Grateloupia jejuensis (Halymeniales, Rhodophyta): a new species previously confused with G. elata and G. cornea in Korea
Grateloupia jejuensis (Halymeniales, Rhodophyta): a new species previously confused with G. elata and G. cornea in Korea
ALGAE. 2013. Sep, 28(3): 233-240
Copyright ©2013, The Korean Society of Phycology
This is an Open Access article distributed under the terms of theCreative Commons Attribution Non-Commercial License ( which permits unrestrictednon-commercial use, distribution, and reproduction in any medium,provided the original work is properly cited.
  • Received : April 12, 2013
  • Accepted : August 21, 2013
  • Published : September 15, 2013
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About the Authors
Su Yeon, Kim
Department of Biology, Chungnam National University, Daejeon 305-764, Korea
Eun Gyu, Han
Department of Biology and Research Institute for Basic Sciences, Jeju National University, Jeju 690-756, Korea
Myung Sook, Kim
Department of Biology and Research Institute for Basic Sciences, Jeju National University, Jeju 690-756, Korea
Jung Kwang, Park
Department of Biology, Chungnam National University, Daejeon 305-764, Korea
Sung Min, Boo
Department of Biology, Chungnam National University, Daejeon 305-764, Korea

Despite specimens’ large size and ease of collection in northeast Asian waters, the species diversity of the genus Grateloupia still needs more research in Korea. We investigated plastid rbc L sequences and carried out detailed morphological observation on flattened halymeniacean red alga collected in twelve locations around Korea and Japan. We describe Grateloupia jejuensis sp. nov. based on the distinct clade with high support in our rbc L tree. Grateloupia jejuensis is characterized by solitary or caespitose habit and flattened thalli with discoid holdfast, cartilaginous texture, and blunt or bifid axis. Grateloupia jejuensis was distantly related to G. elata and G. cornea , which have been morphologically confused with the former, and it formed a sister relationship with Prionitis filiformis from California, USA in the rbc L tree.
The genus Grateloupia was established by C. Agardh (1822) based on G. filicina (J. V. Lamouroux) C. Agardh. Grateloupia is distinguished by Grateloupia -type auxiliary cell ampullae with a single primary ampullar filament and two or three 7- to 13-celled secondary ampullar filaments of a conical out line (Chiang 1970, Kawaguchi 1989, Wang et al. 2000). Recent studies combining morphological and molecular data have resulted in several taxonomic changes. Pachymeniopsis Y. Yamada, Prionitis J. Agardh, Dermocorynus P. L. Crouan & H. M. Crouan, and Phyllymenia J. Agardh have been merged into Grateloupia (Kawaguchi 1997, Wang et al. 2001, De Clerck et al. 2005 b , Wilkes et al. 2005) while G. intestinalis (Harvey) Setchell moved to Glaphyrosiphon Hommersand & Leister (Hommersand et al. 2010).
Grateloupia elata (Okamura) S. Kawaguchi & H. W. Wang was described based on specimens under the name Prionitis elata Okamura (type locality: Shirahama, Chiba Prefecture, located at the Pacific side of Japan). In Korea, Cotton (1906) reported this species (as Prionitis elata ) for the first time at Wonsan on the northeast coast of the Korean peninsula. Afterwards, G. elata was reported between Busan and Gangneung on the east coast of Korea and Gapado on Jeju Island (Lee and Kang 2001).
Grateloupia cornea Okamura (1913) was transferred to the genus Carpopeltis and then changed to Prionitis cornea (Okamura) E. Y. Dawson (Dawson 1958). However, Wang et al. (2001) reinstated the original name, Grateloupia cornea , when they merged Prionitis into Grateloupia . In Korea, Kang (1966) reported this species on the east to south coasts and on Jeju Island. Subsequently, this species has been reported in many floristic reports (Lee and Kang 2001).
The taxonomy of the genus Grateloupia remains confused due to the low taxonomic utility of vegetative morphological characteristics, such as surface morphology, thallus habit and texture, and frequency of proliferations (Wang et al. 2001, De Clerk et al. 2005 a , Wilkes et al. 2005). Recent studies show that the rbc L gene is a suitable marker for taxonomic study of Grateloupia (Wang et al.
