不同区域污水处理厂活性污泥中微生物菌落结构分析杨倩46637061f524ccbff021844

20143312CHEMICALINDUSTRYANDENGINEERINGPROGRESS3329111112212100092210042DGGE16srDNAPCRBlast89β-Bataproteobacteriaγ-GamaproteobacteriaFirmicutesDGGEX508A10006613201412332908DOI10.3969/j.issn.1000-6613.2014.12.033DiversityofbacterialgroupsinactivatedsludgesamplesfromdifferentareasofChinaYANGQian1JIANGYangyue1WANGXiaojun1CHENYingwen1SHENShubao1SHILili2LIUJining21CollegeofBiotechnologyandPharmaceuticalEngineeringNanjingUniversityofTechnologyNanjing210009JiangsuChina2NanjingInstituteofEnvironmentalSciencesMEPNanjing210042JiangsuChinaAbstractThediversityofbacterialgroupsinactivatedsludgesamplesreceivedindifferentsewagetreatmentplantsfromdifferentareasofChinawasinvestigatedbycombiningwithtraditionalpureculturetechniqueanddegenerationgradientgelelectrophoresis(DGGE)approach.AfterthecoloniesweresequencedtheobtainedsequenceresultsbyBlastanalysiswereusedtoconstructthephylogenetictree.MostbacteriawereaffiliatedwiththeBetaproteobacteriaGammaproteobacteriaandFirmicutes.TheDGGEpatternsshowedthatthereweremanycommonbandsinallsystemssuggestingthehighsimilarityofbacterialcommunitiesindifferentsystems.Meanwhiletherewerealsospecificbandsineachsample.Alsothesimilarityinthesameareaisgreaterthanthatindifferentareaswhichismainlyrelatedtodifferentnaturalandeconomicenvironmentandpeople’slivinghabits.KeywordsactivatedsludgebacterialgroupsbacterialidentificationDGGE95%[1][2-3]2014[e-mail]20143333302080[4-8][9]1%15%[10]16SrRNAPCRFISH[11-15]-PCR-DGGE[16-21]PCR-DGGEAbd[22]NicoBoon[23]11.11.1.11−20DNALBLuria-Bertani10.0g5.0g10.0g20.0g1000mLpH7.21.21.2.1GB4789.219941/m3·d−1CD20A2/O2013.3.27CB30Unitanks2013.4.27JXZ64A/O2013.4.27JN4A2/O2013.4.27DTS15A2/O2013.5.31LD20A2/O2013.5.31LR20A2/O2013.5.31SSW302013.9.12XNH40BAF2013.9.12A2/OAnaerobic-Anoxic-OxicUnitanksBAFBiologicalaeratedfilterSN0168921.2.2LB3716SrRNA16SrDNA27F5’-AACTGAAGAGTTTGATCCTGGCTC-3’1492R5’-TACGGCTACCTTGTTACGACTT-3’PCR25μL2×MasterMix12.5μL1μL(10μmol/L)DNA1μL(15ng)9.5μLPCR952min951min551.5min722min297210minPCR1%1.2.3DNAPCRUltraCleanTMSoilDNAIsolationMoBIOLaboratoriesDNA1%123331DNAPCR50μL10×PCR5μLMgCl23.5μL(20mmol/L)dNTP2μL(10mmol/L)518R(5’-ATTACCGCGGCTGCTGG-3’)GC338F(5’-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGGACTCCTACGGGAGGCAGC-3’)1μL(10μmol/L)TaqDNAREGENBIOTEC0.3μL(1.5U)DNA1μL(15ng)36μLPCR1%DGGECBS-DGGECBSScientificCo.8%35%65%100%7mol/L40%1×TAEPCR500ng60100V12hEB20minUVP[24]1.316SrDNANCBIGenbankBlastClustalXBioEditMega4.0DGGEGelcompar[25]UPGMAunweightedpair-groupmethodwitharithmeticaverages22.1SV30MLSSSVI2SV30/%MLSS/mg·L−1SVI/mL·g−1/CFU·mL−1CD644167153.581.3×107CB35752646.51.9×107JXZ46857354.934.9×107JN41818850.072.3×107DTS13.8273450.483.2×106LD10.2191653.242.6×106LR33.4502866.432.2×106SSW12180066.671.4×106XNH70920076.094.6×1062CDSVISVI50120CD2.2CDCBJXZJN10121011DTSLDLR9118SSWXNH98916SrDANNCBIBlastBlast96%[26]PseudomonasAcinetobacterBacillusAeromonasKlebsiella[27]ClustalxMega13FirmicutesProteobacteriaβ-Bataproteobacteriaγ-Gamaproteobacteria[28][29][30]β-γ-γ-20143333321233332014333334116SrDNAActinobacteriaStreptomycetaceaeMicrococcaceaeCDCDSVI2.3DGGE2DGGE216SrDNAV3PCRDGGE1XNH20DTS29CD35CBBODCODpH4567910674CBJXZ81.34%JN58.35%LDLR85.47%DTS1233353LaneCBJXZJNCDLDLRDTSSSWXNHCB100.0081.3466.6755.3161.3550.4551.6248.4730.08JXZ81.34100.0058.3551.7754.4447.7154.1945.0440.11JN66.6758.35100.0058.2760.2148.4146.9347.7041.17CD55.3151.7758.27100.0056.1447.0742.7144.0723.08LD61.3554.4460.2156.14100.0085.4775.7727.5419.29LR50.4547.7148.4147.0785.47100.0081.7224.0715.22DTS51.6254.1946.9342.7175.7781.72100.0028.1920.73SSW48.4745.0447.7044.0727.5424.0728.19100.0045.59XNH30.0840.1141.1723.0819.2915.2220.7345.59100.0075.77%SSWXNH45.59%61.35%50%DGGE293CBJXZJNCDLDLRDTSSSWXNH50%120083“”3DGGE[1]3DGGE38916SrDNAFirmicutesβ-Bataproteobacteriaγ-GamaproteobacteriaDGGE[1]LiuCXZhangYYangMetal.AnalysisofbacterialcommunitystructuresintwosewagetreatmentplantswithdifferentsludgepropertiesandtreatmentperformancebynestedPCR-DGGEmethod[J].JournalofEnvironmentalSciences(China)200719160-66.[2]LiJYHuBLZhengPetal.FilamentousgranularsludgebulkinginalaboratoryscaleUASBreactor[J].BioresourceTechnology20089993431-3438.[3]GulezGDelosReyesIIIFL.Multipleapproachestoassess2014333336filamentousbacterialgrowthinactivatedsludgeunderdifferentcarbonsourceconditions[J].JournalofAppliedMicrobiology20091062682-691.[4]ZieglerMLangeMDottW.Isolationandmorphologicalandcytologicalcharacterizationoffilamentousbacteriafrombulkingsludge[J].WaterResearch199024121437-1451.[5]SoddeJASeviourRJ.Microbiologyoffoaminginactivatedsludgeplants[J].JournalofAppliedMicrobiology1990692145-176.[6]HiraishiAMasamuneKKitamuraH.Characterizationofthebacterialpopulationstructureinananaerobic-aerobicactivatedsludgesystemonthebasisofrespiratoryquinoneprofiles[J].AppliedandEnvironmentalMicrobiology1989554897-901.[7]PrakasamTBSDonderoNC.AerobicheterotrophicbacterialpopulationsofsewageandactivatedsludgeI.enumeration[J].AppliedMicrobiology1967153461-467.[8]IlialetdinovANMendeshevAManasbaevaABetal.Activi