The Nutrient Stoichiometry of Benthic Microalgal Growth: Redfield Proportions are Optimal Author(s): Helmut Hillebrand and Ulrich

Sommer Reviewed work(s): Source: Limnology and Oceanography, Vol. 44, No. 2 (Mar., 1999), pp. 440-446 Published by: American Society of Limnology and Oceanography Stable URL: http://www.jstor.org/stable/2670615 . Accessed: 29/11/2012 08:44
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440
Limtntiol. Oceanogr.,44(2), 1999,440-446 ? 1999,by theAmerican Societyof Limnology and Oceanography, Inc.

Notes

The nutrient stoichiometry of benthic microalgal growth: Redfield are optimal proportions
Abstract-Cellular nutrient ratiosare often appliedas indicatorsof nutrient limitation in phytoplankton studies, especially the so-calledRedfield ratio.For periphyton, similardata are scarce. We investigated the changesin cellularC: N: P stoichiometry of benthic in response microalgae to different levels and typesof nutrient limitation and a variety of abioticconditionsin laboratory withnatural experiments inocula.C: N ratiosincreasedwithdecreasing growth rate,irrespective of the limiting nutrient. At the highestgrowth rates,the C: N ratiorangeduniformly around7.5. N: P ratios<13 indicated N limitation, while N P ratios> 22 indicated P limitation. UnderP limitation, the C P ratiosincreasedat low growth rateand variedaround130 at highest growth rates.For a mediumwithbalancedsupplyof N and P, an optimalstoichiometric ratioof C:N:P = 119:17:1 could be deduced for benthic whichis slightly thantheRedfield microalgae, higher ratio (106:16: 1) consideredtypicalfor optimally growing phytoplankton. The optimalratiowas stableagainstchanges in abioticconditions. In conclusion, cellular nutrient ratiosare proposedas an indicator in periphyton. fornutrient status

The chemicalcomposition of oceanic sestonis knownto be relatively constant at a C: N: P ratioof 106: 16: 1 (Redfield1958; cf. Copin-Montegut and Copin-Montegut 1983). This biogeochemical ratiobecame widelyknownas the "Redfieldratio" and was subsequently inphysiologically foraquatic organisms. terpreted Droop (1974, 1975) investhenutrient of phytoplankton tigated content within different limitation scenariosand developed the cell quota growth model.Internal nutrient ratiosareequivalent to carbon-based cell quotas. Nitrogenand phosphorussupply supporting maximum growth ratewas shownto lead to phytoplankton theRedfield ratio(Goldmanet al. stoichiometry resembling 1979; Elrifi andTurpin 1985), and theinternal nutrient ratios wereproposed as an indicator ofalgal nutrient status (Healey andHendzel 1980; Flynn1990). Despitesomecriticism (Rytherand Dunstan 1971; Tettet al. 1985), biomass stoichihas been widelyappliedto assess nutrient ometry supplyto in marine phytoplankton (Paasche and Erga 1988; Burkhardt and Riebesell 1997) and freshwater studies(Sommer1991a; Hecky et al. 1993). It shouldbe notedthatC: N: P ratios close to the Redfieldratio do not indicatethe absence of limitation that neilight (Tettet al. 1985). Theyjust indicate ther N norP are limiting factors of growth (Goldman1986). The use of nutrient ratiosas an indexof nitrogen or phoswas also recommended for benthicmiphoruslimitation croalgae(Borchardt 1996),because oftheclose phylogenetic and pelagic microalgae. betweenbenthic relationship Howit has seldombeen appliedto date ever,in benthicstudies, (Engle and Melack 1993; Rosemond1993; Rosemondet al. 1993; Hillebrandand Sommer 1997). The infrequent use betweencellular stoichimay be because the relationship and growth rateformarinemicrophytobenthos has ometry

