Algae 2012, 27(2): 83-94http://dx.doi.org/10.4490/algae.2012.27.2.083
Open Access
Research Article
Copyright © The Korean Society of Phycology 83 http://e-algae.kr pISSN: 1226-2617 eISSN: 2093-0860
Phylogenetic relationships and distribution of Gelidium crinale and G. pusillum (Gelidiales, Rhodophyta) using cox1 and rbcL sequences
Kyeong Mi Kim1 and Sung Min Boo1,*1Department of Biology, Chungnam National University, Daejeon 305-764, Korea
The taxonomic distinctiveness and cosmopolitan distributions of the red algae Gelidium crinale and G. pusillum re-
main unclear. Both species were first described in Devon in southwestern England; namely in Ilfracome for G. crinale
and Sidmouth for G. pusillum. We analyzed mitochondrial cox1 and plastid rbcL sequences from specimens collected in
East Asia, Australia, Europe and North America. In all phylogenetic analyses of cox1 and rbcL sequences, G. crinale was
distinct from congeners of the genus. The analyses also revealed a sister relationship with the G. coulteri and G. capense
clade. Nineteen cox1 haplotypes were identified for G. crinale, and they were likely geographically structured. Despite the
distinctiveness in both cox1 and rbcL datasets, the sister relationship of G. pusillum in the genus was not resolved. Our
cox1 and rbcL datasets indicate that G. crinale is a cosmopolitan species, found in East Asia, Australia, Europe and North
America, while the distribution of G. pusillum is restricted to Europe and Atlantic North America. Our results suggest that
infraspecific classification of G. pusillum may be abandoned.
Key Words: cox1; distribution; Gelidium crinale; Gelidium pusillum; phylogeny; rbcL
INTRODUCTION
Gelidium Lamour. is composed of approximately 127
described species distributed globally along tropical, sub-
tropical, and artic shorelines (Freshwater and Rueness
1994, Shimada et al. 2000, Millar and Freshwater 2005,
Kim et al. 2011, in press, Guiry and Guiry 2012). Members
of the genus can be the most abundant organisms within
intertidal algal assemblages. Gelidium is economically
important as food, and one of the most promising agar
sources in rhodophytes. It has recently been used for in-
dustrial paper pulp production in Korea (Seo et al. 2010).
However, identification of individual Gelidium speci-
mens is notoriously difficult because of the high degree of
morphological variation, particularly in the smaller and
medium-sized species (Dixon and Irvine 1977a).
Gelidium crinale (Hare ex Turner) Gallion and G. pusil-
lum (Stackhouse) Le Jolis, which are small and morpho-
logically diverse, are among the most difficult species to
identify in red algae, and their distributions are unclear.
These species are traditionally recognized as two distinct
species (Feldmann and Hamel 1936, Silva et al. 1996), and
Womersley and Guiry (1994) found a difference between
the types of G. crinale and G. pusillum. On the contrary, G.
crinale and G. pusillum were merged by Dixon and Irvine
(1977a, 1977b). Seven to nine varieties or formas have
been described in each of G. crinale and G. pusillum (Silva
et al. 1996, Guiry and Guiry 2012). The name G. pusillum
is commonly used for any small tuft-forming Gelidium
(Silva et al. 1996). Although G. crinale and G. pusillum are
considered as the most widely distributed species in the
genus AlgaeBase (Guiry and Guiry 2012), the occurrence
Received February 27, 2012, Accepted May 28, 2012
*Corresponding Author
E-mail: [email protected]: +82-42-821-6555, Fax: +82-42-822-9690
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://cre-ativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Algae 2012, 27(2): 83-94
http://dx.doi.org/10.4490/algae.2012.27.2.083 84
and Fredericq 2002); and for cox1, cox143F-cox11549R
(Geraldino et al. 2006) and C622F-C880R (Yang et al.
2008).
Ninety-three rbcL sequences including 13 new se-
quences and 65 cox1 sequences including 24 new Gelid-
ium sequences were collated using the Se-Al version
2.0a11 software (Rambaut 1996) and aligned visually.
Outgroup taxa used were representatives from Gelidiella
Feldmann et G. Hamel, Pterocladia J. Agardh, Pterocla-
diella Santelices et Hommersand, and Ptilophora (Suhr)
Kützing (Freshwater et al. 1995, Kim et al. 2011).
