Delesseriaceae, Rhodophyta)

Research Article

Algae 2011, 26(3): 211-219 http://dx.doi.org/10.4490/algae.2011.26.3.211 Open Access

A new Korean red algal species, Haraldiophyllum udoensis sp. nov. (Delesseriaceae, Rhodophyta)

Myung Sook Kim1,* and Jeong Chan Kang1 1Department of Biology, Jeju National University, Jeju 690-756, Korea

The genus Haraldiophyllum comprises seven species worldwide. Six of these are endemics with limited distributions, whereas the type species H. bonnemaisonii has been reported from the Atlantic Ocean. In Korea, H. bonnemaisonii has been previously recorded from the southern coast. During a red algal collection at Udo, Jeju Island, Korea, we found a potentially undescribed Haraldiophyllum species and analyzed its morphology and rbcL sequences. Herein we describe a new species, H. udoensis sp. nov., and compare our Udo specimen to similar congeners. This new species is characterized by one or several elliptical blades on a short cylindrical stipe with fibrous roots, blades that are monostromatic except at the base and on reproductive structures, a lack of network and microscopic veins, entire margins, lack of proliferations, growth through many marginal initials, and two distinct tetrasporangia layers. A phylogenetic rbcL sequence analysis demonstrated H. udoensis was distinct from the United Kingdom’s H. bonnemaisonii, as well as from other species. Morphological and sequence data indicated a previous misidentification of H. udoensis as the type species H. bonnemaisonii. Based on maximum likelihood analysis, Myriogramme formed a sister clade with H. udoensis, with rela- tively low bootstrap support.

Key Words: Delesseriaceae; Haraldiophyllum udoensis sp. nov.; morphology; rbcL; Rhodophyta; taxonomy

INTRODUCTION

Zinova (1981) established the genus Haraldiophyl- infossum A. J. K. Millar (1994); H. mirabile (Kylin) Zino- lum based upon Myriogramme bonnemaisonii Kylin va (1981); H. nottii (Norris & M. J. Wynne) M. J. Wynne (1924), from Atlantic Europe. The Haraldiophyllum type (1983); and H. sinuosum (A. H. S. Lucas) A. J. K. Millar species, H. bonnemaisonii, is characterized by a broad, (1990). erect, foliose thallus with a cylindrical stipe, the absence The type species H. bonnemaisonii is typically dis- of a midrib and veins, monostromatic and polystromatic tributed in the Atlantic Ocean (Maggs and Hommersand blades, and tetrasporangia sori formed upon both sides 1993, Hardy and Guiry 2003, John et al. 2004, Barbara et of the blade (Maggs and Hommersand 1993). To date, al. 2005, Lin et al. 2007). The other six species, however, seven species of Haraldiophyllum have been recognized are endemics with limited distributions. H. mirabile and worldwide (Guiry and Guiry 2011), as follows: H. bonne- H. nottii live only along the western coast of North Amer- maisonii (Kylin) A. D. Zinova (1981); H. crispatum (J. D. ica (Wynne 1983), H. sinuosum and H. infossum occur Hooker & Harvey) Lin, Hommersand & Nelson (2007); H. along the eastern coast of Australia and in the southwest- erosum (Harvey) A. J. K. Millar & J. M. Huisman (1996); H. ern Pacific (Millar 1990, 1994), H. erosum inhabits west-

This is an Open Access article distributed under the terms of the Received May 15, 2011, Accepted August 13, 2011 Creative Commons Attribution Non-Commercial License (http://cre- Corresponding Author ativecommons.org/licenses/by-nc/3.0/) which permits unrestricted * non-commercial use, distribution, and reproduction in any medium, E-mail: [email protected] provided the original work is properly cited. Tel: +82-64-754-3523, Fax: +82-64-756-3541

