全 文 ：第29卷第4期
2010年8月
生态科学
EcologicalS ience
29(4)：318—323
Aug．2010
黄伟伟，蔡卓平，肖群，段舜山．杜氏盐藻和亚心形扁藻在不同接种密度和氮浓度下的细胞群体生长【J】．生态科学，2010，29(4)：
318．323． ．
HUANGWei-wei，CAIZlmo-ping，X1AOQun，DUANShun—shah．ColonialcellgrowthofDunalieUasalinandPlatymonas
subcordiformisunderdifferentinoculationdensityandn trogenconc ration[J]．EcologicalScience，2010，29(4)：318-323．
ColonialcellgrowthofDunaliellasa inandPlatymonassubcordiformis
underdifferentinoculationdensityandnitrogenconcentration
HUANGWd．weil’_，CAIZhuo．pin91⋯，XIAOQunl，DUANShun．shanl+
1．InstituteofHydrobiology,JinanUniversi饥Cruangzhou510632,China
2．CollegeofEnvironmentalScienceandEngineering,Ton∥iUniversity,Shanghai200092，China
3．SouthC inaSeInstituteofOceanology,ChineseAcademyofSci nces,Guangzhou510301。China
Abstract：Weinvestigatedthecolonialcelgrowthofmarineeconomicmicroalgae，DunaliellasalinaandP atymonassubcordiformis
underdifferentc llinoculationdensities(10：0，7：3，5：5，3：7and0 10)andnitrogenco centrations(10wn trogen：1．4mg‘L一，
mediumnitrogen：14mg’L。andhigIInitrogen：140mg’L“)byexaminingsomeparametersofalgalcelldensity,biomass，chlorophylla
andproteinco tent．Theresultsindicatedthatcolonialcelgrowthofthesetwoalgaeincreasedwitlltheincreasingofnitrogen
concentrations．Besides，cellgrowthofmixed—cultureWasuperiortomonoculturewhenathighandmediumnitrogenconcentrations；
especiallywhen廿leyw reculturedataproportionof7：3．Forexample，chlorophyllacontentwasincreasedbyashighas1．17pand7．77folds(P<0．01)，andproteinco tentwasimprovedby19．1％(P<0．05)and195．3％pgrowthofmixed-cultureWasremarkablylimitedbynitrogenconcentrationwhenatlownitrogenconcentration．
Keywords：Dunaliellasalina；Platymonassubcordiform&；mixedculture；inoculationdensity；nitrogen
doi：10．39690．issn．1008-8873．2010．04．003中图分类号：Q178．1 文献标识码：A 文章编号：1008-8873(2010)04·318-06
杜氏盐藻和亚心形扁藻在不同接种密度和氮浓度下
的细胞群体生长
黄伟伟1，一，蔡卓平1，一，肖 群1，段舜山p
1．暨南大学水生生物研究所，广州510632
2．同济大学环境科学与工程学院，上海000000
3．中国科学院南海海洋研究所，广州510301
【摘要】在氮浓度为1．4、14、和140nag·L-1下，对杜氏盐藻(Dunaliellasali )和亚心形扁藻(Platymonassubcordiformis)进行细
胞接种比例为10：0、7：3、5：5、3：7和0：10的培养试验，研究不同海洋经济微藻种间混合培养的细胞群体生长效应。结
果表明，杜氏盐藻与亚心形扁藻细胞群体的生长随着氮浓度的增加而提高；两种藻在氮浓度分别为14和140mg·L’1时混合培养
的细胞群体生长明显优于单独培养。中、高氮下杜氏盐藻与亚心形扁藻以7：3的接种比例混合时，微藻的细胞群体生物量和胞
内物质含量相比单独培养和其他比例均有显著提高，如叶绿素a含量比单独培养分别提高1．17倍和7．77倍；蛋白质含量比单独
培养分别提高19．1％和195．3％等。而低氮浓度下，藻的生长受到氮浓度的限制。
关键词：杜氏盐藻；亚心形扁藻；混合培养；接种比例；氮浓度
收稿日期：2010-04．10收稿，2010-06．30接受
基金项目：国家自然科学基金项目(40876074，30770336)；广东省科技计划重点引导项目(2005833201001)：珠海市科技计划重大项目
(PC20061045)
作者简介：黄伟伟(1983—)，女，博士研究生，主要从事饮用水处理。
串通讯作者，E．mail：tssduan@3nu．edu．∞
万方数据
4期 黄伟伟，等：杜氏盐藻和亚心形扁藻在不同接种密度和氮浓度下的细胞群体生长 319
1 Introduction
Marinemicroalgaeareamajornaturalsourcewith
extremelyrapidgrowth，exceedingrichnessinoil
content,stableproductivityofl pids，hydrocarbonsand
otherbioactivesubstances【IJ．Astheypossesstheability
toconvertsunlight,waterandcarbondioxidentoalgal
biomass，marineicroalgaecanb widelyusedin
bioremediationapplications，seweragetr atment,
nitrogenfixingandproductionofsustainableenergy,
whichhaveimportantsocialndeconomicvalues[21．At
present，numerouslaboratorycommercial
applicationsof microalgaeh vebeenwell
documented【jJ．Alargenumberofstudieshavebeen
carriedoutoexplorethwaytoanincreasingof
microalgaldensityt451．Concerningtheculturemodeof
