J Appl Phycol (2013) 25:1331–1340
DOI 10.1007/s10811-013-9989-3
Compensatory abilities depending on seasonal timing
of thallus excision of the kelp Undaria pinnatifida cultivated
in Matsushima Bay, northern Japan
Xu Gao & Hikaru Endo & Makoto Yamana &
Kazuya Taniguchi & Yukio Agatsuma
Received: 15 November 2012 / Revised and accepted: 10 January 2013 / Published online: 23 January 2013
# Springer Science+Business Media Dordrecht 2013
Abstract Improved cultivation technology for the kelp
Undaria pinnatifida is greatly needed to increase production
to meet increasing commercial demand. A previous cultiva-
tion trial indicated that the crop yield of U. pinnatifida
sporophytes could be increased greatly by thallus excision
in late February due to compensatory growth of the remain-
ing tissues. To develop this potential new cultivation tech-
nology, it is essential to identify the time period during
which this kelp can compensate and its physiological
responses to thallus excision. In this study, U. pinnatifida
sporophytes were excised at about 30 cm above the meri-
stem at the beginning of January, February, March, and
April, respectively. Morphological features, photosynthesis,
nutrient uptake, and carbon and nitrogen contents of excised
kelps were measured and compared with these parameters in
control kelps grown without excision. Both experimental
and control kelps were farmed together in Matsushima
Bay, northern Japan. The kelps excised in January and
February showed significant increases in the lengths and
dry weights of the blade, photosynthetic rates, nutrient up-
take rates, and carbon and nitrogen contents compared with
the control kelps, and the growth phase was prolonged for at
least 1 month. No significant increases were found in dry
weights and carbon and nitrogen contents of sporophylls
until early April, which indicated that the maturation period
was delayed. At the end of this experiment, the nitrogen
contents of sporophyll tissues formed after excisions were
significantly lower than those of tissues formed before exci-
sions. In contrast, the kelps excised in March and April
showed no significant increases in morphological and
X. Gao : H. Endo : M. Yamana : K. Taniguchi : Y. Agatsuma (*)
Laboratory of Marine Plant Ecology, Graduate School of
Agricultural Science, Tohoku University, Sendai, Miyagi
Prefecture 981-8555, Japan
e-mail: agatsuma@bios.tohoku.ac.jp
physiological parameters compared with control kelps.
These results suggest that U. pinnatifida sporophytes
exhibited great compensation when excisions were con-
ducted during the growing phase in January and February
but not in March and April when the maturation phase had
started. The regulation of resource allocation to growth and
maturation after thallus excisions in January and February
likely results in prolongation of the growth phase and mat-
uration phase in excised kelps.
Keywords Compensation . Cultivation . Thallus excision .
Resource allocation . Undaria pinnatifida . Phaeophyta
Introduction
Compensatory growth appears to be a common tolerance
strategy of terrestrial and aquatic plants against damages
(Strauss and Agrawal 1999; Stowe et al. 2000; Thomson
et al. 2003; Vergés et al. 2008; Cerda et al. 2009). Plants
have evolved a diverse suite of tolerance traits including
compensatory growth, increasing photosynthesis, activation
of dormant meristems, and changes in reproductive strate-
gies and resource allocation patterns (Strauss and Agrawal
1999; Tiffin 2000). The ability to tolerate damages is known
to be influenced by the timing of the damage (Maschinski
and Whitham 1989; Trumble et al. 1993; Paige 1994;
Simons and Johnston 1999) or, more precisely, by the de-
velopmental stage at which damage occurs (Bergelson et al.
1996). Many higher plants are more likely to compensate for
damage that occurs early in the growing season, before the
reproduction phase has started (Maschinski and Whitham
1989; Lehtila and Syrjänen 1995; Lennartsson et al. 1998).
In seaweeds, experimental studies of compensation are
rare, and little is known about how these plants compensate
1332
for damages and how damage may affect future fitness by
altering their internal organization. Although some research-
ers have given increasing attention to the compensatory
ability of seaweeds in recent years (Littler et al. 1995;
Steinberg 1995; Honkanen and Jormalainen 2002; Vergés
et al. 2008; Cerda et al. 2009; Hay et al. 2011), no studies
have focused on the effect of the timing of damage on
compensation. Likewise, little information is available about
the compensatory ability of large commercial species and its
possible application to increase aquaculture production.
The annual kelp Undaria pinnatifida (Harvey) Suringar
is one of the most important and popular commercial sea-
weeds endemic to cold temperate coastal regions of Japan
(Akiyama and Kurogi 1982). As an invasive species, U.
pinnatifida has spread to other coastal waters of the world
where populations are now well established (Sanderson
1990; Casas and Piriz 1996; Campbell et al. 1999; Silva et
al. 2002; Aguilar-Rosas et al. 2004). This kelp has been
extensively cultivated on a large scale in Asian countries
(Yamanaka and Akiyama 1993), and it was intentionally
introduced recently for commercial cultivation in European
countries due to its excellent taste and nutritional value (e.g.,
ICES 2007; Peteiro and Freire 2011). In Japan, the crop
yield of this species has not been able to satisfy increasing
commercial demand in recent years. Therefore, the devel-
opment of new cultivation technology that can increase
production is greatly needed.
A previous cultivation trial reported that U. pinnatifida
sporophytes excised in late February showed marked com-
pensatory growth compared with control kelps without ex-
cision, and therefore, the harvest period was prolonged (Gao
et al. 2012). To improve this potential new cultivation tech-
nology, it is essential to identify the time period during
which this kelp can compensate for thallus excision and
the physiological responses that support compensatory
growth. Gao et al. (2012) also indicated that the maturation
period of this kelp was delayed markedly after excision due
to the elongation of its growth phase. Therefore, the alloca-
tion of resources to growth and maturation may have
changed after thallus excision, but this hypothesis requires
testing.
In the present study, we examined seasonal changes of
morphological features of farmed U. pinnatifida sporo-
phytes excised during the growing phase (January and
February) and maturation phase (March and April) to iden-
tify the time period during which this kelp can compensate
for excision. In addition, physiological parameters (photo-
synthesis, nutrient uptake, and resource contents) of the
kelps excised at different times were measured and com-
pared with those of control kelps grown without excision.
Changes in resource allocation between growth and
