全 文 ：第27卷第5期
2008年10月
生态科学
EcologicalS ience
27(5)：351-356
Oct．2008
BacterialcommunitiesnsedimentsofLakeXuanwu
ZENGJin，YANGLiu-yan+，LIANGYi，LIJia．yun，XIAOLin，JIANGLi-juan
StateK yLaboratoryofPollutionControlandResourceReuse,SchooloftheEnvironment,NanjingUnivers ty,Nanjing210093,China
Keywords：bacterialcommunity；lakesedim nt；DGGE；CCA；I．JlkeXuanwu
’
南京玄武湖底泥微生物群落结构研究
曾 巾，杨柳燕’，梁 医，黎嘉韵，肖 琳，蒋丽娟
污染控制与资源化研究国家重点实验室，南京大学环境学院，江苏南京，210093
【摘要】 采用变性梯度凝胶电泳(DGGE)的方法研究了南京玄武湖底泥中的微生物群落结构分布。此外，我们
还采用了典范对应分析(CCA)的方法研究了底泥中一些环境因子对微生物群落结构的影响。在玄武湖三个不同湖
区共选取八个采样点采集底泥样品，测定不同底泥样品中的总氮(TN)、总磷(TP)、有机质(OM)、pH和氧化还
原电位(Eh)，发现不同采样点的环境参数存在差异。其中，采样点s2底泥样品的总氮和有机质含量最高，氧化还
原电位水平最低。对DoGE分离的DNA条带进行测序，测序结果利用BLAST与GenBank数据库中的菌种进行对比，
运用MEGA3．0软件构建了系统进化树。结果表明：玄武湖底泥中分离出的微生物属于Proteobacteria,Actinobacteria,
Verrucomicrobia和Nitrospira，它们都属于富营养化湖泊中常见的微生物类群。典范对应分析(CCA)的结果表明：
底泥样品的pH值和氧化还原电位水平对底泥当中的微生物群落结构有显著影响。
关键词：微生物群落；底泥；DGGE；CCA；玄武湖
中图分类号：Q938 文献标识码：A 文章编号：1008—8873(2008)05．351-06
I Introduction
Organicmaterialsdecompositionandtra sformationof
nutrientelementsis hemostimportantfu ctionofthe
microbialcommunityinfreshwatersediment．Therefore，
informationonthecompositionofthemicrobial
communityisofkeyimportanceforab tterunderstanding
ofthemetabolicpro essesinthisaquaticecosystem【11．
、Ⅳhilebotht environmentalfactorsandthemicrobial
communitystructureinlakehavebeenwell-documented,
thetwoaspectswereoftenconsideredseparately．Inrece t
years，canonicalcorrespondenceanalysis(CCA)hasbeen
adoptedtodemonstrateherelationshipbetweenbacterial
communitycompositionandenvironmentalfac orsnd
proventobemores nsitivehanunivariatemethods[21．
LakeXuanwuinNanjingsafamousre ortlakeof
China,嬲wellasa typicalurban,shallowl ke．
Eutrophicationhasgreatlyaffectedtherecreationalvalue
ofLakeXuanwuandtheoutlookfNanjingcityisalso
influenced．Theaimofthisstudywastoexaminethe
收稿日期：2008-09·10收稿，2008-10-10接受
基金项目：973项目：湖泊蓝藻水华生态灾害形成机理及防治的基础研究(2008C阱18102)
作者简介：曾 巾(1982--)，女，博士研究生，研究方向；湖泊元素生物地球化学循环．
·通讯作者，E-mail：yangly伽ju．edtLen
万方数据
352 生态科学EcologicalScience 27卷
TableI Averagequalityofsedimentofdifferentsamplings tes．Totalnitrogen(T№；totalphosphorus(TP)；organic
matter(oM)；redoxpotential(Eh)
墨星垒i坐!里堡!塑巳!塑
Sites Locationa TN TP TT OM Eh
(翌型g：盟) (型＆塑) ： f型g：塑) f婴12
Sl 32004’49．15’，N118047’05．63”E 3．39+0．0160．47士o．02 7．69 59．4+4．4 ·121
S2 32005’13．83’，N118047’18．59”E 4．69士O．01 0．38士O．19 7．47 85．2士1．5 -174
S3 32004’59．10’，N118047，25．86’，E 3．41士-0．11 0．424--0．oo7．36 55．6士0．8 ·111
S4 32004’49．937，N118047’41．85”E 2．464-0．170．22士O．0l 7．17 21．7士0．4 -116
S5 32004’31．427，N118047’32．21”E 2_30士O．170．414,0．057．68 24．54-1．7 -102
S6 32004706．13’，N118047’58．75”E 2．38士o．08 O．27士O．01 7．73 25．44-1．2 —99
S7 32004’08．34’，N118047’24．70”E 2．24：L-0．13 0．414-0．037．81 22．7士1．9 -124
S8 32004’13．93’，N11804706．64”E 3．85+0．100．46士0．04 7．34 62．8士I．9 -133
％ocationsweredeterminedusingGPS
bValuesaregivenasmean4-standarddeviation(n=3)
spatialdistributionofthesedimentbac erialeommunity
presentindifferentareasofLakeXuanwu．Additionally，
thecontributionsoftotalitrogenfiN)，TP，OM，pHand