Materials used in the present study
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Materials used in the present study
2000, Faye et al. 2004, Lee et al. 2009, Yang et al. 2013). In the present study, we analyzed rbc L from unidentified specimens in Korea as well as other species and observed morphological characters. As a result, we describe Grateloupia jejuensis sp. nov. and discuss the phylogenetic relationships of G. jejuensis with the other species of the genus.
In total, 38 specimens of 13 species in the Halymeniaceae were collected from Korea, China, Japan, Russia, and USA. Field observation and collections of Grateloupia jejuensis were made at intertidal zone of 12 locations around Korea and Japan from 2009 to 2012.
Specimens were sectioned with a freezing microtome (FX-801; Yamato Kohki Industrial Co. Ltd., Tokyo, Japan). Photographs were taken with a DP-71 camera (Olympus, Tokyo, Japan) attached to a microscope (BX 51; Olympus). Voucher specimens are housed at the herbarium of Chungnam National University, Daejeon, Korea (CNUK).
We analyzed 84 sequences representing 48 Halymeniaceae species, including 38 new sequences and 46 GenBank sequences. Information for specimens, their collection data, and GenBank accession numbers of rbc L sequences are given in Table 1 . DNA extraction, PCR amplification, and sequencing follow the procedures in Boo et al. (2010). For amplification and sequencing reaction of the rbc L gene, specific primer pairs were rbc LF145- rbc LR898 and rbc LF762- rbc LR1442 (Kim et al. 2010). Electropherogram outputs from each sample were edited using Chromas version 1.45 ( ). Eighty-nine rbc L sequences were collated and aligned using Se-Al (version 2.0a11), as per Rambaut (2002). Maximum likelihood analyses were conducted using RAxML (Stamatakis 2006) with the GTR + Γ evolutionary model. We performed 200 independent tree inferences using the -# option with default ‑I (automatically optimized SPR rearrangement) and ‑c (25 distinct rate categories) option in the program to identify the best tree. To generate bootstrap values for the best phylogeny, we used 1,000 replications under the same program with the same model settings.
- Grateloupia jejuensis S. Y. Kim, E. G. Han & S. M. Boo sp. nov. (Fig. 1)
Thallus solitariam aut caespitosa, applanatis et dichotomis calami, saepe flabellatae in adumbration, apice cum terminus in obtunsus aut bifida, cartilaginous in textum. Cortex cum 7-8 compactus corticales cellulis, sphaericus in intimus cellulis et angulata superficies cellulis. Medulla cum parvos sphaericus cellulis, 5-7 μm in diametros. Tetrasporic sporophylls, tenuis et parvos, oblongae, acuminatus ad basin, seriatum per duo marginibus. Tetrasporangi is cruciatis dividuus, plasmatio ex corticales cellam iacuit in the proliferous ramulis.
Thallus solitary or caespitose, flattened and dichotomous branches, often flabellate in outline, apex with ending in blunt or bifida, cartilaginous in texture. Cortex with 7-8 compact cortical cells, spherical in innermost cells and angulated surface cells. Medulla with small spherical cells, 5-7 μm in diameter. Tetrasporic sporophylls, thin and small, oblong, tapering at base, seriated along both margins. Tetrasporangia cruciately divided, forming from cortical cell layer in the proliferous branchlets.
Holotype. CNU040218, intertidal zone of Hado (33°31ʹ N 126°54ʹ E), Jeju, Korea, Oct 4, 2009 ( Fig. 1 A), Herbarium of Chungnam National University (CNUK), Daejeon, Korea. Isotypes: CNU040217, -19, -220-1, NIBRAL0000138259- 60.
Etymology. The specific epithet refers to Jeju Island, Korea, where the type was collected.
Distribution. Found from Gangneung on the east coast to Jinhae on the south coast, and from Ilkwari to Sungsan around Jeju Island. Grateloupia jejuensis was frequently found throughout the year on rocks in tide pools and sheltered places in lower tidal zones. Specimens were collected in February, June, July, October, and December; tetrasporophytes were collected in May and October. Gametophytes were not found.