maturation in response to thallus excisions were also
investigated.
J Appl Phycol (2013) 25:1331–1340
Materials and methods
Seedling production and cultivation trial
Gametophytes of U. pinnatifida were obtained by induction of
zoospores from parental plants farmed in Matsushima Bay
(38°32′ N, 141°31′ E) in the early summer of 2008. After
zoospores were released into tanks filled with filtered seawater,
the water was stirred to a homogeneous consistency to make
the zoospores attach to 2-mm diameter plastic fiber threads at
similar initial settlement densities. The seedlings culture was
first conducted indoors under controlled environmental con-
ditions (18 °C, irradiance of 40 μmol photons m−2 s−1, and a
photoperiod of 12:12 h light/dark). The culture medium con-
sisted of sterilized seawater with additional nitrate (PESI;
Tatewaki 1966), to which GeO2 (20 μg mL−1) was added to
inhibit the growth of protozoa and diatoms. Aeration was used
to mix the nitrate and seawater evenly. The medium was
renewed every 15 days. After several months, the seedlings
grew to 1–2 cm long (on average). The threads on which the
young sporophytes had attached were cut into 4-cm sections.
For field cultivation trials, 50 thread sections were individually
tucked into five 50-m long, 15-mm diameter ropes at intervals
of 1 m. Five ropes prepared in this manner were deployed at
the sea surface with floats at intervals of 3 m in Matsushima
Bay in October. In order to standardize the cultivation con-
ditions, cultivation density was adjusted to 15 plants per thread
section in late December. One 4-cm section was considered to
be one bunch. For experimental groups, thalli were excised at
about 30 cm above the meristem at the beginning of January,
February, March, and April, respectively (Fig. 1). For the
control group, no excision was conducted during cultivation.
Measurements of morphological features
To examine the changes in morphological features, three
bunches of each of the four experimental groups were collect-
ed randomly every 2 weeks beginning 2 weeks after thallus
excisions and continuing into late May. Three bunches of
control kelps were collected every 2 weeks from early
January to late May. These plants were immediately trans-
ported to the laboratory and then were initially hung for at
least 15 min to drain excess seawater. Next, measurements of
blade length were taken from 15 plants in sequence starting
with the largest one. Plants then were carved into midrib,
blade, and sporophyll, and each part was weighed after drying
in a convection oven (Yamato DX400, Yamato Scientific Co.)
at 60 °C for 1 week.
Measurements of photosynthesis and nutrient uptake
One bunch from each experimental group was collected
2 weeks after each excision, and one bunch from the control
J Appl Phycol (2013) 25:1331–1340
Fig. 1 Picture of mature U. pinnatifida sporophyte showing exci-
sion site
group was collected at the same time. The eight largest
plants on each bunch were used to measure photosynthesis
and nutrient uptake. Five discs with a surface area of about
2.0 cm2 were cut from each plant using a cork borer (1.6 cm
in diameter) from the margins of the meristem. These discs
were incubated in flowing seawater for 12 h before measure-
ments, to allow recovery from cutting damage.
An improved differential gas-volumeter called “Product–
meter” (Yokohama et al. 1986) was used to measure photo-
synthesis and nutrient uptake. The discs were placed in a
reaction vessel containing 10 mL of filtered seawater (GF/F,
Whatman,, UK) while the control vessel contained only
10 mL of filtered seawater. The reaction and control vessels
were attached to Product-meters, immersed in a water bath
with a thermostat (Taitec CL–150 F, Koshigaya, Japan) to
maintain a constant water temperature and shaken using a
motor drive at 150 rpm min−1. The vessels were illuminated
from below by projector lamps with light reflected by mir-
rors placed under the water bath. Photo slide projectors
(Elmo S–300, Japan) with incandescent lamps (Philips
KP–10s 100V, 300 W; Japan) were used as the light
source. Irradiances were regulated by using neutral den-
sity glass filters (Toshiba TND–50, 25, 12.5, Japan) and
measured with a quantum photon meter (LI–COR LI–
192S, USA).
To measure photosynthetic rate, the discs were incubated
at six temperatures (5, 10, 15, 20, 25, and 30 °C) with a
photon flux intensity of 180 μmol photons m−2 s−1. The
initial nutrient concentrations of the sterilized seawater were
set to 400 μg L−1 for NO3–N and NH4–N, and 80 μg L−1
for PO4–P. The nutrient concentrations in seawater were
almost below the limit of detection. The discs were incubat-
ed for 30 min to allow them to adapt to the experimental
1333
Table 1 The four initial concentrations (μg L−1) designed of NH4–N,
NO3–N, and PO4–P
Combination NH4–N NO3–N PO4–P
1 0 400 80
2 100 400 80
3 200 400 80
4 400 400 80
environments, and the oxygen evolution they produced was
then measured every 3 min six to ten times. Eight replicates
were used for photosynthesis measurement.
The uptake rates of NO3–N and NH4–N were obtained
from the differences between the concentrations analyzed by
the auto-analyzer (TRAACS 800; Bran–Luebbe, Japan) af-
ter a 60-min incubation of the discs and those in the con-
trols. Eight replicates were used for nutrient uptake
measurements. To examine the effect of thallus excision
on nutrient uptake rates more closely, the initial concentra-
tions of NH4–N were set to four levels (Table 1) and mea-
sured respectively. After incubations, the fresh and dry
weights were measured. Nutrient uptake rates were estimat-
ed using the following equation:
VN; P ¼ ðSt  StþT Þ  0:01=ðA  T Þ
where VN,P =uptake rates of NO3–N and NH4–N (micrograms
per square centimeter per hour); t=time (hours); T=time in-
terval (hours); St =initial concentration at time t (micrograms
per liter); St+T =final concentration at time t+T (micrograms
per liter); and A=area of the disc (2.0 cm2).
Analysis of carbon and nitrogen contents
One bunch from each experimental group was collected
4 weeks after each excision, and one bunch from the
control group was collected at the same time. The eight
largest plants on each bunch were used to analyze
carbon and nitrogen contents of blade tissues. One disc
with a surface area of about 2.0 cm2 was cut from each
plant using a cork borer (1.6 cm in diameter) from the
margin of the meristem.
One bunch from each experimental group in addition to
those for morphological measurements was collected every
2 weeks beginning 2 weeks after each excision date and
continuing until late May, and one bunch of control kelp
was collected every 2 weeks from early January to late May.
The eight largest plants on each bunch were selected to
determine the seasonal changes of carbon and nitrogen
contents of sporophyll tissues. One disc with a surface area
of about 2.0 cm2 was cut from each plant using a cork borer
1334 J Appl Phycol (2013) 25:1331–1340

