Professional Documents
Culture Documents
Agren CNPinPlants EcLet04
Agren CNPinPlants EcLet04
REPORT
The C : N : P stoichiometry of autotrophs theory
and observations
Abstract
Goran I. Agren Evolution has set biochemical constraints on the chemical composition of living
Department of Ecology and organisms. These constraints seem to lead to increases in N : C and P : C ratios with
Environmental Research, increasing relative growth rate for all types of organisms. The N : P ratio also seems to
Swedish University of decrease with relative growth rate for heterotrophs whereas autotrophs may show a
Agricultural Sciences, PO Box more complex behaviour. Here I will show that, from biochemical considerations, N : C
7072, SE-750 07 Uppsala,
should increase linearly and P : C quadratically with relative growth rate in autotrophs
Sweden
with the consequence that N : P increases at low relative growth rates, passes a
E-mail: goran.agren@eom.slu.se
maximum and then decreases at high relative growth rates. These predictions are verified
against observations for a freshwater alga (Selenastrum minutum) and a tree seedling (Betula
pendula). Changes in temperature, light or other factors that affect the growth rate of
autotrophs interact with nutrient supply in such a way that there are no simple rules for
as to how N : P will change.
Keywords
Betula pendula, nitrogen, phosphorus, relative growth rate, RNA, Selenastrum minutum,
specific growth rate, stoichiometry.
Depending on the value lmax NP relative to the maximum alga (Selenastrum minutum (Naeg.) Collins, Elrifi & Turpin
relative growth rate, lmax, three possibilities exist: 1985) and one for a tree seedling (Betula pendula Roth.,
Ericsson & Ingestad 1988; Ingestad et al. 1994). Multiple
(1) lmax NP > lmax. In this case, increasing growth rates
regressions of N and P concentrations against l and l2 were
will always lead to increasing N : P. This is contrary to
calculated (cn a + bl + cl2). For both species, the
the observations for heterotrophs.
quadratic term was not significant (P > 0.79) in the
(2) lmax NP lmax. In this case, N : P will decrease with
regression with N concentration whereas the linear term
increasing l for all l of practical interest and the
was not significant (P > 0.37) for P concentration. All other
autotrophs behave as heterotrophs with respect to
terms (except the constant term in the regression for N
C : N : P stoichiometry.
concentration for B. pendula) were highly significant
(3) lmax NP < lmax. In this case, N : P increases for small
(P < 0.001) and r2 values exceed 0.88.
values of l, passes a maximum and then decreases as l
The regressions and the observed data are shown in
increases. Depending on the curvature of the rNP(l)-
Fig. 1 and the resulting parameter estimates are given in
function, the N : P ratio can be more or less sensitive
Table 1. The observations conform well to the theoretically
to changes in l.
predicted linear relationship between N concentration and
relative growth rate and the quadratic relationship (without
COMPARISON WITH OBSERVATIONS linear term) between P concentration and relative growth
rate, respectively. The observed N : C and P : C ratios for
I will now compare the predictions from the theory section
the non-limiting elements are also included in Fig. 1. It is
with observations. As will become clear below, such a
clear that there exists a considerable capacity for excess
comparison requires data from experiments where the
uptake of the non-limiting element, a capacity that is much
organism has been subjected to N and P limitation under
larger for S. minutum than B. pendula. Another way of looking
otherwise similar conditions. It is not difficult to find