not yetbeen testedexperimentally. Kahlert(1998) recently reviewed literature data fromfreshwater periphyton and foundthatC: N: P ratiosare a reliabletool forthe assessmentof the nutrient statusof benthicalgae, proposing an optimum ratioof 158:18: 1. In thepresent study, we wantedto answerthe following questions: a consistent between ben(1) Is there relationship thicmicroalgal ratesand cellularstoichiometry, growth and (2) Is thisrelation independent of abioticconditions? To investigate the responseof internal nutrient ratiosto changesin nutrient regimes, we used a semicontinuous dilution of culture mediacombined withsampling in intervals. We used natural inocula of algae to simplify to comparison natural assemblages. The algae fortheinoculawerescraped from an artificial substrate in theKiel Fjord,Western Baltic were started. Sea, 10 d beforethe experiments The algae werecultured in unenriched filtered seawater under thesame abioticconditions as theexperimental treatments (Table 1). At thebeginning of theexperiments, 1 ml of the inoculum was added to each treatment. The experiments were conductedin flat-bottom, culture flasks transparent, polystyrene with30 ml totalmediumcontent. The algae grewas a biofilmon the bottomof the flasks, whichwere shakenonce was a dense monolayer of cells lacking daily.This biofilm an overstory. It can therefore be assumedthat CO2 was available in excess. The mediaused in theexperiments of organismconsisted free-filtered seawater(0.2-Amcellulose-acetate from filters) the same location,enriched withnutrients and tracemetals. The balanced medium(designatedV) contained80 Amol liter-' N (as NaNO3), 80 Amol liter-' Si (as Na2O3Si X I P (as Na2HPO4X 2H20), resulting and 5 Amolliter5H20), in a mediumN: P ratio of 16. For the N-limited medium thenitrogen (designated concentration was reducedto Niim), 10 Amolliter-'.For the Si-limited media,Si was re(Si,im) duced to 10 Amol liter-'; forthe P-limited medium(Piim), no phosphate in 1.0 Amolliter-'P The was added,resulting were conductedin autumn1997 and spring experiments 1998. In the autumn fourdifferent experiment, media were of an alteration consisted of diapplied,and thetreatments lutionrate (Table 1). In spring1998, threedifferent media were used, and the temperature was altered(Table 1). The twoexperiments in thetaxonomic differed furthermore comwhich positionof the inoculumand in the lightintensity, was measured witha LiCor LI 189 (Table 1). Each treatment was conducted in triplicate, in 24 cultures in auresulting tumn(fourmedia X two dilution rates X threereplicates) and 27 cultures in spring media X three (three temperatures X three replicates). Thrice a week, the algae were countedalive employing an inverted microscope(Leitz DMIRB) at X630 magnification.Up to 400 cells werecounted persample.To compare

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Notes

441

thetypeof dilution algal inocula.The tableliststhecode of theexperiment, in limitation withnatural experiments Table 1. Treatments and themediaused. thetemperature, of theexperiment, thelightintensity, rate,theduration Dilution rate (d-) 0.5 0.07 0.5 0.5 0.5 Light (,umol 21 21 35, 8 34, 7 32, 2 Temp.
(OC)

Code H (highdilution) L (low dilution) C (cold temp.) M (medium temp.) W (warmtemp.)

Time 22 Sep-10 Nov 97 Ditto 19 Jan-9Mar 98 Ditto Ditto

m-2 s-')

Appliedmedia V, Njm, Plimg Si1,,n V, Njj, Pjj, Sii, V, NljM1PlIm V, N,in19Plim V, N,in19Plin,

14 14 5 14 19

the different species, which span several size classes, biovolumewas calculated by fitting appropriate geometric modIn addition, a week, els (Hillebrand et al. submitted). thrice a sample was takenwitha sterilepipette fromthe bottom of the flask.The samplevolumecorresponded to the daily dilution, i.e., 15 ml fortreatments C, M, W,and H (forcodes, see Table 1) and 5 ml fortreatment L. The sampleswere dividedand filtered on precombusted Whatman GF/Cfilters foranalysesofparticulate CN andP,respectively. Particulate phosphate was determined as orthophosphate aftera combineddigestion withheatand acid (Hillebrand and Sommer 1997). Since thisanalysisneeds a highamount of material, thethree of one treatment had to be pooled. Parreplicates ticulatecarbonand nitrogen were measuredwitha Fisons CN-Analyzer (NA 1500N). The experiments were dividedintotwo phases. After 28 dilution was stopped to enforce a strond, thedailymedium ger limitation by decreasingthe supplywith new media.

out once a week. After the Samplingwas carried sampling, totalvolume of the cultures was filledup to 30 ml again. The L treatment was stoppedat day 28, but the otherexperiments were conducted untilday 49. The daily growth ratesA were calculatedfromthe total biovolumeof each replicate according to Eq. 1: ln B2-ln B
t2 t,

