从不同反应器筛选鉴别好氧反硝化菌汪苹

汪苹,项慕飞,翟茜,100037:2AB,,,.A,pH613,Q(DO)212~611mgPL,(Q(C)PQ(N),Q(C)Q(CODCr))9;B,pH618~718,Q(DO)212~310mgPL,Q(C)PQ(N)1512Q(NO3--N)80mgPL.260%,.220BTB,8DNAPCR,16SrRNA.GenBankBlast,,8(Pseudomonas),(Delftia),(Herbaspirillum)(Comamonas).8.:;16SrRNA;;Delftia:X172:A:1001-6929(2007)04-0120-05SelectionandIdentificationofAerobicDenitrifiersfromDifferentReactorsWANGPing,XIANGMu-fei,ZHAIQianSchoolofChemicalandEnvironmentEngineering,BeijingTechnologyandBusinessUniversity,Beijing100037,ChinaAbstract:Byadoptingtwosequencebatchreactors,ReactorsAandB,nitrateassubstrate,usingintermittentaerationmode,theaerotolerantnitrogenremovalsludgewasdomesticatedandaccumulated.InReactorA,pHwasapproximately613,DO212~611mgPL,Q(C)PQ(N)9;andinReactorB,pHwas618~718,DO212~310mgPL,Q(C)PQ(N)15.Bothreactorskepttheinfluentnitrateconcentrationat80mgPL.WhenTINremovalefficiencyofbothreactorsreached60%,theaccumulationofaerobicdenitrifierswasregardedascompleted.TwentyBTBpositivebacteriawereisolatedfromtwoSBRreactorsand8DNAsamplesofthemwereamplifiedbyPCRandprocessedfor16SrRNAsequence.ObtainedresultswereprocessedthroughBlastsimilaritysearchbyGenBank,analyzedthroughgenetictreeandcomparedthroughidentification.Asaresult,the8bacteriabelongtogenusPseudomonas,Delftia,HerbaspirillumandComamonas,respectively.These8bacteriawereallaerobicdenitrifiersbydenitrifyingmeasurement.Atpresent,noDelftiahasbeenreportedasaerobicdenitrifier.Keywords:aerobicdenitrifiers;16SrRNA;phylogeneticanalysis;Delftia:2006-10-24:2007-02-05:(KZ200510011008);(KZ200410011007):(1953-),,,,[e-mail][1](Paracoccuspantotropha)(Pseudomonassp.)(Alcaligenesfaecalis).,Robertson,Frette[2]-16,.Pai[3]3,T6,NH4+-NNO2--N.,(HyphomicrobiumX)[4](Klebsiella)[5](Ochrobactrum)[6](Zoogloea)[3].,.,:¹,,;º,,;»OH-,pH.,,204ResearchofEnvironmentalSciencesVol.20,No.4,2007DOI10.13198/j.res.2007.04.124.wangp.022,,,,.2SBR20,8,4.NO3--N,.11.1NO3--N,,.SBR2,,155mm,800mm,,12L.,,DO.,,,6L,6L.,2(1),A,;B,.2pH.1ABTable1CharacteristicsofacclimatedsludgeofreactorsAandBQ(MLSS)P(g#L-1)Q(MLVSS)P(g#L-1)SVIP(mL#g-1)SVP%A4.554.0661.528B2.502.10)15~20A():pH613,Q(DO)212~611mgPL,Q(NO3--N)80mgPL,Q(C)PQ(N)9,480min,(TIN)60100%~83103%.,,.,(1(a)).1(a),Q(NO3--N),Q(NO2--N),NO3--NNO2--N,NO2--N.B():pH618~718,Q(DO)212~310mgPL,Q(NO3--N)80mgPL,Q(C)PQ(N)15,330min,Q(NO3--N)918mgPL,TIN86%~90%.,,.,.1(b)3.1(a),200min,,NO2--N,NO3--N.1.2BTBTakayaN[5]1A()B()Fig.1ConcentrationsofnitrogencompoundsinliquidofreactorA(controlled)andreactorB(notcontrolled),(BTB).NO3-pH,pH1214:,.ABBTB,