a new korean red algal species, haraldiophyllum...

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

Open Access

Research Article

Copyright © The Korean Society of Phycology 211 http://e-algae.kr pISSN: 1226-2617 eISSN: 2093-0860

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 charac-

terized 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 pro-

liferations, 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-

lum based upon Myriogramme bonnemaisonii Kylin

(1924), from Atlantic Europe. The Haraldiophyllum type

species, H. bonnemaisonii, is characterized by a broad,

erect, foliose thallus with a cylindrical stipe, the absence

of a midrib and veins, monostromatic and polystromatic

blades, and tetrasporangia sori formed upon both sides

of the blade (Maggs and Hommersand 1993). To date,

seven species of Haraldiophyllum have been recognized

worldwide (Guiry and Guiry 2011), as follows: H. bonne-

maisonii (Kylin) A. D. Zinova (1981); H. crispatum (J. D.

Hooker & Harvey) Lin, Hommersand & Nelson (2007); H.

erosum (Harvey) A. J. K. Millar & J. M. Huisman (1996); H.

infossum A. J. K. Millar (1994); H. mirabile (Kylin) Zino-

va (1981); H. nottii (Norris & M. J. Wynne) M. J. Wynne

(1983); and H. sinuosum (A. H. S. Lucas) A. J. K. Millar

(1990).

The type species H. bonnemaisonii is typically dis-

tributed in the Atlantic Ocean (Maggs and Hommersand

1993, Hardy and Guiry 2003, John et al. 2004, Barbara et

al. 2005, Lin et al. 2007). The other six species, however,

are endemics with limited distributions. H. mirabile and

H. nottii live only along the western coast of North Amer-

ica (Wynne 1983), H. sinuosum and H. infossum occur

along the eastern coast of Australia and in the southwest-

ern Pacific (Millar 1990, 1994), H. erosum inhabits west-

Received May 15, 2011, Accepted August 13, 2011

*Corresponding Author

E-mail: [email protected]: +82-64-754-3523, Fax: +82-64-756-3541

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 2011, 26(3): 211-219

http://dx.doi.org/10.4490/algae.2011.26.3.211 212

microscope (ECLIPSE 80i; Nikon) and then imported all

images into PhotoShop 7.0.1 (Adobe Systems, San Jose,

CA, USA) for plate assembly.

Molecular study

We air-dried the piece of cleaned thallus and desic-

cated it using silica gel for the DNA extraction. Using the

DNeasy Plant Mini Kit (Qiagen, Hilden, Germany), and

following the manufacturer’s instructions, we ground the

silica gel-dried thallus in liquid nitrogen and extracted

the total genomic DNA. Extracts were dissolved in 200

mL of distilled water. For amplification and sequencing

reactions of genes, we employed the specific primer pairs

rbcLF7-rbcLR753 and rbcLF645-rbcS start (Lin et al. 2001,

Gavio and Fredericq 2002). For all polymerase chain re-

action (PCR) amplifications, we used Swift MaxPro ther-

mal cyclers (ESCO, Singapore) with an AccuPower PCR

PreMix (Bioneer, Daejeon, Korea). The PCR comprised

an initial denaturation step, at 94°C for 4 min, 35 cycles

1 min at 94°C, 1 min at 50°C, 2 min at 72°C, and a final 7

min extension cycle, at 72°C. To purify the PCR products,

we used an AccuPrep PCR Purification Kit (Bioneer) and

then sequenced them commercially (Macrogen, Seoul,

Korea). Both electropherogram outputs from each sam-

ple were edited using the program Chromas version 1.45.

Using BioEdit (Hall 1999), we collated the total se-

quences and then aligned them visually. No alignments

posed problems, as we observed no gaps. The outgroups

we used were Augophyllum delicatum (AF257400) and

Nitophyllum punctatum (AF257402). To confirm the

taxonomic position of a potentially undescribed species

of Haraldiophyllum from Udo, we conducted a maxi-

mum likelihood (ML) analysis, using PAUP 4.0 (Swofford

2002). We determined the best model for the individual

data sets using Modeltest 3.4 software (Posada and Cran-

dall 1998). The best model was a general time reversible

(GTR) evolutionary model with gamma correction for

among-site variation (Γ) and proportion of invariable

sites (I). To estimate ML trees, we used a heuristic search

with 100 random addition sequence replicates and tree

bisection and reconnection (TBR) branch swapping. To

test the node stability, we performed bootstrap analyses

with 1,000 replicated ML searches, using the same pro-

gram and settings.