ern Australia (Millar and Huisman 1996), and H. crispa- microscope (ECLIPSE 80i; Nikon) and then imported all tum occurs in Australia and New Zealand (Lin et al. 2007). images into PhotoShop 7.0.1 (Adobe Systems, San Jose, In Asia, Nam and Kim (1996) initially identified H. CA, USA) for plate assembly. bonnemaisonii based on the specimen collected from Hadong and Kwangyang along the south coast of Korea. Molecular study They described the Korean specimens as characteristi- cally having monostromatic blades (except for the basal We air-dried the piece of cleaned thallus and desic- portion), a procarp with one carpogonial branch and two cated it using silica gel for the DNA extraction. Using the sterile groups, and tetrasporangial sori on both surfaces DNeasy Plant Mini Kit (Qiagen, Hilden, Germany), and of the blades (Nam and Kim 1996). They also noted that following the manufacturer’s instructions, we ground the Korean specimens reached 10-20 cm in height and con- silica gel-dried thallus in liquid nitrogen and extracted sisted of flabellate blades on a prominent stipe; however, the total genomic DNA. Extracts were dissolved in 200 Korean H. bonnemaisonii exhibits some differences when mL of distilled water. For amplification and sequencing compared to specimens from the British Isles (Maggs and reactions of genes, we employed the specific primer pairs Hommersand 1993), having deeply cleft and non-dichot- rbcLF7-rbcLR753 and rbcLF645-rbcS start (Lin et al. 2001, omously lobed blades. Gavio and Fredericq 2002). For all polymerase chain re- Because the marine algal flora of Udo, Jeju Island, Ko- action (PCR) amplifications, we used Swift MaxPro ther- rea remains very poorly documented, we have collected mal cyclers (ESCO, Singapore) with an AccuPower PCR extensively there to improve our knowledge of red algal PreMix (Bioneer, Daejeon, Korea). The PCR comprised biodiversity. The red algae within the Udo marine forests an initial denaturation step, at 94°C for 4 min, 35 cycles consist of a turf community growing around and between 1 min at 94°C, 1 min at 50°C, 2 min at 72°C, and a final 7 digitate Ecklonia cava holdfasts. While collecting algae min extension cycle, at 72°C. To purify the PCR products, from Udo, we found a potentially undescribed species of we used an AccuPrep PCR Purification Kit (Bioneer) and Haraldiophyllum that belonged to the tribe Myriogram- then sequenced them commercially (Macrogen, Seoul, meae. A careful examination revealed that this alga rep- Korea). Both electropherogram outputs from each sam- resented a new species in the Delesseriaceae. Here, we ple were edited using the program Chromas version 1.45. report on this newly discovered species based on its veg- Using BioEdit (Hall 1999), we collated the total se- etative morphology and rbcL gene sequences, and dis- quences and then aligned them visually. No alignments cuss its taxonomic status among the species of the genus. posed problems, as we observed no gaps. The outgroups To elucidate the phylogenetic relationships of the taxa we used were Augophyllum delicatum (AF257400) and from Udo, Jeju Island, we downloaded the rbcL sequence Nitophyllum punctatum (AF257402). To confirm the data of four Haraldiophyllum species from GenBank. taxonomic position of a potentially undescribed species of Haraldiophyllum from Udo, we conducted a maximum likelihood (ML) analysis, using PAUP 4.0 (Swofford MATERIALS AND METHODS 2002). We determined the best model for the individual data sets using Modeltest 3.4 software (Posada and Cran- Morphology dall 1998). The best model was a general time reversible (GTR) evolutionary model with gamma correction for For our morphological study, we collected the speci- among-site variation (Γ) and proportion of invariable men on a scuba dive, and preserved them in 4% formalin sites (I). To estimate ML trees, we used a heuristic search / seawater, with the exception of a small piece of thallus with 100 random addition sequence replicates and tree to be used for molecular study. The voucher specimen bisection and reconnection (TBR) branch swapping. To was deposited in the herbarium of Jeju National Uni- test the node stability, we performed bootstrap analyses versity (JNUB), Jeju, Korea. We sectioned the preserved with 1,000 replicated ML searches, using the same pro- specimen by hand or using a freezing microtome NK- gram and settings. 101-II (Nippon Optical Works Co. Ltd., Tokyo, Japan). The sectioned preparations were stained with 1% aniline blue acidified with 1% HCl and mounted in 25-30% Karo RESULTS syrup. To photograph the specimen, we used a Digital Sight DS-Fi1 camera (Nikon, Tokyo, Japan) attached to a Haraldiophyllum udoensis M. S. Kim et J. C. Kang, sp. nov.

http://dx.doi.org/10.4490/algae.2011.26.3.211 212 Kim & Kang Haraldiophyllum udoensis sp. nov.