(1)

including B = biovolume. t = time. werecalThe ratiosof C: N, N: P, and C: P, respectively, culatedon a molarbasis. Theywerecompared to thepositive ratesby a three-parameter dailygrowth exponential equation (Eq. 2). The use of ratiosin regression analysisis notwith-

function on ,u.The tableliststreatments and media,ratiofor Table 2. Fit of exponential (Eq. 2) to C: N, C: P, or N: P ratiodependent r2,and parameter estimates, as well as theF-ratio between whichtheregression was calculated, number of data,coefficient of determination level: * P < 0.05; ** P < 0.01; *** P regression explainedmean squaresand residualmean square(Sokal and Rohlf1995). Significance < 0.001; ns, not significant. no regression n.r., (estimation procedure did not converge).
Treatment Med Ratio n
r2

F(2;

3)

All All All All All C M W H L All All. All All All C M W H L All

All V Njim Slim All All All All All All V N,,m Sl,. All All All All All Plini
P,ini
Plim

C:N C:N C:N C: N C: N C: N C:N C:N C:N C:N C:P C:P C:P C: P C:P C:P C:P C:P C: P C:P N: P

322 102 98 96 26 77 63 63 89 30 122 39 37 36 10 23 20 21 26 18 37

0.1669 0.3692 0.362 0.1118 0.1952 0.3847 0.2253 0.2675 0.1796 n.r. 0.0655 0.2380 0.4306 n.r. 0.4067 0.2076 0.2618 0.1655 n.r. 0.1366 0.2668

7.5 7.2 5.9 7.5 8.5 4.4 6.8 7.6 8.4

+ + + + + + + + +

0.7 0.6 0.9 2.6 0.6 2.4 2.3 0.8 0.4

1.8 + 1.9 + 1.8 + 2.0 + 1.8 ? 2.2 + 2.3 + 2.6 + 1.3 +

0.1 0.1 0.2 0.2 0.6 0.2 0.3 0.3 0.2

-6.8 -9.9 -5.0 -3.1 -27.5 -3.3 -6.0 -26.2 -12.1

+ + + + + + + + +

2.6 4.2 2.3 2.5 18.3 1.9 4.3 11.2 7.3

32.04*** 28.97*** 26.96*** 5.78** 10.18** 23.14*** 7.99** 10.96** 9.42*** 4.17* 3.28* 12.85** 2.40 ns 3.54 ns 2.23 ns 1.79 ns 1.17 ns 6.50**

143.6 ? 48.5 119.0 + 50.7 139.2 + 47.8 51.2 + 80.3 + 73.3 + 116.7 + 53.8 61.9 120.4 27.4

5.1 + 0.3 5.1 + 0.4 6.1 + 0.2 4.9 5.0 5.7 4.6 + + + + 0.5 0.5 0.4 0.6

-8.2 ? 10.6 -5.1 + 4.2 -17.5 + 8.0 -8.6 -5.8 -5.4 -20.7 + + + + 10.4 6.9 6.0 22.0

216.1 + 105.0 20.9 + 2.6

5.8 + 0.7 3.1 + 0.3

-15.6 + 15.0 -24.7 + 14.6

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442
4e6 4e6

Notes
4e6

cv

~
5

~ ~

~
4 3e6

~
3e6

WV Wfl

3e

C P1

3e6 NEE

E~
E

EE E
E
2 e6

E E
1e6----

le6

10

20

30

40

50

10

20

30

40

50

E2

time[d]

2_~
4ee
-

time[d]

time[d]

4ee
HV H SI,11 H Pim

3e6

3e6

L N11m L SIj

E E E
o 32e6

32e6

E E

E
0
.

and spring. from autumn le

HNri

'

1e6

0
0 10 20 30 40 50

0~~~~
0 10 20 30 40 50

time [d]

time [d]

ofmeanoftotal biovolume ( Fig. 1. Development and spring. from autumn

ofbenthic overtimeforall treatments andmediaofexperiments tm/M/MM2) microalgae

out difficulties (Sokal and Rohlf 1995), but normaldistri(P > butionof the data of all ratiovariableswas affirmed 5.1). test,Statistica 0.05, Kolmogorov-Smirnov R= a + with R = molarratioof C: N, N: P, and C: P, respectively. rate. A = dailygrowth a, b, c = parameters. With a negativeestimatefor c, the resulting curve deline to a horizontal creases and approachesasymptotically withy = a. Therefore, parameter a can be takenas an estimateof the optimalratio.The validityof the regression model was estimated by an analysisof variancecomparing variance variance (Sothemodel-explained withtheresidual kal and Rohlf1995). The presenceof a global convergence in theregression was affirmed minimum by using procedure different software and different estimation procedures (Statistica5.1 and Statgraphics 6.1). followeda sigmoidal Total biovolumeof the treatments curvein all cases, explaining 87.03% ? 12.62 (mean ? SD) of the variance(Fig. 1). Final biovolumeswere generally at lowertemperature and lowerin theN,i,treatments. higher
e(b+c:1,f)