redoxpotential(Eh)(whichhavebeenproventobekey
factorsforbacterioplanktoncommu itvstructures)on
microbialcommunityd stributionwereassessedusing
multivariateanalysistechniques．
2 MaterialsandMethods
Samplingprocedureandanalysisofphy7sicochemical
parameters
Sedimentsweresampledfromeightsites(S1一s8)inthree
differentlakezonesandthelocationofeachsitewas
recordedwithGPS(Tablel、．S dimentsamplesw re
collectedas pticallyusingasampler(HL·CN，Hengling
technologyLtdCorp．，China)．Threereplicatesofs diment
ateachsitewerecollectedandsamplesforeachsitewere
homogenizedandstoredintoa sterile50mLcollection
tube．
EhandpHweremeasuredinsituwithspecific
electrodes(PUB-5，REX，China)．TN，TPand0M
concentrationsweremeasuredaccordingtoJinandTu【31
afterthesedimentsamplesw redriedwithaFreezeDryer
(1abconco，Cole—ParmerInstru ntCo．，USA)．
DNAextraction．PCRamplificationandDGGEanalysis
Sedimentsamplesw repreparedwithTENPbuffer(50
mmol／LTris，20mmol／LEDTA，100mmol／LNaCl，0．01
g／mLPVPP，pH10．0)andPBSbuffer(8g NaCl，0．2g
KCl，1．44gNa2HP04，0．24gKH2P04，dilutedtol L，pH
7．4)toremovethPCRinhibitingsubstances．DNAwas
extractedfollowingtheinsitulysismethodofGillan【4】．
Theprimerset，341f-GC(5-GC-clamp—CCTACGGGA
GGCAGCAG一35and907r(57-CCGTCAATTCCr兀GA
GTTT-33wasusedinthisstudy．A40-base—pairGC-clamp
(CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCA
CGGGGGG)wasattachedtothe5’endoftheforward
primer(3410．PCRamplificationandDGGEanalysis
wereperformedaccordingtoZhaoetalI川．
AnalysisofDGGEbandingpattemandbandssequencing
Ageldocumentationsystem,GelDoc2000，Quantity-One
4．5．2(Bio·gad,USA)wasemployedtoevaluateth
DGGEbandprofiles．Strongandwell．isolatedDGGE
ba dswereexcisedfromthegelandl6S删Afragments
werere-amplified，exceptthatnoGCclampwas
incorporatedintotheforwardp imer．ThesePCRproducts
werepurifiedusingQiaquickPCRpreps(Qiagen，
Valencia，CA)andsequencedusinganABIPRISM3730
automatedsequencer(AppliedBiosystems，FosterCity,
CA)byShanghaiSangonBiologicalTechnologyCo．Ltd．
BandsequenceswerecomparedtotheGenBank
databaseu ingtheBasicLocalA ignmentS archTool
(BLAST)algorithm．Sequenceswerealignedwith
referencetaxaandavailableenvironmentalcloneswithin
thedatabasesusingtheCLUSTALX program．
Phylogenetictreewasonstructedbytheneighbor-joining
万方数据
5期 曾 巾，等：南京玄武湖底泥微生物群落结构研究 353
methodbasedondistanceestimationscalculatedbythe
methodofJukesandCantor．
Thesequencesgeneratedinthistudyhavebeen
depositedin heGcnBankdatabase．
Multivariablestatisticsanalysis
Toperforma statisticalan ysisoftheDGGEprofiles
versusthesedimentproperties，wechoscanonical
correspondenceanalysis(CCA)‘61．TheanalysisWas
carriedoutusingCANOCO4．5 softwarepackage
(Biometris，Netherlands)andtheordinafioplotsfor
speciesandenvironmentalvariableswerecharacterizedby
biplots．
3 ResultsandDiseussion
Thephysicochemicalparametersofsedimentsamplesare
showninTable1．111econcen仃ationsofTNandOMwere
thehighestatthesiteS2．whilethvalueofEhWasthe
lowesta this ite．Additionally，TNW spositively
correlatedwithorganicmatterp=0．95，甩=8，P<0．01)
whileTPandEhweren gativecorrelated(，=-0．86，刀=8，
P<0．01)。Xueet al I7J investigatedsediment