A B

Fig. 1. Haraldiophyllum udoensis sp. nov. (A) Holotype (JNU-MSK 30601HU, tetrasporophyte) specimen deposited in the Herbarium of Jeju Na- tional University (JNUB), Jeju, Korea. (B & C) Enlarged photos of Fig. 1A to show tetrasporangial sori (B) and the blade attached to a stone (C). Scale bars represent: A, 5 cm; B & C, 3 cm.

Description. Thalli ellipticus, 22 cm longis et 6 cm la- cells divide obliquely; growth is marginal and diffuse, tis, consisto singuli aut aliquot laminae affixus per fibro- without distinct apical cells; tetrasporangia occur in el- sus hapteron; lamina in brevis cylindricus stipitis, 2-3 cm liptical sori 2 mm long on median, formed simultaneous- longis et 1 mm crassus; lamina tenuis, monostromaticis ly on both surfaces of the blade; mature tetrasporangia super, tristromaticis in reproductivus area et polystro- forming two layers, spherical, tetrahedrally divided, to maticis versus basis, microscopicus vena absens, integer 80-100 µm in diameter. margo et marginalis cellulae divisus oblique; crescens Holotype. JNU-MSK30601HU, tetrasporophyte, col- marginalis et diffuses, sine distinctus cullula apicalis; lected on June 14, 2009 (Fig. 1A). tetrasporangia eferens in ellipticus sori 2 mm longis in Type locality. Haumokdong, Udo, Jeju Island, Korea medianas factus simultaneous in superficiebus amba- (33°30´21˝ N, 126°56´09˝ E) bus lamina; tetrasporangia matura factus bistromatica, Korean name. 우도민엷은잎 spherical, tetrahedraliter divisa, ad 80-100 µm diametro. Distribution. Known to occur on the south coast (Nam Thallus elliptical, 22 cm long and 6 cm wide, consisting and Kim 1996) and at Udo, Jeju Island, in this study. of one or several blades attached by a fibrous holdfast; Habitat. We collected Haraldiophyllum udoensis at 12 blade on a short, cylindrical stipe, 2-3 cm long and 1 mm m in the sublittoral zone of Udo, Jeju Island, Korea. The thick; blade thin, monostromatic above, tristromatic in thallus grew as an epilithic blade on a stone covered with reproductive areas, and polystromatic toward the base, Lithophyllum sp. in habitats exposed to strong currents. microscopic veins absent; entire margins and marginal Among the plants collected along the south coast of Ko-