(2)

Moreover,the springexperiments resultedin higherfinal biovolumethantheautumn experiment; thelatter againwas dividedin lowerbiovolumein L treatments to H compared treatments (Fig. 1). Daily growth ratesincreasedin the beginning of theexperiment and decreasedafterwards, varying aroundzero after attainment of the carrying capacity(Fig. 2). In theautumn experiments, 30 specieswerepresent, representing the Bacillariophyceae, Chlorophyceae, and cyanobacteria. In spring, 30 species represented theBacillariophyceae, cyanobacteria, and Rhodophyceae. Almost all treatments were dominated chainby diatoms(unicellular, and tube-dwelling), forming, whereasnonheterocystous cyanobacteria became codominant at 19?C temperatures and highN content. C:N ratios increasedwith time (significant regression P < 0.05, exceptforLV, LN,i,,, slopes overtime, and HSI,i,,), up to 20 in P,in,, up to 23 in V, and up to 45 in N,in, cultures, respectively. This increasereflected the increasing strength of nutrient limitation. The ratioof C: N decreasedexponentiallywithincreasing rate(Fig. 3; Table 2), indepengrowth dentof treatment andlimiting The regression nutrient. model describedthe relationship betweenC: N ratiosand growth ratessignificantly (F-ratio, P < 0.01; Table 2) forall media and treatments, L. The optimal excepttreatment ratioat high growth rates,estimated by parameter a of theregression, is

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Notes
1,5 1,5 15

443

-v

M-WV
M Niim

CV...

..

WNi

1,0

1;0

ijm

M im

10WPim

0,

0,5

-0,

10

20 time[d]

30

40

50

05~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~0 20 10 0 50 40 20 30 10
o time[d] time[d] 1,5

30

40

50

1,5

-HV
10
- .H

H Niim 1,0

H Pli
Slim

L Niim L Pim~

LV

-o,s
70

-o,s'
5

0,

0,0

0,0

-0,5 0

10

20

30

40

50

-0,51 0

10

20

30

40

50

time [d]

tme[d]

to time. rate(Eq. 1) in relation Fig. 2. Mean of dailygrowth

and mediain Fig. 4, comtreatments plotted forthedifferent error(except forthe L treatment). pared to theirstandard The estimatesof the optimalC: N ratios showed widely beintervals (Fig. 4). A difference confidence overlapping treatments could tweenthe optimalC: N ratiosof different thus,C: N ratiosincreasedirretherefore not be sustained; nutrient. spectiveof thelimiting on C: P data regression exponential The three-parameter media,and at at P < 0.01, onlyfortheP,im was significant P < 0.05 also forV and thepooled data (Table 2). In an Sirelation betherewas no consistent situation, or N-limited rate.Onlyunder tweenC: P ratiosand growth P,im conditions with decreasing did the N: P ratio increase exponentially phase (1u rate(Table 2). N: P ratiosin thestationary growth < 0.1) weresignificantly different whencomparing different analysisof variance[ANmedia (P < 0.05; Kruskal-Wallis by Dunn's comparison OVA] on ranks;all pairwisemultiple abioticconmethod, Sigma Stat) but not betweendifferent ditions (Fig. 5). The meanof theN: P ratiowas significantly lower at N,i, (10.69) comparedto P,ir,(33.62), while the at balancedmedium N: P (for cellularN: P was intermediate
V 20.58, for Si,im 14.22).

