Phycologia (2013) Volume 52 (6), 461471

Published 5 December 2013

Differential nutrient uptake by two segregated red algae

in an estuarine intertidal zone
NCHEZ
RAQUEL SA

DE

PEDRO*, F.X. NIELL

AND

RAQUEL CARMONA

Departamento de Ecologa, Facultad de Ciencias, Campus de Teatinos, Universidad de Malaga, 29071 Malaga, Spain
SA NCHEZ DE PEDRO R., NIELL F.X. AND CARMONA R. 2013. Differential nutrient uptake by two segregated red algae in an
estuarine intertidal zone. Phycologia 52: 461471. DOI: 10.2216/13147.1
Bostrychia scorpioides (Rhodophyta: Ceramiales) and Catenella caespitosa (Rhodophyta: Gigartinales) were segregated
but slightly overlapping in the narrow intertidal of the Palmones River Estuary (Southern Spain). This study investigated
whether there were interspecic differences in nutrient uptake capabilities and how meaningful these were to zonation.
Uptake kinetics of ammonium, nitrate, and phosphate were measured over successive time intervals across a range of
environmentally realistic concentrations. The interactions between ammonium and nitrate in mixed N source uptake were
also tested. Using internal N and P content, turnover rates were calculated for each species for periods of emersion and
submersion. Nutrient uptake kinetics were saturable for almost all nutrients except for ammonium in B. scorpioides,
which showed surge uptake responses through linear kinetics. Vmax and afnity progressively decreased over 3 h for all
nutrients in C. caespitosa and only for phosphate in B. scorpioides. At high dissolved inorganic nitrogen concentrations
both species exhibited greater uptake rates for ammonium than nitrate; although, similar N uptake rates were obtained at
the low ones. Ammonium contributed to the total N uptake in B. scorpioides to a greater extent; whereas, in C. caespitosa,
this only occurred at high N concentrations. In mixed N source uptake experiments, ammonium and nitrate uptake rates
by B. scorpioides decreased when the other N source was equimolarly supplied, while in C. caespitosa these interactions
were absent. Catenella caespitosa showed a Vmax and afnity for phosphate fourfold higher than for B. scorpioides, with
greater interspecic differences than those found for N uptake. Catenella caespitosa renewed internal nutrients faster and
was more dependent on the external nutrient supply than B. scorpioides, which can withstand longer periods without
nutrient supply due to its slower nutrient turnover rate. Different capabilities for nutrient acquisition in the two species
suggested an adaptation to the nutrient availability at each intertidal position.
KEY WORDS: Ammonium, Bostrychia, Catenella, Nitrate, Phosphate, Time dependency, Zonation

INTRODUCTION
Zonation in intertidal ecosystems has been related to the
physiology of macroalgae, based on their abilities to survive
under emersion conditions (Dring & Brown 1982; Davison &
Pearson 1996; Zuccarello et al. 2001; Skene 2004). Most
research on macroalgae zonation has been performed on
rocky shores, but spatial segregation patterns have also been
reported in estuaries and mangroves. These areas are mainly
represented by cosmopolitan species of the genera Bostrychia, Catenella, Caloglossa, Stictosiphonia, Gracilaria, and
Polysiphonia (Davey & Woelkerling 1985; Phillips et al. 1996;
Pena et al. 1999; Melville & Pulkownik 2007).
Previous studies have associated the differences in nutrient
uptake of co-occurring species to their differential tidal
zonation. For example, Fucus species from the middle and
upper intertidal zones enhance their uptake rates of nitrate
and ammonium in water following emersion (Thomas &
Turpin 1980; Thomas et al. 1987) but not that of phosphate
(Hurd & Dring 1990, 1991). This relationship between higher
N uptake rates and the upper position in the intertidal zone
has been reported for several intertidal species of New
Zealand (Phillips & Hurd 2003, 2004) and for some Porphyra
species from the upper eulittoral zone (Kim et al. 2008) and
high-positioned seaweed assemblages (Bracken et al. 2011).
* Corresponding author (rsdpc@uma.es).

These uptake patterns can even occur between high and low
shore specimens of the same species (Phillips & Hurd 2004;
Hays 2007; Bracken et al. 2011) and are considered a
phenotypic acclimation to the environmental conditions.
As a general rule, physical factors control the upper
zonation of intertidal habitats; whereas, biological interactions play a major role in the lower zones (Lewis 1964).
Interspecic competence has been described for several
marine benthic organisms (Connell 1972), including intertidal macroalgae (Chapman & Johnson 1990; Karez 2003).
Species coexisting in a habitat compete for light, substrate,
and nutrients (Lobban & Harrison 1994). Davison &
Pearson (1996) hypothesised that the intertidal distribution
limits are controlled by competitive exclusion, although the
stress caused by nutrient limitation or desiccation mediates
these interspecic interactions. Since nutrients are intermittently available for upper intertidal species, their nutrient
uptake abilities would be presumably different than for
lower intertidal ones. In fact, the ability to take up nutrients
effectively during submersion could be decisive in determining the shore position (Hurd & Dring 1990).
Ecophysiological studies of macroalgae have revealed
different nutrient uptake strategies that allow species to
survive when nutrients are exhausted. Species with high
uptake rates and rapid growth have a competitive advantage
to sequester transient pulses of nutrients (Rosenberg et al.
1984; Pedersen & Borum 1997). Likewise, species with slow
nutrient uptake but high storage capability might compen461

462

Phycologia, Vol. 52 (6), 2013

and over a common submersion period for these species (3
h). In this study, we dene environmental nutrient concentrations as the most frequent concentrations of ammonium,
nitrate, and phosphate over the period of study. Furthermore, since both nitrogen sources are present in the natural
environment, interactions between ammonium and nitrate
uptake are also tested in this study.
The aim of this research is to address the following
questions: (a) What interspecic differences exist in the
nutrient uptake kinetics over time? (b) What is the effect of
one nitrogen source in the uptake of the other one? (c) How
might differential nutrient uptake capability at environmental concentrations and real submersion times affect the
zonation of the two species?

MATERIAL AND METHODS

Fig. 1. Tidal regime in Palmones River Estuary and intertidal
distributional limits of Bostrychia scorpioides and Catenella
caespitosa. Data extracted from tide heights in 2011 measured in
Algeciras Port (3687.1 0 N, 5826.6 0 W) by the Spanish Institute of
Oceanography (Ministerio de Defensa, 2010).

sate for this lack of uptake afnity (Ryther et al. 1981;