(1.6 cm diameter) from the middle part of sporophyll with
the maximum width.
On the final day of this experiment, one bunch from
each experimental group that had never been previously
sampled was collected, and the eight largest plants on
each bunch were used to compare the carbon and nitro-
gen contents of sporophyll tissues formed before and
after excision. One disc with a surface area of about
2.0 cm2 from each part was cut from each plant using a
cork borer (1.6 cm diameter).
After drying, the dry weight of each sample was
measured, and the discs then were mashed up and
analyzed for carbon and nitrogen contents using an
Elemental Analyzer (EAGER200, Fisons Instruments,
USA).
Statistical analysis
Differences in morphological features and carbon and nitro-
gen contents of sporophyll tissues among months were ana-
lyzed using one-way analysis of variance (ANOVA). All of
the variables were tested for normality and homogeneity prior
to conducting ANOVA. When a significant difference was
identified by ANOVA, Tukey’s multiple comparison test was
used for comparison of means. Significant differences in
morphological and physiological features between each ex-
perimental group and the control, and significant differences
in carbon and nitrogen contents between sporophyll tissues
formed before and after excision were analyzed using
the Student’s t test. Differences were considered to be
significant at a probability of 5 % (P<0.05).

Table 2 Seasonal blade length,

dry weights of blade, and sporo- Time C JE FE ME AE
phyll of U. pinnatifida sporo-
phytes excised in January, Blade length, cm
February, March, April, and Early January 89.5±10.7a
control kelps cultivated in
Late January 117.4±16.8b 58.4±12.6a*
Matsushima Bay
Early February 146.2±14.3c 75.9±11.3b*
Late February 143.1±15.5c 109.3±14.4c* 61.5±12.6a*
Early March 141.0±14.1c 139.7±12.2d 91.0±13.5b*
Late March 142.4±12.4c 138.7±15.4d 120.6±14.6c* 29.3±5.3a*
Early April 139.5±13.3c 135.8±10.8d 117.9±13.3c* 27.9±5.4a*
Late April 122.9±11.2b 116.0±11.5c 100.1±10.7b* 24.6±4.3a* 29.6±4.4a*
Early May 120.1±13.6b 113.3±14.8c 93.8±13.1b* 25.7±3.1a* 26.7±3.4a*
Late May 115.9±11.5b 109.1±16.7c 95.5±13.8b* 22.1±4.5a* 23.5±4.1a*
Dry weights of blade, g
Early January 20.4±3.7a
Late January 31.0±5.8b 19.4±2.5a*
Early February 45.7±5.3c 31.5±4.0b*
Late February 60.7±6.9d 44.8±5.4c* 18.2±3.1a*
Early March 56.8±5.2d 56.5±5.3d 30.7±4.3b*
Late March 58.3±5.8d 60.3±6.6d 42.5±4.8c* 10.0±1.3a*
Early April 57.2±5.3d 57.7±4.3d 45.0±4.1c* 11.5±2.0a*
Late April 43.8±5.0c 42.1±4.1c 31.6±3.0b* 9.6±1.2a* 10.5±1.1a*
Early May 45.2±4.6c 45.9±4.8c 29.7±3.8b* 10.3±1.4a* 9.8±1.2a*
Late May 42.8±4.1c 40.4±5.4c 27.9±3.5b* 7.4±1.5a* 7.1±1.0a*
Dry weights of sporophyll, g
Early January 5.7±0.67a
Late January 6.0±0.58a 4.8±0.56a
Data are the means±SE. Values Early February 10.2±1.73ab 5.9±0.63a
with different letters in the same Late February 14.8±2.59b 6.4±0.43a* 9.9±0.56a
column are significantly differ-
Early March 15.5±2.53b 6.1±0.23a* 10.9±1.45a
ent from each other (P<0.05);
N=15 for all kelps Late March 29.8±3.48c 7.2±0.92a* 9.8±0.68a* 23.6±2.56a
c b a
JE January, FE February, ME Early April 26.9±4.34 15.8±2.83 * 9.5±0.71 * 26.8±2.49a
March, AE April, C control kelps Late April 25.1±3.52c 23.1±3.45c 20.0±2.27b 24.4±3.33a 24.0±2.45a
*P < 0.05 indicates difference Early May 27.7±3.36c 33.3±4.58d 31.7±3.71c 25.7±2.41a 22.3±2.40a
level between each excised kelp Late May 24.9±4.51c 28.5±2.29cd 29.4±2.58c 21.5±2.50a 20.1±1.93a
and control
J Appl Phycol (2013) 25:1331–1340 1335

70
January 146.2 cm in early February (P<0.05). No significant changes
60 * in blade length were found from early February to early April
*
50 (P>0.05), but it decreased significantly thereafter (P<0.05).
* *
