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Richard W. Eppley, Jane N. Rogers, and James J. Mccarthy
Richard W. Eppley, Jane N. Rogers, and James J. Mccarthy
ABSTRACT
Uptake rate of nitrate and ammonium was studied as a function of nitrate or ammonium
concentration with cultures of 16 species of marine phytoplankton. Half-saturation con-
stants (the concentration supporting an uptake rate one-half the maximum rate) were
computed as a measure of the ability of a species to use low levels of nitrate and ammonium.
The half-saturation constant (K,) varied approximately in proportion to cell size and
inversely with specific growth rate. Small-celled oceanic species, such as Coccolithus
huxleyi, showed the lowest K, values (GO.5 PM) and Gonyaulux polyeclra the highest
(K, > 5 PM). The K, values are considered to be important characteristics of organisms
living in nitrogen-limited environments.
Some apparent success was achieved in predicting the competitive advantage of one
species over another by calculating specific growth rates as functions of nitrate and ammo-
nium concentration for species with known growth response to irradiance, temperature,
and daylength. For example, at about 20C and 16 hr daylength C. huxleyi should be able
to compete successfully with Skeletonema costatum when nitrate, ammonium, and irradi-
ance are low, Skeletonema costatum should grow faster than C. huxleyi only when irra-
diance and nitrate or ammonium are fairly high. Rock pool fonns appear to be relatively
unsuccessful in the ocean because of a sluggish response to irradiance and not because of
an inability to use low levels of nitrate and ammonium. The success of G. polyedru, often
the dominant dinoflagellate in local red water blooms, appears to be due to its vertical
migration from the sea surface in daylight to nitrate-rich water at 10-15-m depth at night,
rather than to any special ability to grow rapidly or use low levels of nitrate and ammonium.
TABLE 2. Half-saturation constants for uptake (K, ) of nitrate and ammonium by cultured marine
phytophznkton at 18C. K, units in ,umotes,‘liter
Nitrate Ammonium
- - Cell
-c95% k95% diam*
Organism l-c.9 conf. limit Ku conf. limit (PI
Oceanic species
Coccolithus huxleyi BT-6 0.1 0.3.1 0.1 0.7 5
C. huxleyi F-5 0.1 1.6 0.2 0.9 5
Chaetoceros gracilis 0.3,O.l 0.5,0.2 0.5,0.3 0.5,0.3 5
Cyclotella nanu 13-1 0.3,0.7 0.4,o.s 0.4 0.3 5
Neritic diatoms
Skeletonema costatumz 0.5,0.4 0.4, 0.1 3.6,0.8,0.8 0.8,0.7,0.5 8
Leptoc ylindrus danicus 1.3, 1.2 0.5,O.l 3.4,0.9,0.5 1.4,0.2,0.4 21
Rhixosolenia stolterfothii 1.7 0.4 0.5,0.5 0.9,0.4 20
R. robustaf 3.5, 2..5 1.0, 1.0 5.6,9.3 2.0, 1.5 85
Ditylum brightwellii 0.6 1.7 1.1 0.6 30
Coscinodiscus lineatus 2.4,2.8 OS, 0.6 2.8,1.2 2.6, 1.0 50
C . wailesii 2.1,5.1 0.3, 1.8 4.,3,5.5 5.4,2.0 210
Asterionella japonica 0.7,1.3 0.3,0.5 1,5,0.6 1.2,O.B 10
Neritic or littoral flagellates
Gonyaulux polyedra 8.6, 10.3 -, 2.4 5.7, 5.3 0.6, 1.1 45
Gymnodinium splendens 0.9 1.1 1.0 47
Monochrysis lutheri E 0.3 0.5 0.4 5
Isochrysis galbana 0:1,0.1 0.2,0.2 -- - 5
Dunaliella tertiolecta 1.4 1.1 0.1 0.6 8
Natural marine communities (from MacIsaac and Dugdale 1969)
Oligotrophic GO.2 ( 6 expts ) Ol.l-0.6 ( 3 expts )
Eutrophic 21.0 ( 3 expts) I..3 ( 1 expt)
* Geometric mean diameter rounded off to the nearest micron.
t This notation means that -0.2 < KS <0.4. Negative KS values have no physical interpretations.