RESULTS

Haraldiophyllum udoensis M. S. Kim et J. C. Kang, sp. nov.

ern Australia (Millar and Huisman 1996), and H. crispa-

tum occurs in Australia and New Zealand (Lin et al. 2007).

In Asia, Nam and Kim (1996) initially identified H.

bonnemaisonii based on the specimen collected from

Hadong and Kwangyang along the south coast of Korea.

They described the Korean specimens as characteristi-

cally having monostromatic blades (except for the basal

portion), a procarp with one carpogonial branch and two

sterile groups, and tetrasporangial sori on both surfaces

of the blades (Nam and Kim 1996). They also noted that

Korean specimens reached 10-20 cm in height and con-

sisted of flabellate blades on a prominent stipe; however,

Korean H. bonnemaisonii exhibits some differences when

compared to specimens from the British Isles (Maggs and

Hommersand 1993), having deeply cleft and non-dichot-

omously lobed blades.

Because the marine algal flora of Udo, Jeju Island, Ko-

rea remains very poorly documented, we have collected

extensively there to improve our knowledge of red algal

biodiversity. The red algae within the Udo marine forests

consist of a turf community growing around and between

digitate Ecklonia cava holdfasts. While collecting algae

from Udo, we found a potentially undescribed species of

Haraldiophyllum that belonged to the tribe Myriogram-

meae. A careful examination revealed that this alga rep-

resented a new species in the Delesseriaceae. Here, we

report on this newly discovered species based on its veg-

etative morphology and rbcL gene sequences, and dis-

cuss its taxonomic status among the species of the genus.

To elucidate the phylogenetic relationships of the taxa

from Udo, Jeju Island, we downloaded the rbcL sequence

data of four Haraldiophyllum species from GenBank.

MATERIALS AND METHODS

Morphology

For our morphological study, we collected the speci-

men on a scuba dive, and preserved them in 4% formalin

/ seawater, with the exception of a small piece of thallus

to be used for molecular study. The voucher specimen

was deposited in the herbarium of Jeju National Uni-

versity (JNUB), Jeju, Korea. We sectioned the preserved

specimen by hand or using a freezing microtome NK-

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

syrup. To photograph the specimen, we used a Digital

Sight DS-Fi1 camera (Nikon, Tokyo, Japan) attached to a

Kim & Kang Haraldiophyllum udoensis sp. nov.

213 http://e-algae.kr

cells divide obliquely; growth is marginal and diffuse,

without distinct apical cells; tetrasporangia occur in el-

liptical sori 2 mm long on median, formed simultaneous-

ly on both surfaces of the blade; mature tetrasporangia

forming two layers, spherical, tetrahedrally divided, to

80-100 µm in diameter.

Holotype. JNU-MSK30601HU, tetrasporophyte, col-

lected on June 14, 2009 (Fig. 1A).

Type locality. Haumokdong, Udo, Jeju Island, Korea

(33°30´21˝ N, 126°56´09˝ E)

Korean name. 우도민엷은잎

Distribution. Known to occur on the south coast (Nam

and Kim 1996) and at Udo, Jeju Island, in this study.

Habitat. We collected Haraldiophyllum udoensis at 12

m in the sublittoral zone of Udo, Jeju Island, Korea. The

thallus grew as an epilithic blade on a stone covered with

Lithophyllum sp. in habitats exposed to strong currents.

Among the plants collected along the south coast of Ko-

Description. Thalli ellipticus, 22 cm longis et 6 cm la-

tis, consisto singuli aut aliquot laminae affixus per fibro-

sus hapteron; lamina in brevis cylindricus stipitis, 2-3 cm

longis et 1 mm crassus; lamina tenuis, monostromaticis

super, tristromaticis in reproductivus area et polystro-

maticis versus basis, microscopicus vena absens, integer

margo et marginalis cellulae divisus oblique; crescens

marginalis et diffuses, sine distinctus cullula apicalis;

tetrasporangia eferens in ellipticus sori 2 mm longis in

medianas factus simultaneous in superficiebus amba-

bus lamina; tetrasporangia matura factus bistromatica,

spherical, tetrahedraliter divisa, ad 80-100 µm diametro.