30e,,.AB20BTB,WXZ-1~WXZ-20.20,,,WXZ-11,19.,.1.316SrRNAPCR,,100LL,5min,PCR.16SrRNAPCR,f27(5c-AGAGTTGATCCTGGCTCAG-3c)r1492(5c-GGTTACCTTGTTACGACTT-3c)..PCR(50LL):10@Buffer5LL,dNTPs4LL,f27r14921LL,39LL,DNA0.5LL,Taq0.5LL.PCR:¹94e5min;º94e1min;»50e1min;¼72e2min,25;½72e10min.1.4pH,BTB.12h,10%50mLDM250mL(DM:KNO30.6g;KH2PO41g;MgSO4#7H2O1g;2.4g;Q(C)PQ(N)=10,pH710).,8.30e,160rPmin(DO66%~92%).2.4d,N-(1-)-Q(NO3--N)Q(NO2--N).,,(NOx--NTIN).22.116SrRNA20BTB8PCR,A5,B3.1%,16SrRNA,GenBankBlast(),2.28PCRTable2PCRamplificationproductsofeightstrainsPbpWXZ-41362WXZ-71392AWXZ-91406WXZ-101401WXZ-131359WXZ-141370BWXZ-171359WXZ-181419DNAMAN28,.:8,4:(Pseudomonas),(Delftia),(Herbaspirillum)(Comamonas)(2);WXZ-4,WXZ-7,WXZ-10100%,PseudomonasaspleniiPseudomonasputida99%,,;WXZ-13,Pseudomonassp.ADPP.japonica99%,,WXZ-4,WXZ-7,WXZ-1098%,.,AB,APseudomonasDelftia,BHerbaspirillumComamonas(3).PseudomonasComamonas2,DelftiaHerbaspirillum212220216SrRNAFig.2Unrootedphylogenetictreebasedonthe16SrRNAsequences.HerbaspirillumBaldani1986,.Herbaspirillum,();NO3--N;NO3-NO2-,N2O[7].Delftia.1(Microcystins,MCs),DelftiaacidovoransUSTB04[8],.,WXZ-9Delftiaacidovorans99%.2.22,NO3--N,NO2--N,.2:NO3--NNO3--N;NOx--N(NOx--N=NO3--N+NO2--N).82,4.4,8NO3--NNO2--N,5NO3--N56%,1316SrRNATable3Resultsof16SrRNAsequencesimilarityanalysisGENBANK()ABWXZ-4DQ356895PseudomonasputidaP1WXZ-7DQ356900PseudomonasputidaP16WXZ-10DQ356902PseudomonasputidaP19WXZ-13DQ356896Pseudomonassp.P3WXZ-9DQ356901DelftiatsuruhatensisP18WXZ-14DQ356897Herbaspirillumsp.P4WXZ-17DQ356899ComamonastestosteroneP6WXZ-18DQ356898ComamonasstestosteroneP7Pseudomonasputida(99%)Pseudomonassp.(99%)Delftiasp.(100%)Herbaspirillumsp.(100%)Comamonastestosterone(100%)WXZ-4,WXZ-7,WXZ-10100%WXZ-4,WXZ-7,WXZ-10WXZ-1398%WXZ-17,WXZ-1895%WXZ-17,WXZ-18WXZ-988%WXZ-18100%30.5%,2.[9]Microvirgulaaerodenitrificans(),8510~710mgPL(30e,66%~92%),Microvirgulaaerodenitrificans(64%~96%,3dNO3--N513%~6115%).1234:4(4d)Table4ResultsofdenitrificationcapabilitymeasurementoffourdaysQ(NO3--N)P(mg#L-1)QP(mg#L-1)P%NO2--NNO3--NNOx--NNO3--NNOx--NWXZ-4114.54.148.652.757.654.0WXZ-7114.54.947.051.956.054.7WXZ-9115.58.945.654.560.552.8WXZ-10115.50.193.293.319.319.2WXZ-13115.53.130.433.573.771.0WXZ-1491.81.063.864.830.529.4WXZ-1791.80.935.636.561.260.2WXZ-1881.45.968.474.316.08.78.3a.BTB220BTB,PCR,16SrRNA,,8Pseudomonas,Delftia,HerbaspirillumComamonas,.,16SrRNA(16SrDNA),,.b.,66%~92%,5NO3--N56%,130.5%,216%,.c.2:NO3--N,.d.pH,,,NO3--N.ANO3--NNO2--N,NO2--N;PseudomonasDelftia.BA,NO3--N,NO2--N;Herbaspi