these resultsshow thathigh C: N ratios Conclusively, in general,while C : P and were due to nutrient limitation N: P ratiosincreasedsignificantly onlywithP limitation. is an theview that biomassstoichiometry Our data support

as forphyforbenthic microalgae indicator of nutrient status and toplankton, forwhichit was developedexperimentally conceptually (Droop 1974, 1975; Healey 1978; Healey and theC: P andN: P ratios Hendzel1980). Similar to ourresults, increased withdecreasing rate growth of pelagic microalgae underP limitation, while the C: N ratioincreasedwithdeP and N limitation. the Moreover, rateunder creasing growth rateswas higherunderN increaseof C: N at low growth in our data and in phytolimitation thanunderP limitation plankton studies(Perry1976; Sakshaug and Holm-Hansen et al. 1979;HealeyandHendzel1980).Under 1977; Goldman theC: P and N: P ratios werefound to decrease N limitation, rates(Elrifi and Turpin1985). It can withdecreasing growth be concluded from thesedata thatC: N ratiosare applicable whileC: P andN: P ratios as general indicators of limitation, allow indication of P vs. N limitation. The criticism theindicator value of thenutriconcerning (Tettet entratiospointedmainlyat light-limited conditions al. 1985; Wynneand Rhee 1986), leadingto theconclusion thatlightlimitation was not reflected by biomass stoichiIn agreement withGoldman(1986), theoptimal raometry. our autumn withlowerlight experiment tios emerging from theratiosfrom thespring exfrom intensities did notdiffer Wynneand Rhee (1986), periment (Fig. 4). This contradicts who described N: P ratiosin planktonic changesin optimum and wavelength. algae caused by changesin lightintensity

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444
v 45 40 35 30

Notes
Plim 45 40 35 30 0 25

25 o 15

z 20
+0 10

z 20

+ + +

+ ++0

o 15 + +

t+

+ 1 0 0,0

5++

+ 0,6

++ + 0,8 1,0

++ 0,2 0,4

4+
0,6

0 0,0

0,2

0,4

1,2

1,4

1,6

1,8

2,0

2,2

0,8

1,0

1,2

1,4

1,6

p [d J]
Slum
45 40 35 30
.025
CU 0.