Maximum likelihood (ML) phylogenetic analysis of
rbcL was performed using the GTR + Γ + I model imple-
mented in RAxML software (Stamatakis 2006). We used
200 independent tree inferences with the “number of
run” option, with default optimized subtree pruning and
regrafting (SPR) rearrangement and 25 distinct rate cate-
gories to identify the best tree. Statistical support for each
branch was obtained from 1,000 bootstrap replications
using the same substitution model and RAxML program
settings.
Bayesian analyses (BA) were performed for combined
and individual datasets with MrBayes v.3.1.1 (Ronquist
and Huelsenbeck 2003) using the Metropolis-coupled
Markov chain Monte Carlo (MC3) with the GTR + Γ + I
model. For each matrix, one million generations of two
independent runs were performed with four chains and
sampling trees every 100 generations. The burn-in peri-
od was identified graphically by tracking the likelihoods
at each generation to determine whether they reached
a plateau. The 15,001 trees for rbcL and 22,501 trees for
cox1 sampled at the stationary state were used to infer the
Bayesian posterior probability.
A statistical parsimony network of cox1 haplotypes was
created using TCS version 1.21 software (Clement et al.
2000). Haplotype and nucleotide diversity measurements
were performed using DnaSP software (Rozas and Rozas
1999).
RESULTS
Molecular analyses
A total of 93 sequences from Gelidium and outgroups
were aligned using a 1,266-nucleotide (nt) portion of
rbcL. Variable sites were found at 492 portions (38.9%),
and 393 portions (31%) were parsimoniously informa-
tive. All GenBank accessions of G. crinale from nine coun-
tries formed a single monophyletic group with maximum
of both species in many countries should be reassessed.
In this study we characterized two species, namely G.
crinale and G. pusillum, using two molecular markers.
To evaluate the relationship and distribution of G. crinale
and G. pusillum, we analyzed plastid rbcL, and mitochon-
drial cox1 including type locality materials. Plastid rbcL is
commonly used for Gelidium phylogeny (Freshwater and
Rueness 1994, Freshwater et al. 1995, Shimada et al. 2000,
Millar and Freshwater 2005, Nelson et al. 2006, Kim et al.
2011). Recent studies have revealed that mitochondrial
cox1 is useful for both DNA bar-coding of gelidioid red
algae and to examine their distribution patterns (Fresh-
water et al. 2010, Wiriyadamrikul et al. 2010, Kim et al.
2012). In this study, we included published rbcL sequenc-
es analyzed from G. crinale type material (Freshwater et
al. 2010) and samples collected in the G. pusillum type
locality.
MATERIALS AND METHODS
Taxon sampling and morphological observation
A total of 18 specimens of G. crinale were obtained for
this study: 17 collected from 10 locations including Chi-
na, Hong Kong, Korea, Spain and the UK, and one strain
from the culture collection of the University of Texas,
UTEX (Appendix A). Ten G. pusillum field collections
were made at five locations in France, Spain and the UK.
Materials for morphological observations were mounted
on herbarium sheets, while clean apical parts of the spec-
imens were desiccated in silica gel for DNA extraction.
Tissues were sectioned using a freezing microtome (FX-
802A; Coper Electronics Co., Ltd., Kanagawa, Japan), and
sectioned preparations were stained with 1% aqueous
aniline blue. Photographs were taken with an FX-35DX
camera (Nikon, Tokyo, Japan) attached to a Vanox AHBT3
microscope (Olympus, Tokyo, Japan). Voucher specimens
were deposited at the herbarium of Chungnam National
University (CNUK), Daejeon, Korea.
DNA extraction, sequencing and phylogenetic analyses
Twenty-eight specimens were available for DNA extrac-
tion (Appendix A). DNA extraction, PCR amplification,
and sequencing are described in Geraldino et al. (2010).
