J Appl Phycol (2013) 25:567–574
DOI 10.1007/s10811-012-9891-4
Genetic differentiation of high-temperature tolerance in the kelp
Undaria pinnatifida sporophytes from geographically separated
populations along the Pacific coast of Japan
Xu Gao & Hikaru Endo & Kazuya Taniguchi &
Yukio Agatsuma
Received: 3 July 2012 / Revised and accepted: 3 August 2012 / Published online: 23 August 2012
# Springer Science+Business Media B.V. 2012
Abstract The kelp Undaria pinnatifida has a widespread
latitudinal range in Japan, with populations exposed to very
different temperature regimes. To test the hypothesis that U.
pinnatifida exhibits genetic differentiation in its temperature
response, juvenile sporophytes from a warmer location
(Naruto, southern Japan) and two colder locations (Okirai
Bay and Matsushima Bay, northern Japan) were collected
and transplanted to long lines, cultivated under the environ-
mental conditions in Matsushima Bay. These plants were
bred using successive self-crossing methods for three gen-
erations and the characteristics of photosynthesis, growth,
survival, and nitrogen contents of the third-generation juve-
nile sporophytes (2–3 cm) then were measured and com-
pared. The plants from Naruto showed significantly higher
photosynthetic activities and respiration than those from the
northern populations at warmer temperatures of 20–35°C.
The juvenile sporophytes from all three locations had sim-
ilar growth rates below 18°C, but significant differences
were observed at 18–24°C. The optimum temperatures for
growth were 14–16°C in plants that originated from Okirai
Bay and Matsushima Bay and 18°C in plants that originated
from Naruto. These results reflected the differences in lati-
tude. Dead plants were observed at high temperatures of 22
and 24°C in the northern population plants, whereas no
plants from Naruto died. Juvenile sporophytes from Naruto
exhibited the greatest capacity to accumulate high nitrogen
reserves. These results suggest that the differences in high-
temperature tolerance in juvenile U. pinnatifida sporophytes
from geographically separated populations are due to genet-
ic differentiation rather than phenotypic plasticity.
X. Gao : H. Endo : K. Taniguchi : Y. Agatsuma (*)
Laboratory of Marine Plant Ecology,
Graduate School of Agricultural Science, Tohoku University,
1-1, Tsutsumidori-amamiya, Aoba,
Sendai, Miyagi 981-8555, Japan
e-mail: agatsuma@bios.tohoku.ac.jp
Keywords Ecotype . High temperature tolerance . Growth .
Survival . Nitrogen content . Undaria pinnatifida
Phaeophyta
Introduction
Ecotype is defined as the genetic differences in intraspecific
populations that reflect the regional responses of organisms
to local environmental conditions, and it occurs in many
plants (Turresson 1922; Clausen et al. 1940). The existence
of ecotypes has been demonstrated by the transplantation of
plants from different regions to a given location, their pro-
gression through more than one life cycle under the same
ambient conditions, and, finally, a comparison of their char-
acteristics (Begon et al. 1990).
Temperature is one of the primary abiotic factors responsi-
ble for the geographical distribution of marine plants (Lüning
and Freshwater 1988; Fields et al. 1993). Many macroalgae
have the ability to adjust and optimize their physiological
characteristics to the prevailing temperature conditions in the
short term (Raven and Geider 1988; Davison et al. 1991;
Falkowski and Laroche 1991; Fairhead and Cheshire 2004).
In the long term, however, the phenotypes of plants acclima-
tized to the temperature conditions of different habitats will be
separated by natural selection and will differentiate into eco-
types with different temperature tolerances (Russell 1986;
Breeman and Pakker 1994). In Laminarian species, there are
many reports of the existence of ecotypes adapted to various
ambient temperature regimes (Kain 1969; Espinoza and
Chapman 1983; Novaczek 1984; Bolton and Anderson
1987; Gerard and DuBois 1988; Serisawa et al. 2004; Graham
et al. 2007). For example, Gerard (1997) reported that greater
heat tolerance of Saccharina latissima was demonstrated by a
greater capacity for nitrogen accumulation that was not attrib-
utable to greater thermal stability of the photosynthetic
568
apparatus. Although the internal nutrient contents of algae
differ significantly among collection locations (Gao and
Nakahara 1990; Staehr and Wernberg 2009), no study to date
has compared the cellular nutrient content of plants from
different locations after they have been transplanted to the
same location and cultured there for a long period of time.
The annual kelp Undaria pinnatifida (Laminariales;
Phaeophyta) is one of the major commercial seaweeds na-
tive to cold temperate coastal areas of Japan (Akiyama and
Kurogi 1982). It has been extensively cultivated as one of
important and popular seafood in Asian countries (Saito
1972; Yamanaka and Akiyama 1993) even though it is
known to be an invasive species (Hay and Luckens 1987;
Hay 1990; Fletcher and Manfredi 1995; Campbell et al.
1999; Stuart et al. 1999). U. pinnatifida gametophytes have
greater tolerance to high temperatures than related taxa of
the northern hemisphere (tom Dieck 1993), possibly be-
cause it is adapted to the Japanese coastline, which is char-
acterized by strong temperature fluctuations between
seasons and latitudes (Funahashi 1973; van den Hoek and
Breeman 1990; Lüning 1990). The critical upper tempera-
ture for growth of U. pinnatifida sporophytes collected from
Mugisaki Bay, central Japan was 27°C (Morita et al. 2003),
which is higher than the upper limit of 25°C in plants from
Matsushima Bay, northern Japan (Akiyama and Kurogi
1982). This result agreed with previous reports that plants
from the south generally have greater tolerance to high
temperatures than those from north, although it is unknown
whether this difference is due to genetic differentiation or
phenotypic plasticity.
In recent years, global warming-induced increases in
surface seawater temperature in kelp farming areas have
caused problems such as failed seedling production, prema-
ture deterioration of adult plants in the sea, and unexpected
algal blooms (Lignell et al. 2003). To counteract these
negative effects, researchers have proposed that specimens