40 * The blade length of kelps excised in January increased signif-
30 icantly from 58.4 cm in late January to 139.7 cm in early
20 March (P<0.05). From late January to late February, blade
10 length differed significantly between kelps excised in January
and control kelps (P<0.05), but significant differences were
0
not detected from early March to late May (P>0.05). The
70
February blade length of kelps excised in February increased signifi-
60
*
* cantly from 61.5 cm in late February to 120.6 cm in late March
50 * (P<0.05), but they had significantly lower values than those
*
40 * of the control kelps until the end of the experiment (P<0.05).
Oxygen evolution (µLO2 cm–2 h–1)

30 No significant variations occurred in blade lengths of kelps

20 excised in March and April (P>0.05), and excised kelps had
10 significantly lower values than control kelps until the end of
0 the experiment (P<0.05).
70
The dry weight of blade of control kelps increased signif-
March icantly from 20.4 g in early January to 60.7 g in late February
60
(P<0.05). No significant changes in blade weight occurred
50 from late February to early April (P>0.05), but it decreased
40 significantly thereafter (P<0.05). The dry weight of blades of
30 kelps excised in January increased significantly from 19.4 g in
20 late January to 60.3 g in late March (P<0.05). From late
10 January to late February, blade weight differed significantly
between kelps excised in January and control kelps (P<0.05),
0
but this difference did not exist from early March to late May
70
April (P>0.05). The dry weight of blades of kelps excised in
60 February increased significantly from 18.2 g in late February
50 to 45.0 g in early April (P<0.05), but the weights were
40 significantly lower than those of the control kelps until the
30
end of the experiment (P<0.05). In contrast, no significant
changes were found in kelps excised in March and April
20
(P>0.05), and excised kelps had significantly lower values
10 than control kelps until the end of the experiment (P<0.05).
0 The dry weight of sporophylls of control kelps increased
0 5 10 15 20 25 30 35 significantly from 5.7 g in early January to 29.8 g in late March
Temperature ( C) (P<0.05), but no significant variation occurred thereafter
(P>0.05). There was little change in dry weight of sporophyll
Fig. 2 Gross photosynthetic rates of blade discs taken from near the tissues of kelps excised in January from late January to late
meristem in each excised kelp (open circles) and control kelp (filled
circles) at different temperatures 2 weeks after excisions. Asterisks March (P>0.05), but dry weights increased significantly from
indicate a significant difference at P<0.05 between excised and control 7.2 g in late March to 33.3 g in early May (P<0.05). From late
kelps. N=8 for all kelps. Black bars indicate standard deviations February to early April, sporophyll weight differed significant-
ly between kelps excised in January and control kelps
(P<0.05), but such differences were not found from late
Results April to late May (P>0.05). Similarly, few changes in dry
weight of sporophyll of kelps excised in February occurred
Morphological features from late February to early April (P>0.05). However, the kelps
excised in February showed a significant increase in dry
Seasonal morphological features in all excised and control weight of sporophylls from 9.5 g in early April to 31.7 g in
kelps are shown in Table 2 (seasonal blade length, dry weights early May (P<0.05). In late March and early April, sporophyll
of blade, and sporophyll). The blade length of control kelps weight differed significantly between kelps excised in
increased significantly from 89.5 cm in early January to February and control kelps (P < 0.05), but significant
1336 J Appl Phycol (2013) 25:1331–1340