p [d-1]
Niim
45 40 35 30 25
+

z 20
++0++

z 20
-a +

~~~~~~~~~~~~~~~~~~++ +i ++
+ +
+ + ++ +

10
5 0 0,0

10

i.

+.t5+#++ 0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6 0 0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6

p [dJ

p[1

on growth nonlinear of Eq. 2. fordifferent media.Lines represent fits Fig. 3. C: N ratiosof benthic microalgae dependent rate,plotted are presented in Table 2. Parameter estimates

we approachour data in the same way. The minimalcell quotas (qo) of N and P can be calculatedfromEq. 2 by substituting the estimates of a and b 1u = 0 and inserting from Table 2. The mean ratioof qON:q0P was 18.56 forall treatments, whereasthe ratio forPlin, was 48.8. The latter ratioreflects thenutrient-limited situation and notan optimal ratio. Besides, these calculationsshow thatcell quotas of benthicmicroalgae(ranges of qo are 0.060-0.083 mol N are mol-'C and 0.002-0.008 mol P mol-'C, respectively) within the same orderof magnitude as thosereported from freshwater phytoplankton (Sommer1991a,b). Based on theworkof Healey and Hendzel (1980), Hecky et al. (1993) suggested limitsof indicating values forphywas indinutrient ratios.ModerateN limitation toplankton > 14.6, whereas catedby C: N > 8.3 and severelimitation P limitation moderate was indicated by C: P > 129 and severe limitation by C: P > 258 and N: P > 22. Fromourdata,an optimal ratioof fast-growing periphyton in analogyto the optimal can be determined phytoplankton ratios, based on the estimates of a (Eq. 2) forthebalanced in a C: N: P ratioof experiments (mediumV). This results

phase of cultures (1u = 0). This is a contradictionto the conceptof optimal ratios(1tt = Atmax), as becomesevident, if

However, theycalculatedtheoptimum minN: P ratiofrom imal cell quotas,whichrefer to the stationary (i.e., limited)

119: 17: 1. However, itseemsto be moreuseful to giveranges of ratiosin nonlimited These rangeswere calcuconditions.

it from theminimum. estimate and subtracting The optimum rangesare 5-10 forC:N and 90-185 forC:P. N:P ratios between13 and 22 indicatebalance betweennitrogen and These rangesallow the establishment phosphate. of C: N: P ratiosas indicators of nutrient limitation (Fig. 6). Withan N: P ratio< 13 and a C: N ratio> 10, theperiphyton can be assigned N limited. Withan N: P ratio> 22 and a C: P ratio > 180, themicrobenthic is P limited. assemblage These ratiosare moresimilar to theRedfield ratioand the limitsgivenby Hecky et al. (1993) forphytoplankton than to the optimalratioproposedforfreshwater periphyton by Kahlert(1998). Based on a literature she foundan survey, ratioforfreshwater than optimum periphyton slightly higher theRedfield ratio(C: N: P = 158: 18: 1) andproposed much higher indicators of limitation thanHecky et al. (1993): C: P > 369 and N: P > 32 forP limitation and C: N > 11 and N: P < 12 forN limitation. The reasonforthediscrepancy is probablybecause macrophytes (e.g., Cladophora) were includedin the data survey, which are knownto differ in their Atkinson C: N: P ratios. and Smith(1983) found a median of C: N: P of 550:30: 1 in a comparisonof marine

lated fromthe estimatesfor the different media (V, P,i,, N,i,,, and Si,im; Table 2), addingthestandard error to themaximum

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Notes
14 12 10

445

C:N ratio
50 40L_
lm Pj U

C:P ratio
- 500

8[I z
CL 0

estimate
lj

30
20

N-Iimi-4 tation

, P-limitation

- 400

- 3 00
200

treatment

treatments (M-L) and Fig. 4. OptimalC:N ratiosof different media (N,in,-Si,im). OptimalC: N ratiosare presented as estimate of parameter a (Eq. 2), its standard error, and confidence interval. For treatment L, no optimal ratiocould be fitted. "All" represents the optimal ratiofitted on all data.

10 0 no li 0 O 10

no or other

>'100 0

limitation 20 30 40 N:Pratio

whichare supposedto containmorestructural macroalgae, carbon.In theirdata, however, the Redfieldratioposed a lowerlimitof thefoundratios, whichmaybe reachedonly in verynutrient-rich localitieslike Antarctic waters(Weykam et al. 1996). C: N: P ratioswerereported from studies dealing withmicrophytobenthos before,but theywere rewithout them to growth were ported relating rates.The ratios in the rangereported here (Reuteret al. 1986; Engle and Melack 1993; Rosemond1993; Rosemondet al. 1993; Turner et al. 1994; Vymazal and Richardson1995; Hillebrand and Sommer1997). Therefore, the limitsfornutrient ratios proposedby us can be appliedforperiphyton dominated by diatomsand cyanobacteria. A notehas to be addedin proof: Biomassstoichiometry of natural benthic communities maybe influenced by a highproorbycarbon of detritus bothofwhich were portion limitation, of detritus excludedin our experiments. The influence was to be less thanexpected forphytoplankton found (Healeyand Hendzel 1980), butit has to be evaluated forperiphyton (cf.
50

Fig. 6. Schematicdiagramon the use of nutrient ratiosas inor phosphorus dicator of nitrogen limitation. Shaded barsrepresent therangeof optimal is givenin thetext. ratios. Further explanation

Kahlert was shownto changeC: N: 1998). Carbonlimitation P ratiosin a marine pelagic diatom, by decreasing C: P and N: P ratiosand increasing and RieC: N ratios(Burkhardt it shouldbe notedthatin dense besell 1997). Furthermore, the algae could have different access to nutrients. biofilms, This dependson whether thesestemfrom thewatercolumn or from so thatemerging stoisediment nutrient pore water, chiometry maybe themeanof a gradient. In conclusion, cellularnutrient ratiosarea useful approach forthedetection of nitrogen orphosphorus limitation in benthicmicroalgaeas well as in phytoplankton. The following constraints shouldbe observed:The cellularC: P ratiois an indexof P limitation, thecellularC: N ratioindicates limitationin general,and the N: P ratiodistinguishes between N or P limitation.
Helmut Hillebrand Ulrich Sommer

40

0 cu

301
Z20

0-

Institut Meereskunde fuir Meeresbotanik Abteilung Dustembrooker Weg 20 D-24105 Kiel Germany

Acknowledgments withMaria Kahlert, This study was initiated Uniby discussions of Uppsala. Financialsupport was granted from theInstitut versity H L C M W V NLIM PLIM Slum furMeereskunde and theDeutscheForschungsgemeinschaft (grant treatment theCN analysis S0145/15-1).The help of ThomasHansen during of different treatments is gratefully The manuscript cultures fromcomFig. 5. N: P ratiosof stagnant acknowledged. profited and nutrient ments Rolf Karez,Carlos by MonikaFeiling,UlrikeG. Beminger, content, depictedas mean ? SE of all sample dates M. Duarte,Stephen V. Smith, and one anonymous reviewer. with ,u< 0.1.

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446 References

Notes
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