Primer pairs for amplification and sequencing of each
gene were as follows: for rbcL, F7-R753 and F645-RrbcS
start (Freshwater and Rueness 1994, Lin et al. 2001, Gavio
Kim & Boo Gelidium crinale and G. pusillum
85 http://e-algae.kr
Fig. 1. Maximum likelihood tree of Gelidium using 93 rbcL sequences calculated using the GTR + Γ + I evolution model (-lnL = 10511.506124; substitution rate matrix RAC = 0.951940, RAG = 5.742494, RAT = 1.371355, RCG = 1.258690, RCT = 10.348932, RGT = 1; shape parameter [α] = 1.575834). Maximum likelihood bootstrap values and Bayesian posterior probabilities are shown for each clade. Only bootstrap values ≥50% and ≥0.95 Bayesian posterior probabilities are shown.
Algae 2012, 27(2): 83-94
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Fig. 2. Maximum likelihood tree of Gelidium using 65 cox1 sequences calculated using the GTR + Γ + I evolution model (-lnL = 9341.027999; substitution rate matrix RAC = 1.153854, RAG = 10.881554, RAT = 1.077417, RCG = 0.000017, RCT = 20.342285, RGT = 1; shape parameter [α] = 1.101521). Maximum likelihood bootstrap values and Bayesian posterior probabilities are shown for each clade. Only bootstrap values ≥50% and ≥0.95 Bayesian posterior probabilities are shown.
Kim & Boo Gelidium crinale and G. pusillum
87 http://e-algae.kr
terete to flattened distally (up to 550 µm wide). Spanish
specimen appears to have cylindrical branches (Fig. 4D).
Large dome-shaped apical cells are evident at the apices
and project over the cortical margin (Fig. 4E). Axes and
branches consist of cortex and medulla (Fig. 4F). Three
to five layered cortex consist of small, pigmented, rect-
angular surface, and oval inner cortical cells. Medulla is
composed of large, colourless medullary cells that were
intermixed with internal rhizoidal filaments.
G. pusillum thalli are cartilaginous and prostrate axes
are terete, quite regular in diameter, and erect branches
(Fig. 5A-C). Erect axes arise from the dorsal sides of inde-
terminate, prostrate axes, are compressed and up to 1.5
cm in height. Branches develop irregularly branching of
up to three orders. Dome-shaped apical cells are present
at the apices (Fig. 5D). Axes and branches consist of cor-
tex and medulla (Fig. 5E). The cortex consists of three to
four layers of small pigmented cells, whereas the medulla
is composed of large colorless cells.
DISCUSSION
Taxonomy of Gelidium crinale and G. pusillum
Based on both mitochondrial cox1 and plastid rbcL
datasets, Gelidium crinale was distinct from G. pusil-
lum and other congeners of the genus. All G. crinale rbcL
sequences from East Asia, Europe, Australia and North
America formed a monophyletic clade with a published
sequence (AF308786) from the type material (Freshwater
et al. 2010). G. crinale specimens from East Asia, Australia
and North America corresponded to the description by
Feldmann and Hamel (1936), and were identified based
on the thalli being caespitose and having cylindrical to
compressed prostrate and cylindrical erect axes with fili-
form branches. G. crinale formed a clade with G. capense
from South Africa and G. coulteri from USA. It was diffi-
cult to identify a synapomorphic characteristic for these
three species.
The intraspecific divergence (0.00-2.74%) of G. crinale
from Asia, Australasia, Europe and North America is simi-
lar to that (up to 2.65%) of the species in previous studies
(Freshwater et al. 2010) and that (0.00-2.45%) of Hypnea
flexicaulis Yamaighi et Masuda (Geraldino et al. 2006).
Seven varieties or formas have been described in G.
crinale; f. luxurians Collins (type locality, Pacific Beach,
San Diego Co. California) (Collins et al. 1906), var. lu-
bricum (Kützing) Hauck, var. spathulatum (Kützing)
Hauck (Adriatic Sea) (Womersley and Guiry 1994), var.
support (Fig. 1). The G. crinale clade consisted of three
subgroup; Asian, Australian, and European / American
group. Piarwise divergence of G. crinale was up to 2.74%.
G. crinale was sister to the clade of G. coulteri Harvey and
G. capense (S. G. Gmelin) P. C. Silva (93% for ML and 1.0
for BA). Eleven of G. pusillum sequences from France,
Norway, Spain, and UK formed a monophyletic group
(100% for ML and 1.0 for BA).