Korean name. 댓잎도박
Morphology. Plants are solitary or caspitose, arising from discoid holdfast, and cartilaginous in texture. The thallus is dark purple to red in color and bright red in the upper portion. The thallus is 8-16 cm high, 0.2-0.4 cm wide, and 200-350 μm thick, with a short stipe (0.5-1 cm long). Branches are divided up to eight times dichotomously, and apex endings are blunt or bifid. The marginal branches produce proliferous branchlets that are often irregular and small (up to 5 mm long and 1 mm wide), scattered over the middle part of the thallus except for the stipe and basal part. The internal thallus is composed of compacted cortex and dense medulla. The cortex is 7-8 cell layers and is 5-20 μm in diameter. The outer cortex is composed of 3-4 layers with narrowly ellipsoidal cells, and the inner cortex is composed of 3-4 layers with irregular or rounded transparent cells. The medulla is composed of filamentous cells (5-7 μm in diameter). Tetrasporangia are formed from the cortical cell layer in the proliferous branchlets. The mature tetrasporangia are cruciately divided, narrowly ellipsoidal in shape, and 30-40 μm long by 10-15 μm wide.
Phylogeny of rbcL. 1,094-nucleotide portions of the rbc L gene were aligned for 84 sequences (38 new sequences),
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Grateloupia jejuensis S. Y. Kim, E. G. Han & S. M. Boo sp. nov. (A) Holotype (CNU040218, tetrasporophyte) of Grateloupia jejuensis collected from Hado, Jeju, Korea on Oct 4, 2009, deposited in the Herbarium of Chungnam National University (CNUK), Daejeon, Korea. (B) Close-up view of tetrasporic sporophylls (ts). (C) Cross section of thallus showing cortex (c) and medullar layers (m). (D) Longitudinal section of thallus showing cortex (c) and medullar layers (m). (E) Cross section of tetrasporic sporophylls. (F) Close-up view of a tetraspore (t). Scale bars represent: A, 2 cm; B, 5 mm; C, 100 μm; D & E, 50 μm; F, 20 μm.
representing 48 species of Halymeniaceae. All sequences of G. jejuensis from 12 sites in Korea and Japan were identical. The rbc L difference of G. jejuensis with Prionitis filiformis Kylin was 14-15 bp (1.28-1.37%), while G. jejuensis differed by 27-29 bp (2.28-2.45%) from G. elata and 63 bp (5.48%) from G. cornea .
In the phylogenetic tree ( Fig. 2 ), Grateloupia jejuensis was placed in a single clade and clearly separated from the other species of the genus. The sister species of G. jejuensis was Prionitis filiformis (86% for maximum likelihood). Grateloupia cornea was resolved as a sister taxon of G. chiangii from Japan with 91% bootstrap support. Grateloupia angusta shows sister relationships with G. elliptica and G. lanceolata .
The main finding of the present study is the discovery of a new species, Grateloupia jejuensis , in Korea based on rbc L sequence data and morphological evidence. It had previously been misidentified as G. elata (Lee 2008) or G. cornea (Lee 1987) because of similarity in cartilaginous texture, and erect, linear, compressed habit. In this context, the existence of G. elata and G. cornea in Korea may be questionable.
The comparative morphology of Grateloupia jejuensis with similar species is summarized in Table 2 . Grateloupia elata is much larger in thallus size than G. jejuensis (20- 50 cm vs. 8-16 cm). Kawaguchi et al. (2001) reported that the difference in size is the one of main characteristics to distinguish G. asiatica from G. filicina (10-30 cm vs. 9-12 cm). The width of G. elata is less than that of G. jejuensis (1.5-2.5 mm vs. 2-4 mm). The difference in habitat is also a remarkable characteristic between the two species: G. elata
Morphological comparison between Grateloupia jejuensis and the morphologically similar species
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Morphological comparison between Grateloupia jejuensis and the morphologically similar species
occurs in deep waters (Okamura 1899), whereas G. jejuensis usually occurs in tide pools and / or intertidal zone. Grateloupia cornea is also similar to G. jejuensis in having cartilaginous texture and linear habit. However, G. jejuensis has 7-8 cortical cell layers, while G. cornea has 13-15 cortical cell layers. The structure of medullary cells of G. jejuensis is more densely constructed than those of G. cornea .
Okamura (1899) stated that G. elata resembles G. angusta (Okamura) S. Kawaguchi & H. W. Wang in structure, habit, and substance, but differed in fruit-bearing (tetraspores and cystocarps) portion and cells of the intermediated layer. G. angusta is distinguished from G. jejuensis by composition of medulla and number of cortical cells. G. jejuensis has a densely composed medulla with 7-8 cortical cells, while G. angusta has a very densely composed medulla with 14-16 cortical cells.