2
NH4 N January Febuary March April
1.5 *
* *
*
* *
1
Uptake rates (µg L–1 cm–2 h–1)

0.5

2
NO3 N
1.5
*
* * * * *
* *
1

0.5

0
0 100 200 400 0 100 200 400 0 100 200 400 0 100 200 400
Initial concentration of NH4–N (µg L–1)

Fig. 3 Uptake rates of NH4–N and NO3–N of blade discs taken from 2 weeks after excisions. Asterisks indicate a significant difference at
near the meristem in each excised kelp (open columns) and control kelp P<0.05 between excised and control kelps. N=8 for all kelps. Black
(filled columns) at four different initial concentrations of NH4–N bars indicate standard deviations

differences did not exist from late April to late May (P>0.05).
In contrast, no significant changes were detected in dry weights 2.5
of sporophylls of kelps excised in March and April (P>0.05). Carbon
2
Photosynthesis and nutrient uptake *
*
1.5
Two weeks after excisions, the gross photosynthetic rates of
blade tissues of kelps excised in January and February were
Resource contents (mg cm–2)

1
significantly higher at temperatures of 10–30 °C than those
of control kelps (P<0.05) (Fig. 2). However, no significant 0.5
differences in gross photosynthetic rates were observed at
each temperature in kelps excised in March and April com- 0
pared with the control kelps (P>0.05).
The uptake rates of NH4–N increased with an increase in 0.25
Nitrogen
initial concentration of NH4–N in all excised and control kelps *
(Fig. 3). The uptake rates of NH4–N in kelps excised in *
0.2
January and February were significantly higher than those of
the control kelps at initial concentrations of 100–400 μg L−1 0.15
(P<0.05). Similarly, the uptake rates of NO3–N in kelps
excised in January and February at all initial concentrations 0.1
of NH4–N were significantly higher than those of the control
kelps (P<0.05). In contrast, the uptake rates of both NH4–N 0.05
and NO3–N of kelps excised in March and April were lower
than those of the control kelps (P>0.05). 0
JE FE ME AE
Carbon and nitrogen contents Fig. 4 Carbon and nitrogen contents of blade discs taken from near the
meristem in each excised kelp (open columns) and control kelp (filled
Four weeks after excisions, the carbon and nitrogen contents columns) 4 weeks after excisions. Asterisks indicate a significant
difference at P<0.05 between excised and control kelps. N=8 for all
of blade tissues of kelps excised in January and February kelps. Black bars indicate standard deviations. JE, FE, ME, and AE
were significantly higher than those of control kelps represent four experimental kelps excised at the beginning of January,
(P<0.05) (Fig. 4). However, no significant differences were February, March, and April, respectively
J Appl Phycol (2013) 25:1331–1340 1337

7 0.8

c C
6 c C 0.7 c C
c C

Nitrogen content (mg cm–2)

c
Carbon content (mg cm–2)

C c
c c
C

b b B
A* A*
5 A* A* 0.6
b A* B*
A* A* A*
B* A* B A*
B* A*
A*
a a B ab
a ab
A* A* c A* A
A* a A A A* a
4 A A B 0.5 a
A* B A* A
A* A A A* B
A* A* A* B

3 0.4
Jan. Feb. Mar. Apr. May Jan. Feb. Mar. Apr. May

Fig. 5 Seasonal carbon and nitrogen contents of sporophylls of U. large letters indicate a significant difference at P<0.05 among
pinnatifida excised at the beginning of January (filled squares), months. Asterisks indicate a significant difference at P<0.05 be-
February (open squares), March (filled triangles), April (open tween excised and control kelps. N=8 for all kelps. Black bars
triangles), and control kelps (open circles). Different small and indicate standard deviations

found between control kelps and kelps excised in March and 0.5
Carbon
April (P>0.05).
The carbon and nitrogen contents of sporophylls of 0.4
control kelps increased significantly from early January
to late March (P<0.05), and then no significant varia- 0.3
tions occurred until early May (P>0.05) (Fig. 5). From
late January to late March, no significant changes in 0.2
Resource contents (mg mg–1)