at the excess uptake is by comparing the critical N : P ratio
observations of either N or P limitation but studies where
with those observed. As can be seen in Fig. 1, observed
both elements have been made limiting are rare. I have used
N : P ratios all fall well above the critical line under P
two such studies for a detailed analysis: one for a freshwater
N : C (mg g1)
N : C (mg g1)
200 50
100 25
0 0
100
P : C (mg g1)
P : C (mg g1)
75 10
50
5
Redfield ratio
25
0 0
40
15
Figure 1 N : C, P : C, and N : P ratios as
N : P (g g1)
30
N : P (g g1)
/CN (g mg)1 day)1) /NP (mg mg)1 day)1) bN (mg g)1) bP (mg g)1) lmax NP (day)1) rNP(lmax NP) (mg mg)1)
Table 2 Comparison of adjusted r2 for linear and quadratic regressions of N : C and P : C on relative growth rate. The number of
observations is given after the r2-value. The largest r2-values for N and P limited growth, respectively, are marked in bold
N limited P limited
Selenastrum minutum 0.939/18 0.897/18 0.826/21 0.898/21 Elrifi & Turpin 1985
Betula pendula 0.931/10 0.896/10 0.831/16 0.881/16 Ericsson & Ingestad 1988;
Ingestad et al. 1994
Eucalyptus globulus 0.981/9 0.958/9 0.830/8 0.890/8 Ericsson 1994
Scenedesmus acutus 0.955/6 0.888/6 0.831/6 0.704/6 Sterner 1995
Cyclotella meneghiana 0.924/6 0.974/6 0.826/6 0.938/6 Sterner 1995
Cyclotella meneghiana* 0.853/6 0.779/6 0.897/4 0.943/4 Shafik et al. 1997
Aureoumbra lagunensis 0.902/5 0.983/5 0.957/4 0.900/4 Liu et al. 2001
*The values from the three lowest relative growth rates with P limitation were excluded from the analysis because of decreasing P : C with
increasing relative growth rate.
limitation and well below under N-limitation, with the but they combine multiplicatively leading to the quadratic
exception of some P-limited points for B. pendula, but the relation between P and relative growth rate. The observed
classification of these points as N-limited is questionable. linear relationship between N concentration and relative
The excess uptake decreases with increasing relative growth growth is the N productivity (Agren 1985a) but its extension
rate, which is most clearly seen in the decreased deviation to other elements is, on the basis of this analysis, less
between actual N : P and the critical N : P at high values of l. obvious. I have previously suggested that a linear rela-
In addition to the two studies discussed above, I have tionship between autotroph nutrient concentration and
tested regressions of the form cn a + bl and relative growth rate also should apply to elements other than
cn a + bl2 (n N or P) on five other experiments N (Agren 1988). This might still be a relevant approximation
(Table 2). In all cases, both the linear and the quadratic because the curvature in the P concentrationrelative
regression fit the data well but a linear regression model fits growth rate curve is small and it will require higher quality
the data best in five of the seven cases under N limitation data to observe it than is available in many cases. Although
and the quadratic regression model fits the data best in five the experiments included in Table 2 indicate a quadratic
of the seven cases under P limitation. The five additional rather than a linear relationship for P, the linear regression is
experiments are only represented by a few data points and also a good description. Other experiments (eucalyptus,
whether the linear or the quadratic models will give the best Kirschbaum et al. 1992; three grass species, Ryser et al. 1997)
fit can in several cases depend on a single data point. reveal no curvature or have a clear curvature (Pavlova lutheri,
Terry 1980) in the relationship between relative growth rate
and P concentration (data not shown).