Wang et al. (2001) merged Prionitis to Grateloupia ; however, the formal proposals were not for all species of Prionitis . The genus Prionitis is still used in the Halymeniaceae. In our rbc L tree, P. filiformis was the sister of G. jejuensis and grouped together with all other Grateloupia species with high bootstrap support. It is therefore suggested that P. filiformis can be transferred to Grateloupia . Before taxonomic revision, a more detailed morphological study of P. filiformis is needed.
In the phylogenetic tree ( Fig. 2 ), ten Korean species were grouped into four different subclades with strong bootstrap support. Grateloupia jejuensis was included in subclade I. Prionitis filiformis , P. sternbergii (C. Agardh) J. Agardh, Grateloupia americana from the northeast Pacific and Grateloupia acuminata Holmes, G. asiatica , G. divaricata , G. elata , G. jejuensis , G. livida (Harvey) Yamada, G. patens (Okamura) S. Kawaguchi & H. W. Wang, G. schmitziana (Okamura) S. Kawaguchi & H. W. Wang from
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Maximum likelihood (ML) tree inferred from rbcL sequences calculated using the GTR + Γ evolution model. Values above branches refer to ML bootstrap values. Bolded samples show the species which was newly analyzed in this study.
the northwest Pacific, were mixed within this subclade, suggesting that many species were likely to have diverged in the north Pacific Ocean. Grateloupia cornea and G. angusta formed subclade II. Given the morphological similarity to G. jejuensis , the habit similarity is a result of convergence. This result is consistent with previous studies (Wang et al. 2001, De Clerk et al. 2005 a , Wilkes et al. 2005, Lee et al. 2009). Thallus habits are homoplasious within Grateloupia ; thus, additional taxon sampling will provide a better understanding of phylogenetic relationships of the species.
We thank Shigeo Kawaguchi for comments on the presentwork. This research was supported by MarineBio21program grants from the Ministry of Maritime Affairs &Fisheries, Korea to S. M. Boo and the National Instituteof Biological Resources (NIBR), funded by the Ministryof Environment (MOE), Korea to M. S. Kim for collectingspecimens.
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Agardh C. A. 1822 Species algarum rite cognitae, cum synonymis,differentiis specificis et descriptionibus succinctis.Volumen primum pars posterior ex officina Berlingiana, Lundae [Lund] 169 - 398
Boo S. M. , Kim S. Y. , Hong I. S. , Hwang I. K. 2010 Reexaminationof the genus Pterocladiella (Gelidiaceae,Rhodophyta) in Korea based on morphology and rbcLsequences. Algae 25 1 - 9    DOI : 10.4490/algae.2010.25.1.001
Chiang Y. -M. 1970 Morphological studies of red algae of thefamily Cryptonemiaceae. Univ. Calif. Publ. Bot. 58 1 - 95
Cotton A. D. 1906 Marine algae from Corea. Bull. Misc. Inform. Kew. 1906 366 - 373
Dawson E. Y. 1958 Notes on Pacific coast marine algae, VII. Bull. South. Calif. Acad. Sci. 57 65 - 80
De Clerck O. , Gavio B. , Fredericq S. , Barbara I. , Coppejans E. 2005a Systematics of Grateloupia filicina (Halymeniaceae,Rhodophyta), based on rbcL sequence analysesand morphological evidence, including the reinstatementof G. minima and the description of G. capensis sp.nov. J. Phycol. 41 391 - 410
De Clerck O. , Gavio B. , Fredericq S. , Cocquyt E. , Coppejans E. 2005b Systematic reassessment of the red algal genus Phyllymenia (Halymeniaceae, Rhodophyta). Eur. J. Phycol. 40 169 - 178