carbon and nitrogen contents of sporophylls of kelps

excised in January were detected (P>0.05), but from 0.1
late March to early May, significant increases occurred
(P<0.05). For kelps excised in February, no significant 0
changes in carbon and nitrogen contents of sporophylls 0.05
Nitrogen
were observed from late February to early April, but * *
they increased significantly thereafter until early May. In 0.04
contrast, no significant changes in carbon and nitrogen
contents of sporophyll of kelps excised in March and 0.03
April were detected from the time of excision to early
May (P>0.05). All excised and control kelps showed a 0.02
significant decrease in carbon and nitrogen contents in
May (P<0.05). 0.01
The carbon and nitrogen contents of sporophyll tissues
formed before and after excisions are shown in Fig. 6. The 0
carbon contents of new tissues of all excised kelps were JE FE ME AE
slightly higher than those of the old tissues (P>0.05). The Fig. 6 Carbon and nitrogen contents of sporophyll discs taken from
nitrogen contents of new tissues of kelps excised in January tissues formed before (filled columns) and after (open columns) exci-
and February were significantly lower than those of the old sions in each kelp at the end of the experiment. Asterisks indicate a
significant difference at P<0.05 between excised and control kelps.
tissues (P<0.05). No significant differences between old N=8 for all kelps. Black bars indicate standard deviations. JE, FE, ME,
and new tissues were detected in kelps excised in March and AE represent four experimental kelps excised at the beginning of
and April (P>0.05). January, February, March, and April, respectively
1338
Discussion
This study is the first to evaluate the influence of the timing
of damage on the compensatory capability of large seaweed.
U. pinnatifida exhibited quite different compensatory
responses to thallus excision at different developmental
stages. As an important method for increasing production,
this finding may have a great potential for wide application
in commercial cultivation of this kelp in Japan and other
parts of the world.
In the present study, the blade length and dry weight of
kelps without excision increased significantly by early and
late February, respectively, but then did not increase there-
after (Table 2, seasonal blade length and dry weights of
blade). These results indicate that U. pinnatifida sporo-
phytes cultivated in Matsushima Bay stop vegetative growth
in late February. The reproductive phase of sporophytes of
Laminariales starts after growth has stopped (Markhan
1973; Kain 1975). Thus, this kelp starts to enter the matu-
ration phase after late February. The kelps excised at the
beginning of January and February exhibited a marked
increase in morphological features, which agrees with the
previous finding that the growth of this kelp was promoted
greatly by thallus excision in late February (Gao et al. 2012).
In contrast, morphological features of the kelps excised at
the beginning of March and April did not increase signifi-
cantly. These results suggest that U. pinnatifida sporophytes
farmed in Matsushima Bay are not capable of great com-
pensatory growth in response to thallus excisions conducted
after March when the maturation phase has begun. Similar
results have been reported in some terrestrial plants, indi-
cating that plants damaged early in the growing season tend
to compensate better than plants damaged late in the repro-
ductive season (Maschinski and Whitham 1989; Paige
1994; Lennartsson et al. 1998).
The photosynthetic capacity of kelps excised in January
and February was greater than that of the control kelps (Fig. 2).
This response may be partially due to the enhanced external
light environment, as happens with higher plants (Jãremo et al.
1996; Mabry and Wayne 1997). Photosynthesis in the remain-
ing tissues was stimulated by greater light penetration, which
may have resulted in an increase in pigment concentration and
photosynthesis efficiency. Intracellular nitrogen content great-
ly influences pigments such as chlorophyll (Chapman et al.
1978; Bird et al. 1982; Lingnell and Pedersen 1987; Lapointe
1981) and photosynthetic activity (Levy and Gantt 1990). The
greater capacity for nutrient accumulation may also partly
increase the concentration of pigments and support increased
photosynthesis and compensatory growth (Figs. 3 and 4).
However, there were no significant increases in photosynthe-
sis, nutrient uptake, and resource contents of kelps excised in
March and April. Therefore, compensatory responses of ex-
cised kelps may be due not only to the improvement of
J Appl Phycol (2013) 25:1331–1340
external environmental conditions but also to internal physio-
logical regulation. Plant hormones play a critical role in stim-
ulating or inhibiting many physiological processes in
seaweeds (Bradley 1991; Basu et al. 2002; Nimura and
Mizuta 2002; Stirk et al. 2003; Tarakhovskaya et al. 2007;
Wiesemeier et al. 2008) including U. pinnatifida (Abe et al.
1972; Yan 1984). Kai et al. (2006) reported that growth and
reproduction of Laminarian species are regulated by internal
auxin level, as is true for terrestrial plants. Therefore, we