$ At 28C, KS for nitrate uptake was 1.0 + 0.5; at SC, it was 0.0 -C 0.5.
fi An oceanic species according to Cupp ( 1943).
NITRATE AND AMMONIUM UPTAKE 915
1 I , I I I I I 4
5 IO0
costatum
L! briglrlwellii
C. huxleyi
0. tertiolecta
2 IO -2
3 s
-!
% / x
K
'r-3 /
22 -I
Skeletonema costatum
/
8 OC -
i -II III1 11 0.15
0
0 5 IO IRRADIANCE (lylminl
general, phytoplankton growth rates de- earlier observations that N-deficient cells
crease with increasing cell size (Williams have high rates of nitrogen uptake (Fitz-
1965; Eppley and Sloan 1966). The rock gerald 1968; Syrett 1962).
pool and estuarine species, ‘Monochrysis
lutheri, Isochrysis galbanu, and Dunaliella DISCUSSION
tertiolectu, show low K, values in spite of It is difficult to be certain that the K,
their abundance in rock pools which one for growth is accurately measured by such
would expect to be rich in nitrogenous short-term uptake experiments. For ex-
nutrients. Other characteristics possibly ample, Droop (1968) calculated a 20-fold
limiting their distribution are discussed greater K, for short-term uptake of vitamin
later. Br2 by M. Zutheri than for growth in a
Uptake rates of cultures grown on nitrite chemostat culture; variation of growth rate
to N-depletion were high compared with with the nutrient concentration in the cell,
growth rates, both for nitrate and ammo- independent of that in the medium, was
nium. For example, the specific growth rate noted in Droop’s vitamin study, in phos-
during culturing was 0.7-0.9 doublings/ day phate-limited cultures of Phaeodactylum
for R. robustu, but the corresponding rates tricornutum ( Kuenzler and Ketchum 1962))
calculated from rates of short-term uptake and in nitrate-limited chemostat cultures of
and the nitrogen content of the cells would I. galbana (Caperon 1968). However, K,
have been 8 and 13 doublings/dav for for uptake in our experiments agreed well
nitrate and ammonium uptake, respectively. with independent measurements of K, cal-
Similarly, the observed specific growth rate culated from exponential growth rates of
of Gonyaulux polyedra was 0.4-0.5 dou- batch cultures limited by nitrate (Eppley
blings/day, although 1.5-4 doublings/day and Thomas 1969) and are consistent with
were caldulated from uptake rates.- Thus the values observed with 24-hr incubations
these experiments to determine K, give no of samples of natural communities of oce-
useful information on exponential -growth anic and neritic phytoplankton based on
rates of N-sufficient cells but only confirm uptake of l”lV-labeled nitrate and ammo-
NITRATE AND AMMONIUM UPTAKE 917
nium ( MacIsaac and Dugdale 1969). A according to these calculations, both neritic
more detailed discussion is given elsewhere diatoms are favored by high irradiance and
( Eppley and Thomas 1969). nitrate. DunaZieZZa tertiolecta generally
If these measurements are to be of any fares poorly because of its relatively slug-
ecological value, other than emphasizing gish response to irradiance. Apparently this
the obvious fact that oceanic phytoplankton is not an artifact of poor measurements since
possess mechanisms for using very low Jitts ct al. ( 1964) found a light response
levels of nitrogenous nutrients, they must curve for D. tertiolecta similar to ours.
help explain observed phytoplankton species Another rock pool flagellate, M. Zutheri,
distribution, for the uptake K, values have with a maximum specific growth rate of 1.2
little meaning for ecology in themselves. doublings/day at 2OC, has a growth rate vs.
Hence we have used the values to calculate irradiance response curve like that for D.
specific growth rates of different species in tertiozecta (Jitts et al. 1964). Thus our
idealized conditions that we think manifest comments for D. tertiolecta may apply also
the essential features of natural waters. For to M. Zutheri.
this we needed not only the K, for nitrogen The calculations suggest further that C.
uptake but measurements of specific growth huxleyi is the best competitor with any
rate, with adequate nutrients, as influenced ammonium level and low irradiance (Fig.
by irradiance, daylength, and temperature. SC). Because of its low K, for ammonium,
Few studies have been extcnsivc enough D. tertiolecta would grow faster than the
for such purposes, and none are above two diatom species at low ammonium con-
criticism. In what follows we have used centration (<0.5 PM) and low irradiance.