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rea by Nam and Kim (1996) and by us for this study, tetra- terspecific p distance) values for the rbcL region within sporophytes were found from March to September. Nam Haraldiophyllum ranged from 31 bp (2.7%) between H. and Kim (1996) collected male and female gametophytes mirabile from the United States and Haraldiophyllum in July and September, respectively, but we collected no sp. from Chile to 87 bp (6.7%) between H. bonnemaisonii gametophytes for this study. and Haraldiophyllum sp. from Chile. The most-related Etymology. We have chosen the specific epithet to genus, Myriogramme, differed by 109-142 bp (8.3-11%) represent the name of the islet collection site-Udo, Jeju from H. udoensis and by 103-169 bp (8-13%) from the rest Island. of the genera we tested in the present study. Morphology. The plant is a greenish red-brown, 22 cm In the phylogenetic tree from rbcL, the new species high, 6 cm broad, foliose, and erect on a cylindrical stipe formed an independent clade with strong support (99% up to 2-3 cm long and 1 mm thick (Fig. 1A). The thallus for ML) and a sister group with H. mirabile from the Unit- consists of one or several blades, attached by a small fi- ed States and Haraldiophyllum sp. from Chile (Fig. 4). brous holdfast, that lack midrib and microscopic veins The ML analysis of rbcL data indicated that H. udoensis (Fig. 1B & C). The blade is thin and monostromatic above, showed clear separation from H. bonnemaisonii and H. becoming tristromatic in reproductive areas and polys- crispatum from New Zealand. The ML tree showed two tromatic toward the base with evanescent midribs (Fig. groups within Haraldiophyllum, and the bootstrap sup- 2A-D), and increasing in thickness from 35 µm near the ports were quite strong (100% and 91%). The first clade apices to 100 µm basally. The blade comprises cells that included H. udoensis as the new species, H. mirabile and are elongate-polygonal in surface view (Fig. 3A) and 45- Haraldiophyllum sp., and the second clade contained H. 70 µm long × 15-45 µm wide, linked by secondary pit con- crispatum and H. bonnemaisonii. The genus Haraldio- nections that increase in abundance in the polystromatic phyllum monophyly had definitive bootstrap support regions. The blade has entire margins, and marginal cells (100%). The genus Myriogramme formed a sister rela- divide anticlinally or obliquely, growing marginal meri- tionship with the genus Haraldiophyllum. stems without distinct apical cells (Fig. 3A). Tetrasporangia are produced in small, numerous, circular to elliptical sori, formed randomly on both sides DISCUSSION of the blade, and scattered throughout the blade except along the margins and the lowermost parts. Tetrasporan- Haraldiophyllum udoensis is newly described from gial sori are 2 mm long × 0.6 mm wide, and protrude on Udo, Jeju Island, Korea, based on our detailed morpho- either side of the blade, approximately 200 µm thick (Fig. logical observations and molecular analysis. This new 3B). In the transverse section of the sori, the tetrasporan- species is characterized by the following features: 1) one gia appear in two layers between both surface cells (Fig. or several elliptical blades on a short, cylindrical stipe 3C). The tetrasporangia are produced from inner corti- with fibrous roots; 2) monostromatic blades, except at the cal cells but occasionally also from primary cells. Mature base and on reproductive structures; 3) a lack of network tetrasporangia are spherical, 80-100 µm in diameter, and and microscopic veins; 4) entire margins with a lack of divided tetrahedrally (Fig. 3D). proliferations; 5) growth by means of many marginal ini- Molecular analysis of rbcL. To align the five Haraldio- tials; and 6) an arrangement of two distinct tetrasporan- phyllum species with putative relative genera in the fami- gia layers. Molecular phylogenetic analysis based on rbcL ly Delesseriaceae, we used a 1,303 nucleotide (nt) portion sequences demonstrated H. udoensis’s distinctness from of rbcL gene and two outgroups. Variable sites occurred its congeners, and separated it clearly from the European at 464 positions (35.6%), and 350 positions (26.9%) were H. bonnemaisonii. The genus Haraldiophyllum contains parsimoniously informative. In the ML analysis, we es- two clusters of species, one from the United Kingdom timated the –ln likelihood score as 7,899.6647 under and New Zealand and the other occurring along the Pa- the GTR + Г + I model. The sequences differed by up to cific Coasts of Korea, the United States, and Chile. Our a 72 bp (5.5%) pairwise distance between H. udoensis molecular analysis produced similar results to those Lin from Korea and H. bonnemaisonii from the United King- et al. (2007) reported. The new species is epilithic in the dom. The new species differed from H. crispatum from sublittoral zone at depths up to 12 m. It was previously New Zealand, H. mirabile from the United States, and identified in Korea asH. bonnemaisonii (Nam and Kim Haraldiophyllum sp. from Chile by 64, 32, and 39 bp, 1996). Although H. udoensis closely resembles the type respectively. The uncorrected sequence divergence (in- species of Haraldiophyllum (H. bonnemaisonii from the

http://dx.doi.org/10.4490/algae.2011.26.3.211 214 Kim & Kang Haraldiophyllum udoensis sp. nov.

A D

Fig. 2. Haraldiophyllum udoensis sp. nov. (A-D) Transverse section of the blade, which is monostromatic (A & B), tristromatic (C), and polystromatic toward the base (D). Scale bars represent: A, 50 μm; B & C, 30 μm; D, 100 μm.