Campbell 1999; Young et al. 2009).
In this study, we examine the nutrient uptake rates and
kinetics of two estuarine rhodophyceans: Bostrychia scorpioides (Hudson) Montagne ex Ku tzing (Rhodophyta,
Ceramiales) and Catenella caespitosa (Withering) L.M.
Irvine (Rhodophyta, Gigartinales). These salt marsh epiphytic species are attached and entangled on the lower
branches of halophylic macrophytes of the Palmones River
Estuary (36810 0 13.4076 00 N, 5826 0 27.5742 00 W) located in
Algeciras Bay, Cadiz (Southern Spain). This partially mixed
estuary has very weak tides that range between 0.6 and 2 m
(Izquierdo 2001), and water renewal time changes seasonally
from 0.25 to 1.47 days (Niell et al. 2005). Salinity varies over
a tide cycle between 29 and 35 (Clavero et al. 1997).
Seasonally, mean water temperature in winter is 148C and
238C during summer. The ecophysiological interest of this
study is related to the small-scale spatial zonation that exists
between these species. Bostrychia scorpioides grows in the
upper intertidal zone and C. caespitosa is found in the lower
zone, even over the mudat sediment. Furthermore, both
species coexist and overlap in the middle part. This is the rst
record of the small-scale spatial segregation between these
species in the estuary.
Nutrient uptake characterisation of B. scorpioides and C.
caespitosa will allow identication of the competitive
advantages that cause them to overcome the intermittent
tidal and seasonal nutrient supply in the estuarine intertidal
zone. Ammonium is currently the major form of dissolved
inorganic nitrogen (DIN) in the Palmones River Estuary
(Niell et al. 2005) and its concentration is highly variable, as
in other estuaries (Sharp et al. 1982; Kennison et al. 2011). In
contrast, nitrate and phosphate concentrations remain more
stable over the year and in lower concentrations (Niell et al.
2005). According to the high variability of nutrient
concentration within seasons and over a tidal cycle, the
nutrient uptake kinetics are examined in a range of nutrient
concentrations beyond the mean nutrient concentrations,

Algal material
Distributional limits of B. scorpioides and C. caespitosa in the
middle estuary were veried using surveying techniques
related to established tidal baselines (Fig. 1). From March
2010 to May 2011, specimens of B. scorpioides and C.
caespitosa were collected at low tide from identical tidal
elevations, in the zone of distributional overlap (1.101.20 m),
to prevent variations within species related to tidal position,
despite the narrow horizon (2030 cm height). In parallel,
water samples were taken to determine nutrient concentrations (n 3). Plants were carefully removed from the lignied
stems of salt marsh vegetation, placed in plastic containers
with estuary water, and transported in a cooler to the
laboratory. Vegetative thalli of both species were selected to
reduce physiological variability (Harrison & Hurd 2001), then
rinsed in natural nonltered seawater and gently cleaned of
sediment and epiphytes.
Plants were acclimated in nutrient-free medium for 5 to 6
days, to examine surge uptake capability after a nutrient
deprivation period. Algae were kept under constantly
submerged conditions, in aerated 1.5 l perspex cylinders with
ltered seawater (Whatman GF/C lter, Whatman International Ltd, Maidstone, UK), at pH 8.2 (adjusted by adding
NaOH or HCl), salinity 34. Cultures were maintained in a
walk-in cold room at 15 6 18C, and irradiance was set at
about 45 lmol photons m2 s1, provided by uorescent lamps
(cool daylight, F-18W/54765 Sylvania) under 12:12 h light:
dark photoperiod. This irradiance corresponded to the zone of
distributional overlap of these species in their natural habitat
and was not limiting for growth. All irradiances in this study
were measured using a spherical sensor, connected to a
radiometer (LI-CORt LI-250A, Li-Cor Biosciences, Lincoln,
Nebraska, USA).
Nutrient uptake experimental design
The time-course of uptake was obtained by a combination of
the multiple ask and perturbation method as recommended
by Pedersen (1994). The multiple perturbation technique
follows nutrient depletion over successive time intervals in
several asks with different initial nutrient concentrations.
This method provides information on all uptake phase

Sanchez de Pedro et al.: Differential nutrient uptake by two estuarine macroalgae

responses (surge uptake, externally and internally controlled
uptake) and the time dependency of nutrient uptake.
Intraspecic variability between individuals was also considered (different specimens were used).
Ammonium, nitrate, and phosphate uptake were analysed
in both species in single nutrient addition experiments (as
NH4Cl, NaNO3, and Na2HPO4). In an additional experiment,
nitrate and ammonium were supplied together (as NH4NO3),
to study the interaction between both nitrogen sources. Four
initial concentrations were set for each nutrient (5, 10, 20, and
40 lM for nitrate and ammonium, in both single and mixed
addition experiments, and 2, 5, 10, and 20 lM for phosphate).
Articial seawater, pH 8.2 and salinity 34, was spiked with
adequate volumes of nutrient solutions to obtain the initial
concentrations. The lower concentration range for phosphate
rather than for N sources was chosen, based on its lower
concentration values in the Palmones River Estuary (Izquierdo 2001; Niell et al. 2005).
Uptake experiments were performed at an irradiance of 230
lmol photons m2 s1, saturating light for photosynthesis
(Sanchez de Pedro et al. pers. comm.). All incubations were
conducted in Falcon tubes lled with 50 ml medium of each
treatment as indicated above and 200 mg FW of algal
biomass. Four replicates were performed for each initial
nutrient concentration. Incubation tubes were placed on an
orbital shaker (SSL1 STUARTt, Camlab Ltd, Cambridge,
UK) at 2 3 g, to prevent boundary layer formation around
thalli. Nutrient depletion in each ask was followed by taking
3 ml aliquots of the incubation medium at 15, 30, 45, 60, 120,
and 180 min, which were frozen and stored at 208C until
analysis. Water samples were analysed in an automated
nutrient analyser QuAAtro AQ2 AACE (Seal Analytical Ltd,
Fareham, UK), using standard methods for ammonium
(Slawyk & MacIsaac 1972), nitrate (Shinn 1941; Wood et al.
1967), and phosphate (Murphy & Riley 1962).
Nutrient uptake rates (lmol nutrient g 1 DW h1) were
calculated from depletion of the nutrient concentration in the
incubation medium over time using the equation:
V S S0  St 3 V0 =B 3 t

where V is the net uptake rate, S0 and St are the nutrient

concentrations (lM) at the initial and at the end of the
interval, V0 is the volume during the incubation time interval, t
is the time elapsed between two successive samplings (h), and
B is the dry weight biomass (g DW). Uptake rates at
analogous time intervals were plotted against nutrient
concentration, as indicated by Pedersen (1994), and six plots
for each nutrient treatment were obtained.
Nutrient uptake rates at the environmental nutrient
concentrations in the estuary were calculated as the mean of
raw nutrient uptake rates values around that concentration,
regardless of the time interval in which they were obtained.
Total N and P uptake per tidal cycle was estimated by
multiplying mean uptake rates per submersion time in a 12 h
tidal cycle (2.5 and 5 h for B. scorpioides and C. caespitosa,
respectively). Turnover rates for N and P were estimated by
the following formula:
Turnover rate N ; P uptake 3 mN ; P=% N ; P 3 104