characteristiesatfivedifferentsitesofLakeXuanwuand
foundthatthetotalphosphorusc ncen廿afionsrangedfrom
0．25t0O。39mg／g．Thechangesoftotalni仃ogen．organic
mattersandpHwere2．51-2．89mg／g，18．6—61．2mg／g，
7．15—7．87．respectively．‘111einfluenceofdomesdc
wastewaterdischargedfroma hotelseal"siteS2may
explainthelevatednutrientconcentrationsat h sstation．
111eDGGEpattemsofsedimentsfromdifferent
samplingsitesareshowninFig．1，andtheprominent
bandshavebeenindicated．Station2hadthelower
microbiald versityinsedimentssample．Twenty-three
prominentbandswereexcisedfromthegelsandsubjected
tosequencing．Identifiedbacteriaclusteredintofivemajor
phylogeneticgroups，includingtheVerrucomicrobia(B4，
B8，B10，B14，B18and821)，Actinobacteria(B2，B6，B13
andB16)，Nitrospira(BI，B3，B7，B15，B19and822)，
a-(B9)，andp-Proteobacteria(B5，B11，B12，B17，B20
andB23)(Fig．2)．
Thebacterialgroupswedetectedbysequencea alysis
飚．1DGGEbandprofilesfromdifferents dimentsamples．
TheiabebofSI-$8atthetopofthelanesrepresent
correspondingsamplingsites．Serialnumbersinthefigure
representsthepredominantbands．
areamongthecommongroupsfoundin freshwater
environments[引．Previousres archndicatedhat
Verrucomicrobiaarerelativelymoreprevalentin u rophic
lakesthaninoligo—ormesotrophiclakes【91．InourCCA
result，Verrucomicrobia(B4，8，10，14，18 nd21)were
gatheredpositivecorrelatedwi也OMandTNinthe
sedim nt,whichwasconsistentwithpreviousreports．
Actinobacteriaareprev lentinfreshwatersndestuaries
wimvaryinghydrologicalandlinmologicalchar cteristics
andfromdistantgeographiclocationst91．InourCCA
analysis，theidentifiedActinobacteriaclusterednegatively
withTN，whichsupportsthenotionthatActinobacteriaare
lesscommoninthelakesofhightrophicstatusthani the
lakesoflowtrophicstatus【9】．Anothergroupofbacteria
commonlypresentinoursampleswaProteobacteria．Six
ofoursequencesbelongedtop-Proteobacteriasubg oup
andoneto小subgroup．Previousstudyshowedthatthe弘
万方数据
Gram+．HGC
Nitrospira
F喀2Phylogenetictr eofsequencedDGGEbands．Thetrewasconstructedbyusingtheneighbor-joiningmethod．Abootstrap
analysiswaperformedwlth1000repetitions．andonlyv luesabove50areshown．
andy-Proteobacteriaa豫of锄ldominantgroupsinhigher
salinityenvironment,whereasB．Proteobacteriappearsto
bepredominantindigenousc mmunityinfreshwater
environments【21．Ashowni Fig．3，Proteobacteria
assembledinthenegatived r ctionofvectorTP，suggesting
thisgroupprefershigherredoxpotentialandlowerTP
万方数据
5期 曾 巾，等：南京玄武湖底泥微生物群落结构研究 355
concentration．Thec mmonoccl卫q"erlceofNitrospirainou
studysuggeststhatNitrospiraisa predominantgroup
existedinsedimentsofeutrophicationlakes．Thisorganism
canutilizenitriteundermicroaerophilicconditions．Italso
growsautotrophicallybyusingnitrateasallelectron
acceptorunderanOXlCsettings．
Fig．3Canonicalcorrespondenceanalysis(CCA)ordination
diagramofDGGEdata,withenvironmentalfac ors嬲
arrowsandmostabundantbac eria．TotalnitrogenO’№；
totalphosphorus(TP)；organicmatter(oM)；redoxpotential
(Eh)．Blacktriangle,sequencedband；opentriangle，bandnot
sequenced．GroupsIII,HIand Vofphyiotypesale
indicatedbyn旷aybackground．
