INDUSTRIALWATER&WASTEWATERVol.42No.6Dec.,2011α-1,1,2(1.,300072;2.,430010)MBR,,,。[1],[2-3],:PCR-DGGE4α-。,α-,,。,,MBR,,MBR。MBR,79.6%77.7%,50%~60%。,/,80%。MBR,47.5%~84.1%,。:;PCR-DGGE;16SrDNA;;;:X703.1:A:%1009-2455(2011)06-0020-04Analysisofcommunitystructuresofα-proteobacteriaindifferentphosphorusremovingsystemsLIUQian1,YANGXi-long1,2(1.CollegeofEnvironmentalScienceandEngineering,TianjinUniversity,Tianjin300072,China;2.CentralandSouthernChinaMunicipalEngineeringDesignandResearchInstitute,Wuhan430010,China)Abstract:Thecommunitystructuresofα-proteobacteriainfourdifferentphosphorusremovingsystemsunderthenormalrunningconditionwereinvestigatedthroughPCR-DGGEtechnology.Theresearchindicatedthat,thedemandofα-proteobacteriaaboutwaterqualityandtechnicalconditionwasnothigh,almostallofthebacteriastrainswhichconstitutethemainbodyofthephosphorusremovingbacteriacommunityinthesystemweredominantspecies.However,onthewhole,therewereobviousdifferencesamongvarioustreatmentsystems,MBRsystemcontainedmoreabundantα-proteobacteriacommunitythantraditionalactivatedsludgesystem;andmaybeduetothepresenceofspecificbacteria,thediversityindexoftheformeronewasgreaterthanthatofthelaterone.Thecommunitystrcturesimilaritieswere79.6%and77.7%separatedinaerobicandanoxicstagesofMBRsystem;however,thesimilarityratewasonly50%-60%betweenthetwostages.Thecommunitystructuressimilarityratewasabove80%inaerobicandanoxic/anaerobicseparatedwastewatertreatmentsystems.BetweenthetraditionaltreatmentsystemandtheMBRsystem,thecommunitysimilaritycoefficientwasatabout47.5%-84.1%,andthecommunitysimilarityofaerobicstagewasbetter.Keywords:proteobacteria;PCR-DGGE;16SrDNA;phosphorusremovingbacteria;microbialcommunitystructure;membranebioreactor:(07JCZDJC02100):2011-09-16;:2011-10-25·20·1Tab.1OperationconditionsandnumerationofsamplesofwastewatertreatmentplantsAA/O--C1:,C2:,C3:B(),C4:CMBR,(PVDF),0.2μm,,8min,2min,M1:,M2:DMBRM3:,M4:2Tab.2OperationconditionsofsamplingplantsA150~2502015~40520~45152~51~2B200~3506040~501545~6020~303~51~2CMBR140~20010100101001050.9DMBR130~2505025~40530~5081.5~41ρ(CODCr)/(mg·L-1)ρ()/(mg·L-1)ρ(TN)/(mg·L-1)ρ(TP)/(mg·L-1)3Tab.3Operationconditionsofsamplingstagesρ(DO)/(mg·L-1)ρ(MLSS)/(mg·L-1)SV/%CODCrTNTPC14.330∶158206880~9361~7688~9565C21.3332025C30.8240019C44.030∶135304765~8025~5065~7580~95M14.824∶177309885~9385~9490~9681~86M23.0661097M34.624∶175607265~8078~8585~9550~65M41.8463031/%[4]。PCR-DGGE4(MBR)α-,MBR,,。11.14,MBR2。4、1~3。8,41,3,。,DNA。1.2DNA1.2.1500mL,30min,,50mL50mL,6~10℃、9165×g10min;,30mL,10min;,30mLTE,10min;15mL,:α-·21·INDUSTRIALWATER&WASTEWATERVol.42No.6Dec.,20114PCRTab.4PCRprimersandamplificationprocedure//℃/s/℃/s/℃/s1PCRF203α,R1378r3094605760721202PCRR518,F357-GC20109494606065~55556060727260601α-DGGEFig.1DGGEprofileofα-proteobacteriaindifferentsamplingplacesC1C2C3C4M1M2M3M4ABJCDEFGHI5α-DGGETab.5ClassificationofDGGEprofileofα-proteobacteriaα-A、B、C、D、EF、IG、H,5mLDNA[5]。1.2.2DNADNA-//DNA。BioerTechnologyBioFluxDNA。1.3DNA16SrDNAV3PCRα-PCR[6],95℃10min,72℃12min。PCR[7],94℃5min,72℃8min,200.5℃。4。1.4PCRDGGEC.B.S.SCIENTIFICDGGE-2001PCR,8%,30%~60%。,1×TAE,10%1×TAEPCR20μL,150V,60℃6.5h[5-6]。1.5Shannon-WienerQuantityOneDGGE,Shannon-Wiener(H)。H:H=-Si=1ΣPilogPi=-Si=1Σ(ni/N)log(ni/N)(1):Pi=ni/N,ni,N。22.1α-DGGEα-DGGE1,5。51,5A、B、C、DE,CDα-,,,。,、E,。MBR,B,C、DE。2,A。Fα-M1M2,,[8]。I,,α-,。GHα-MBR,,,MBR,,,MBR。·22·6α-ShannonTab.6Shannonindexesofα-proteobacteriacommunityC1C2C3C4M1M2M3M4H0.4560.5480.5140.5400.6050.6140.5470.506C1C2C3C4M1M2M3M4C1C2C3C4M1M2M3M410074.086.759.281.167.078.451.210086.872.668.160.384.154.410069.780.364.982.058.010047.560.059.149.810052.779.662.810060.777.710066.21007α-Tab.7Similarityofα-proteobacteriacommunityindifferentsamplingplaces%2.2α-α-Shannon(H)6。6,α-。Aα-。MBRM1、M2M3、M4,,。,MBR,。2.3α-DGGEQuantityone、,7。7,C3C1,C2C1C2。,α-,。α-MBRM1M3、M2M480%,MBR,MBRα-。C1、M1、M3,C4,C4/。MBR,47.5%~84.1%,,,。3(1)A、B、CD4α-,5,,。α-,,MBR。(2)DGGE,α-47.5%~86.8%,,。MBR,,α-。:[1],,,.[J].,2008,29(2):474-481.[2]AtkinsonBW,MudalyDD,BuxF.ContributionofPseudomonasspptophosphorusuptakeintheanoxiczoneofananaerobic-anoxic-aerobiccontinuousactivatedsludgesystem[J].WaterScienceandTechnology,2001,43(1):139-146.[3].16SrDNA[D].:,2007.[4],,,.[J].,2006,37(6):4-7.[5],,,.MBR[J].,2010,30(1):69-70.[6]GomesNCM,HeuerH,SchonfeldJ,etal.Bacterialdiversityoftherhizosphereofmaize(Zeamays)grownintropicalsoilstudiedbytemperaturegradientgelelectrophoresis[J].PlantandSoil,2001,232(1-2):167-180.[7]MuyzerG,WaalEC,UitterlindenAG.Profilingofcomplexmicrobialpopulationsbydenaturinggradientgelelectrophoresisanalysisofpolymerasechainreaction-amplifiedgenescodingfor16SrRNA[J].AppliedandEnvironmentalMicrobiology,1993,59(3):695-700.[8].MBR[D].:,2010.:(1985-),,,,,()022-87402126()luckyliuqian@126.com。,:α-·23·