where N,P uptake is the total nitrogen or phosphorous

463

amount taken up over a 24 h tidal cycle (lmol N,P g 1 DW

day 1), m (N, P) is the molecular mass (mol g 1 DW), and %
N, P is the total content of N or P per DW.
Thalli from nutrient uptake experiments were dried at 608C
overnight and then powdered with a mixer mill (model
MM400, Retsch UK Ltd, Whitford, UK). Internal ammonium and nitrate were analysed according to Corzo & Niell
(1991): about 0.035 g of homogenised dry powder was
dissolved in 40 ml of deionised water and incubated in a
shaking bath at 308C for 30 min. After centrifugation for 15
min at 2151 3 g, at 258C (refrigerated model 5804R,
Eppendorf UK Ltd, Stevenage, UK), the supernatants were
measured colourimetrically, and internal ammonium and
nitrate concentrations were determined by the automated
procedures already described. All internal nutrient concentrations were expressed as millimolar, by referring the number of
moles to the cell water content.
Elemental C and N content was determined in dry
powdered samples using a CHN elemental auto-analyser
(Model 2400C; Perkin Elmer Analytical Instruments, Waltham, Massachusetts, USA) by DOI method (Kristensen &
Andersen 1987). Total P content was analysed using the
procedure of Pardo et al. (1999) and then determined by the
green malachite method (Fernandez et al. 1985). Total C, N,
and P content of algae was reported as % DW, and tissue C:N
and N:P values were expressed as molar ratios. Dry weight
percentage in B. scorpioides was 28.7% and 15.3% in C.
caespitosa.
Surface area:volume (SA:V) ratio was determined on 28
individuals of each species by the procedure described by
Rosenberg & Ramus (1984). Flat images were obtained form
the thalli by scanning them in a CanoScan LiDE 25 (Canon
USA, Melville, New York, USA), and thallus area was
quantied using ImageJ (Image Processing and Analysis in
Java, National Institutes of Health, Bethesda, Maryland,
USA).
Statistical and data analysis
Corrected Akaikes Information Criterion (AICc, Akaike,
1973; Burnham & Anderson 2002) was used to test whether
linear or saturation ts were more likely to have generated the
relationships between uptake rates and substrate concentrations. This index is the most used information-theoretic
approach to model comparison, and it has been recently used
in marine ecology (Beninger et al. 2012), including comparisons among nutrient uptake kinetic models (Bracken et al.
2011).
Data were tted to linear models by least squares linear
regression when uptake rates increased linearly with concentration. The slope of the function was obtained as a kinetic
parameter. Likewise, saturating uptake responses were tted
to the MichaelisMenten function by least squares nonlinear
regression analyses, and Vmax and Ks parameters were also
calculated. The coefcient Vmax/Ks, reported as a, was also
calculated as the estimation of effective uptake at low nutrient
concentrations (Bracken et al. 2011). Alternatively, data were
tted to the modied MichaelisMenten equation, when the
intercept of V on the abscise axis was greater than zero
(Martinez & Rico 2004), as follows:

464

Phycologia, Vol. 52 (6), 2013

33% of samples, it ranged from 10 to 15 lM NO3, related to
winter values. Total DIN concentrations in the estuary ranged
from 0 to 20 lM in 50% samples and were greater than 60 lM
in 25% samples (Fig. 2). Phosphate concentration varied
between 0 and 2 lM in 69% of samples, and only in 12% of
samples it slightly exceeded 4 lM PO43, with a maximum
mean value of 4.26 6 0.12 lM PO43 (Fig. 2).
Nutrient uptake kinetics

Fig. 2. Environmental nutrient concentrations expressed as the

observed frequency of each concentration range over the study
period.

V Vmax 3 S  X0 =Ks S

where Vmax is the maximum uptake rate, Ks is the half

saturation constant, and X0 is the substrate compensation
point or the concentration at which the uptake rate is zero.
Once a kinetic model was selected, lines or curves across
time intervals were compared. When kinetic parameters were
the same for successive time intervals (tested by AICc), global
shared parameters were provided, and data from these time
intervals were pooled.
As Pedersen (1994) indicated, saturating responses can be
masked by the surge uptake that occurs in the rst 15 min, and
data tted better to linear responses. Thus, if removing uptake
rates from that rst interval gave lower AICc values and better
parameter estimates for the saturation model, these rates
would be plotted as outliers.
When the same uptake kinetic models were obtained, we
used one-way ANOVAs to evaluate (1) the effect of single or
mixed N source addition on the kinetic parameters for each
species, and (2) the interspecic differences for each nutrient
treatment, within the same time intervals.
Differences in uptake rates at environmental concentrations
were tested by two-way ANOVAs using Tukeys HSD-test for
post hoc comparisons. As neither treatments nor concentrations had an effect on internal nutrient content (two-way
ANOVA), data were pooled for each species, and interspecic
differences were analysed by t tests. The signicance level was
set at a 0.05 for all statistical analyses. Data management
and statistical analysis were performed using GraphPad Prism
6.01 (GraphPad Software) and SigmaPlot 11.0 (Systat
Software Inc., Chicago, Illinois, USA).

RESULTS
Environmental nutrient concentrations
Ammonium concentration uctuated over the studied period
and showed two modal values (Fig. 2): in 54% of samples,
concentration ranged between 0 and 20 lM NH4, and in
27%, between 40 and 60 lM NH4. Higher ammonium
concentrations were detected in early winter (85.3 lM NH4).
Nitrate concentration also presented two modal values (Fig.
2). In 40% of samples, this nutrient ranged from 0 to 5 lM
NO3, corresponding with the last spring samples, whereas in

Both species exhibited higher uptake rates for ammonium

rather than for nitrate when each N source was supplied
separately (Fig. 3). Bostrychia scorpioides took up ammonium
and nitrate at a constant rate across time intervals, whereas in
Catenella caespitosa uptake rates decreased progressively over
time (Fig. 3, Table 1). For B. scorpioides, ammonium uptake
kinetics overlapped and described a common linear response
(Fig. 3). In this species, uptake rates at 40 lM NH4 at the
initial interval (015 min) were outside the axis limits and
reached a rate of 104 6 19 lmol NH4 g 1 DW h1. Since
ammonium was completely exhausted in the incubation
medium at 60 min by B. scorpioides, the uptake rates at the
last two time intervals were not calculated (Fig. 3). In contrast,
ammonium uptake kinetics in C. caespitosa saturated and
tted the MichaelisMenten modied model better, due to the
substrate compensation point for ammonium (Fig. 3, Table 1).
Uptake rates for ammonium in C. caespitosa were time
dependent since Vmax diminished gradually from 45 to 180
min, and afnity (as a values) decreased as long as Ks values
remained unchanged (Table 1).
Nitrate uptake kinetics tted the MichaelisMenten model
better than linear model in B. scorpioides, for which common
parameter estimates were obtained after pooling the rates
from all time intervals (Table 1, Fig. 3). Although nitrate
uptake in C. caespitosa also saturated and tted the
MichaelisMenten model, the greatest nitrate uptake rates
occurred in the 015 time interval (Table 1, Fig. 3), and Vmax
and Ks values declined progressively over time (P , 0.05).
Both kinetic parameters decreased to the same extent, thus
afnity (a) for nitrate remained constant over the studied
period (P . 0.05).
By comparing the uptake responses at low and high
environmental concentrations within each nitrogen source,
ammonium uptake rates at 40 lM were 2.3-fold greater than
at 20 lM in B. scorpioides and 1.8-fold in C. caespitosa (Fig.
4). Nitrate uptake rates at 10 lM NO3 were 1.7-fold the rates
at 5 lM, for both species (Fig. 4). The greatest interspecic
differences were found at 40 lM NH4, when uptake rates in
B. scorpioides were fourfold higher than in C. caespitosa (Fig.
4); however, when both N sources were supplied, no
signicant differences were observed between them (Fig. 4).
In mixed addition experiments, ammonium and nitrate
uptake rates by B. scorpioides decreased when the other
nitrogen source was supplied equimolarly (Fig. 4). For this
species ammonium uptake rates declined by 36% at 20 lM and
48% at 40 lM; whereas, nitrate uptake rates were 33% and
51% lower than the obtained in single N source experiments
(Fig. 4). In contrast, in C. caespitosa, ammonium or nitrate did
not affect the uptake of the other N source.
In terms of total DIN, both species showed the same uptake
rate at 20 lM DIN (18 lmol DIN g 1 DW h1), while at 70