BiplotscalingofCCAwithinterspeciesdistancesis
showninFig．3．Accordingtotheresultofmanualforward
selection(Table2)，pHandEhplayedimportantrolesin
themicrobialcommunitycompositionin hesediment．The
analysisba edoninterspeciesdistancesrevealedfour
groupsofbacterialcommunitiesn hsediments(Fig．31．
Ehdisplayeda majorcorrelation、玩也communities
gatheredingroupI．Group11waspositivelycorrelated
withpH．Group111wereminorinfluencedbyTP．Positive
correlationswereshownbyTNandorganicmatterin
groupIV．
In0111"CCAanalysis，physicochemical(pH，Eh)
parametersemergedassignificantenvironmentalfac ors
affectinghemicrobialcommunity．Acorrelationbetween
pHandbacterialcommunitystructttrehasbeenreported
elsewhere[101．pHmaymerelyreflectchangesinother
environmentalfactors，suchastheavailabilityofionsand
tracemetals，whichCanhavebothinhibitoryand
growth-enhancingeffe ts，butafluctuationinpHmayalso
influencemicrobialcommunityd rectly．Fahyeta／f11】
reportedashiftinthemicrobialcommunitydrivenbythe
changesinredoxpotential，whichwascausedbybenzene
metabolism．Thebiodegradationofbenzenedepl tesoxygen，
profoundlyaffectingtheredoxpotential，whichinturn
resultsinasuccessionofmicroorganismsabletodegrade
benzenemicroaerophilicallyorwithdifferentterminal
electronacccptors，suchasnitrate，metalionsorsulfate．
Inconclusion,thisstudyemonstratedthevariabilitiesof
microbialcommunitystructureatdifferentsitesofLake
Xuanwu．Additionally,relationshipsofseveralenv ronmental
factorswithebacterialcommunitywereshown．pHandEh
contributedsignificantlytothebacterialcommunityshiftsin
thesediment．Thedominantb c eriald visionswedetected
羽胃commonlyfoundinfreshwaterecosystems．ToOUrbest
knowledge，thisisthefirsttimetoexplaintherelationship
bel阳，eenoverallmicrobialcommunityandphysicochemical
factorsinthesedimentoffreshwaterecosystembasedOllthe
combinationofDGGEandmultivariateanalysis．Theresultin
thistudyisimportant,bt七ausemicrobialcommunitiesn
lakeplaycrucialrolesinnutrientscycling,whichare
necessarytomaintainecosystemhealthandthepresenceof
particularbacterialcommunitycouldindicatethnutrient
statusofalake．
Table2MarginalandconditionaleffectsofforwardlyselectedenvironmentalvariablesproducedbyCCA．
Environmentalvariab e
Marginaleffects———————Co—n—ditio．nale—ff—ec．．．—t—s
Lambdal LambdaA P-valueF-factor
interspect髂mstan⋯fsea㈣誊 ：耋 篡 ：芝 篡
万方数据
Referenees
11l NixdorfB，JanderJ．2003．Bacterialactivitiesinshallow
lakes--Acomparisonbetweenextremelyacidicand
alkalineeutrophichardwatcrlakes[J】．Hydrobiologia，
506．509：697—705．
12J HenriquesIS，Alves^Tac矗oM．2006．Seasonalnd
spatialvariabilityoffree··livingbacterialcommunity
compositionalongallestuarinegradient(RiadeAveiro，
Portugal)【J】．EstuarineCoastalandShelfScience,
68：139—148．
13l JiIlXC，TuQY1990．SurveySpecificationforLake
Eutrophication【M】．Beijing：EnvironmentalSciencePress．
208．230．
【414 GillanDC．2004．Theeff ctofanacutecopperexposure
011thediversityofamicrobialcommunityinNorthSea
sedimentsasrevealedbyDGGEanalysis-theimportanceof
theprotocol【J】．MarinePollutionBulletin，49：504．513．
151ZhaoXQ，YangLY，YuZYeta1．2008．Characterization