Thallus elliptical, 22 cm long and 6 cm wide, consisting

of one or several blades attached by a fibrous holdfast;

blade on a short, cylindrical stipe, 2-3 cm long and 1 mm

thick; blade thin, monostromatic above, tristromatic in

reproductive areas, and polystromatic toward the base,

microscopic veins absent; entire margins and marginal

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.

A B

C

Algae 2011, 26(3): 211-219


terspecific p distance) values for the rbcL region within

Haraldiophyllum ranged from 31 bp (2.7%) between H.

mirabile from the United States and Haraldiophyllum

sp. from Chile to 87 bp (6.7%) between H. bonnemaisonii

and Haraldiophyllum sp. from Chile. The most-related

genus, Myriogramme, differed by 109-142 bp (8.3-11%)

from H. udoensis and by 103-169 bp (8-13%) from the rest

of the genera we tested in the present study.

In the phylogenetic tree from rbcL, the new species

formed an independent clade with strong support (99%

for ML) and a sister group with H. mirabile from the Unit-

ed States and Haraldiophyllum sp. from Chile (Fig. 4).

The ML analysis of rbcL data indicated that H. udoensis

showed clear separation from H. bonnemaisonii and H.

crispatum from New Zealand. The ML tree showed two

groups within Haraldiophyllum, and the bootstrap sup-

ports were quite strong (100% and 91%). The first clade

included H. udoensis as the new species, H. mirabile and

Haraldiophyllum sp., and the second clade contained H.

crispatum and H. bonnemaisonii. The genus Haraldio-

phyllum monophyly had definitive bootstrap support

(100%). The genus Myriogramme formed a sister rela-

tionship with the genus Haraldiophyllum.

DISCUSSION

Haraldiophyllum udoensis is newly described from

Udo, Jeju Island, Korea, based on our detailed morpho-

logical observations and molecular analysis. This new

species is characterized by the following features: 1) one

or several elliptical blades on a short, cylindrical stipe

with fibrous roots; 2) monostromatic blades, except at the

base and on reproductive structures; 3) a lack of network

and microscopic veins; 4) entire margins with a lack of

proliferations; 5) growth by means of many marginal ini-

tials; and 6) an arrangement of two distinct tetrasporan-

gia layers. Molecular phylogenetic analysis based on rbcL

sequences demonstrated H. udoensis’s distinctness from

its congeners, and separated it clearly from the European

H. bonnemaisonii. The genus Haraldiophyllum contains

two clusters of species, one from the United Kingdom

and New Zealand and the other occurring along the Pa-

cific Coasts of Korea, the United States, and Chile. Our

molecular analysis produced similar results to those Lin

et al. (2007) reported. The new species is epilithic in the

sublittoral zone at depths up to 12 m. It was previously

identified in Korea as H. bonnemaisonii (Nam and Kim

1996). Although H. udoensis closely resembles the type

species of Haraldiophyllum (H. bonnemaisonii from the

rea by Nam and Kim (1996) and by us for this study, tetra-

sporophytes were found from March to September. Nam

and Kim (1996) collected male and female gametophytes

in July and September, respectively, but we collected no

gametophytes for this study.

Etymology. We have chosen the specific epithet to

represent the name of the islet collection site-Udo, Jeju

Island.

Morphology. The plant is a greenish red-brown, 22 cm

high, 6 cm broad, foliose, and erect on a cylindrical stipe

up to 2-3 cm long and 1 mm thick (Fig. 1A). The thallus

consists of one or several blades, attached by a small fi-

brous holdfast, that lack midrib and microscopic veins

(Fig. 1B & C). The blade is thin and monostromatic above,

becoming tristromatic in reproductive areas and polys-

tromatic toward the base with evanescent midribs (Fig.

2A-D), and increasing in thickness from 35 µm near the

apices to 100 µm basally. The blade comprises cells that

are elongate-polygonal in surface view (Fig. 3A) and 45-

70 µm long × 15-45 µm wide, linked by secondary pit con-

nections that increase in abundance in the polystromatic

regions. The blade has entire margins, and marginal cells

divide anticlinally or obliquely, growing marginal meri-

stems without distinct apical cells (Fig. 3A).