Sanchez de Pedro et al.: Differential nutrient uptake by two estuarine macroalgae

465

Fig. 3. Uptake kinetics of ammonium and nitrate separate source experiments by Bostrychia scorpioides and Catenella caespitosa. Uptake
rates at each time interval (min) are shown as different symbols.

scorpioides but changed between concentrations in C.

caespitosa. For B. scorpioides, ammonium contributed about
75% to its total DIN uptake. In contrast, C. caespitosa took up
ammonium and nitrate to the same extent at low concentra-

lM DIN they were signicantly greater for B. scorpioides (65.5

vs 43.3 lmol DIN g 1 DW h1). The contribution of
ammonium or nitrate to the total N uptake at 20 or 70 lM
DIN, in mixed addition experiments, was the same in B.

Table 1. Kinetic parameters of ammonium and nitrate uptake curves measured during successive time intervals in Bostrychia scorpioides and
Catenella caespitosa. Values are x 6 s for parameters.
Nutrient

Species

NH4
NH4

B. scorpioides
C. caespitosa 1

NO3
NO3

B. scorpioides
C. caespitosa

Time interval
(min)
060
015
1530
3045
4560
60120
120180
0180
015
1530
3045
4560
60120

Substrate compensation point at X0 6.0 6 0.4.

Vmax (lmol g1 DW h1)

76
60
53
37
24
7.8
7.4
52.9
26.5
16.3
10.5
6.1

6
6
6
6
6
6
6
6
6
6
6
6

7.9
4.5
12
7.3
6.9
6.9
0.35
10
4.3
2.5
1.6
0.8

Ks (lM)

V(S) 2.4  S
5.1
6.5
7.9
4.8
8.1

6
6
6
6
6

0.33 6
49.5 6
20.3 6
11.5 6
6.7 6
4.9 6

3.7
2.6
7.3
5.0
7.7
0.5
15
6.6
4.1
2.8
1.8

r2

14.9
9.2
6.7
7.7
3.0

22.4
1.1
1.3
1.4
1.6
1.2

0.97
0.96
0.99
0.93
0.95
0.96
0.83
0.82
0.95
0.90
0.86
0.79
0.82

76
16
16
16
16
16
16
78
16
16
16
16
16

466

Phycologia, Vol. 52 (6), 2013

Fig. 4. Uptake rates at the most frequent concentrations of ammonium and nitrate in the estuary by Bostrychia scorpioides and Catenella
caespitosa in separate (open bars) and mixed (lled bars) N source experiments. Different letters indicate signicant differences observed with
the one way ANOVA (P , 0.05).

tions (48% NO3 vs 52% NH4), while at high concentrations

ammonium accounted for 77% of total N taken up as DIN.
Catenella caespitosa displayed greater Vmax and Ks than B.
scorpioides at all time intervals, and interspecic differences
were more evident for phosphate than for the N sources (Fig.
5). Kinetic parameters could be only compared between
species among the time intervals from 0 to 45 min, when Vmax
and afnity were from four- to vefold greater in C. caespitosa
(Table 2). Since phosphate was exhausted after 60 min in the
incubation medium by C. caespitosa, kinetics parameters and
uptake rates between 60 and 180 min were not calculated for
this species (Table 2). For both species, Vmax values declined

over time while afnity (a) decreased six times for C.

caespitosa and was maintained for B. scorpioides.
Internal nutrient content and SA:V
Neither concentration nor treatments affected internal nutrient content in short-term uptake experiments over 3 h (P .
0.05, Table 3). Bostrychia scorpioides had higher values of all
internal nutrients and ratios (P , 0.001) except for N:P (P
0.699), that was greater for C. caespitosa. The greatest
differences between species were found in internal ammonium,
being 2.8-fold higher in B. scorpioides (Table 3). Surface area

Fig. 5. Uptake kinetics of phosphate by Bostrychia scorpioides and Catenella caespitosa. Uptake rates at each time interval (min) are shown as
different symbols.

Sanchez de Pedro et al.: Differential nutrient uptake by two estuarine macroalgae

467

Table 2. Kinetic parameters of phosphate uptake curves measured during successive time intervals in Bostrychia scorpioides and Catenella
caespitosa. Values are x 6 s for parameters.
Species

Time interval
(min)

B. scorpioides

C. caespitosa

Vmax (lmol g1 DW h1)

015
1530
3045
4560
60120
120180
015
1530
3045
4560

5.5
3.6
2.5
1.9
1.1
0.53
22.5
15.0
13.4

6
6
6
6
6
6
6
6
6

0.4
0.35
0.35
0.27
0.16
0.07
1.7
1.2
2.0

to volume ratio (SA:V) obtained for C. caespitosa was around

twofold higher than the found in B. scorpioides (Fig. 3).
Total N and P taken up over a tidal cycle and nutrient turnover
times
Considering submersion times and the environmental nutrient
uptake rates, the total nutrient amount incorporated by each
species was calculated. Total N taken up per tidal cycle was
two times higher in C. caespitosa than in B. scorpioides, at low
N concentrations and 1.3-fold higher at the higher ones (Table
4). Likewise, total P taken up was 7.4-fold greater in C.
caespitosa than in B. scorpioides. Turnover times for N and P
at low concentrations (obtained form turnover rates by
Equation 2) were 35 and 17 days in B. scorpioides; whereas,
in C. caespitosa, N and P were renewed after 16 and 2 days,
respectively (Table 4). At high concentrations, turnover times
for N decreased by around fourfold in B. scorpioides and 3fold in C. caespitosa (Table 4).

DISCUSSION
This is the rst attempt to describe the uptake kinetics for
major nutrients involved in the nutrition of the two
rhodophyceans in the intertidal zone of the Palmones River
Estuary. We found clear differences in P uptake between
species, with a greater ability for P acquisition by Catenella
caespitosa. In contrast, interspecic differences in N uptake
Table 3. Internal nutrient content and surface area:volume of
Bostrychia scorpioides and Catenella caespitosa in nutrient uptake
experiments. Values are x 6 s (n 4080 except for N:P molar ratio
where n 20), pooled from all replicates of each uptake experiment
(NH4, NO3, NH4NO3, and PO43).
Internal nutrient content
Total C (% DW)1
Total N (% DW)1
Total P (% DW)1
C:N (molar ratio)1
N:P (molar ratio)
Total internal NO3 (mM)1
Total internal NH4 (mM)1
SA:V (cm1)1
1

B. scorpioides
31.2
4.25
0.58
8.64
17.7
2.59
17.4
4.8

6
6
6
6
6
6
6
6

0.99
0.24
0.03
0.83
2.44
1.96
3.43
1.3

C. caespitosa
27.3
3.90
0.48
8.20
18.0
1.97
6.33
11.7

6
6
6
6
6
6
6
6

1.03
0.22
0.09
0.32
2.43
1.08
1.64
1.8

Signicant differences between Bostrychia and Catenella (t test,

P , 0.05).