microalgae，relatedrese rchesarmainlyfocusedon
theinterrelationshipofredtidemicroalgae，suchas
AsterionellayormosaandMicrocystisaeruginosa【6J．
There、verealsostudiesindicatingthatthe
interrelationshipofdifferentalgaevariesalot，withcell
densityofmicroalgalcoloniesinmixed—culturebeing
increasedord creasedscomparedwithmonoculture[71．
Nitrogenplaysanimportantrolein hegrowthofalgae，
andliRleinformationisavailableonthculturemodeof
marineeconomicmicroalgae．Therefore，thespecific
objectiveofthisstudyistoevaluatethecolonialgrowth
ofmarinemicroalgaeinmixed-culturendedifferent
cellinoculationdensitiesandnitrogenconcentrations，
withourhypothesisthatphotosyntheticorganismsmay
havethesimilarityinsharingthenaturalesourcesor
interactpositivelynaco-existinggrowthsystem．
2 Materials&Methods
2．1 Algalspeciesandculturecondition
ThemarinemicroalgaeD salinaandP
subcordiformiswe eobtainedfromtheInstituteof
Hydrobiology,JinanUniversity,Guangzhou，China，and
cultivatedunderstandardizedconditionattheconstant
irradiance(5000Ix)andtemperature(20℃)ina12h：
12h(1ight：dark)photoperiodcycleund an
irradianceofabout90pmol‘m-Ls‘1providedby
cool—whitefluorescentligh ．Thegrowthmediumwas
artificialse waterenrichedwithf／2enrichmentsolu ion
formicroalgalcultures．Salinityoftheartificial
seawaterwas30％0andtheinitialpHoftheculturewas
6．5。7．O．
2．2 Experimentald sign
FivetreatmentsweresetupasshowninTable1．
Mixed．cultureofD．salinaa dPsubcordiformiswas
establishedbyusingdifferentc llinoculationdensities
(DlO：SO，D7：S3，D5：S5，D3：S7andD0：S10)．
E chtreatmentwascarriedoutintriplicate．By
comparingthemaximumcapacityofenvironment，it
wasindicatedthatconsumptionofonecelIofP
subcordiformisequaledtothatof2．5cellsofnsalina．
UsingthecelldensityofPsubcordiform括ascriterion．
theinitialtotalcelldensityofmixed-culturewasetat
1．62x103cell·mL～．
TablelMixed-cultureofAsalinaand只subcordiformiin
thexperiment
Nitrogen
concentration
Proportionofmixed-culture
Note：DforD．salina；SforPsubcordiformis．
2．3 Determinationofceildensityandbiomass
Algalcelldensitywasalsomonitoredandrecorded
withthehelpofamicroscopeusingahemocytometer．
Algalbiomasswasdeterminedusinganelectronic
balanceinthe1abafterd yingthefiltersandcollected
algalpelletfor72hatconstant90℃．
2．4 Measurementsofbiochemicalcompositions
Chlorophyllaandproteinco tentswereassayedon
the15mdayatHNandMNconditionsand11mdayat
万方数据
LNconditionaccordingtothemethodsde cribedby
CongandLit8驯．
2·5 Dataanalysis
GrowthcurvefittingwascalculatedbyLogistic
equation[10】onthem ocultureofD．salinaa dP
subcordiformis．ThegrowthregressionequationofD．
sa6nawas"Nt=666．303／(1+e2．68723·062833木0
僻220．987)，themaximumcarryingcapacitywas
6．7x100cell·mL。：thegrowthregressionequationofP．
subcordiformiswasNt=266．368／(1+e3．14476-0．59794书n
∥=0．995)，andthemaximumcarryingcapacitywas
2．66x100cell·mL一．Soconsumptionofoneellof户
subcordiformiscouldbegotbyequalingtothatof2．5
cellsofD．salinaaRertheirmaximumcapacitiesof
environmentwerecompared．
3 Results
3．1 Growthofcelldensityandbiomassduring
microalgalculfivafion
Colonialeeldensitiesmcreasedwithincreasingof
nitrogenco centrations．11圮observedcelldensitiesin
HNappearedtohavethefastestgrowthrateAnditwas
thetreatmentofinoculationproportionD7：S3t11athad
thegreatestvariabilityofeelldensitieswheni MNand