Tetrasporangia are produced in small, numerous, cir-

cular to elliptical sori, formed randomly on both sides

of the blade, and scattered throughout the blade except

along the margins and the lowermost parts. Tetrasporan-

gial sori are 2 mm long × 0.6 mm wide, and protrude on

either side of the blade, approximately 200 µm thick (Fig.

3B). In the transverse section of the sori, the tetrasporan-

gia appear in two layers between both surface cells (Fig.

3C). The tetrasporangia are produced from inner corti-

cal cells but occasionally also from primary cells. Mature

tetrasporangia are spherical, 80-100 µm in diameter, and

divided tetrahedrally (Fig. 3D).

Molecular analysis of rbcL. To align the five Haraldio-

phyllum species with putative relative genera in the fami-

ly Delesseriaceae, we used a 1,303 nucleotide (nt) portion

of rbcL gene and two outgroups. Variable sites occurred

at 464 positions (35.6%), and 350 positions (26.9%) were

parsimoniously informative. In the ML analysis, we es-

timated the –ln likelihood score as 7,899.6647 under

the GTR + Г + I model. The sequences differed by up to

a 72 bp (5.5%) pairwise distance between H. udoensis

from Korea and H. bonnemaisonii from the United King-

dom. The new species differed from H. crispatum from

New Zealand, H. mirabile from the United States, and

Haraldiophyllum sp. from Chile by 64, 32, and 39 bp,

respectively. The uncorrected sequence divergence (in-



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

A

C

D

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.

A

C

D

B

Algae 2011, 26(3): 211-219


nottii are 2-7 cm smaller than other species (Millar 1994,

Womersley 2003, Lin et al. 2007). In particular, H. infos-

sum from Australia differs from all other species in the

genus due to its diminutive size (up to 1.9 cm). Haraldio-

phyllum sinuosum is by far the largest of the genus with

plants reaching lengths of 20-35 cm. Haraldiophyllum

erosum and H. mirabile are of medium size at around 10

cm (Millar and Huisman 1996), but the new species H.

udoensis is approximately 20 cm in length. The shapes of

the base and margins have been demonstrated as valid

characteristics for delineating some species of Haral-

diophyllum, as for example Millar and Huisman (1996)

showed changes of the Haraldiophyllum species’ mar-

gins display a continuum from species with entire mar-

gins to those with fringed margins. Most of Haraldiophyl-

lum species including the new species H. udoensis have

entire or dentate margins, except for two: H. erosum and

H. infossum (Millar 1994). According to Millar and Huis-

man (1996), H. erosum from western Australia has an

unusual marginal fringe with numerous multicellular

British Isles), H. bonnemaisonii has a distinct thallus that

divides regularly and dichotomously into overlapping

lobes, frequently proliferating on the blade and forming

a single tetrasporangia layer in a zigzag manner (Maggs

and Hommersand 1993, Lin et al. 2007). In addition, the

reproductive structure in H. udoensis is larger; the cysto-

carp diameter of > 700 µm contrasts with the < 600 µm in

H. bonnemaisonii, and tetrasporangia in H. udoensis are

> 80 µm in diameter, as opposed to < 70 µm in H. bonne-

maisonii (Table 1).

In recognizing the species of Haraldiophyllum, which

currently contains seven species, the vegetative char-

acteristics are viewed as important diagnostic features,

such as whether thallus size is diminutive, whether shape

of the base and margins are dentate or entire, whether

the branching pattern is dichotomous, whether the

lobed blade is deeply cleft, and the presence or absence

of stipes (Table 1). Thallus size is useful for identifying

species of Haraldiophyllum; H. bonnemaisonii and H.

crispatum reach 20-25 cm, whereas H. infossum and H.

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.



Tabl

e 1.

Mor

phol

ogic

al c

ompa

rison

s of

spe

cies

in th

e ge

nus

Har

aldi

ophy

llum

H. u

doe

nsi

ssp

. nov

.H

. bon

nem

aiso

nii

(K

ylin

) A. D

. Z

inov

a

H. c

risp

atu

m (

J. D

. H

oo

ker

& H

arve

y)

Lin

, Ho

mm

ersa

nd

&

Nel

son

H. e

rosu

m

(Har

vey)

A. J

. K

. Mil

lar

& J

. M.