Ks (lM)

5.6
3.2
2.0
1.5
1.2
0.6
1.6
2.7
7.2
V(S) 0.44  S

6
6
6
6
6
6
6
6
6

1.1
1.0
1.1
0.9
0.8
0.4
0.5
0.7
2.5

r2

1.0
1.1
1.3
1.3
0.9
1
13.8
5.6
1.9

0.94
0.85
0.66
0.61
0.59
0.58
0.86
0.90
0.92
0.95

15
16
16
16
16
16
15
16
16
16

were minor but notable, and depended on the time interval in

which they were compared, the nutrient concentration, and
the nitrogen source.
Time dependency of uptake rates was only observed for C.
caespitosa for all nutrients analysed, mostly affecting Vmax,
as has been reported in other macroalgae (Pedersen 1994;
Pedersen & Borum 1997; Lotze & Schramm 2000) and might
relate to the gradual lling of the internal nutrient pools.
When a single nutrient pulse is provided, three uptake
phases can be distinguished by using the perturbation
method (Fujita 1985; Harrison et al. 1989). We obtained
saturation kinetics for almost all nutrients in both species,
except for ammonium in Bostrychia scorpioides. This type of
response corresponds to the internally and externally
controlled uptake phases (Harrison et al. 1989). In contrast,
the surge uptake phase that we found for ammonium in B.
scorpioides is a general attribute of the perturbation method
design (Pedersen 1994), and it is found in the rst 15 min, as
in other studies (Fujita 1985; Dy & Yap 2001). Although
linear uptake kinetics for ammonium by B. scorpioides
reected a high surge uptake capability and greater uptake
rates than C. caespitosa, the interspecic comparisons were
limited to the rst hour due to the ammonium depletion in
the incubation medium by B. scorpioides. Biphasic uptake
kinetics have been reported in macroalgae for ammonium
(Campbell 1999; Martinez & Rico 2004), nitrate (Conolly &
Drew 1985; Lavery & McComb 1991), and phosphate
(Gordillo et al. 2002; Martinez & Rico 2004). Many studies
exclude the surge uptake phase, because data t better to
saturation models at lower concentrations (usually more
environmentally frequent), or because they cannot statistically prove the existence of a second phase (Martinez & Rico
2004). Nitrate concentrations in the Palmones estuary
Table 4. Estimates of total N and P taken up over a 12 h tidal cycle
at environmental concentrations (see Fig. 5). Turnover times (days)
are indicated in brackets.
B. scorpioides
Total N uptake per tidal cycle (lmol N g
20 lM
44 (35)
. 60 lM
164 (9)

C. caespitosa
1

DW)

Total P uptake per tidal cycle (lmol P g1 DW)

2 lM
5.7 (17)
5 lM

88 (16)
217 (6)
42.4 (2)

468

Phycologia, Vol. 52 (6), 2013

seldom exceed 15 lM, thus, nitrate uptake rates at high

concentrations were not included (Fig. 3); however, ammonium concentrations above 40 lM were frequent in the
estuary and therefore were considered in the ammonium
uptake kinetic for B. scorpioides.
Regardless of the existence of a second kinetic phase, there
is a common concern that species exhibiting surge uptake
responses are able to take up nutrient pulses faster than
species with saturation kinetics (Martinez & Rico 2004).
Rhodophyceans in this study were able to take up nutrients
at concentrations exceeding the frequent values found in the
estuary. It has been suggested that macroalgae from
eutrophic estuaries tend to utilise surge uptake mechanisms
at environmental ammonium concentrations above 40 lM, if
these concentrations are likely to occur (Kenninson et al.
2011). At that concentration we found that B. scorpioides
had greater ammonium uptake than C. caespitosa due to its
surge uptake responses. Only when ammonium concentrations are high in the estuary would B. scorpioides have an
advantage over C. caespitosa in acquiring nutrients. Furthermore, the surge uptake responses might be enhanced as
long as the ammonium is more discontinuously available, i.e.
after neap equinoctial tides (when species can be isolated
from the N source for up to 10 days) or after a period of
nutrient shortage. Since these local events often occur in the
estuary, the surplus capability can be considered a potential
benet to B. scorpioides, as it has been suggested for other
macroalgae species inhabiting the upper intertidal (Lotze &
Schramm 2000; Rees 2003).
The Ks values for nitrate uptake in C. caespitosa must be
interpreted carefully, because they might be an artefact of
the MichaelisMenten modied model. If the plotted curves
were shifted to the abscise origin, Ks would provide lower
values and afnity would be greater.
The combination of nitrate and ammonium in the mixed
addition uptake experiments led to a decrease in both N
sources uptake rates in B. scorpioides but not in C.
caespitosa. Lower nitrate uptake rates caused by the
interference of ammonium have been reported in several
intertidal macroalgae (DElia & DeBoer 1978; Haines &
Wheeler 1978; Thomas & Harrison 1987; Rees et al. 2007),
but it has not been observed for some other species (Bird
1976; Harrison et al. 1986; Abreu et al. 2011). Ammonium,
as a reduced N form, is taken up preferentially by many
macroalgae (DElia and DeBoer 1978; Pedersen & Borum
1997; Luo et al. 2012). This preference might be due to its
lower metabolic cost to be transported and assimilated
compared to nitrate, which might cause the inhibition of
nitrate uptake by ammonium (Syrett 1981). However, the
inhibition of ammonium uptake by nitrate that we found for
B. scorpioides cannot be supported by this statement and it
has not been previously reported. Nonetheless, some macroalgae can take up nitrate in preference to ammonium, if the
former is the main N source in their natural habitat
(Martinez and Rico 2004; Kennison et al. 2011), or take
up both N forms at similar rates (Runcie et al. 2003;
Kraemer et al. 2004). At environmental nutrient concentrations B. scorpioides and C. caespitosa exhibited higher uptake
rates for ammonium than for nitrate, regardless of the
interactions between N sources. Furthermore, ammonium
had a greater relative importance to total N uptake for B.