A total of 65 sequences from Gelidium and three out-
groups were aligned using a 1,200 nt region of the cox1
gene. Among the 447 (37.3%) variable sites, 400 portions
(33.3%) were parsimoniously informative. The topology
based on cox1 sequences was congruent with the rbcL
phylogeny (Fig. 2). The cox1 ML tree showed that all G.
crinale from eight countries were monophyletic (99% for
ML and 1.0 for BA). Twelve G. pusillum from France, Nor-
way, Spain, UK, and USA were monophyletic with maxi-
mum support.
Because of short sequences of cox1 in GenBank, in
haplotype analyses, we used 618 nt cox1 fragment of G.
crinale from 28 individuals collected in Australia, China,
Hong Kong, Korea, Puerto Rico, Spain, UK, and USA. A to-
tal of 19 haplotypes among 33 polymorphic sites (5.3%)
were found. Haplotype and nucleotide diversities of cox1
within G. crinale were 0.912 ± 0.049 (H) and 0.014 ± 0.005
(π), respectively. The 19 haplotypes were placed in three
geographically distinct groups; Asian, Australian, and Eu-
ropean / American groups (Fig. 3). All haplotypes in Asia
were closely related. However, H7 from Hainan, China
was linked to H5 (with six missing haplotypes) and to H3
and H4 (with seven missing haplotypes). Haplotypes H9-
H11 were found in Australia, and haplotypes H12-H19
were found in Puerto Rico, Spain, UK, and USA.
Eleven G. pusillum sequences from France, Norway,
Spain, and UK formed a monophyletic group with maxi-
mum support. Pairwise divergence of G. pusillum was
0.24%. Four haplotypes were found from nine individu-
als of G. pusillum from France, Spain and UK (data not
shown). Four specimens from France and UK shared
same haplotype. Three short cox1 sequences (450 nt long)
from NCBI (HQ412445-7) from France, Norway and USA
were also dentical.
Morphology of Gelidium crinale and G. pusillum
Specimens of G. crinale confirmed by cox1 and rbcL are
shown in Fig. 4. Thalli (Fig. 4A & B) from Jeoncheon, Ko-
rea and Qingdao, China are purple-red, cartilaginous and
composed of terete prostrate axes and erect axes (up to 2
cm high). Specimen from Seoko, Hong Kong (Fig. 4C) are
Algae 2012, 27(2): 83-94
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lum, three are considered as synonyms of the species; var.
conchicolum Piccone et Grunow (type locality, Massawa,
Eritrea, Ethiopia) (Piccone 1884), f. foliaceum Okamura
(Type locality, Shiso-dima, Seto, Kii Province, Japan)
(Okamura 1934), and var. minusculum Weber-van Bosse
(type locality, Daram Inlet, East coast of Misoöl Island,
Indonesia) (Weber-van Bosse 1921) (see AlgaeBase, Guiry
and Guiry 2012). However, six have still been flagged in
the AlgaeBase; var. cylindricum W. R. Taylor (type locality,
Bahia San Francisco, Ecuador) (Taylor 1945), var. mucro-
natum P. J. L. Dangeard, var. pacificum W. R. Taylor (Isla
Santa Maria, Galapagos Islands, Ecuador) (Taylor 1945),
f. pakistancium Afaq-Husain et Shameel (type locality,
Gadani, Karachi, Pakistan) (Afaq-Husain and Shameel
1999), var. pulvinatum (C. Agardh) Feldmann (type local-
ity, Cadíz, Spain) (Feldmann and Hamel 1936), and var.
simplex P. J. L. Dangeard (type locality, Marocco) (Dan-
geard 1949).
The monophyly and low genetic variation (up to 0.24%)
of G. pusillum reveal that G. pusillum is a species with
less genetic diversities and indicate the above six varieties
or formas may be synonyms of the species or belong to
other species. For example, four varieties of G. pusillum
(e.g., var. conchicola, var. cylindricum, var. pacificum, and
var. pulvinatum) reported in Korea (Lee 1994, Lee and
Kim 1995) have not been found in our recent studies de-
spite many trips in their collection sites (Kim et al. 2011,
corymbosum (Kützing) Feldmann et G. Hamel (type lo-
cality, Mediterranean Sea, Italy) (Feldmann and Hamel
1936), var. perpusillum Piccone et Grunow (Eritrea, Mas-
sawa, Italy) (Piccone 1884), var. platycladum W. R. Taylor
(type locality, Port Aransas, Texas, USA) (Taylor 1943) and
var. polycladum (Kützing) Hauck. Of these, var. corymbo-
sum, var. platycladum, and var. perpusillum have been
accepted taxonomically in AlgaeBase (Guiry and Guiry
2012).