DISCUSSION
However, the choice between functions to describe data
The first implication of these analyses is that the relationship should not be made only on the basis of goodness-of-fit to
between organism N and relative growth rate are different data but also with regard to underlying principles and
than those between organism P concentration and relative explanatory power. In that respect, because the parameters
growth rate. The relation for N is linear whereas the relation in eqns 14 can be related to underlying biological
for P is quadratic. This agrees with the theoretical properties, the equations suggested in this paper offer more
predictions based on the assumption that autotroph growth prospects than a phenomenological function like the Droop
requires protein (N based) and proteins presuppose rRNA equation. Moreover, it is possible to extend the present
(P based). Both of these growth-related processes are linear analysis in logical ways. It is, for example, possible to
explore the consequences of partitioning the N and P pools A second observation relates to the problems of
in another way. As ribosomes are also rich in N (N : P 3 identifying the requirements of different elements for
on a mass basis, Sterner & Elser 2002) increasing allocation growth. As can be seen from the sometimes large distances
to ribosomes in the organism results also in increasing between the critical N : P ratio and the observed values for
allocations of protein N. Let N : P in ribosomes be bNri. the non-limiting element in Fig. 1, autotrophs have large
Equation 3 is then replaced by capacities for excess uptake, a capacity that varies both
between species and between elements. Element ratios
N Np bNri Pri bN C 9 obtained under the limitation of one of the elements are
therefore likely to lead to exaggerated expected require-
and a derivation along the lines in the Appendix shows that
ments of the other element. It is only when using results
eqn 5 should be replaced by
from conditions where both elements in the ratio are from
bNri 1 limiting conditions that proper limiting ratios can be
rNC l l2 l bN 10 obtained. In the case of N and P, the N : P ratios derived
/CN /NP /CN
from P-limited conditions with an excess of N are closer to
The equation for rPC remains unchanged. The conse- the critical N : P ratio than N : P ratios obtained under N
quences of introducing this additional N pool is thus to also limitation. This makes sense because the excess N that
make the rNC function quadratic in l. However, the term S. minutum takes up under P limitation and low growth rates
bNril//NP is at most c. 0.2 and the quadratic term will corresponds to an uptake of c. 50 mg g)1 of N and this
probably not be distinguishable from the linear term with approximately doubles the N : P ratio relative to what is
current quality of data. required at the critical level. On the contrary, the decrease in
The numerical values for the parameters found with data N : P by more than an order of magnitude under N
analysed in this paper all point to case 3, i.e. there should be limitation corresponds to an excess uptake of c. 100 mg g)1
a clear maximum in the relationship between N : P and of P. Expressed on a molar basis, the excess uptakes are
relative growth rate. This is in contrast to what seems to be approximately the same, which indicates that the limitation
the case for heterotrophs, where N : P decreases mono- on excess uptake might be a question of handling ion
tonically with relative growth rate. The parameters descri- balances.
bing the interactions between the elements (/CN and /NP) A further complication in understanding the critical N : P
can also be compared with what can be predicted from in autotrophs is variability between tissues. For example, a
biochemical properties. Agren (1985b) estimated from more detailed analysis of the data for B. pendula shows that
observed rates of the catalysing capacity of Rubisco that the critical N : P is considerably higher in leaves than in
/CN should have a value of 0.02 gC (mgN))1 day)1, which stems and roots (data not shown). The relative contribution
is just a factor of 2 higher than observed for S. minutum. of different tissues will change with the size of the plant and
Sterner & Elser (2002) give values for the rate of protein the whole-plant critical N : P will then be changing not only
synthesis by ribosomes. From their values we get /NP of with growth rate but also with plant size. The possibility of
1.1 mgN (mgP))1 day)1. This is a factor of two lower than truly identifying the critical N : P in any given situation may
the value given in Table 1 for B. pendula and a factor of therefore be limited and a single critical N : P might
seven lower for S. minutum. Given the crude approximations therefore serve as an approximate substitute.
used to arrive at these estimates, the agreement between the The variations in stoichiometry shown in Fig. 1 are all
values obtained from biochemical considerations and whole accomplished by manipulating the rate of nutrient supply
organism observations is not unreasonable, although this and thereby the relative growth rate at otherwise constant
point deserves further evaluation. environmental conditions. If the other environmental
20 20
Selenastrum minutum Betula pendula
CN /2
N : P (g g 1)
N : P (g g 1)
15 15
10 10
Figure 2 Critical N : P ratios calculated for
S. minutum and B. pendula for conditions 5 CN /2 & NP /2 5
corresponding to estimated parameter values
(solid lines), when /CN has been halved 0 0
0.0 0.5 1.0 1.5 0.0 0.1 0.2 0.3
(broken lines), and when both /CN and /NP
have been halved (dotted lines). Relative growth (day1)
conditions change, so will all the stoichiometric relations. Agren, G.I. (1988). The ideal nutrient productivities and nutrient
Factors, other than supply rates of nutrients, that decrease proportions. Plant, Cell Environ., 11, 613620.
growth rates tend to increase N : C and P : C (Sterner & Agren, G.I. & Bosatta, E. (1998). Theoretical Ecosystem Ecology
Understanding Element Cycles. Cambridge University Press, Cam-
Elser 2002), but effects on N : P are not well studied.
bridge.