scorpioides than for C. caespitosa, for which it only occurred

at high concentrations, below which nitrate uptake rates
were greater. Therefore, the preference for ammonium at
high concentrations by these rhodophyceans can be related
to the most frequent ammonium concentrations they are
exposed to in the estuary (Niell et al. 2005).
When comparing uptake parameters with those of other
estuarine species, Ks for ammonium in B. scorpioides and for
phosphate in C. caespitosa, were similar to those of some
Chlorophyceans, although Vmax for both nutrients were
lower than for nitrophilous species (Pedersen 1994; Campbell 1999; Runcie et al. 2004; Liu et al. 2012). As in the case
of Ks for ammonium in Caulerpa cupressoides (Williams
1984), the Ks values for phosphate uptake in both species
were in the range of phosphate concentrations in the estuary,
suggesting that these species are well acclimated to the
environmental P concentrations.
Ecological implications of nutrient uptake in the macroalgal
zonation
To determine whether nutrient uptake is a key factor to
understand the distinct distribution of the two species in the
intertidal zonation, it is crucial to determine how often
nutrients are available for the two species of the estuary and
how efcient these species are in fullling their nutritional
requirements during submersion periods (Hurd and Dring
1990). We found that Bostrychia scorpioides takes longer than
Catenella caespitosa to renew its internal N and P content.
This should give B. scorpioides the ability to withstand longer
periods without a supply of nutrients. At neap equinoctial
tides, B. scorpioides species can be emersed for up to 10 days
(calculated from tide tables) and its growth would need to be
supported by stored nutrients. In contrast, C. caespitosa might
be limited by P under these conditions due to the shorter
turnover time for P than for N. We found that internal
nutrient pools in C. caespitosa were low, making this species
more dependant on external nutrient supply relative to B.
scorpioides.
The observed zonation in this study is consistent with the
frequent higher shore positions of Bostrychia species in
estuaries and mangroves (Mann & Steinke 1988; Pena et al.
1999; Zuccarello et al. 2001) and the lower ones in Catenella
species (Davey & Woelkerling 1985; Batelli 2004; Melville et
al. 2005). Distributional limits of nonestuarine intertidal
species have also been shown to be related to N and P uptake
rates (Thomas et al. 1987; Phillips & Hurd 2003, 2004;
Bracken et al. 2011). Our ndings also showed a correspondence between the higher N uptake rates with the upper shore
species, B. scorpioides, but not in the case of phosphate
uptake, whose greater values were observed in C. caespitosa.
Nonetheless, the lower turnover rates of N and P in B.
scorpiodes does support the observed zonation pattern.
It is well known that the N source with a major contribution
to total N uptake can vary within ecosystems (DElia &
DeBoer 1978; Thomas & Harrison 1987). Phillips & Hurd
(2003, 2004) found greater nitrate uptake rates in the upper
shore species for an habitat where the nitrate was more
abundant than ammonium. However, we found higher uptake
rates for ammonium instead of nitrate in B. scorpioides,
probably due to the greater concentration in the Palmones

Sanchez de Pedro et al.: Differential nutrient uptake by two estuarine macroalgae

estuary compared to nitrate concentrations. At the most
frequent DIN concentrations, N uptake was similar between
both species and the interactions between N sources were less
evident. Therefore, the less frequent concentrations above 40
lM might contribute decisively to the dominance of B.
scorpioides over C. caespitosa in the zonation.
Previous studies reported an inverse relationship between
phosphate uptake rates and tidal elevation (Hurd & Dring
1991). These authors attributed these differences to morphological properties, indicating that species with a higher SA:V
ratio show higher phosphate uptake rates. This is also
supported by our ndings, since C. caespitosa had greater
phosphate uptake rates and also a higher SA:V ratio. The
lower internal P content in C. caespitosa might also increase P
uptake capability to prevent from P deciency. This internally
controlled positive feedback has only been reported for N
sources (Fujita et al. 1988; McGlathery et al. 1996), which
actually was observed in our study, except for ammonium in
B. scorpiodies. The higher ammonium concentration in the
estuary appears to explain the greater ammonium uptake
capability in both species rather than the internal nutrient
requirements, particularly in B. scorpioides, which displayed
ammonium surge uptake.
We can conclude that the greater storage capability and
surge uptake for ammonium in B. scorpioides and the higher
dependence of external nutrients in C. caespitosa are consistent
with their differential emersion and submersion periods in the
estuarine intertidal zone. Taken together, our results support
that the species with higher uptake rates and afnity for the
most abundant N source and less dependence on external
nutrient supply would have a competitive advantage in the
upper position in the intertidal zone. Differential nutrient
uptake may play a fundamental role in setting the distributional limits of these rhodophyceans, since it relates well to the
observed zonation. Despite the differential nutrient uptake
responses relate well to the observed zonation, we cannot rule
out that the partial spatial segregation might have been driven
by a competitive interaction for other resources, such as light,
stem surface, or susceptibility to grazers, or by distinct
capacities of physiological recovery following emersion.
Further research is necessary to unravel the nature of their
ecological relationships and the role of desiccation.

ACKNOWLEDGEMENTS
We are grateful to J.A. Fernandez for his helpful comments
during the experiments. We gratefully acknowledge comments from two anonymous reviewers which substantially
improved this manuscript. This study was supported by the
Ministry of Science and Innovation of Spain, within the
project CICYT CTM 200804453/MAR. This research
represents part of an Honour Thesis undertaken by R.
Sanchez de Pedro (Universidad de Malaga, 2011).

REFERENCES
ABREU M.H., PEREIRA R., BUSCHMANN A.H., SOUSA-PINTO I. &
YARISH C. 2011. Nitrogen uptake responses of Gracilaria