Our cox1 haplotype network revealed geological struc-
ture of the G. crinale populations, but pairwise diver-
gence (up to 2.74% in cox1) is in a range of other species,
as mentioned in above. It is therefore difficult to conclude
whether our molecular data support infraspecific classifi-
cation or not. Further sampling is necessary for confirm-
ing the infraspecific categories.
G. pusillum specimens from France, Norway, Spain,
and USA formed a monophyletic clade with those collect-
ed from the type locality (Sidmouth, Devon, England) in
both the cox1 and rbcL trees. G. pusillum is characterized
by tufted thalli having compressed, oval, or lanceolate
branches (Feldmann and Hamel 1936, Silva et al. 1996).
All specimens from UK, France and Spain are similar in
having compressed thalli and oval or lanceolate branch-
es. The sister relationship of G. pusillum in the genus was
not resolved in the cox1 and rbcL trees.
Of nine varieties or formas described within G. pusil-
Fig. 3. Nineteen cox1 haplotypes (H1-H19) network of Gelidium crinale from 22 localities in Australia, China, Hong Kong, Korea, Puerto Rico, Spain, UK, and USA. Small grey circles correspond to missing haplotypes and the size of each circle is proportional to the number of individuals analyzed. Numerals in parentheses refer to the number of specimens with identical sequences. AU, Australia; CH, China; ES, Spain; HK, Hong Kong; KR, Korea; PR, Puerto Rico; UK, United Kingdom; USA, United States.
Kim & Boo Gelidium crinale and G. pusillum
89 http://e-algae.kr
A C
D
B
E F
Jeoncheon, KoreaSep 25, 2010
Cadíz, SpainApr 12, 2010
Qingdao, ChinaOct 11, 2010
Seoko, Hong KongApr 23, 2008
Fig. 4. Morphology of Gelidium crinale. (A) Specimen collected in Jeoncheon, Korea. (B) Specimen collected in Qingdao, China. (C) Specimen collected in Seoko, Hong Kong. (D) Specimen collected in Cadíz, Spain. (E) A dome-shaped apical cell at the tip of a branchlet. (F) Transverse section of an axis. Scale bars represent: A-D, 5 mm; E, 10 μm; F, 50 μm.
Fig. 5. Morphology of Gelidium pusillum. (A) Specimen collected in type locality (Sidmouth, Devon, UK). (B) Specimen collected in Cadíz, Spain. (C) Specimen collected in Caen, France. (D) A dome-shaped apical cell at the tip of a branchlet. (E) Transverse section of an axis. Scale bars represent: A-C, 5 mm; D, 10 μm; E, 50 μm.
A C
D
B
E
Sidmouth, UKApr 4, 2010
Cadíz, SpainApr 11, 2010
Caen, FranceNov 17, 2009
Algae 2012, 27(2): 83-94
http://dx.doi.org/10.4490/algae.2012.27.2.083 90
Freshwater 2005, Freshwater et al. 2010)
Contrary to previous studies on its global distribution
(e.g., Silva et al. 1996, Guiry and Guiry 2012), our cox1 and
rbcL datasets revealed that G. pusillum is likely restricted
to Europe and Atlantic North America. Despite basic lo-
cal alignment search tool (BLAST) search of all sequences
registered in GenBank as well as sequences generated in
the present study, no accession of G. pusillum from Asia,
Australia and Pacific North America matched those from
Europe. However, G. pusillum specimens from Spain and
France formed a clade with those from Sidmouth, UK.
We suggest that records of G. pusillum from Europe and
Atlantic North America may be the result of misiden-
tifications, and the specimens filed under G. pusillum
should be treated with caution. We agree with the views
of Millar and Freshwater (2005) that G. pusillum, reputed
to be cosmopolitan (see Silva et al. 1996, Guiry and Guiry
2012), may be limited to areas around Europe and Atlan-
tic North America.