Hu

ism

an

H. i

nfo

ssu

m A

. J. K

. M

illa

rH

. mir

abi

le (K

ylin

) Z

inov

aH

. not

tii (

No

rris

&

M. J

. Wyn

ne)

M. J

. W

ynn

e

H. s

inu

osu

m

(A. H

. S. L

uca

s) A

. J.

K. M

illa

r

Loca

lity

Ud

o, Je

ju Is

lan

d,

Ko

rea

Shet

lan

d Is

lan

d,

UK

; Ire

lan

dC

amp

bel

l Isl

and

, N

ew Z

eala

nd

Gar

den

Isla

nd,

W

este

rn A

ustr

alia

New

So

uth

Wal

es,

Au

stra

liaA

lask

a, U

SAW

ash

ingt

on

, USA

New

So

uth

Wal

es,

Au

stra

lia

Hab

itat

12 m

in s

ub

litto

ral

zon

eE

xtre

me

low

wat

er

to 2

1 m

24 m

, dre

dge

d a

nd

d

rift

Dre

dge

d fr

om 4

1 m

, dri

ft, e

pip

hyt

ic1-

20 m

, tu

rf c

om

-m

un

ity

-

Shal

low

to d

eep

w

ater

-

Size

(h

eigh

t × w

idth

, cm

)22

× 6

15

× 2

0 25

13

1.

98-

10

2-7

20-3

5

Hab

itO

ne

to s

ever

al

ellip

tica

l bla

de

wit

h fi

bro

us

roo

ts, n

ot d

ich

ot-

om

ou

sly

bla

de

On

e to

sev

eral

fan

-sh

aped

bla

de,

d

ich

oto

mo

usl

y d

ivid

ed

Del

icat

e to

ro

bu

st,

sin

gle

or

seve

ral

bla

des

Dic

ho

tom

ou

sly

or

irre

gula

rly

lob

edO

vate

to e

lon

gate

, ov

erla

p in

im-

bri

cate

arr

ange

-m

ent,

cre

epin

g

-D

ich

oto

mo

usl

y b

ran

chin

g-

Bla

de

Th

in, m

emb

ra-

no

us,

mo

no

-st

rom

atic

an

d

po

lyst

rom

atic

Mo

no

stro

mat

ic

wh

en y

ou

ng,

in

crea

sin

g th

ick-

nes

s

Exc

eed

ingl

y va

ri-

able

in s

hap

e,

elo

nga

te o

r fa

n-

shap

ed, d

ivid

ed

Irre

gula

rly

shap

ed

ho

les

in d

ivid

e su

rfac

es, m

on

o-

stro

mat

ic

Mo

no

stro

mat

ic,

irre

gula

rly

bra

nch

ed b

lad

es

aris

ing

fro

m a

st

olo

nif

ero

us

bas

e

Ro

un

dis

h to

d

eep

ly o

r cl

eft

Th

in, f

ork

ed, fl

at-

bra

nch

ed, m

on

o-

stro

mat

ic

Sho

rt, b

lun

t lo

bes

Stip

eSh

ort

cyl

ind

rica

lC

ylin

dri

cal

Cyl

ind

rica

l Sh

ort

dis

tan

ce-

Less

pro

min

ent

Sho

rt, n

arro

wSh

ort

(6-

7 m

m)

Vein

sA

bse

nt

Ab

sen

tA

bse

nt

Ab

sen

tA

bse

nt

-A

bse

nt

-

Mar

gin

sE

nti

reE

nti

re o

r d

enta

teSm

oo

th o

r cr

isp

ed, e

nti

re o

r d

enti

cula

te

Frin

ged

by

mu

l-ti

cellu

lar

pro

-ce

sses

Pro

min

entl

y d

en-

tate

-Sm

oo

th, e

nti

re, a

fe

w m

inu

te te

eth

En

tire

Pro

lifer

atio

ns

Ab

sen

tP

rese

nt

Som

etim

es p

rese

nt

--

--

-

Cle

ft b

lad

eA

bse

nt o

r d

eep

ly

clef

tSo

met

imes

to

bas

eD

isse

cted

irre

gu-

larl

y in

to b

ran

ch-

--

-A

bse

nt

Gro

wth

Mar

gin

al g

row

thD

iffu

se in

mar

-gi

nal

mer

iste

ms

Mar

gin

al g

row

thM

eris

tem

atic

re

gio

nM

argi

nal

init

ials

-M

argi

nal

wit

h in

ter-

cala

ry d

ivis

ion

s-

Pro

carp

Bo

th s

urf

aces

of

up

per

par

tB

oth

sid

es o

f b

lad

e n

ear

mar

-gi

ns

Th

rou

gho

ut t

he

bla

de

Bo

th s

urf

aces

of

the

bla

de

Bo

th s

ides

of t

he

pro

xim

al b

lad

e su

rfac

es

-O

ne

sid

e-

Size

of c

ysto

carp

(µ

m)