A B

Fig. 3. Haraldiophyllum udoensis sp. nov. (A) Surface view of blade showing elongate-polygonal cells. (B) Small elliptical tetrasporangial sori. (C) Transverse section of sori forming two tetrasporangia layers. (D) Mature tetrasporangia, divided tetrahedrally. Scale bars represent: A & D, 30 μm; B & C, 50 μm.

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Fig. 4. Phylogenetic tree for Haraldiophyllum and relatives, derived from maximum likelihood using plastid-encoded rbcL sequence data. The bootstrap values shown above branches are from bootstrap analyses employing 1,000 replicated maximum likelihood searches.

British Isles), H. bonnemaisonii has a distinct thallus that nottii are 2-7 cm smaller than other species (Millar 1994, divides regularly and dichotomously into overlapping Womersley 2003, Lin et al. 2007). In particular, H. infos- lobes, frequently proliferating on the blade and forming sum from Australia differs from all other species in the a single tetrasporangia layer in a zigzag manner (Maggs genus due to its diminutive size (up to 1.9 cm). Haraldio- and Hommersand 1993, Lin et al. 2007). In addition, the phyllum sinuosum is by far the largest of the genus with reproductive structure in H. udoensis is larger; the cysto- plants reaching lengths of 20-35 cm. Haraldiophyllum carp diameter of > 700 µm contrasts with the < 600 µm in erosum and H. mirabile are of medium size at around 10 H. bonnemaisonii, and tetrasporangia in H. udoensis are cm (Millar and Huisman 1996), but the new species H. > 80 µm in diameter, as opposed to < 70 µm in H. bonne- udoensis is approximately 20 cm in length. The shapes of maisonii (Table 1). the base and margins have been demonstrated as valid In recognizing the species of Haraldiophyllum, which characteristics for delineating some species of Haral- currently contains seven species, the vegetative char- diophyllum, as for example Millar and Huisman (1996) acteristics are viewed as important diagnostic features, showed changes of the Haraldiophyllum species’ mar- such as whether thallus size is diminutive, whether shape gins display a continuum from species with entire mar- of the base and margins are dentate or entire, whether gins to those with fringed margins. Most of Haraldiophyl- the branching pattern is dichotomous, whether the lum species including the new species H. udoensis have lobed blade is deeply cleft, and the presence or absence entire or dentate margins, except for two: H. erosum and of stipes (Table 1). Thallus size is useful for identifying H. infossum (Millar 1994). According to Millar and Huis- species of Haraldiophyllum; H. bonnemaisonii and H. man (1996), H. erosum from western Australia has an crispatum reach 20-25 cm, whereas H. infossum and H. unusual marginal fringe with numerous multicellular

http://dx.doi.org/10.4490/algae.2011.26.3.211 216 Kim & Kang Haraldiophyllum udoensis sp. nov.

Australia H. sinuosum H. A. Lucas) S. H. (A. Millar K. J. New Wales, South - 20-35 - blunt lobes Short, (6-7 mm) Short - Entire - Absent ------1913 Lucas 1990 Millar - - (Norris & (Norris water branching mono branched, stromatic few minute teeth calary divisions in shape irregular H. nottii H. J. M. Wynne) J. M. Wynne USA Washington, to deep Shallow 2-7 Dichotomously flat- Thin, forked, narrow Short, Absent a entire, Smooth, - - with inter Marginal side One 60-1,000 elongate, Slightly to elongate Ovate 35-55 Single 2003 Womersley (Kylin) (Kylin) Alaska, USA - - to Roundish deeply or cleft - - - - H. mirabile H. Zinova 8-10 Less prominent ------1925 Kylin 1983 Wynne - - - - - A. J. K. Australia munity in im overlap bricate arrange ment, creeping irregularly blades branched arising a from stoloniferous base tate blade proximal surfaces patches regularly H. infossum H. Millar New Wales, South 1-20 m, turf com 1.9 to elongate, Ovate Monostromatic, - Absent den Prominently - - initials Marginal sides of the Both - central Elongate, or ir Rounded 50 Single 1994 Millar - - -