Figure 2 shows variations in N : P when environmental Caperon, J. (1967). Population growth in micro-organisms limited
factors change. If light intensity changes, this should only by food supply. Ecology, 48, 715722.
affect the rate of carbon fixation per N, i.e. /CN. However, Droop, M.R. (1974). The nutrient status of algal cells in continuous
if temperature changes, all rate processes should change, i.e. culture. J. Marine Biol. Ass. Unit. Kingdom, 54, 825855.
/CN and /NP. To illustrate this, I have halved /CN alone as Elrifi, I.R. & Turpin, D.H. (1985). Steady-state luxury consumption
well as /CN and /NP together. Both changes result in and the concept of optimum nutrient ratios: A study with
phosphate and nitrate limited Selenastrum minutum (Chlorophyta).
increased critical N : P at low relative growth rates and
J. Phycol., 21, 592602.
decreased critical N : P at high relative growth rates with the Elser, J.J., Fagan, W.F., Denno, R.F., Dobberfuhl, D.R., Folarin,
effects of /CN most pronounced at low relative growth rates A., Huberty, A. et al. (2000). Nutritional constraints in terrestrial
and those of /NP at high relative growth rates. What is less and freshwater food webs. Nature, 408, 578580.
trivial is that changing only the interaction between C and N Elser, J.J., Acharya, K., Kyle, M., Cotner, J., Makino, W., Markow,
(/CN) affects N : P at low relative growth rates, whereas T. et al. (2003). Growth rate-stoichiometry couplings in diverse
changing equally the interaction between C and N and N biota. Ecol. Lett., 10, 936943.
Ericsson, T. (1994). Nutrient requirement of Eucalyptus globulus
and P has an effect on N : P over the entire span of relative
seedlings. In: Eucalyptus for Biomass Production (eds Pereira, J.S. &
growth rates. There are, therefore, no simple relationships Pereira, H.). Commission of the European Communities,
between N : P and growth rates when nutrient supply rates Brussels, pp. 224234.
interact with other rate determining factors. The changes in Ericsson, T. & Ingestad, T. (1988). Nutrition and growth of birch
/CN and /NP used here are fairly large in order to clearly seedlings at varied relative phosphorus addition rates. Physiol.
illustrate qualitative effects. What is further required is an Plant., 72, 227235.
incorporation of different feedback mechanisms that might Geider, R.J. & La Roche, J. (2002). Redfield revisited: variability of
C:N:P in marine microalgae and its biochemical basis. Eur.
offset changes in one single process.
J. Phycol., 37, 117.
The results in Fig. 2 also emphasize the conclusion Hessen, D.O., Agren, G.I., Anderson, T.R., Elser, J.J. & de Ruiter,
from Elser et al. (2000) that autotrophs from terrestrial P. (2004). Carbon sequestration in ecosystems: the role of
and aquatic environments differ with respect to nutri- stoichiometry. Ecology, 85, in press.
ent : C ratios but not with respect to N : P ratios. Ingestad, T. (1982). Relative addition rate and external concentra-
Eliminating the variability resulting from excess uptake tion; driving variables used in plant nutrition research. Plant, Cell
shows that the fundamental biochemistry of the two types Environ., 5, 443453.