data for an oceanic (ubiquitous) species, At higher irradiance ( Fig. SD), the diatoms
CoccoZit7zus huxleyi clone F (Paasche 1967), would grow faster than C. huxleyi only if
the coastal diatoms D. brightwellii (Paasche ammonium is >1.5 PM for S. costatum or
1968) and S. costatum (Jitts et al. 1964), >3 PM for D. brightwellii. Such levels are
and a rock pool form, D. tertiolecta (Epplcy high but are often found in polluted coastal
and Coatsworth 1966). Specific growth rate water and in declining phytoplankton
data are given in the papers cited, or can blooms. We noted numerous D. bright-
be calculated from them, as a function of wellii when nitrate was very low in a declin-
irradiance and temperature with 16-hr light ing bloom of Ceratium furca in November
and 8-hr darkness each 24-hr day. 1967 off Huntington Beach, California.
Variation in specific growth rate (p) with When irradiance is about 0.05 ly/min and
nitrate and ammonium concentration ( S) temperature 20C (Fig. 4), C. huxleyi, S.
was calculated from K, values (Table 2) costatum, and D. brightwellii have essen-
and p,,%taken from growth rate vs. irradi- tially the same pnZ; only at this irradiance
ance curves (Fig. 4) given in the papers would the K, values alone determine the
cited above: supposed competitive advantage of one
species over the others.
According to the laboratory results, D.
Results of the calculations are shown in tertioZecta (a rock pool form) would be an
Fig. 5. effective competitor of diatoms for nitrate
At low irradiance (0.025 ly/min), the or of C. huxleyi for ammonium only with
calculations suggest that C. huxleyi is the bright light, long days (see Eppley and
better competitor and D. tertiolecta the Coa tsworth 1966) and appreciable ammo-
poorest at all nitrate concentrations ( Fig. nium levels. It would probably be an effcc-
5A). With higher irradiance (Fig. 5B) tive competitor where its broad salinity
( 0.1 ly/min-about 20% of full sunlight) tolerance ( McLachlan 1960; Eppley 1963)
the situation is complex, with C. huxleyi would be advantageous. Shallow rock pools
growing fastest with nitrate up to about probably best meet these criteria. The tem-
0.8 j-N; with higher nitrate, S. costatum perature optimum of this organism ( 34C,
becomes the better competitor, In fact, Eppley 1963 ) is also well suited for such
918 Il. W. EI’PLEY, J. N. ROGERS, AND J. J. McCARTIIY
4
NITRATE CONCENTRATION
(PM)
b
C
0 l+F? 0I I I I
0 I 2 0 I 2
AMMONIUMCONCENTRATION(PM)
FIG. 5. Calculated speciFic growth rates vs. nitrate and ammonium concentration at two irradiance
1eveIs (approx l/20, A and C, and l/5, B and D, of surface sunlight irradiance for wavelengths of 400-
700 nm). a = Coccolithus huxleyi, b = Ditylum brightwellii, c = Skeletonema costatum, and d = Dun-
aliella tertiolecta.
habitats. The other species of Fig. 5 would on the basis of differences in their tempera-
presumably be excluded from rock pools turc and salinity optima.
because of low tempcraturc optima and GonyuuZux ~~oZye&a, often the predom-
narrow salinity tolerances. Ryther ( 1954) inant species during local red tides, has a
could account for seasonal changes in the unique strategy that may account for its
relative abundance of diatoms and uni- success despite a low growth rate (about
cellular Chlorophyta in Long Island bays 0.5 doublings/day at 20C and optimal ir-
NITRATE AND AM :MONIUM UPTAKE 919
radiance, in the laboratory) and very high may have widespread importance. Physio-
K, values measured for both nitrate and logical characteristics of phytoplankton
ammonium: its diurnal migration, This measurable with existing methods may be
species, and several other local dinoflagcl- just as important to species distribution as
late species ( Holmes, Williams, and Epplcy the more exotic features remaining for
1967) swim to the surface in daylight, future study.
where nutrients are usually very low, and
to a depth of lo-15 m at night (Eppley, IUZFERENCES
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920 R. W. EPPLEY, J. N. ROGERS, AND J. J. McCARTIIY