Western Australia Western m, drift, epiphytic lobed irregularly holes in divide mono surfaces, stromatic ticellular pro cesses region the blade patches marginal H. erosum H. J. A. (Harvey) M. & J. Millar K. Huisman Island, Garden 41 from Dredged 13 or Dichotomously shaped Irregularly distance Short Absent mul by Fringed - - Meristematic surfacesBoth of - sub- Elongate, or oval Rounded 60 Two and Millar 1996 Huisman - - (J. D. New Zealand New drift single or several blades able in shape, elongate or fan- shaped, divided or crisped, entire denticulate larly into branch blade H. crispatum H. & Harvey) Hooker Hommersand Lin, & Nelson Campbell Island, Island, Campbell and 24 m, dredged 25 to robust, Delicate vari Exceedingly Cylindrical Absent or Smooth present Sometimes irregu Dissected growth Marginal the Throughout - to ovate Circular to ovate Circular 40-85 Single Lin et al. 2007 Haraldiophyllum

------UK; Ireland to 21 m shaped blade, dichotomously divided when young, thick increasing ness base ginal meristems blade near mar gins cal cal mersand 1993 H. bonnemaisonii H. D. A. (Kylin) Zinova Island, Shetland water low Extreme 15 × 20 fan- several to One Monostromatic Cylindrical Absent or dentate Entire Present to Sometimes in mar Diffuse sides of Both 430-600 to ellipti Circular to ellipti Circular 50-70 Single and Hom Maggs - - - elliptical Korea zone elliptical blade with fibrous not dichot roots, omously blade mono nous, and stromatic polystromatic cleft upper part 1996 H. udoensis H. nov. sp. Island, Jeju Udo, 12 m in sublittoral 22 × 6 to several One Thin, membra cylindricalShort Absent Entire Absent or deeply Absent growth Marginal surfacesBoth of 500-900 rounded Small to Circular 80-100 Two This study and Kim Nam - Morphological of species in the genus comparisons

cm) (µm) gia (µm) Table 1. Table Locality Habitat (height × width, Size Habit Blade Stipe Veins Margins Proliferations Cleft blade Growth Procarp of cystocarp Size sori Spermatangial sori Tetrasporangial of tetrasporan Size gia Tetrasporan layer Reference