469

vermiculophylla (Ohmi) Papenfuss under combined and single

addition of nitrate and ammonium. Journal of Experimental
Marine Biology and Ecology 407: 190199.
AKAIKE H. 1973. Information theory and an extension of the
maximum likelihood principle. In: Second international symposium on information theory (Ed. by B.N. Petrov & F. Csaki), pp.
267281. Akademiai Kiado, Budapest.
BATELLI C. 2004. Phytocoenological study of the Catenella
caespitosa (Withering) L. M. Irvine (Gigartinales, Rhodophyceae)
community from the Slovenian coast, northern Adriatic Sea. Acta
Adriatica 45: 121130.
BENINGER P.G., BOLDINA I. & KATSANEVAKIS S. 2012. Strengthening
statistical usage in marine ecology. Journal of Experimental
Marine Biology and Ecology 426427: 97108.
BIRD K.T. 1976. Simultaneous assimilation of ammonium and
nitrate by Gelidium nudifrons (Gelidiales: Rhodophyta). Journal
of Phycology 12: 238241.
BRACKEN M.E.S., JONES E. & WILLIAMS S.L. 2011. Herbivores, tidal
elevation, and species richness simultaneously mediate nitrate
uptake by seaweed assemblages. Ecology 92: 10831093.
BURNHAM K.P. & ANDERSON D.R. 2002. Model selection and
multimodel inference: a practical information-theoretic approach,
2nd Edition. Springer, New York. 488 pp.
CAMPBELL S.J. 1999. Uptake of ammonium by four species of
macroalgae in Port Phillip Bay, Victoria, Australia. Marine and
Freshwater Research 50: 515522.
C HAPMAN A.R.O. & JOHNSON C.R. 1990. Disturbance and
organization of macroalgal assemblages in the Northwest
Atlantic. Hydrobiologia 192: 77121.
CLAVERO V., FERNANDEZ J.A. & NIELL F.X. 1997. The inuence of
tidal regime on phosphate accumulation in a shallow small
estuary (Palmones River, southern Spain). Scientia Marina 61:
3743.
CONNELL J.H. 1972. Community interactions on marine rocky
intertidal shores. Annual Review of Ecology and Systematics 3:
169192.
CONOLLY N.J. & DREW E.A. 1985. Physiology of Laminaria IV.
Nutrient supply and daylength, major factors affecting growth of
L. digitata and L. saccharina. Marine Ecology 6: 299320.
CORZO A. & NIELL F.X. 1991. Determination of nitrate reductaseactivity in Ulva rigida C. Agardh by the in situ method. Journal of
Experimental Marine Biology and Ecology 146: 181191.
DAVEY A. & WOELKERLING W.J. 1985. Studies on Australian
mangrove algae. III. Victorian communities: structure and
recolonization in western port bay. Journal of Experimental
Marine Biology and Ecology 85: 177190.
DAVISON I.R. & PEARSON G.A. 1996. Stress tolerance in intertidal
seaweeds. Journal of Phycology 32: 197211.
DELIA C.F. & DEBOER J.A. 1978. Nutritional studies of two red
algae. II. Kinetics of ammonium and nitrate uptake. Journal of
Phycology 14: 266272.
DRING M.J. & BROWN F.A. 1982. Photosynthesis of intertidal brown
seaweeds during and after periods of emersion: a renewed search
for physiological causes of zonation. Marine Ecology Progress
Series 8: 301308.
DY D.T. & YAP H.T. 2001. Surge ammonium uptake of the cultured
seaweed, Kappaphycus alvarezii (Doty) Doty (Rhodophyta:
Gigartinales). Journal of Experimental Marine Biology and
Ecology 265: 89100.
FERNANDEZ J.A., NIELL F.X. & LUCENA J. 1985. A rapid and
sensitive automated determination of phosphate in natural
waters. Limnology and Oceanography 30: 227230.
FUJITA R.M. 1985. The role of nitrogen status in regulating transient
ammonium uptake and nitrogen storage by macroalgae. Journal
of Experimental Marine Biology and Ecology 92: 283301.
FUJITA R.M., WHEELER P.A. & EDWARDS R.L. 1988. Metabolic
regulation of ammonium uptake by Ulva rigida (Chlorophyta): a
compartmental analysis of the rate-limiting step for uptake.
Journal of Phycology 24: 560566.
GORDILLO F.J.L., DRING M.J. & SAVIDGE G. 2002. Nitrate and
phosphate uptake characteristics of three species of brown algae
cultured at low salinity. Marine Ecology-Progress Series 234: 111
118.

470

Phycologia, Vol. 52 (6), 2013

HAINES K.C. & WHEELER P.A. 1978. Ammonium and nitrate uptake
by the marine macrophytes Hypnea musciformis (Rhodophyta)
and Macrocystis pyrifera (Phaeophyta). Journal of Phycology 14:
319324.
HARRISON P.J. & HURD C.L. 2001. Nutrient physiology of seaweeds:
application of concepts to aquaculture. Cahiers De Biologie
Marine 42: 7182.
HARRISON P.J., DRUEHL L.D., LLOYD K.E. & THOMPSON P.A. 1986.
Nitrogen uptake kinetics in three year-classes of Laminaria
groenlandica (Laminariales: Phaeophyta). Marine Biology 93:
2935.
HARRISON P.J., PARSLOW J.S. & CONWAY H.L. 1989. Determination
of nutrient uptake kinetic parameters. A comparison of methods.
Marine Ecology Progress Series. 52: 301312.
HAYS C.G. 2007. Adaptive phenotypic differentiation across the
intertidal gradient in the alga Silvetia compressa. Ecology 88: 149
157.
HURD C.L. & DRING M.J. 1990. Phosphate uptake by intertidal
algae in relation to zonation and season. Marine Biology 107:
281289.
HURD C.L. & DRING M.J. 1991. Desiccation and phosphate uptake
by intertidal fucoid algae in relation to zonation. British
Phycological Journal 26: 327333.
IZQUIERDO J.J. 2001. Aspectos del ciclo de los nutrientes no
conservativos en el estuario del ro Palmones. Ph.D. dissertation.
University of Malaga, Spain. 231 pp.
KAREZ R. 2003. Competitive ranks of three Fucus spp. (Phaeophyta)
in laboratory experiments testing of Keddys competitive
hierarchy model. Helgoland Marine Research 57: 8390.
KENNISON R.L., KAMER K. & FONG P. 2011. Rapid nitrate uptake
rates and large short-term storage capacities may explain why
opportunistic green macroalgae dominate shallow eutrophic
estuaries. Journal of Phycology 47: 483494.
KIM J.K., KRAEMER G.P. & YARISH C. 2008. Physiological activity
of Porphyra in relation to eulittoral zonation. Journal of
Experimental Marine Biology and Ecology 365: 7585.
KRAEMER G.P., CARMONA R., CHOPIN T., NEEFUS C., TANG X. &
YARISH C. 2004. Evaluation of the bioremediatory potential of
several species of the red alga Porphyra using short-term
measurements of nitrogen uptake as a rapid bioassay. Journal
of Applied Phycology 16: 489497.
KRISTENSEN E. & ANDERSEN F.. 1987. Determination of organic
carbon in marine sediments: a comparison of two CHN-analyzer
methods. Journal of Experimental Marine Biology and Ecology
109: 1523.
LAVERY P.S. & MCCOMB A.J. 1991. The nutritional eco-physiology
of Chaetomorpha linum and Ulva rigida in Peel Inlet, Western
Australia. Botanica Marina 34: 251260.
LEWIS J.R. 1964. The ecology of rocky shores. English University
Press, London. 323 pp.
L OBBAN C.S. & HARRISON P.J. 1994. Seaweed ecology and
physiology. Cambridge University Press, Cambridge. 366 pp.
LOTZE H.K. & SCHRAMM W. 2000. Ecophysiological traits explain
species dominance patterns in macroalgal blooms. Journal of
Phycology. 36: 287295.
LUO M.B., LIU F. & XU Z.L. 2012. Growth and nutrient uptake
capacity of two co-occurring species, Ulva prolifera and Ulva
linza. Aquatic Botany 100: 1824.
MANN F.D. & STEINKE T.D. 1988. Photosynthetic and respiratory
responses of the mangrove-associated red algae, Bostrychia
radicans and Caloglossa leprieurii. South African Journal of
Botany 54: 203207.
MARTINEZ B. & RICO J.M. 2004. Inorganic nitrogen and phosphorus
uptake kinetics in Palmaria palmata (Rhodophyta). Journal of
Phycology 40: 642650.
MCGLATHERY K.J., PEDERSEN M.F. & BORUM J. 1996. Changes in
intracellular nitrogen pools and feedback controls on nitrogen
uptake in Chaetomorpha linum (Chlorophyta). Journal of
Phycology 32: 393401.
MELVILLE F. & PULKOWNIK A. 2007. Seasonal and spatial variation
in the distribution of mangrove macroalgae in the Clyde River,
Australia. Estuarine, Coastal and Shelf Science 71: 683690.
MELVILLE F., PULKOWNIK A. & BURCHETT M. 2005. Zonal and
seasonal variation in the distribution and abundance of mangrove