of a high-temperature-tolerant ecotype should be used as
parental plants in kelp farming (Pang et al. 2007; Liu and
Pang 2010; Gao et al. submitted). However, it should be
pointed out that temperature tolerance is an inherited feature
and possibly could not be altered in short-term selection by
artificial treatment during cultivation (Pang et al. 2007).
Thus, it is necessary to test the temperature tolerance char-
acteristics of natural populations. Furthermore, the need for
plants to survive and grow well in farming area calls for a
practical method for judging and selecting the desired pa-
rental plants for seedling production. Therefore, whether or
not high-temperature tolerance can be maintained over sev-
eral generations needs to be tested. Although Uwai et al.
(2006) found that plants collected from Naruto and those
from northeastern Japan exhibited similar molecular genetic
characteristics, possibly due to the introduction of plants
from northern Japan through cultivation since the 1960s
J Appl Phycol (2013) 25:567–574
(Ohno et al. 1999), it remains unclear whether the two
populations differ in their temperature tolerance.
In the present study, we examined physiological differ-
ences of juvenile sporophytes of U. pinnatifida transplanted
from natural populations in Naruto and Matsushima Bay
after cultivation in Matsushima Bay for 3 years. We also
examined whether the differences in temperature tolerance
could be maintained after 3 years of acclimation. Because
plants from the natural population in Matsushima Bay may
be uniquely adapted to native waters, plants from natural
population in Okirai Bay, which is located near Matsushima
Bay, were also transplanted to the study site for compar-
ison with the Naruto plants cultivated under the same
ambient conditions. The results may indicate whether the
physiological differences among populations, such as
high-temperature tolerance, are due to genetic differenti-
ation or phenotypic plasticity. Additionally, the results of
this study are expected to provide important information
for improving the cultivation of U. pinnatifida in Japan
and possibly in other parts of the world.
Materials and methods
Sporophyte transplantation
In November 2006, healthy juvenile U. pinnatifida sporo-
phytes less than 50 cm in total length were collected from
natural populations in Naruto (34°13′ N, 135°37′ E),
Tokushima Prefecture, in southern Japan and in Okirai
Bay (39° 05′ N, 142° 51′ E), Iwate Prefecture, and in
Matsushima Bay (38° 32′ N, 141° 31′ E), Miyagi Prefec-
ture, in northern Japan (Fig. 1). They were transplanted to
the long lines in Matsushima Bay at a depth of 0.5 m and
were cultivated. The range of year-round seawater temper-
ature in Matsushima Bay is 7.2–24.4°C (Miyagi Fisheries
Technology Center 1995–2004), which is slightly higher
than that of Okirai Bay (6.3–23.3°C) (Iwate Fisheries
Technology Center 1995–2004) but lower than that of
Naruto (9.3–26.2°C) (Tokushima Agriculture, Forestry
and Fisheries Technology Support Center 1995–2004). In
order to maintain their respective genetic characteristics,
these plants were bred by use of successive self-crossing
methods for three generations (Feng et al. 2005; Pang and
Xu 2005). More than 30 sporophytes that survived and
grew well in the farming area were selected as the desired
parental plants, and gametophyte clones were cultured exten-
sively for seedling production.
Collection and maintenance
Juvenile sporophytes of U. pinnatifida from each parental
stock were collected from the long lines in Matsushima Bay
J Appl Phycol (2013) 25:567–574
Fig. 1 Map of Japan showing the three collected locations of juvenile
sporophytes of U. pinnatifida
in October 2009 and transported quickly to our laboratory
using insulated cool boxes filled with seawater. The average
lengths of these collected juvenile sporophytes were 1.01±
0.29 mm (n0800) from Okirai Bay, 0.97±0.31 mm (n0800)
from Matsushima Bay, and 0.90±0.32 mm (n0900) from
Naruto, respectively. The seedling strings with juvenile
sporophytes were cultured at 17°C (corresponding to the
seawater temperature in October along the coast of Matsush-
ima Bay) in large flasks containing 4 L of enriched 25 %
PESI medium (Tatewaki 1966). The PESI medium was
made using sterilized seawater with a salinity 32 psu and pH
7.9 from the coast of Ishinomaki, which was near the study
site and experienced environmental conditions similar to those
in Matsushima Bay. A photoperiod of 12:12 h (light/dark) and
a light intensity of 40 μmol photons m−2 s−1 were employed.
These juvenile sporophytes remained in these conditions for
several days until they reached a mean length of 2–3 cm which
was large enough for following experiments. The medium
was renewed every day. More than 400 healthy juvenile
sporophytes from each location were separated from seedling
strings at the base of the stipe by using a razor blade for
subsequent experiments.
Juvenile culture
The culture experiment was conducted at eight tempera-
tures: 10, 12, 14, 16, 18, 20, 22, and 24°C. During the
experiment, light intensity of 180 μmol photons m−2 s−1
and neutral day (12:12 h, light/dark) conditions were main-
tained. Light was provided by 40-W cool-white fluorescent
tubes. A time clock was used for each incubator to regulate
the light and dark cycle. A total of 72 flasks were used, with
each population replicated in three separate flasks at each of
the eight temperatures, and each flask contained 500 mL of
seawater enriched with 25 % PESI medium, to which
569
1.5 mL GeO2 (3 μg L−1) was added to inhibit the growth
of diatoms and protozoa. Ten juvenile sporophytes from a
given U. pinnatifida population were put into each flask;
these flasks, which received gentle aeration, were then
placed within eight incubators and cultured for 12 days.
The culture medium was renewed every 3 days. GeO2 was
also added every time after medium replenishment. The
growth situation was observed every day and the propor-
tions of dead individual were calculated. Fresh weights of
all juvenile sporophytes prior to the experiments and at the
end were measured after being blotted dry, and the relative
growth rates (RGR, % day−1) were calculated with the mean
fresh weight for each replicate using the following equation:


RGR % day
1 ¼ 100 lnðWt =Wo Þ=t
where Wo is the initial fresh weight, Wt is the final fresh
weight after the experiment, and t is the number of days.
Nitrogen content analysis
Nitrogen contents were examined among juvenile spor-
ophytes derived from the three populations at the end of
the 12-day experiment. For each temperature, eight sur-
viving juvenile sporophytes from each population after
culture were selected and placed into screw bottles and
then dried in a convection oven (NDO–600ND, Eyela,
Tokyo, Japan) at 60°C for 24 h. After measurement of
dry weight, the samples were crushed and used for
analysis of nitrogen content by an Elemental Analyzer
(EAGER200, Fisons Instruments, USA).
Measurement of photosynthesis
In a separate set of experiments, photosynthesis of the
plants was measured under different conditions. An
improved differential gas-volumeter called “Product-me-
ter” (Yokohama et al. 1986) was used to measure pho-
tosynthetic rates. Each juvenile sporophyte from each
population was placed in a reaction vessel containing
10 mL sterilized 25 % PESI seawater (GF/F, Whatman,
Maidstone, UK) (Tatewaki 1966), while a compensation
vessel contained only 10 mL sterilized seawater. The
reaction and compensation vessels were attached to prod-
uct meters and immersed in a water bath with a thermostat
(Taitec CL–150 F, Koshigaya, Japan) maintained at a
constant water temperature and shaken by means of a
motor drive at 150 rpm min−1. The vessels were illumi-
nated from below with photo slide projector lamps (Elmo
S-300, Nagoya, Japan) and with incandescent lamps (Phi-
lips KP-10s 100V, 300 W; Japan), whose light was
reflected with mirrors placed under the water bath.
Irradiances were regulated by using neutral density glass
570 J Appl Phycol (2013) 25:567–574