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processes. They report these marginal processes are of- group cutting off from the supporting cell prior to the ten polysiphonous and divaricately branched, and they initial carpogonial branch. The production of the second always terminate single apical cell in a transversely-di- sterile cell group delays until after carpogonial branch viding (Millar and Huisman 1996). H. infossum often has formation. The same sequence occurs in H. crispatum, H. a stoloniferous base and prominently dentate blade mar- erosum, and H. nottii, but that of H. infossum differs in gins (Millar 1994). forming the two sterile cell groups before initiating the The branching pattern in relation to the presence or carpogonial branch (Millar 1994). Millar and Huisman absence of lobed blades and stipes can also be an impor- (1996), therefore, concluded these differences appear to tant distinguishing feature. H. bonnemaisonii comprises be stable within the Haraldiophyllum species. In the new one to several fan-shaped blades, borne on cartilaginous species, H. udoensis, procarp formation and cystocarp stipes and dividing dichotomously into overlapping development follow a pattern similar to those previous lobes with rounded apices (Maggs and Hommersand studies describe (Millar 1994, Millar and Huisman 1996, 1993, Lin et al. 2007). H. crispatum has an exceedingly Nam and Kim 1996, Lin et al. 2007). variable shape, dividing partially into broad lobes with While researchers often use the female reproductive rounded margins or cleaving into irregular, dichoto- organ’s structure to distinguish species within the genus mously-branched segments with proliferations (Lin et al. and genera in the family, they have not considered the 2007). H. erosum’s blades have dichotomous or irregular number of layers in the tetrasporangia arrangement to lobes and often have irregularly-shaped holes in the sur- be of any significance in this genus. The type speciesH. faces (Millar and Huisman 1996). H. infossum has ovated bonnemaisonii has only a single layer of tetrasporangia to elongated individual blades that often overlap in an arranged in a zigzag manner, and the same feature oc- imbricate arrangement with the creeping stoloniferous curs in H. crispatum, H. infossum, and H. nottii. However, parts compressed (Millar 1994). H. mirabile from Alaska two species, H. udoensis sp. nov. and H. erosum, form two has less prominent stipes, and the blades are roundish layers of tetrasporangia. To verify the importance of the to deeply lobed or cleft (Kylin 1925). The blades of H. number of layers in the tetrasporangia arrangement from nottii are complicated and subdichotomously branched a phylogenetic perspective, further observation and mo- with lobed, narrow stipes (Norris and Wynne 1968). The lecular analysis for all seven species are necessary. blades of H. sinuosum are generally entire and not deep- The genus Haraldiophyllum superficially resembles ly cleft but have short, blunt lobes, and the plants have two genera, Myriogramme and Nitophyllum, although short stipes that do not persist into the lower blades (Lu- Haraldiophyllum clearly belongs to the tribe Myrio- cas 1913). The new species H. udoensis appears to differ grammeae based on vegetative and reproductive devel- from other species mostly by its one to several elliptical, opmental characteristics (Hommersand and Fredericq non-lobed blades. 1997). The genus Haraldiophyllum is readily separated Researchers have documented the diagnostic char- from the genus Myriogramme by carposporangia orga- acteristics of the genus Haraldiophyllum’s reproductive nization, which is terminal solitary in Haraldiophyllum structures for the female reproductive organs, such as the but terminal chains in Myriogramme (Wynne 1983, Lin procarp, carposporangia, and cystocarp. The procarps et al. 2007). The new species H. udoensis forms a separate contain two groups of sterile cells, one group of cover cells clade from Myriogramme based on molecular analysis of and two periaxial cells parallel to the frond’s longitudinal rbcL sequence data. However, the new species is linked axis (Millar 1994). The carposporangia are borne singly to Myriogramme as a sister clade with low bootstrap and terminally on gonimoblast filaments. The cystocarp support in the ML analysis. Haraldiophyllum is distin- has a fusion cell incorporating gametophytic cells at the guished from the genus Nitophyllum by the number of floor of the cystocarpic cavity. Kylin (1925) noted this fea- sterile cell groups in the procarps, with two for Haraldio- ture of the cystocarp fusion cell is probably useful for ge- phyllum and only one for Nitophyllum, as well as by the neric recognition of the genus Haraldiophyllum. Maggs two periaxial cells’ alignment with respect to the frond’s and Hommersand (1993) and Millar (1994) also included longitudinal axis, showing a parallel manner in Haral- this characteristics in their generic circumscription of diophyllum and a perpendicular manner in Nitophyllum Haraldiophyllum, while Millar and Huisman (1996) rec- (Zinova 1981, Wynne 1983). In the molecular phylogenet- ognized two different sequences of cell formation lead- ic tree, the Haraldiophyllum clade is distantly separate ing to the procarp’s final structure in Haraldiophyllum. from the Nitophyllum clade including its type species, N. They observed H. bonnemaisonii has only one sterile cell punctatum from Spain.

http://dx.doi.org/10.4490/algae.2011.26.3.211 218 Kim & Kang Haraldiophyllum udoensis sp. nov.