macroalgae in the Parramatta River, Australia. Estuarine, Coastal

and Shelf Science 64: 267276.
Ministerio De Defensa. 2010. Anuario de mareas 2011 que
comprende la Pennsula Iberica, Ceuta, Islas Canarias, Tanger,
Ifni, Sahara occidental y Antartica. Ministerio de Defensa,
Secretaria General Tecnica, Spain. 447 pp. I.S.S.N. 0213 1897.
MURPHY J. & RILEY J.P. 1962. A modied single solution method
for the determination of phosphate in natural waters. Analytica
Chimica Acta 27: 3136.
A. & PALOMO L. 2005. Palmones
NIELL F.X., CLAVERO V., AVILES
River-Spain. In: Modeling nutrients loads and response in river and
estuary systems (Ed. by R.C. Russo & R. Lekevicius). NATO/
CCMS Pilot Study. 271: 911.
PARDO P., SANCHEZ J.F.L., RAURET G., RUBAN V., MUNTAU H. &
QUEVAUVILLER P. 1999. Study of the stability of extractable
phosphate content in a candidate reference material using a
modied Williams extraction procedure. Analyst 124: 407411.
PEDERSEN M.F. 1994. Transient ammonium uptake in the macroalga Ulva lactuca (Chlorophyta): nature, regulation, and the
consequences for choice of measuring technique. Journal of
Phycology 30: 980986.
PEDERSEN M.F. & BORUM J. 1997. Nutrient control of estuarine
macroalgae: growth strategy and the balance between nitrogen
requirements and uptake. Marine Ecology Progress Series 161:
155163.

E.J., ZINGMARK R. & NIETCH C. 1999. Comparative

PENA
photosynthesis of two species of intertidal epiphytic macroalgae
on mangrove roots during submersion and emersion. Journal of
Phycology 35: 12061214.
PHILLIPS A., LAMBERT G., GRANGER J.E. & STEINKE T.D. 1996.
Vertical zonation of epiphytic algae associated with Avicennia
marina (Forssk.) Vierh. pneumatophores at Beachwood Mangroves Nature Reserve, Durban, South Africa. Botanica Marina
39: 167176.
PHILLIPS J.C. & HURD C.L. 2003. Nitrogen ecophysiology of
intertidal seaweeds from New Zealand: nitrogen uptake, storage
and utilisation in relation to shore position and season. Marine
Ecology-Progress Series 264: 3148.
PHILLIPS J.C. & HURD C.L. 2004. Kinetics of nitrate, ammonium,
and urea uptake by four intertidal seaweeds from New Zealand.
Journal of Phycology 40: 534545.
REES T.A.V. 2003. Safety factors and nutrient uptake by seaweeds.
Marine Ecology Progress Series 263: 2942.
REES T.A.V., DOBSON B.C., BIJL M. & MORELISSEN B. 2007. Kinetics
of nitrate uptake by New Zealand marine macroalgae and
evidence for two nitrate transporters in Ulva intestinalis L.
Hydrobiologia 586: 135141.
ROSENBERG G. & RAMUS J. 1984. Uptake of inorganic nitrogen and
seaweed surface area: volume ratios. Aquatic Botany 19: 6572.
ROSENBERG G., PROBYN T.A. & MANN K.H. 1984. Nutrient uptake
and growth kinetics in brown seaweeds: response to continuous
and single additions of ammonium. Journal of Experimental
Marine Biology and Ecology 80: 125146.
RUNCIE J.W., RITCHIE R.J. & LARKUM A.W.D. 2003. Uptake
kinetics and assimilation of inorganic nitrogen by Catenella nipae
and Ulva lactuca. Aquatic Botany 76: 155174.
RUNCIE J.W., RITCHIE R.J. & LARKUM A.W.D. 2004. Uptake
kinetics and assimilation of phosphorus by Catenella nipae and
Ulva lactuca can be used to indicate ambient phosphate
availability. Journal of Applied Phycology 16: 181194.
RYTHER J.H., CORWIN N., DEBUSK T.A. & WILLIAMS L.D. 1981.
Nitrogen uptake and storage by the red alga Gracilaria tikvahiae
(McLachlan, 1979). Aquaculture 26: 107115.
SHARP J.H., CULBERSON C.H. & CHURCH T.M. 1982. The chemistry
of the Delaware estuary. General considerations. Limnology and
Oceanography 27: 10151028.
SHINN J.A. 1941. Ind. Eng. Chem. (Annual edition), 13:33. In: A
practical approach handbook of seawater analysis (Ed. by J.D.H.
Strickland and T.R. Parson). Ottawa. 310 pp.
SKENE K.R. 2004. Key differences in photosynthetic characteristics
of nine species of intertidal macroalgae are related to their
position on the shore. Canadian Journal of Botany-Revue
Canadienne De Botanique 82: 177184.

Sanchez de Pedro et al.: Differential nutrient uptake by two estuarine macroalgae

SLAWYK G. & MACISAAC J.J. 1972. Comparison of two automated
ammonium methods in a region of coastal upwelling. Deep Sea
Research and Oceanographic Abstracts 19: 521524.
S YRETT P.J. 1981. Nitrogen metabolism of microalgae. In:
Physiological bases of phytoplankton ecology (Ed. By T. Platt).
Canadian Journal of Fisheries and Aquatic Sciences 210: 182209.
THOMAS T.E. & HARRISON P.J. 1987. Rapid ammonium uptake and
nitrogen interactions in ve intertidal seaweeds grown under eld
conditions. Journal of Experimental Marine Biology and Ecology
107: 18.
THOMAS T.E. & TURPIN D.H. 1980. Desiccation enhanced nutrient
uptake rates in the intertidal alga Fucus distichus. Botanica
Marina 23: 479481.
THOMAS T.E., TURPIN D.H. & HARRISON P.J. 1987. Desiccation
enhanced nitrogen uptake rates in intertidal seaweeds. Marine
Biology 94: 293298.
WILLIAMS S.L. 1984. Uptake of sediment ammonium and translocation in a marine green macroalga Caulerpa cupressoides.
Limnology and Oceanography 29: 374379.

471

WOOD E.D., ARMSTRONG F.A.J. & RICHARDS F.A. 1967. Determination of nitrate in sea water by cadmium-copper reduction to
nitrite. Journal of the Marine Biological Association of the United
Kingdom 47: 2331.
YOUNG E.B., BERGES J.A. & DRING M.J. 2009. Physiological
responses of intertidal marine brown algae to nitrogen deprivation and resupply of nitrate and ammonium. Physiologia
Plantarum 135: 400411.
ZUCCARELLO G.C., YEATES P.H., WRIGHT J.T. & BARTLETT J. 2001.
Population structure and physiological differentiation of haplotypes of Caloglossa leprieurii (Rhodophyta) in a mangrove
intertidal zone. Journal of Phycology 37: 235244.

Received 19 February 2013; accepted 26 June 2013

Associate Editor: Allen J. Milligan