35
filters (Toshiba TND-50, 25, 12.5, Tokyo, Japan) and mea-
30 dA
sured with a quantum photon meter (LI-COR LI-192 S, USA). cA cA
cA cA
The juvenile sporophytes were incubated for 30 min to allow 25 cA bB

RGR (% day–1)
cA
bA
them to adapt to each experimental temperature, and then the 20 bA bB bA
bA aB
oxygen evolution produced by the juveniles was measured six aA
aA aA
15 aA aB
to ten times every 3 min.
dB eA
Photosynthesis versus light (P–L) and temperature 10
dB
(P–T) curves were generated separately for each light 5
eB
intensity and temperature per unit area. For the P–L eB
0
10 12 14 16 18 20 22 24
curve determination, juvenile sporophytes from each
Temperature (ºC)
population were subjected to eight different light inten-
sities (0, 5, 10, 20, 50, 100, 180, and 360 μmol photons Fig. 2 The relative growth rates of juvenile sporophytes of U. pinna-
m−2 s−1) at four different seawater temperatures (10, 15, tifida from three populations cultured for 12 days at various temper-
atures. Black, gray, and white bar charts indicate the relative growth
20, and 25°C). For the P–T curve determination, differ- rates of juvenile sporophytes from Okirai Bay, Matsushima Bay, and
ent sporophytes were used and factor levels were set as Naruto, respectively. Different small and large letters indicate statistical
follows: temperature at 5, 10, 15, 20, 25, 30, and 35°C; differences (p<0.05) of juvenile sporophytes among temperature and
light intensity at 0 and 180 μmol photons m−2 s−1. Eight among three populations at each temperature. N030 individuals. Black
bars indicate standard deviations
replicates were used in all experimental combinations.

Statistical analysis Juvenile sporophytes exhibited different survival percen-

A two-way analysis of variance (ANOVA) was used to
compare the photosynthesis versus temperature (P–T)
curves, growth rates, and nitrogen contents among temper-
atures and among juvenile sporophytes from the three pop-
ulations. In addition, a one-way ANOVA was used to
compare photosynthesis versus light (P–L) curves from the
three populations at different temperatures at each light
intensity. All of the variables were tested for normality and
homogeneity prior to conducting ANOVA. To identify dif-
ferences among temperatures and among juvenile sporo-
phytes from the three populations, Tukey’s HSD post hoc
comparison was applied. Differences were considered to be
significant at a probability of 5 % (p<0.05).
tages among the populations at high temperatures (Fig. 3).
Survival percentages of 100 % were observed in all three
populations from 10 to 20°C. However, dead plants from
Okirai Bay and Matsushima Bay appeared, respectively, after
2 days and 5 days at higher temperatures of both 22 and 24°C.
At 24°C, the survival percentages were 60 and 70 % for the
22ºC
100
80
60
40
Survival percentage (%)