Our ongoing studies of the Delesseriaceae from Jeju Is- sitets Årsskrift, Ny Följd, Andra Afdelningen 21:1-87. land, Korea have thus far clarified the generic placement Lin, S. M., Fredericq, S. & Hommersand, M. H. 2001. System- of Haraldiophyllum and led to the recognition of a new atics of the Delesseriaceae (Ceramiales, Rhodophyta) species, H. udoensis. On Jeju Island, many delesseria- based on large subunit rDNA and rbcL sequences, in- ceous taxa remain that require critical reexamination in cluding the Phycoryoideae, subfam. nov. J. Phycol. light of recent taxonomic developments within the family. 37:881-899. Lin, S. -M., Hommersand, M. H. & Nelson, W. A. 2007. An assessment of Haraldiophyllum (Delesseriaceae, Rho- ACKNOWLEDGEMENTS dophyta), including H. crispatum (J. D. Hooker et Har- vey) comb. nov. from New Zealand based on rbcL and This work was supported by a National Research Foun- LSU sequence analysis and morphological evidence. dation of Korea grant, funded by the Korean Government Eur. J. Phycol. 42:391-408. (2011-0003792), and as part of the Survey of Indigenous Lucas, A. H. S. 1913. Notes on Australian marine algae, I. Biology of Korea by the National Institute of Biological Proc. Linn. Soc. N. S. W. 38:49-60. Resources (NIBR). Maggs, C. A. & Hommersand, M. H. 1993. Seaweeds of the British Isles. Vol. 1. Rhodophyta. Part 3A. Ceramiales. HMSO, London, 444 pp. REFERENCES Millar, A. J. K. 1990. Marine red algae of the Coffs Harbour region, northern New South Wales. Aust. Syst. Bot. 3:293- Bárbara, I., Cremades, J., Calvo, S., López-Rodríguez, M. C. 593. & Dosil, J. 2005. Checklist of the benthic marine and Millar, A. J. K. 1994. Haraldiophyllum infossum sp. nov. (De- brackish Galician algae (NW Spain). Anales del Jardín lesseriaceae, Rhodophyta), a diminutive turf-forming Botánico de Madrid 62:69-100. red alga from the south-western Pacific. Bot. Mar. Gavio, B. & Fredericq, S. 2002. Grateloupia turuturu (Halyme- 37:125-132. niaceae, Rhodophyta) is the correct name of the non- Millar, A. J. K. & Huisman, J. M. 1996. Haraldiophyllum ero- native species in the Atlantic known as Grateloupia do- sum comb. nov. (Delesseriaceae, Rhodophyta) from ryphora. Eur. J. Phycol. 37:349-359. Southern and Western Australia. Aust. Syst. Bot. 9:61-69. Guiry, M. D. & Guiry, G. M. 2011. AlgaeBase. Worldwide Nam, K. W. & Kim, Y. S. 1996. Haraldiophyllum (Rhodophy- electronic publication, National University of Ireland, ta): a Delesseriaceous genus newly recorded in Korea. Galway. Available from: http://www.algaebase.org. Ac- Algae 11:101-106. cessed Mar. 3, 2011. Norris, R. E. & Wynne, M. J. 1968. Notes on marine algae of Hall, T. A. 1999. BioEdit: a user-friendly biological sequence Washington and Southern British Columbia, III. Syesis alignment editor and analysis program for Windows 1:133-146. 95/98/NT. Nucleic Acids Symp. Ser. 41:95-98. Posada, D. & Crandall, K. A. 1998. Modeltest: testing the Hardy, F. G. & Guiry, M. D. 2003. A check-list and atlas of the model of DNA substitution. Bioinformatics 14:817-818. seaweeds of Britain and Ireland. British Phycological So- Swofford, D. L. 2002. PAUP*: phylogenetic analysis using par- ciety, London, 435 pp. simony (*and other methods). Version 4.0b10. Sinauer Hommersand, M. H. & Fredericq, S. 1997. Characterization Associates, Sunderland, MA. of Myriogramme livida, Myriogrammeae trib. nov. (De- Womersley, H. B. S. 2003. The marine benthic flora of south- lesseriaceae, Rhodophyta). J. Phycol. 33:106-121. ern Australia, Rhodophyta, Part IIID. Ceramiales: Deles- John, D. M., Prud'homme van Reine, W. F., Lawson, G. W., seriaceae, Sarcomeniaceae, Rhodymelaceae. Australian Kostermans, T. B. & Price, J. H. 2004. A taxonomic and Biological Resources Study & State Herbarium of South geographical catalogue of the seaweeds of the western Australia, Canberra & Adelaide, 533 pp. coast of Africa and adjacent islands. Nova Hedwigia Wynne, M. J. 1983. The current status of genera in the Deles- Beih. 127:1-339. seriaceae (Rhodophyta). Bot. Mar. 26:437-450. Kylin, H. 1924. Studien über die Delesseriaceen. Lunds Uni- Zinova, A. D. 1981. De positione systematica Nitophylli versitets Årsskrift, Ny Följd, Andra Afdelningen 20:1- (Myriogrammes) yezoensis (Yamada et Tokida) Mikami 111. (Delesseriaceae). Novit. Syst. Plant. non Vasculariume Kylin, H. 1925. The marine red algae in the vicinity of of the 18:10-15. biological station at Friday Harbor, Wash. Lunds Univer-

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