500-

900

430-

600

--

--

60-1

,000

-

Sper

mat

angi

al s

ori

Smal

l ro

un

ded

Cir

cula

r to

elli

pti

-ca

lC

ircu

lar

to o

vate

Elo

nga

te, s

ub

-m

argi

nal

pat

ches

Elo

nga

te, c

entr

al

pat

ches

-Sl

igh

tly

elo

nga

te,

irre

gula

r in

sh

ape

-

Tetr

asp

ora

ngi

al s

ori

Cir

cula

r to

el

lipti

cal

Cir

cula

r to

elli

pti

-ca

lC

ircu

lar

to o

vate

Ro

un

ded

or

oval

Ro

un

ded

or

ir-

regu

larl

y-

Ova

te to

elo

nga

te

-

Size

of t

etra

spo

ran

-gi

a (µ

m)

80-1

00

50-7

0 40

-85

60

50

-35

-55

-

Tetr

asp

oran

gia

laye

r Tw

o

Sin

gle

Sin

gle

Two

Si

ngl

e -

Sin

gle

-

Ref

eren

ceT

his

stu

dy

Nam

an

d K

im

1996

Mag

gs a

nd

Ho

m-

mer

san

d 1

993

Lin

et a

l. 20

07M

illar

an

d

Hu

ism

an 1

996

Mill

ar 1

994

Kyl

in 1

925

Wyn

ne

1983

Wo

mer

sley

200

3Lu

cas

1913

Mil

lar

1990

Algae 2011, 26(3): 211-219


group cutting off from the supporting cell prior to the

initial carpogonial branch. The production of the second

sterile cell group delays until after carpogonial branch

formation. The same sequence occurs in H. crispatum, H.

erosum, and H. nottii, but that of H. infossum differs in

forming the two sterile cell groups before initiating the

carpogonial branch (Millar 1994). Millar and Huisman

(1996), therefore, concluded these differences appear to

be stable within the Haraldiophyllum species. In the new

species, H. udoensis, procarp formation and cystocarp

development follow a pattern similar to those previous

studies describe (Millar 1994, Millar and Huisman 1996,

Nam and Kim 1996, Lin et al. 2007).

While researchers often use the female reproductive

organ’s structure to distinguish species within the genus

and genera in the family, they have not considered the

number of layers in the tetrasporangia arrangement to

be of any significance in this genus. The type species H.

bonnemaisonii has only a single layer of tetrasporangia

arranged in a zigzag manner, and the same feature oc-

curs in H. crispatum, H. infossum, and H. nottii. However,

two species, H. udoensis sp. nov. and H. erosum, form two

layers of tetrasporangia. To verify the importance of the

number of layers in the tetrasporangia arrangement from

a phylogenetic perspective, further observation and mo-

lecular analysis for all seven species are necessary.

The genus Haraldiophyllum superficially resembles

two genera, Myriogramme and Nitophyllum, although

Haraldiophyllum clearly belongs to the tribe Myrio-

grammeae based on vegetative and reproductive devel-

opmental characteristics (Hommersand and Fredericq

1997). The genus Haraldiophyllum is readily separated

from the genus Myriogramme by carposporangia orga-

nization, which is terminal solitary in Haraldiophyllum

but terminal chains in Myriogramme (Wynne 1983, Lin

et al. 2007). The new species H. udoensis forms a separate

clade from Myriogramme based on molecular analysis of

rbcL sequence data. However, the new species is linked

to Myriogramme as a sister clade with low bootstrap

support in the ML analysis. Haraldiophyllum is distin-

guished from the genus Nitophyllum by the number of

sterile cell groups in the procarps, with two for Haraldio-

phyllum and only one for Nitophyllum, as well as by the

two periaxial cells’ alignment with respect to the frond’s

longitudinal axis, showing a parallel manner in Haral-

diophyllum and a perpendicular manner in Nitophyllum

(Zinova 1981, Wynne 1983). In the molecular phylogenet-

ic tree, the Haraldiophyllum clade is distantly separate

from the Nitophyllum clade including its type species, N.

punctatum from Spain.

processes. They report these marginal processes are of-

ten polysiphonous and divaricately branched, and they

always terminate single apical cell in a transversely-di-

viding (Millar and Huisman 1996). H. infossum often has

a stoloniferous base and prominently dentate blade mar-

gins (Millar 1994).