propose that the compensatory responses to thallus excision
in U. pinnatifida may be due to the stimulation of plant
hormones. In future studies, the variation of plant hor-
mone levels should be compared between excised kelps
and the control.
In this study, the kelps without excision exhibited a
significant increase in sporophyll growth from January to
late March (Table 2, sporophyll). In the kelps excised in
January and February, this rapid sporophyll development
was markedly delayed. However, these sporophylls grew
significantly thereafter, which likely was due to the rapid
increase of carbon and nitrogen contents (Fig. 5). By the end
of this experiment, there were no significant differences in
carbon content between sporophyll tissues formed before
and after excisions in January and February; however, the
nitrogen contents of new tissues formed after excisions were
significantly lower than those of old tissues formed before
excisions (Fig. 6). In contrast, there were no significant
variations in dry weight and resource contents of sporo-
phylls of kelps excised in March and April. These results
indicate that a large shift in resource allocation to growth
and maturation occurred following the thallus excisions that
took place before the vegetative growth had stopped.
Almost all accumulated resources might be allocated to the
blade to compensate for the mass loss of reserved resources,
therefore resulting in compensatory growth of the blade
and delay of sporophyll development. Similarly, the
compensatory response is achieved through the modular
structure of terrestrial plants, which is efficiently inte-
grated through specialized tissues for long-distance in-
ternal translocation of stored and photo-assimilated
compounds (Trumble et al. 1993; Taiz and Zeiger
2002). Thus, plants can quickly re-allocate resources to
damaged parts when required.
Due to changes in resource allocation after thallus exci-
sion, the growth phase of the kelps excised in January and
February was prolonged for at least 1 month, which resulted
in extension of the harvest period. This means that the
harvest of high-quality blade and sporophyll tissues will
become possible during periods when they previously were
unavailable. Compared with kelps without excision, the
kelps excised in January and February experienced an al-
most 50 % increase in biomass production because of their
strong compensatory abilities. These conclusions provide
J Appl Phycol (2013) 25:1331–1340
important new information that can be used to improve the
aquaculture production of this kelp.
Acknowledgments We sincerely thank Mr. M. Ise of the Shiogama
Daiichi Fisheries Cooperative Association for providing the seedlings
used in the present study and for his kind cooperation with kelp
cultivation and collection; the staffs of the Miyagi Prefecture Fisheries
Technology Institute and Akita Prefecture Fisheries Promotion Institute
for using seawater for culture experiment; Professor O. Nishimula and
Mr. Chiba for helping with the analysis of nutrient concentration; and
assistant professor K. Ito for helping with the measurement of carbon
and nitrogen content.
References
Abe H, Uchiyama M, Sato R (1972) Isolation and identification of
native auxins in marine algae. Agric Biol Chem 36:2259–2260
Aguilar-Rosas R, Aguilar-Rosas LE, Avila-Serrano G, Marcos-
Ramirez R (2004) First record of Undaria pinnatifida (Harvey)
Suringar (Laminariales, Phaeophyta) on the Pacific coast of
Mexico. Bot Mar 47:255–258
Akiyama K, Kurogi M (1982) The decrease in crops from natural
plants following crop increase from cultivation. In: Cultivation
of Undaria pinnatifida (Harvey) Suringar. Bull Tohoku Natl Fish
Res Inst 44:91–100
Basu S, Sun H, Brian L, Quatroano RL, Muday GK (2002) Early
embryo development in Fucus distichus is auxin sensitive. Plant
Physiol 130:292–302
Bergelson J, Juenger T, Crawley MJ (1996) Regrowth following her-
bivory in Ipomopsis aggregate: compensation but not overcom-
pensation. Am Nat 148:744–755
Bird KT, Habig C, DeBusk T (1982) Nitrogen allocation and storage
patterns in Gracilaria tikvahiae (Rhodophyta). J Phycol 18:344–
348
Bradley PM (1991) Plant hormones do have a role in controlling
growth and development of algae. J Phycol 27:317–321
Campbell SJ, Bité JS, Burridge TR (1999) Seasonal patterns in the
photosynthetic capacity, tissue pigment and nutrient content of
different developmental stages of Undaria pinnatifida
(Laminariales, Phaeophyta) in Port Phillip Bay, South-Eastern
Australia. Bot Mar 42:231–241
Casas GN, Piriz ML (1996) Surveys of Undaria pinnatifida
(Laminariales, Phaeophyta) in Golfo Nuevo, Argentina.
Hydrobiologia 326/327:213–215
Cerda O, Karsten U, Rothäusler E, Tala F, Thiel M (2009)
Compensatory growth of the kelp Macrocystis integrifolia
(Phaeophyceae, Laminariales) against grazing of Peramphithoe
femorata (Amphipoda, Ampithoidae) in northern-central Chile. J
Exp Mar Biol Ecol 377:61–67
Chapman ARO, Markham JW, Lüning K (1978) Effects of nitrate
concentration on the growth and physiology of Laminaria sac-
charina (Phaeophyta) in culture. J Phycol 14:195–198
Gao X, Endo H, Yamana M, Taniguchi K, Agatsuma Y (2012)
Compensation of the brown alga Undaria pinnatifida