20
Results
0

Growth and survival 24ºC

100
The RGRs of juvenile sporophytes from the three U. pin- 80
natifida populations at various temperatures are shown in
Fig. 2. The RGRs were affected significantly by tempera- 60
ture and population (two-way ANOVA, temperature effect,
40
F053.517, p<0.05; population effect, F032.273, p<0.05).
Juvenile sporophytes from Naruto grew well at all temper- 20
atures, with the fastest growth at 18°C. The fastest growth
0
of juvenile sporophytes from Okirai Bay and Matsushima 0 1 2 3 4 5 6 7 8 9 10 11 12
Bay was observed at 14–16°C. There were no differences Culture period (days)
in the RGRs among the populations at low temperatures of
Fig. 3 Survival percentages of juvenile sporophytes of U. pinnatifida
10–16°C, but the RGRs from Naruto at higher temper- from Okirai Bay (squares), Matsushima Bay (triangles), and Naruto
atures of 18–24°C were significantly higher than those (circles) cultured for 12 days at high temperatures of 22 and 24°C.
from the northern populations. Black bars indicate standard deviations
J Appl Phycol (2013) 25:567–574 571

100
Okirai Bay and Matsushima Bay samples, respectively, at the 10 C
80
end of experiment. In contrast, no juvenile sporophytes from
60
Naruto died at these high temperatures during this experiment.
40
20
Nitrogen content
0
-20
The nitrogen content was population-dependent, and tem-
-40
perature did not significantly affect nitrogen contents for all 100
juvenile sporophytes (two-way ANOVA, population effect, 15 C
80

Gross photosynthesis rate (µLO2 cm–2 s–1)

F013.381, p<0.05; temperature effect, F01.837, p>0.05; 60
Fig. 4). Juvenile sporophytes from Naruto had higher nitro- 40
gen contents at all temperatures compared to those from 20

northern populations, and significant differences between 0

the Naruto population and the northern populations were -20

-40
found at the relatively higher temperatures of 18–24°C. The 100
nitrogen content of juvenile sporophytes from Okirai Bay and 20 C a
80
a a
Matsushima Bay were not significantly different (p>0.05). 60
b b
40 b
b b
Photosynthesis and respiration 20
b

0
b
Gross photosynthetic rates calculated per area of juvenile -20 b
a
sporophytes increased rapidly with increasing light intensity -40
up to 180 μmol photons m−2 s−1 at 10–25°C in sporophytes 100
25 C a
a
80 a
from all three populations (Fig. 5). In addition, for all
60 b
populations, compensation photosynthetic rates were in al- b
b

most the same irradiance range of 0–5 μmol photon m−2 s−1, 40
b
b b

and the highest photosynthetic rates were observed at 20

180 μmol photons m−2 s−1. At 10 and 15°C, significant 0

0 bb 50 100 150 200 250 300 350 400
differences in gross photosynthetic rates were not found at -20
a
-40 Light intensity (µmol photons m-2 s-1)
all light intensities among these populations (one-way
ANOVA, p>0.05). Although there were also no differences Fig. 5 Photosynthesis–light curves at various temperatures per unit
at 5–50 μmol photons m−2 s−1, significant differences were area of juvenile sporophytes of U. pinnatifida collected from Okirai
found at 100–360 μmol photons m−2 s−1 employed at 20 and Bay (triangle), Matsushima Bay (square), and Naruto (circle). Different
25°C between juvenile sporophytes from Naruto and those small letters indicate statistical differences (p<0.05) of juvenile spor-
ophytes among three populations. N08 individuals. Black bars indicate
from the northern populations (one-way ANOVA, p<0.05). standard deviations

0.05

a a
Similarly, dark respiration rates of juvenile sporophytes from
Nitrogen (mg/mg d.w.)