The branching pattern in relation to the presence or

absence of lobed blades and stipes can also be an impor-

tant distinguishing feature. H. bonnemaisonii comprises

one to several fan-shaped blades, borne on cartilaginous

stipes and dividing dichotomously into overlapping

lobes with rounded apices (Maggs and Hommersand

1993, Lin et al. 2007). H. crispatum has an exceedingly

variable shape, dividing partially into broad lobes with

rounded margins or cleaving into irregular, dichoto-

mously-branched segments with proliferations (Lin et al.

2007). H. erosum’s blades have dichotomous or irregular

lobes and often have irregularly-shaped holes in the sur-

faces (Millar and Huisman 1996). H. infossum has ovated

to elongated individual blades that often overlap in an

imbricate arrangement with the creeping stoloniferous

parts compressed (Millar 1994). H. mirabile from Alaska

has less prominent stipes, and the blades are roundish

to deeply lobed or cleft (Kylin 1925). The blades of H.

nottii are complicated and subdichotomously branched

with lobed, narrow stipes (Norris and Wynne 1968). The

blades of H. sinuosum are generally entire and not deep-

ly cleft but have short, blunt lobes, and the plants have

short stipes that do not persist into the lower blades (Lu-

cas 1913). The new species H. udoensis appears to differ

from other species mostly by its one to several elliptical,

non-lobed blades.

Researchers have documented the diagnostic char-

acteristics of the genus Haraldiophyllum’s reproductive

structures for the female reproductive organs, such as the

procarp, carposporangia, and cystocarp. The procarps

contain two groups of sterile cells, one group of cover cells

and two periaxial cells parallel to the frond’s longitudinal

axis (Millar 1994). The carposporangia are borne singly

and terminally on gonimoblast filaments. The cystocarp

has a fusion cell incorporating gametophytic cells at the

floor of the cystocarpic cavity. Kylin (1925) noted this fea-

ture of the cystocarp fusion cell is probably useful for ge-

neric recognition of the genus Haraldiophyllum. Maggs

and Hommersand (1993) and Millar (1994) also included

this characteristics in their generic circumscription of

Haraldiophyllum, while Millar and Huisman (1996) rec-

ognized two different sequences of cell formation lead-

ing to the procarp’s final structure in Haraldiophyllum.

They observed H. bonnemaisonii has only one sterile cell



sitets Årsskrift, Ny Följd, Andra Afdelningen 21:1-87.

Lin, S. M., Fredericq, S. & Hommersand, M. H. 2001. System-

atics of the Delesseriaceae (Ceramiales, Rhodophyta)

based on large subunit rDNA and rbcL sequences, in-

cluding the Phycoryoideae, subfam. nov. J. Phycol.

37:881-899.

Lin, S. -M., Hommersand, M. H. & Nelson, W. A. 2007. An

assessment of Haraldiophyllum (Delesseriaceae, Rho-

dophyta), including H. crispatum (J. D. Hooker et Har-

vey) comb. nov. from New Zealand based on rbcL and

LSU sequence analysis and morphological evidence.

Eur. J. Phycol. 42:391-408.

Lucas, A. H. S. 1913. Notes on Australian marine algae, I.

Proc. Linn. Soc. N. S. W. 38:49-60.

Maggs, C. A. & Hommersand, M. H. 1993. Seaweeds of the

British Isles. Vol. 1. Rhodophyta. Part 3A. Ceramiales.

HMSO, London, 444 pp.

Millar, A. J. K. 1990. Marine red algae of the Coffs Harbour re-

gion, northern New South Wales. Aust. Syst. Bot. 3:293-

593.