删．Increasedeelldensitieswere19．1％and14％higher
thanmonocultureatMNp<0．05)；whileinthelatter
case(I-IN)，itWas69％and90．6％higherp<0．01，Fig．1)．
Alongwiththeincreasingof nitrogen
concentration，algalbiomassincreasedgradually,
confirmingthathighernitrogenco centrationscould
promotethcellcoloniesgrowthandfinalyieldof
microalgae(Fig．2)．Additionally,biomassatm xed-
cultureofdifferenti oculatedproportionwas
comparativelyh gherthanmonocultureatHNandMN
conditions．ForHN，increasedbiomassofthetreatment
D7：S3was13．7％p<0．05)or50．7％p<0．01)higher
incomparisontom noculture，butclosetobiomassof
thetreatmentD3：S7∞>0．05)．ForMN，thisbehavior
wasalsoobserved，withtheh g estbiomassbeingatD7：
S3；then xtWasD5：S5andD3：S7，followedby
monocultured户subcordiformisandD．salina．However,
noobviousincreasewasdemonstratedatlownitrogen
conditioninwhicht ebiomasswaaveragedatabout
0．8g．L-1．
Fig．1TotalcelldensityofD．salina∞)and只subcordiformis
(S)underdifferentc llinoculationproportionsandLN(a)，
MNCo)andHN(c)conditions
3．2 Biochemicalcompositionsduringmicroalgal
cultivation
It Wasevidentthathehighestcontentsof
万方数据
4期 黄伟伟，等：杜氏盐藻和贬心形扁藻在不同接种密度和氮浓度下的细胞群体生长 321
1．6
1．4
，、1．2
皂1
io．8
g0．6
∞0．4
O．2
O
2．5
2
1．5
l
O．5
O
DlO：soDO：S10‘D7：岛D5：s5D3：s7
D1O：∞DO：S10 D7：岛D5：S5D3：s7
DlO：∞DO：S1O D7：∞ D5：S5D3：s7
Proportiontreatments
Fig．2Biomassofcellcoloniesof及salina(D)and只
subcordiformis(S)underdifferentcellinoculation
proportionsandLN(a)，MN(”andHN(c)conditions
chlorophylla(24．4mg‘L-1)andprotein(339．6ug’L-1)
perkedatHNtreatmentincontrasttoMNandLN，and
thesedifferencewereverysignificantp<0．01)，
demonstratingthatchlorophyllaandproteinco tents
werelargelyinfluencedbythenitrogenco centration
forallthetreatmentexamined(Fig．3andFig．41．
MonoculturedD．salinaatHNconminedsignificantly
morechlorophyllaandproteinco tentsi comparison
toMNandLN．只subcordiform8varieda lotin
chlorophyllacontentandproteinco tentsamong
differentNlevelsp<0．01)．Theseresultswereingood
agreementwiththecelldensitiesandbiomass
investigatedabov ．
Thecontentsofchlorophyllawererelativelyhi曲
forthetreatmentofD7：S3，D5：S5，andD3：S7．In
thesethreecases，themaximumchlorophyllacontentin
thetreatmentofcellinoculationproportionofD7：S3
wasincreasedpproximately37％and89．8％p<0．01)
inMN，1．17and7．77pcomparisonwithmonoculture，andchlorophylla
contenti D7：S3treatmentWas12．58％and23．4％
higherthanD5：S5andD3：S7p<0．05)atMN，and
55．4％，68．7％higherthanD5：S5andD3：S7atHN(P
nitrogen，contentsofchlorophyllawereconsiderably
lowerinthemonoculturedPs bcordiform&．
Manifestvariationinproteinco tentWasalsofound
in mixed-cultureunderdifferentcellinoculation
proportionsandnitrogenconcentrations．Therewereno
strikingdifferencesi mixed-cultureandmonoculture
intheaccumulationofchlorophyllaandprotein．All
theseresultssuggestedhathighernitrogen
concentrationandifferentc llinoculationproportion
mightaveapotentialin ncreasingonthebiosynthesis
andmetabolismofproteinandchlorophylla．
4 Discussions
Previousstudieshaveindicatedthatnutrientssuch
asnitrogenorphosphorusaressentialforthegrowthof
phytoplankton⋯1．Caifoundthath ghernitrogen
concentrationtreatmentcouldsignificantlyenhancethe
algalgrowth[12J．Wangfoundthat尸donghaienseW s
moretolerantto utrientdeficiencya dsustainedth