ofdepth-relatedmicrobialcommunitiesnlakesedimentby
denaturinggradientgelelectrophoresisofamplified16S
rRNAfragmentsif]． JournalofEnvironmental
Science-China，20：224．230．
16l TotBraakC J E 1987．Theanalysisof
vegetation-environmentrelationshipsby canonical
correspondencea alysis[J】．Vegetation，69：69-77．
【7】XueLQ，LuXW：WuL．2004．Simulatingstudyonthe
innersourcenutrientxchangebetweenwater-sediment
interfaceintheXuanwuLake啊．ChineseE vironmental
‘PollutionControl,26：163-166．
嘲8 HaukkaKKolrnonenE，HyderReta1．2006．Effecto
nutrientloadingOB bacterioplanktoncommunity
compositioninLakeMesocosms【J】．MicrobialEcol gy,
51：137-146．
【919 KolmonenE，SivonenkRapalaJ．2004．Diversityof
cyanobacteriaandheterotrophicbacteriancyanobacterial
bloomsinLakeJoutikas，Finland[r1．AquaticMicrobial
Ecology，36：201-211．
【10lStepanauskasRMoranMA，BergamasehiBAeta1．2003．
Covarianceof baeterioplanktoncomp sitionand
environmentalvari blesinatemperatedeltasystem叨．
AquaticMicrobialEcology．3l：85-98．
IIJFahyALethbridgeGEarleReta1．2005．Effectso
long-termb lL瑚epollutionOilbacterialdiversityand
communitystructureingroundwater叨．Environmental
Microbiology．7：1192·1199．
万方数据
南京玄武湖底泥微生物群落结构研究
作者： 曾巾， 杨柳燕， 梁医， 黎嘉韵， 肖琳， 蒋丽娟， ZENG Jin， YANG Liu-yan， LIANG
Yi， LI Jia-yun， XIAO Lin， JIANG Li-juan
作者单位： 污染控制与资源化研究国家重点实验室,南京大学环境学院,江苏南京,210093
刊名： 生态科学
英文刊名： ECOLOGICAL SCIENCE
年，卷(期)： 2008,27(5)

参考文献(11条)
1.Jin X C;Tu Q Y Survey Specification for Lake Eutrophication 1990
2.Henriques I S;Alves A;Tacao M Seasonal and spatial variability of free-living bacterial community
composition along an estuarine gradient (Ria de Aveiro,Portugal)[外文期刊] 2006
3.Fahy A;Lethbridge G;Earle R Effects of long-term benzene pollution on bacterial diversity and
community structure in groundwater[外文期刊] 2005(8)
4.Stepanauskas R;Moran M A;Bergamaschi B A Covariance of bacterioplankton composition and
environmental variables in a temperate delta system[外文期刊] 2003
5.Nixdorf B;Jander J Bacterial activities in shallow lakes-A comparison between extremely acidic and
alkaline eutrophic hard water lakes[外文期刊] 2003
6.Kolmonen E;Sivonen K;Rapala J Diversity of cyanobacteria and heterotrophic bacteria in
cyanobacterial blooms in Lake Joutikas,Finland[外文期刊] 2004
7.Haukka K;Kolmonen E;Hyder R Effect of nutrient loading on bactarioplankton community composition
in Lake Mesocosms[外文期刊] 2006
8.Xue L Q;Lu X W;Wu L Simulating study on the inner source nutrient exchange between water-sediment
nterface in the Xuanwu Lake 2004
9.Ter Braak C J E The analysis of vegetation-environment relationships by canonical correspondence
analysis[外文期刊] 1987
10.Zhao X Q;Yang L Y;Yu Z Y Characterization of depth-related microbial communities in lake sediment
by denaturing gradient gel electrophoresis of amplified,16S rRNA fragments[外文期刊] 2008(2)
11.Gillan D C The effect of an acute copper exposure on the diversity of a microbial community in
North Sea sediments as revealed by DGGE analysis-the importance of the protocol[外文期刊] 2004(5/6)

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