Millar, A. J. K. 1994. Haraldiophyllum infossum sp. nov. (De-

lesseriaceae, Rhodophyta), a diminutive turf-forming

red alga from the south-western Pacific. Bot. Mar.

37:125-132.

Millar, A. J. K. & Huisman, J. M. 1996. Haraldiophyllum ero-

sum comb. nov. (Delesseriaceae, Rhodophyta) from

Southern and Western Australia. Aust. Syst. Bot. 9:61-69.

Nam, K. W. & Kim, Y. S. 1996. Haraldiophyllum (Rhodophy-

ta): a Delesseriaceous genus newly recorded in Korea.

Algae 11:101-106.

Norris, R. E. & Wynne, M. J. 1968. Notes on marine algae of

Washington and Southern British Columbia, III. Syesis

1:133-146.

Posada, D. & Crandall, K. A. 1998. Modeltest: testing the

model of DNA substitution. Bioinformatics 14:817-818.

Swofford, D. L. 2002. PAUP*: phylogenetic analysis using par-

simony (*and other methods). Version 4.0b10. Sinauer

Associates, Sunderland, MA.

Womersley, H. B. S. 2003. The marine benthic flora of south-

ern Australia, Rhodophyta, Part IIID. Ceramiales: Deles-

seriaceae, Sarcomeniaceae, Rhodymelaceae. Australian

Biological Resources Study & State Herbarium of South

Australia, Canberra & Adelaide, 533 pp.

Wynne, M. J. 1983. The current status of genera in the Deles-

seriaceae (Rhodophyta). Bot. Mar. 26:437-450.

Zinova, A. D. 1981. De positione systematica Nitophylli

(Myriogrammes) yezoensis (Yamada et Tokida) Mikami

(Delesseriaceae). Novit. Syst. Plant. non Vasculariume

18:10-15.

Our ongoing studies of the Delesseriaceae from Jeju Is-

land, Korea have thus far clarified the generic placement

of Haraldiophyllum and led to the recognition of a new

species, H. udoensis. On Jeju Island, many delesseria-

ceous taxa remain that require critical reexamination in

light of recent taxonomic developments within the family.

ACKNOWLEDGEMENTS

This work was supported by a National Research Foun-

dation of Korea grant, funded by the Korean Government

(2011-0003792), and as part of the Survey of Indigenous

Biology of Korea by the National Institute of Biological

Resources (NIBR).

REFERENCES

Bárbara, I., Cremades, J., Calvo, S., López-Rodríguez, M. C.

& Dosil, J. 2005. Checklist of the benthic marine and

brackish Galician algae (NW Spain). Anales del Jardín

Botánico de Madrid 62:69-100.

Gavio, B. & Fredericq, S. 2002. Grateloupia turuturu (Halyme-

niaceae, Rhodophyta) is the correct name of the non-

native species in the Atlantic known as Grateloupia do-

ryphora. Eur. J. Phycol. 37:349-359.

Guiry, M. D. & Guiry, G. M. 2011. AlgaeBase. Worldwide

electronic publication, National University of Ireland,

Galway. Available from: http://www.algaebase.org. Ac-

cessed Mar. 3, 2011.

Hall, T. A. 1999. BioEdit: a user-friendly biological sequence

alignment editor and analysis program for Windows

95/98/NT. Nucleic Acids Symp. Ser. 41:95-98.

Hardy, F. G. & Guiry, M. D. 2003. A check-list and atlas of the

seaweeds of Britain and Ireland. British Phycological So-

ciety, London, 435 pp.

Hommersand, M. H. & Fredericq, S. 1997. Characterization

of Myriogramme livida, Myriogrammeae trib. nov. (De-

lesseriaceae, Rhodophyta). J. Phycol. 33:106-121.

John, D. M., Prud'homme van Reine, W. F., Lawson, G. W.,

Kostermans, T. B. & Price, J. H. 2004. A taxonomic and

geographical catalogue of the seaweeds of the western

coast of Africa and adjacent islands. Nova Hedwigia

Beih. 127:1-339.

Kylin, H. 1924. Studien über die Delesseriaceen. Lunds Uni-

versitets Årsskrift, Ny Följd, Andra Afdelningen 20:1-

111.

Kylin, H. 1925. The marine red algae in the vicinity of of the

biological station at Friday Harbor, Wash. Lunds Univer-

a new korean red algal species, haraldiophyllum...

Documents