a a a a
0.04 a a
aa aa a
bb
Naruto were significantly higher than those from the northern
aa a bb b
bb b populations at 20 and 25°C, but significant differences were
0.03
not found at 10 and 15°C (one-way ANOVA, p>0.05).
0.02
The gross photosynthetic rate of juvenile sporophytes dif-
0.01
fered among the three populations, and was influenced signif-
icantly by temperatures (two-way ANOVA, temperature
0
10 12 14 16 18 20 22 24 effect, F073.281, p<0.05; population effect, F041.697,
Temperature (ºC) p<0.05; Fig. 6). The gross photosynthetic rates increased
Fig. 4 Nitrogen contents of juvenile sporophytes of U. pinnatifida
with the rise in temperature up to 25°C but decreased
from three populations cultured for 12 days at various temperatures. thereafter for all populations. Although there were no
Nitrogen content represent as dry weight (mg) per 1 mg thallus dry significant differences in gross photosynthetic rates of
weight. Black, gray, and white bar charts indicate the nitrogen contents juvenile sporophytes among the populations at low tem-
of juvenile sporophytes from Okirai Bay, Matsushima Bay, and Naruto,
peratures of 5–15°C (two-way ANOVA, p>0.05), signifi-
respectively. Different small letters indicate statistical differences
(p<0.05) of juvenile sporophytes among three populations. N08 individ- cantly higher values from Naruto were found at higher
uals. Black bars indicate standard deviations temperatures of 20–35°C compared to the northern
572
120
100
Oxygen changes (µLO2 cm–2 s–1)
80
60 a b
a a
40 a a
20 a a
a a
0 b
-20
a a
-40
a a
-60
0 5 10
Fig. 6 Photosynthesis–temperature curves at 180 μmol photons m−2
s−1 (open) and dark respiration temperature curves (solid) per unit area
of juvenile sporophytes of U. pinnatifida collected from Okirai Bay
(triangle), Matsushima Bay (square), and Naruto (circle). Different
small letters indicate statistical differences (p<0.05) of juvenile spor-
ophytes among three populations. N08 individuals. Black bars indicate
standard deviations
populations (two-way ANOVA, p<0.05). The dark respi-
ration rate of juvenile sporophytes also differed among the
three populations, but significant differences were not
found among temperatures (two-way ANOVA, temperature
effect, F01.183, p>0.05; population effect, F020.257,
p<0.05; Fig. 6). Dark respiration of juvenile sporophytes
from all three populations increased slightly with a rise tem-
perature from 5 to 35°C (two-way ANOVA, p>0.05). These
sporophytes did not significantly differ in respiration rate per
area at 5–15°C, but significantly higher values were found in
the specimens from Naruto compared to those from the north-
ern populations at 20–35°C (two-way ANOVA, p<0.05).
Discussion
This study showed that temperature ecotypes exist in U.
pinnatifida between a southern natural population (Naruto)
and two northern natural populations (Okirai Bay and Mat-
sushima Bay) in waters along the coasts of Japan. The
optimum temperatures for growth of third-generation juve-
nile sporophytes from different populations exhibited obvi-
ous latitudinal dependence. This conclusion was further
supported by the significant differences in physiological
characteristics between southern and northern populations
when exposed to high temperatures.
Juvenile U. pinnatifida sporophytes sampled from Mie
Prefecture, central Japan have a 2°C higher critical upper
temperature for growth than those from Miyagi Prefecture,
northern Japan (Akiyama and Kurogi 1982; Morita et al.
2003). However, these results are inadequate to demonstrate
the genetic differentiation of U. pinnatifida in Japan because
the plants were not transferred to and cultivated in the same
environmental conditions and then compared phenotypical-
ly (Begon et al. 1990). In this study, after transplantation and
J Appl Phycol (2013) 25:567–574
a three generations of acclimation, the optimal growth tem-
perature for juvenile sporophytes from Naruto was still 2–4°C
a
a
a
a b higher than those of juvenile sporophytes from northern pop-
b
b ulations; moreover, the optimal temperature coincided with
b b
a b the latitudinal gradient of their distributions. Thus, this is the
b
a
first study to document the existence of temperature ecotypes
b b b
a b
of U. pinnatifida and its genetic stability. Additionally, juve-
b b b
a nile sporophytes from Naruto exhibited significantly higher
a a a
a RGRs and survival percentages at high temperatures of 18–
15 20 25 30 35 40
24°C than sporophytes from northern populations, indicating
Temperature ( C)
greater heat tolerance.
Gerard (1997) reported that greater high-temperature tol-
