100mL).015TBE.115PCRPCR,Parafilm,-70oC,4h,30L(pH=716)TE,-20.116DGGEPCRDGGE().30%60%.150V,1TAE,60,5h.,DNA.,,30LTE(pH=716).PCR,113.PCR.117PCR[5],16SrDNAV3:P152ATTACCGCGGCTGCTGG23,P252CGC2CCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGCCCCATACGGGAGGCAGCAG23,GCDGGE..MarkerBebcoDNA3000-100bpMarkerDL2000.118:(GL-20B)(ThermoIECMicromaxRF)PCR(AMPLITRONR)(DYB21)DGGE(BIO2RADDcodeTM)LGJ(FR2200)SmartView2001.2825gL-1,.(=260nm),10d,3733..50mL6000rmin-1,DNAPCR2DGGE.211DNADNA16SrDNAPCR,1%,DNA220240bp(basepair),16SrDNAV3.DNADNA,,RNA.212DGGE16SrDNA21211.,10dDNA,16SDNAPCR.DGGE1,11,2.2Table2Theanalysisofinfluenceofhighconcentrationofnitrateionsonmicrobialecosysteminmineralsubstrate1,A1,.2A2,.3A1A2,,A3,A4A5.43,A1A2,.5,A1A5.6A1-A5,.76,A1-A5,6.8A1A5,.9A1A5,.10,A1-A5,.11A1A5,,.1DGGEFig.1Theinfluenceofhighconcentrationofnitrateionsonmicrobialecosysteminmineralsubstrate(byDGGEelectrophoreses)2DGGEFig.2Theinfluenceofhighconcentrationofnitrateionsonmicrobialecosysteminbasicsubstrate(byDGGEelectrophoreses)473252.2.2.10d,DNA,16SDNAPCR.DGGE2.,.10(2).3.316SrDNATable3Thesequencesof16SrDNAsegmentsofpartofmicroorganismandcomparisonwithdatabaseNCBINCBITTAGCCGGGGCTTCTTCTCCGGGTACCGTCATTATCGTCCCCGGTGAAAGAATTTTACAATCCTAAGACCTTCATCATTCACGCGGCATGGCTGCGTCAGGCTTTCGCCCATTGCGCAAGATTCCCCACTGCTGBrevundimonasbacteroides(100%);Brevundimonassp.dcm7A(100%);Brevundimonassp.GOBB32110(100%);Brevundimonasdiminuta(100%);Brevundimonasvesicularis(100%)AntarcticbacteriumR28358(100%);GlacialicebacteriumMS2T3202(100%)BlackwaterbioreactorbacteriumBW23(100%)Caulobactersp.(100%);CaulobactercrescentusCB15(100%);glacierbacteriumFXI47(100%)Alphaproteobacteriumzj26(100%)10779016122288092163009722807.BLASTQ3TTAGCCGGGGCTTTCTGCTCAGATACCGTCAACCAAAGAGCAGTGTACTCTCCTTAGCTTTCTTCTCTGGCAACAGAATTTTACGAACCGAAAACCTTCTTCACTCACGCGGCGGTGCTCGGTCAGAATTCCGTCCATTGGCGAAGATTCCCTACTGCTGPaenibacillussp.R26928(95%);Paenibacillussp.MB2039(95%)10779016402-30298293992969774.BLASTQ3GGCTTTCTGGTTAGGTACCGTCAAGGTACCGACTTATTCGGTCGATACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAGACCTTCATCACTCACGCGGCGTCGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGBacillusoleronius(96%);Bacillussporothermodurans(96%);Ba2cillussp.IPso2(96%);Bacillussp.IPso1(96%);BacteriumLMG18435(95%)Virgibacilluscarmonensis(95%);Virgibacillusnecropolis(95%)10779016792297482160979853606.BLASTQ3GGCTTTCTGGTTAGGTACCGTCAAGGTGCCAGCTTATTCAACTGGTACTTGTTCTTCCCTAACAACAGAGTTTTACGACCCGAAAGCCTTCATCACTCACGCGGCGTCGCTCCGTCAGACTTTCGGCCATTGCGGAAGATTCCCTACTGCTGBacillussp.112(97%);Bacillussp.Tf253(97%);Bacillussp.Tf1-41(97%);Bacillussp.ICS20418(97%)BacillaceaebacteriumPH27B(97%)unculturedsoilbacterium(97%)1077901739230396231578988497.BLASTQ3AGCCGGGGCTTTCTCGTTAGGTACCGTCAGACCGGGAGGTCATCCCGGCGGTTCTTCCCTAACAACAGAACTTTACGATCCGAAAACCTTCATCGTTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGAneurinibacillusaneurinilyticus(100%);Aneurinibacillusmigulanus(100%);Aneurinibacillusmigulanus(100%);Aneurinibacillusan2eurinolyticus(100%);Aneurinibacillusthermoaerophilus(94%);An2eurinibacillussp.DB4(96%