Faye E. J. , Wang H. W. , Kawaguchi S. , Shimada S. , Masuda M. 2004 Reinstatement of Grateloupia subpectinata(Rhodophyta, Halymeniaceae) based on morphologyand rbcL sequences. Phycol. Res. 52 59 - 67
Hommersand M. H. , Leister G. L. , Ramírez M. E. , Gabrielson P. W. , Nelson W. A. 2010 A morphological and phylogeneticstudy of Glaphyrosiphon gen. nov. (Halymeniaceae,Rhodophyta) based on Grateloupia intestinaliswith descriptions of two new species: Glaphyrosiphonlindaueri from New Zealand and Glaphyrosiphonchilensis from Chile. Phycologia 49 554 - 573    DOI : 10.2216/09-106.1
Kang J. W. 1966 On the geographical distribution of marinealgae in Korea. Bull. Pusan Fish. Coll. 7 1 - 125
Kawaguchi S. 1989 The genus Prionitis (Halymeniaceae,Rhodophyta) in Japan. J. Fac. Sci. Hokkaido Univ. Ser. V (Bot.) 14 193 - 257
Kawaguchi S. 1997 Taxonomic notes on the Halymeniaceae(Gigartinales, Rhodophyta) from Japan. III. Synonymizationof Pachymeniopsis Yamada in Kawabata withGrateloupia C. Agardh. Phycol. Res. 45 9 - 21    DOI : 10.1111/j.1440-1835.1997.tb00057.x
Kawaguchi S. , Wang H. W. , Horiguchi T. , Sartoni G. , Masuda M. 2001 A comparative study of the red alga Grateloupiafilicina (Halymeniaceae) from the NorthwesternPacific and Mediterranean with the description ofGrateloupia asiatica, sp. nov. J. Phycol. 37 433 - 442
Kim M. S. , Kim S. Y. , Nelson W. 2010 Symphyocladialithophila sp. nov. (Rhodomelaceae, Ceramiales), a newKorean red algal species based on morphology and rbcLsequences. Bot. Mar. 53 233 - 241    DOI : 10.1515/bot.2010.031
Lee H. B. , Ph.D. disserration 1987 A systematic study of Halymeniaceae (Rhodophyta,Cryptonemiales) in Korea. Seoul National University Seoul, Korea Ph.D. disserration 412 -
Lee J. I. , Kim H. G. , Geraldino P. J. L. , Hwang I. K. , Boo S. M. 2009 Molecular classification of Grateloupia (Halymeniaceae, Rhodophyta) in Korea. Algae 24 231 - 238
Lee Y. 2008 Marine algae of Jeju. Academy Publication Seoul 477 -
Lee Y. , Kang S. 2001 A catalogue of the seaweeds in Korea. Cheju National University Press Jeju 473 - 474
Okamura K. 1899 Contributions to the knowledge of JapanIII. Bot. Mag. Tokyo 13 4 - 5
Okamura K. 1912 Icones of Japanese algae. Vol. 2. Kazamashobo Tokyo 66 - 68
Okamura K. 1913 Icones of Japanese algae. Vol. 3. Kazamashobo Tokyo 63 - 64
Rambaut A. 2002 Se-Al: Sequence alignment editor. Available from:
Stamatakis A. 2006 RAxML-VI-HPC: maximum likelihoodbased phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22 2688 - 2690    DOI : 10.1093/bioinformatics/btl446
Wang H. W. , Kawaguchi S. , Horiguchi T. , Masuda M. 2000 Reinstatement of Grateloupia catenata (Rhodophyta, Halymeniaceae) on the basis of morphology and rbcL sequences. Phycologia 39 228 - 237    DOI : 10.2216/i0031-8884-39-3-228.1
Wang H. W. , Kawaguchi S. , Horiguchi T. , Masuda M. 2001 A morphological and molecular assessment of the genus Prionitis J. Agardh (Halymeniaceae, Rhodophyta). Phycol. Res. 49 251 - 261    DOI : 10.1111/j.1440-1835.2001.tb00255.x
Wilkes R. J. , McIvor L. M. , Guiry M. D. 2005 Using rbcL sequence data to reassess the taxonomic position of some Grateloupia and Dermocorynus species (Halymeniaceae, Rhodophyta) from the north-eastern Atlantic. Eur. J. Phycol. 40 53 - 60    DOI : 10.1080/09670260400024634
Yang M. Y. , Han E. G. , Kim M. S. 2013 Molecular identification of Grateloupia elliptica and G. lanceolata (Rhodophyta) inferred from plastid rbcL and mitochondrial COI genes sequence data. Genes Genomics 35 239 - 246