Conclusions
Identifications of G. crinale and C. pusillum based on
morphology alone is problematic and usually requires
molecular analyses for confirmation. Thus, reports of
G. pusillium outside from of Europe and Atlantic North
America should be treated with caution, and herbarium
specimens identified as G. pusillum in East Asia, Austra-
in press). Instead, at least one new species in Korea are
likely previously misidentified as variant of G. pusillum
(Kim et al. in press). Specimen (Fig. 5B) from the type
locality of G. pusillum var. pulvinatum, Cadíz, Spain did
not reveal variation in molecular data and morphology.
Therefore, we suggest that infraspecific classification of
G. pusillum may be abandoned.
Distribution of Gelidium crinale and G. pusillum
A distribution map of G. crinale and G. pusillum is
shown in Fig. 6. Two interesting biogeographic patterns
emerge when considering the overall distribution of both
species and their distinct lineages revealed by phyloge-
netic analyses. The parsimony network of cox1 haplo-
types revealed a geographic structure (Fig. 3); Asian, Aus-
tralian, and European / American groups. In the Asian
group, seven haplotypes were found and closely related.
Australian group is quite distinct from Asian, and Europe-
an / American groups. Specimens from North Carolina,
USA and Europe made one group with a single missing
haplotype. The genetic connectivity between Europe and
north America may be caused by similar oceanographic
conditions. Our results indicate that G. crinale is a cosmo-
politan species, although there are high morphological
variations and intraspecific divergences. This result is in
agreement with previous studies showing that G. crinale
is distributed globally (Shimada et al. 1999, Millar and
Fig. 6. Map showing current geographic distribution of Gelidium crinale and G. pusillum based on the present study and previous publications (Freshwater and Rueness 1994, Freshwater et al. 1995, 2010, Shimada et al. 1999, Millar and Freshwater 2005, Kim et al. 2011).
Kim & Boo Gelidium crinale and G. pusillum
91 http://e-algae.kr
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ACKNOWLEDGEMENTS
We thank Jeong Kwang Park for preparing plates of
morphology, Juliet Brodie and Keith Hiscock for help in
collection trip in Devon, UK and Dr. José Lucas Perez in
Cadíz, Spain, respectively. This work was supported by
Marine Biotechnology Grants from the Ministry of Land,
Transportation Maritime Affairs and Basic Science Grant
(2012-0704) of Korean Research Foundation to SMB.
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Appendix A. Gelidium crinale and G. pusillum samples used in the present study
Collection sites (date)GenBank accession no.
SourcerbcL cox1
Gelidium crinale (Hare ex Turner) Gaillon
Australia; Fish Hook Bay, Rottnest Is. AF308788 - Freshwater et al. 2010
Australia; Fish Hook Bay, Rottnest Is. AY350780 - Freshwater et al. 2010
Australia; Green Is., Rottnest Is. (Aug 14, 2002) HQ412492 HQ412464 Freshwater et al. 2010
Australia; Green Is., Rottnest Is. (Aug 14, 2002) HQ412493 HQ412465 Freshwater et al. 2010
Australia; Old Gulch, Lord Howe Is. - HQ412466 Freshwater et al. 2010
Australia; Summer Cloud Bay, Jervis Bay AY350781 HQ412467 Freshwater et al. 2010
China; Donghai road, Qingdao (Oct 11, 2010) JX096509 JX096528 In this study
China; Donghai road, Qingdao (Oct 11, 2010) JX096510 - In this study
China; Hainan (May 11, 2009) JX096511 JX096529 In this study