populationgr wth，whilecellmultiplicationof
Skeletonemacost tumrequiredamuchhighernitrogen
concentrationt¨J．ItWasalsowelldocumentedthatwhen
hegrowthofalgaewashighlyr．cs倒ctedbynitrogen．
somephysiobiologicalresponsesuchasenergy
l
9
8
7
6
5
4
3
2
l
O
0
0
O
O
O
O
0
O
O
万方数据
!丝 生查型堂量1212星!￡苎!墼堕翌!! 兰竺鲞
transformationandcarbonfixationc uldbeseriously
influenced[14J．Inthisresearch，eelld nsity，biomass，
proteinandchlorophyllacontentswereextremely
influencedbylOWnitrogen．D．salinaisaun cellular
ChlorophytaalgaofChlorophyceaeclassand
Volvocalesorder．Themainmorphological
characteristicthatdistinguishesthisalgafromtheother
1．2
l
O．8
0．6
0．4
O．2
O
3．5
3
2．5
2
1．5
l
O．5
O
DlO：∞DO：S10D7：岛D5：KSD3：s7
DlO：S0D0：S10D7：岛D5：s5D3：s7
D10：S0D0：S10D7：S3D5：KS D3：S7
Proportiontreatments
Fig．3ChlorophyllncontentofcellcoloniesofAsa／／na㈣
and卫subcordiformls(S)underdifferenteeHinoculation
proportionsandLN(_)，MNCo)andFIN(c)conditions
Volvocalesistheabsenceofapolysaccharidecellwall．
Forthisreason，D．salinacouldbceasilydigestedby
humansandanimals．Besides，itisal oamicroalgae
withighefficiencyofconversionoflight，energy，
nutrientsi obiomass．SOwhenD．salinaSunder
160
140
120
100
80
60
40
20
0
400
350
300
250
200
150
t00
50
O
D10：SOD0：S10D7：S3 D5：S5 D3：s7
D10：SOD0：S10D7：s3 D5：s5 D3：s7
DlO：8D DO：SlOD7：整D5：S5D3：b7
Proportiontreatments
Fig．4Proteinco tentofcellcoloniesof及salina(D)and卫
subcordiformis(s) underdifferentcellinoculation
proportionsandLN(-)，MN(b)andHN(c)conmUons
O
O
O
O
0
O
O
如
”
加
”
m
．n‘1．1．M；暑口3口ou眉SoJld
{_1．一．M—口旨口3口ou再凶=h声△口．Io一8
如
笱
加
：2
m
，
O
万方数据
4期 黄伟伟，等：杜氏盐藻和亚心形扁藻在不同接种密度和氮浓度下的细胞群体生长 323
appropriatecul ivationconditions，itCallaccumulate
morethan10％ofitsdryweightaswellasc盯oteneand
chlorophylla．Besides，whenthisalgawasgrownin
highernitrogen，presenceofhighNimposesastresson
theanapleroticmetabolismIl5】．Inthepresentstudy,
colonialce lgrowthofnlarinemicroalgaein
mixed-cultureunderdifferentcellinoculation
proportionswashighlighted．Theresultsindicatedthat
mixed—cultureofRsubcordiformisandD．salinaHN
andMNcouldsignificantlyi f uencegrowth
performanceofcolonialcel s，thusresultedinincreased
celldensity,cellbiomass，contentsfchlorophyllaand
protein．Ourres ltswereinconsistencewiththose
obtainedbyCai[16 ．However,inthecaseoflow
nitrogen，contentsofchlorophyllawereconsiderably
lowerinthemonoculturedPsubcordiformis．Marine
microalgaecouldcontinuouslyreleasemetabolitesto
thenvironments．andtheseproductionscouldrestrict
orpromoteth growthofotheralgaefl71．Dueto
nutritioncompetition，differentalgaemayformcertain
predominancewhen也eyaremixedtogether,butthere
arealsotherfactorsinfluencingthisformation，so
mixed—culturingof microalgaewithdifferent
inoculationdensitymayprobablyshowcertainpotential
inaccumulatingbioactivesubstancesll81．
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万方数据
杜氏盐藻和亚心形扁藻在不同接种密度和氮浓度下的细胞群
体生长
作者： 黄伟伟， 蔡卓平， 肖群， 段舜山， HUANG Wei-wei， CAI Zhuo-ping， XIAO Qun，
DUAN Shun-shan
作者单位： 黄伟伟,HUANG Wei-wei(暨南大学水生生物研究所,广州,510632;同济大学环境科学与工程学
院,上海,000000)， 蔡卓平,CAI Zhuo-ping(暨南大学水生生物研究所,广州,510632;中国科
学院南海海洋研究所,广州,510301)， 肖群,段舜山,XIAO Qun,DUAN Shun-shan(暨南大学水
生生物研究所,广州,510632)
刊名： 生态科学
英文刊名： ECOLOGICAL SCIENCE
年，卷(期)： 2010,29(4)

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