Ingestad, T., Hellgren, O. & Lund Ingestad, A.B. (1994). Data base
of autotrophs should be quite similar. If the two taxa used
for birch plants at steady-state. Performance of birch plants
for Fig. 2 are representative for aquatic and terrestrial (Betula pendula Roth.) under non-limiting conditions and under
environments, and this seems reasonable, the growth limitation by N and light. Department of Ecology and Envi-
environment has at least two major effects that need to be ronmental Research, Swedish University of Agricultural
explained: the higher maximum relative growth rate and Sciences, Report 75. Uppsala.
the larger excess uptake in aquatic taxa than in terrestrial Kirschbaum, M.U.F., Bellingham, D.W. & Cromer, R.N. (1992).
taxa. A deeper understanding of these facts will probably Growth analysis of the effect of phosphorus nutrition on
seedlings of Eucalyptus grandis. Aust. J. Plant Physiol., 19, 5566.
require a detailed biochemical analysis along the lines of,
Knecht, M.F. & Goransson, A. (2004). Terrestrial plants require
for example, Sterner & Elser (2002) and Geider & La nutrients in similar proportions. Tree Physiol., 24, 447460.
Roche (2002). Liu, H., Laws, E.A., Villareal, T.A. & Buskey, E.J. (2001). Nutrient-
limited growth of Aureoumbra lagunensis (Pelagophyceae), with
implications for its capability to outgrow other phytoplankton
ACKNOWLEDGEMENT species in phosphate-limited environments. J. Phycol., 37, 500508.
I appreciate comments from Magnus Knecht on this Makino, W., Cotner, J.B., Sterner, R.W. & Elser, J.J. (2003). Are
bacteria more like plants or animals? Growth rate and resource
manuscript.
dependence of bacterial C:N:P stoichiometry. Func. Ecol., 17,
121130.
REFERENCES Rhee, G-Y. (1980). Continuous culture in phytoplankton ecology.
Adv. Aquat. Microbiol., 2, 151203.
Agren, G.I. (1985a). Theory for growth of plants derived from the Ryser, P., Verduyn, B. & Lambers, H. (1997). Phosphorus alloca-
nitrogen productivity concept. Physiol. Plant., 64, 1728. tion and utilization in three grass species with contrasting
Agren, G.I. (1985b). Limits to plant production. J. Theor. Biol., 113, response to N and P supply. New Phytol., 137, 293302.
8992.
Shafik, H.M., Herodek, S., Presing, M., Vorors, L. & Balogh, K.V. Terry, K.L., Laws, E.A. & Burns, D.J. (1985). Growth rate varia-
(1997). Growth of Cyclotella meneghiniana Kutz. II. Growth and tion in the N:P requirement ratio of phytoplankton. J. Phycol., 21,
cell composition under different growth rates with different 323329.
limiting nutrient. Ann. Limnol., 33, 223233.
Sterner, R.W. (1995). Elemental stoichiometry of species in eco- Editor, J. Elser
systems. In: Linking Species and Ecosystems (eds Jones, C.G. & Manuscript received 22 October 2003
Lawton, J.H.). Chapman & Hall, New York, pp. 240252. First decision made 2 December 2003
Sterner, R.W. & Elser, J.J. (2002). Ecological Stoichiometry The Biology
of Elements from Molecules to the Biosphere. Princeton University
Second decision made 17 December 2003
Press, Princeton. Third decision made 23 December 2003
Terry, K.L. (1980). Nitrogen and phosphorus requirements of Manuscript accepted 5 January 2004
Pavlova lutheri in continuous culture. Bot. Mar., 23, 757764.
APPENDIX. DERIVATION OF EQNS 5 AND 6 Divide through with C and use the l 1/C(dC/
dt) 1/N(dN/dt)
Substitute Np from eqn 3 into eqn 1 and divide through
with C 1 dN N 1 dN
rNC l /NP rPC bP bN l
C dt C N dt
1 dC N
l /CN /CN bN rNC /CN /CN bN A3
C dt C
A1 Solving eqn A3 for rPC and using eqn 5 to eliminate rNC
gives eqn 6.
and solve for rNC to obtain eqn (5).
Differentiate eqn 3 with respect to time and use eqn 2 to
eliminate dNp/dt and eqn 4 to eliminate Pri
dN dNp dC dC
bN /NP Pri bN
dt dt dt dt A2
dC
/NP P bP C bN
dt