连续式白腐真菌膜生物反应器处理染料废水_微生物群落结构及其与脱色

46320105ActaScientiarumNaturaliumUniversitatisPekinensisVol.46No.3May20102006AA06Z331502780342009-10-202010-01-07200062[e-mail]-DGGE100MDSDGGE。18SrDNANCBIPhanerochaetechrysosporiumP.chrysosporiumShannon-Wiener1.920.92。DGGEMDS82.5%93.2%。PCR-DGGEMDSX703MicrobialCommunityStructuresinContinuousFMBRTreatingDyestuffWastewaterRelationshipbetweenCommunityStabilityandReactorPerformanceGAOShangMALihuaHUANGMinshengTAOFangCHENChengShanghaiKeyLabofUrbanizationandEco-restorationSchoolofResourceandEnvironmentScienceEastChinaNormalUniversityShanghai200062[e-mail]hainreactiondenaturinggradientgelelectrophoresisPCR-DGGEwasusedtoanalyzethestabilityofmicrobialcommunitystructureincontinuouswhiterotFungalMembraneBio-ReactorFMBRandtheresultswereinterpretedusingsophisticatedmultidimensionalscalingMDSanalysis.Theexperimentwascarriedoutfor100dayscontinuously.TheresultsindicatethatthePhanerochaetechrysosporiumP.chrysosporiumwasidentifiedbysequencinganditwasthepredominantspeciesinmicrobialcommunity.Thenwiththetimeincreasingthepredominantwasmoreandmoreobvious.TheShannon-Wienerindicesofthemicrobialcommunityattheendoftheexperimentdecreasedto0.92lowerthantheaverageof1.92.TheMDSanalysisshowthatthechangesamongthemicrobialcommunitieswasrelativelystableduringthecontinuousFMBRoperationstable.Theaveragerateofdecolorationmaintainedat82.5%andthemaximumratecouldreachto93.2%.ThecontinuousFMBRcoulddecolorizedyestuffwasterwaterefficientlyduringtheoperation.Keywordswhiterotfungalmembranebio-reactorFMBRpolymerasechainreactiondenaturinggradientgelelectrophoresisPCR-DGGEmicrobialcommunitystructuresmultidimensionalscalingMDSdecolorationrate104461-2。3。。whiterotfungalmembranebio-reactorFMBRPCR-DGGEpolymerasechainreactiondenaturinggradientgelelectrophoresesmulti-dimensionalscalingMDS。11.1X-3B1。Takara。1X-3BFig.1ThechemicalstructureofreactivebrilliantredX-3BC19H10O7N6S2Cl2Na21.2PhanerochaetechrysosporiumP.chrysosporium。PDA37℃74℃。PDAL－1200g20gKH2PO43gMgSO4·7H2O1.5gB10.01g20gpH6.5～7.0。4。L－110g0.2g0.54gKH2PO42.56gMgSO4·7H2O0.71gB10.001g70mL10mmol/LpH4.5。L－10.6gMnSO4·H2O0.5gNaCl1gFeSO4·7H2O0.1gCoCl2·H2O0.19gCaCl2·2H2O1.56gZnSO4·7H2O0.1gCuSO4·5H2O0.1gKAlSO42·12H2O0.01gHBO30.01gNa2MoO4·2H2O0.01g。1.3、20.425mm。100mm×300mm×250mm10100mm5L0.2μm150mm×300mm×250mm5L。24。81cm。2Fig.2SchematicdiagramofwhiterotfungicontinuousMBRfordyewastewatertreatment2043。1∶4X-3B0.2L/h100mg/L。。1.4DNA18SrDNAV31.4.1ITS3-ITS451。1ITS3-ITS4Table1TheprimerITS3-ITS4offungi5'→3'/bpITS3GCATCGATGAAGAACGCAGCITS3-ITS43506ITS4TCCTCCGCTTATTGATATGCITS3-ITS4350GCCGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGG1.4.2PCRPCR2。PCR95℃3min94℃1min55℃1min72℃30s3572℃7min。2%。2PCRTable2ThePCRamplificationV/μLdNTPs10mmol/L1MgCl225mmol/L410×Mg2+65'-20μmol/L12.5μmol/L13'-20μmol/L12.5μmol/L1DNA100μg/mL1TaqDNA5U/μL0.5ddH2O35.5501.5DGGEBioRadDcodeTMPCR。8%30%60%PCR5μL10。150V60℃4。。UMAX。1.6Shannon-WienerShannon-WienerH′=－ΣPilnPiPi=Ni/N。NiiN。1.7540nm7A124A2η=A1－A2/A1×100%。22.1DGGEPCR-DGG8。1005DGGE53。3。35P.chrysosporium。CandidatropicalisTrichospo-riellasp.GeotrichumcandidumHypocrealixii4。3Table3Strainsidentifiedbymeansof18SrDNAsequencingfragmentsfromDGGEband/bpID/%1371Phanerochaetechrysosporium99AF475147280Geotrichumcandidum90AJ8768933248Trichosporiellasp.90DQ0690494378Hypocrealixii98AY8572505287Candidatropicalis100AF219001IDGenBank。30446M=Markerp=Phanerochaetechrysosporiumh=Hypocrealixiic=Candidatropicalist=Trichosporiellasp.g=Geotrichumcandidum3DGGEFig.3DGGEprofilesofdifferentPCRsamplesinthecontinuousMBR2.2Shannon-WienerShannon-Wiener。75P.chrysosporium435%～50%2055Trichosporiellasp.3。75P.chrysosporium65%～70%。5Shannon-Wiener1.5～2.51.92。Shannon-Wiener7510℃P.chrysosporium4。CandidatropicalisTrichosporiellasp.Geotrichum4P.chrysosporiumFig.4ThepeakareapercentageofcharacteristicbandsofP.chrysosporiumcandidumHypocrealixii4pHP.chrysosporium、pH75Shannon-WienerP.chrysosporium40435Shannon-WienerFig.5TheShannon-Wienerindexofmicrobialcommunitystructure。2.3100100mg/LX-3B82.5%693.2%20～30。3576.5%。35P.chrysosporium35P.chrysosporium75P.chrysosporium6。2.4MDSDGGEMDSDGGE910SPSS15.0MDS7。100。20559095100P.chrysosporium。5P.chrysosporiumP.chrysosporium、pH。3118SrDNAP.chryso-sporiumCandidatropicalisTrichosporiellasp.GeotrichumcandidumHypocrealixii4。275P.chrysosporium35%～50%20554。Shannon-Wiener1.920.92P.chrysosporium6Fig.6DyewastewaterdecolorizationofthecontinuousFMBR50446d7MDSFig.7MDSordinationdiagramofthechangesincontinuousMBRmicrobialcommunitiesduring100daysP.chrysosporium。3DGGEMDS20559095100P.chrysosporium。82.5%93.2%。P.chrysosporium。1ZhangFMJeremySKKelvinNT.DevelopmentofbioreactorsystemsfordecolorizationofOrangeⅡusingwhiterotfungus.EnzymeandMicrobialTechnology1999241/248-532GaoSChenCTaoFetal.VariationofperoxidaseisoenzymeandbiofilmofPhanerochaetechrysosporiumincontinuousmembranebioreactorforReactiveBrilliantRedX-3Btreatment.JournalofEnvironmentalSciences2009217940-9473.pH.2005266173-1794TienMKirkTK.LigninperoxidaseofPhanerochaetechrysosporium.MethodsinEnzymology1988161238-249