(Laminariales; Phaeophyta) after thallus excision under culti-
vation in Matsushima Bay, northern Japan. J Appl Phycol.
doi:10.1007/s10811-012-9925-y
Hay KB, Poore AGB, Lovelock CE (2011) The effects of nutrient
availability on tolerance to herbivory in a brown seaweed. J Ecol
99:1540–1550
Honkanen T, Jormalainen V (2002) Within-alga integration and
compensation: effects of simulated herbivory on growth and
1339
reproduction of the brown alga Fucus vesiculosus. Int J Plant
Sci 163:815–823
ICES (2007) Alien Species Alert: Undaria pinnatifida (Wakame or
Japanese kelp). ICES Cooperative Research Reports no. 283,
36pp
Jãremo J, Nilsson P, Tuomi J (1996) Plant compensatory growth:
herbivory or competition? Oikos 77:238–247
Kai T, Nimura K, Yasui H, Mizuta H (2006) Regulation of sorus
formation by auxin in Laminariales sporophyte. J Appl Phycol
18:95–101
Kain JM (1975) The biology of Laminaria hyperborea. VII.
Reproduction of the sporophyte. J Mar Biol Assoc UK 55:567–
582
Lapointe BE (1981) The effects of light and nitrogen no growth,
pigment content and biochemical composition of Gracilaria foli-
fera V. Angustissima (Gigartinales, Rhodophyta). J Phycol
17:90–95
Lehtila K, Syrjänen K (1995) Compensatory responses of two
Melampyrum species after damage. Funct Ecol 9:511–517
Lennartsson T, Nilsson P, Tuomi J (1998) Induction of overcompensa-
tion in the field Gentian, Gentianella campestris. Ecology
79:1061–1072
Levy I, Gantt E (1990) Development of photosynthetic activity in
Porphyridium purprueum (Rhodophyta) following nitrogen star-
vation. J Phycol 26:62–68
Lingnell A, Pedersen M (1987) Nitrogen metabolism in Gracilaria
secundata (Harv.). Hydrobiologia 151/152:431–441
Littler MM, Littler DS, Taylor PR (1995) Selective herbivore increases
biomass of its prey—a chiton–coralline reef-building association.
Ecology 76:1666–1681
Mabry CM, Wayne PW (1997) Defoliation of the annual herb Abutilon
theophrasti—mechanisms underlying reproductive compensa-
tion. Oecologia 111:225–232
Markhan JW (1973) Observation on the ecology of Laminaria sin-
clairii on three northern Oregon beaches. J Phycol 9:336–341
Maschinski J, Whitham TG (1989) The continuum of plant responses
to herbivory: the influence of plant association, nutrient availabil-
ity and timing. Am Nat 134:1–19
Nimura K, Mizuta H (2002) Inducible effects of abscisic acid on
sporophyte discs from Laminaria japonica Areschoug. J Appl
Phycol 14:159–163
Paige KN (1994) Herbivory and Ipomopsis aggregate: difference in
response, difference in experimental protocol: a reply to
Bergelson and Crawley. Am Nat 143:739–749
Peteiro C, Freire Ó (2011) Effect of water motion on the cultivation of
the commercial seaweed Undaria pinnatifida in a coastal bay of
Galicia, Northwest Spain. Aquaculture 314:269–276
Sanderson JC (1990) A preliminary survey of the distribution of the
introduced macroalga Undaria pinnatifida (Harvey) Suringer on
the east coast of Tasmania. Bot Mar 33:153–157
Silva PC, Woodfield RA, Cohen AN, Harris LH, Goddard JHR (2002)
First report of the Asian kelp Undaria pinnatifida in the north-
eastern Pacific Ocean. J Appl Phycol 4:333–338
Simons AM, Johnston MO (1999) The cost of compensation. Am Nat
153:683–687
Steinberg PD (1995) Interaction between the canopy dwelling echinoid
Holopneustes purpurescens and its host kelp Ecklonia radiata.
Mar Ecol Prog Ser 127:169–181
Stirk WA, Novák O, Strnad M, Van Staden J (2003) Cytokinins in
macroalgae. Plant Growth Regul 41:13–24
Stowe KA, Marquis RJ, Hochwender CG, Simms EL (2000) The
evolutionary ecology of tolerance to consumer damage. Annu
Rev Ecol Evol Syst 31:565–595
Strauss SY, Agrawal AA (1999) The ecology and evolution of plant
tolerance to herbivory. Trend Ecol Evol 14:179–185
Taiz L, Zeiger E (2002) Plant physiology. Sinauer, Massachusetts
1340
Tarakhovskaya ER, Maslov YI, Shishova MF (2007) Phytohormones
in algae. Russ J Plant Physiol 54:163–170
Tatewaki M (1966) Formation of a crustose sporophyte with
unilocular sporangia in Scytosiphon lomentaria. Phycologia
6:62–66
Thomson VP, Cunningham SA, Ball MC, Nicotra AB (2003)
Compensation for herbivory by Cucumis sativus through in-
creased photosynthetic capacity and efficiency. Oecologia
134:167–175
Tiffin P (2000) Mechanisms of tolerance to herbivore damage: what do
we know? Evol Ecol 14:523–536
Trumble JT, Kolodney-Hirsh DM, Ting IP (1993) Plant compensation
for arthropod herbivory. Annu Rev Entomol 38:93–119
J Appl Phycol (2013) 25:1331–1340
Vergés A, Pérez M, Alcoverro T, Romero J (2008) Compensation and
resistance to herbivory in seagrasses: induced responses to simu-
lated fish consumption. Oecologia 155:751–760
Wiesemeier T, Jahn K, Pohnert G (2008) No evidence for the induction
of brown algal chemical defense by the phytohormones jasmonic
acid and methyl jasmonate. J Chem Ecol 34:1523–1531
Yamanaka R, Akiyama K (1993) Cultivation and utilization of
Undaria pinnatifida (wakame) as food. J Appl Phycol 5:249–253
Yan ZM (1984) Studies on tissue culture of Laminaria japonica and
Undaria pinnatifida. Hydrobiologia 116/117:314–316
Yokohama Y, Katayama N, Furuya K (1986) An improved type of
“Productmeter”, a differential gas-volumeter, and its application
to measuring photosynthesis of seaweeds. Jpn J Phycol 34:37–42