China; Hainan (May 11, 2009) JX096512 JX096530 In this study
China; Oriental Hotel, Yantai (Sep 12, 2011) JX096513 JX096531 In this study
Hong Kong; Seoko (Apr 23, 2008) - JX096532 In this study
Hong Kong; Seoko (Apr 23, 2008) JX096514 JX096533 In this study
Hong Kong; Seoko (Apr 23, 2008) JX096515 JX096534 In this study
Japan; Awaji Is., Hyogo Pref. (Oct 1, 1996) AB017679 - Shimada et al. 1999
Korea; Bumseom, Jeju (Aug 23, 2009) JX096516 - In this study
Korea; Bumseom, Jeju (Jul 5, 2009) JX096517 JX096535 In this study
Korea; Dugok, Namhaegun (2010) - JX096536 In this study
Korea; Dugok, Namhaegun (2010) - JX096537 In this study
Korea; Dugok, Namhaegun (2010) - JX096538 In this study
Korea; Jeoncheon (Nov 25, 2010) JX096518 JX096539 In this study
Korea; Muchangpo, Boryeong (Jul 29, 2007) HM629823 HM629863 Kim et al. 2011
Korea; Pyeongdae, Jeju (May 29, 2010) - JX096540 In this study
Puerto Rico; Playa Esperanza, Manati U00983 HQ412463 Freshwater et al. 1995, 2010
Spain; Aramar, Asturias AF308791 - Freshwater et al. 2010
Spain; Aramar, Asturias AF308792 - Freshwater et al. 2010
Spain; La Arana, Malaga AF308789 - Freshwater et al. 2010
Spain; La Garita, Canary Is. AF308793 - Freshwater et al. 2010
Spain; Tarifa, Cadíz (Apr 12, 2010) JX096519 JX096541 In this study
UK; Ilfracombe, Devon AF308786 - Freshwater et al. 2010
UK; Sidmouth, Devon (Apr 4, 2010) JX096520 JX096542 In this study
USA; Beaufort Inlet, North Carolina AF308795 HQ412458 Freshwater et al. 2010
USA; Bogue Sound, North Carolina (Feb 16, 1991) HQ412489 HQ412460 Freshwater et al. 2010
USA; Fort Fisher, North Carolina (May 14, 1991) - HQ412461 Freshwater et al. 2010
USA; Marineland, Florida AF308794 - Freshwater et al. 2010
USA; Masonboro Inlet, North Carolina U00981 HQ412457 Freshwater et al. 1995
USA; North Carolina AF308790 - Freshwater et al. 2010
USA; Port Aransas, Texas U00982 - Freshwater et al. 1995
USA; Port Aransas, Texas (UTEX culture collection) JX096521 JX096543 In this study
USA; Radio Is., North Carolina (Feb 16, 1991) HQ412488 HQ412459 Freshwater et al. 2010
USA; Stump Sound, North Carolina (Jul 11, 2003) HQ412491 HQ412462 Freshwater et al. 2010
Algae 2012, 27(2): 83-94
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Appendix A. Continued
Collection sites (date)GenBank accession no.
SourcerbcL cox1
Gelidium pusillum (Stackhouse) Le Jolis
France; Cancale, Brittany U01000 HQ412446 Freshwater and Rueness 1994, Freshwater et al. 2010
France; Lion sur-mer, Caen (Nov 17, 2009) HM629832 HM629872 Kim et al. 2011
France; Lion sur-mer, Caen (Nov 17, 2009) JX096522 - In this study
France; Wimereux U01001 - Freshwater and Rueness 1994
Norway; Fedje, Hordaland U00999 HQ412445 Freshwater et al. 1995, 2010
Spain; El Chato, Cadíz (Apr 11, 2010) JX096523 JX096544 In this study
Spain; El Chato, Cadíz (Apr 11, 2010) JX096524 JX096545 In this study
UK; Ilfracombe, Devon (Apr 3, 2010) - JX096546 In this study
UK; Ilfracombe, Devon (Apr 3, 2010) - JX096547 In this study
UK; Ilfracombe, Devon (Apr 3, 2010) - JX096548 In this study
UK; Ilfracombe, Devon (Apr 3, 2010) - JX096549 In this study
UK; Penmon, Anglesey U01002 - Freshwater and Rueness 1994
UK; Sidmouth, Devon (Apr 4, 2010) JX096525 JX096550 In this study
UK; Sidmouth, Devon (Apr 4, 2010) JX096526 - In this study
UK; Sidmouth, Devon (Apr 4, 2010) JX096527 JX096551 In this study
USA; Masonboro Inlet, North Carolina (Dec 30, 2006) - HQ412447 Freshwater et al. 2010