erance is due to a higher nitrogen and protein content in the
algae, and he suggested that the content of reserved nitrogen
inside the algae plays a key role in determining the response of
the species when exposed to elevated temperatures. Similar
results were also obtained in our study. Juvenile sporophytes
from Naruto showed a greater capacity to accumulate higher
nitrogen reserves than sporophytes from northern populations,
especially at high temperatures. It is generally known that
intracellular nitrogen contents determine the quantity of pig-
ments such as chlorophyll (Chapman et al. 1978; Bird et al.
1982; Lingnell and Pedersen 1987) and the photosynthetic
activity of algae (Levy and Gantt 1990). Indeed, sporophytes
from Naruto had significantly higher photosynthetic rates at
high temperatures of 20–35°C compared to specimens from
the northern populations. This phenomenon was also reported
for S. latissima (Gerard and DuBois 1988) and Ecklonia cava
(Serisawa et al. 2004). Thus, high internal nitrogen reserves
may contribute to maintenance of photosynthetic activity and
consequent rapid growth and a high survival percentage. It
should be noted that the juvenile sporophytes derived from
three different populations were cultured in the same light,
temperature, and enriched nutrient conditions; thus, the influ-
ence from those abiotic factors could be excluded. Therefore,
the observed physiological differences, especially the different
capacities for nitrogen accumulation, likely are attributable to
genetic differentiation among the populations rather than phe-
notypic plasticity.
Water-temperature tolerance is at least partly responsible
for the geographic distribution patterns of marine plants
(Lüning 1990). Due to natural selection, genetic differenti-
ation has been demonstrated in a number of macroalgal
species in relation to a variety of environmental factors
derived from differences in latitude (Russell 1986). In Ja-
pan, juvenile sporophytes of U. pinnatifida from both
farmed and natural populations have been shown to exhibit
genetic variation in high-temperature tolerance along the
latitudinal gradient (Gao et al. 2012; this study). Due to
the rapid change of temperature conditions from north to
south along the coasts of Japan (van den Hoek and Breeman
1990), selection pressure probably has consistently operated
in adapting U. pinnatifida to various temperature conditions.
J Appl Phycol (2013) 25:567–574
The ability to acclimate to a wide range of temperatures
partially accounts for the success of U. pinnatifida over a
large geographic range, and genetic adaptation appears to be
critical to long-term persistence of the populations exposed
to suboptimal temperature conditions.
The different high-temperature tolerance of U. pinnati-
fida ecotypes and their genetic stabilities demonstrated in
this study may have important implications for cultivation.
U. pinnatifida farming in Japan can be divided into two
principal phases: seedling production, which is conducted
in cooled tanks from late spring to early autumn, and sub-
sequent open sea cultivation until the following spring
(Ohno 2004). The success of summer-seedling production
in cooled water makes it possible to cultivate plants that will
reach marketable size. More tolerance to high temperature
would allow an earlier shift of production and transfer of
juvenile sporophytes from cooled tanks to the open sea, there-
by reducing cooling costs, which in turn would benefit Unda-
ria farmers. It has been proposed that the high-temperature-
tolerant strains/ecotypes are a more ideal source of parental
plants for use in commercial cultivation of Saccharina japon-
ica (Pang et al. 2007; Liu and Pang 2010). Boyer (1982)
reported that populations adapted to unfavorable or stressful
environments may be more productive under suboptimal con-
ditions. Therefore, plants collected from the Naruto natural
population should be used as the parental stock in mariculture
of U. pinnatifida in Japan.
Acknowledgments We sincerely thank the staff members of the
Miyagi Prefecture Fisheries Technology Center, Iwate Fisheries Tech-
nology Center, and Tokushima Agriculture, Forestry, and Fisheries
Technology Support Center for providing the juvenile sporophytes
and long-term seawater temperature data used in the present study
and for their help with kelp cultivation and collection. We also thank
the staff of the Miyagi Fisheries Technology Center for allowing us to
use their seawater for the culture experiment.
References
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