Hyung-Seop Kim

PHYLOGENETIC RELATIONSHIPS AMONG LINEAGES OF THE CERAMIACEAE (CERAMIALES, RHODOPHYTA) BASED ON NUCLEAR SMALL SUBUNIT rDNA SEQUENCE DATA1

Phylogenetic relationships among 69 species of the Ceramiales (51 Ceramiaceae, six Dasyaceae, seven Delesseriaceae, and five Rhodomelaceae) were determined based on nuclear SSU rDNA sequence data. We resolved five strongly supported but divergent lineages among the included Ceramiaceae: (i) the genus Inkyuleea, which weakly joins other orders of the Rhodymeniophycidae rather than the Ceramiales in our analyses; (ii) the tribe Spyridieae, which is sister to the remainder of the included ceramialean taxa; (iii) the subfamily Ceramioideae, weakly including the tribe Warrenieae; (iv) the subfamily Callithamnioideae; and (v) the subfamily Compsothamnioideae, which emerges as sister to the Dasyaceae/Delesseriaceae/Rhodomelaceae complex, thus rendering the Ceramiaceae sensu lato unequivocally paraphyletic, as has been argued separately on anatomical grounds by Kylin and Hommersand. Our data support a restricted concept of the Ceramiaceae that includes only one of the five lineages (Ceramioideae) that we have resolved. In addition to failing to ally with the Ceramiales in our molecular analyses, species of Inkyuleea differ substantially from other Ceramiaceae sensu lato in details of pre‐ and postfertilization development. The genus Inkyuleea is here assigned to the Inkyuleeaceae fam. nov., which we provisionally retain in the Ceramiales. Species of Spyridia also differ from the remaining Ceramiaceae in their postfertilization development, and, in light of our molecular data, the genus Spyridia is assigned to the Spyridiaceae. The Callithamnioideae is strongly monophyletic (100% in all analyses), which, in combination with key anatomical differences, supports elevation to family status for this lineage as the Callithamniaceae. Similarly, the Compsothamnioideae is solidly monophyletic in our molecular trees and has a unique suite of defining anatomical characters that supports family status for a complex that we consider to include the tribes Compsothamnieae, Dasyphileae, Griffithsieae, Monosporeae, Ptiloteae, Spermothamnieae, Sphondylothamnieae, Spongoclonieae, and Wrangelieae, for which the reinstated family name Wrangeliaceae is available.

Taxonomic Reappraisal of Dilophus Okamurae (Dictyotales, Phaeophyta) from The Western Pacific Ocean

I.-K. Hwang, W.J. Lee, H.-S. Kim and O. De Clerck. 2008. Taxonomic reappraisal of Dilophus okamurae (Dictyotales, Phaeophyta) from the western Pacific Ocean. Phycologia 48: 1–12. DOI: 10.2216/07-68.1 Dilophus okamurae, a common species of brown alga in the warm temperate western Pacific Ocean (Philippines, Taiwan, China, Korea and Japan), is characterized by a typical vegetative morphology that would support its traditional placement in the genus Dilophus. The species has a unilayered cortex, and the medulla is multilayered at least near the margins of the thallus. Sporangia on a two-celled stalk, without an involucrum, however, indicate a close relationship with the recently described genus Rugulopteryx. Molecular phylogenetic analyses, based on three separate loci, large subunit rDNA (LSU), rbcL and psbA confirm the close phylogenetic relationship between D. okamurae and R. radicans, and R. marginatus and R. suhrii. A transfer of D. okamurae to the genus Rugulopteryx is therefore proposed. Contrary to the expected diplobiontic life cycle, typically encountered in Dictyotales, the species exhibits an asexual life cycle consisting of sporophytes only in the populations studied on the east and south coast of Korea. Gametophytes were only obtained by culturing field collected specimens.

Confirmation on Taxonomic Status of Spatoglossum pacificum Yendo (Dictyotaceae, Phaeophyceae) Based on Morphology and Plastid Protein Coding rbcL, rbcS, psaA, and psbA Gene Sequences

Phonological, morphological and molecular characteristics of Spatoglossum pacificum Yendo are examined. S. pacificum has an annual life cycle composed of saprophytes with apparent absence of male and female gametophytes in Korea. The seasonal growth of this species explains that the annual growth is closely related to the monthly variation of water temperature. S. pacificum has protruding reproductive structures above the outmost cortical layer. Although this observation is restricted to several species, reproductive structures on the thallus can make S. pacificurn distinguishable from S. crassum and S. lacturn. The morphogenesis of a midrib at the base of S. pacificum in this study is the same as those of Dictyopteris but different from those of S. crassum and S. lacturn, suggesting that S. pacificum is closely related to Dictyopteris. In the comparison of plastid gene sequences among species of Spatoglossum and Dictyopteris, S. pacificum is more similar to D. divaricata and D. undulate than those of S. crassum in rbcL, rbcS, psbA and psaA. This result is congruent with the anatomical characteristic of a midrib at the base of the thallus and the protrusion of reproductive organs on the thallus. The phylogenetic relationship based on these plastid genes also shows that S. pacifism is included in Dictyotpteris Glade and separated from S. crassum. We propose the new combination of Dictyopteris pacifica (Yendo) I.K. Hwang, H.S. Kim et W.J. Lee, comb. nov. based on the differences of anatomical characteristics of the midrib, the existence of reproductive organs on thallus and the molecular analyses.

REGENERATION AND SEXUAL DIFFERENTIATION OF GRIFFITHSIA JAPONICA (CERAMIACEAE, RHODOPHYTA) THROUGH SOMATIC CELL FUSION1

Hybrid cells were obtained from somatic cell fusion among male, female, and tetrasporangial plants in Griffithsia japonica Okamura by a wound‐healing process. Isolated fusion cells regenerated new mature plants with mixed reproductive structures. The plants regenerated from hybrid cells between male and female plants developed into 1) spermatangiate, 2) carpogonial, 3) bisexual with spermatangia and carpogonial branches, 4) mixed‐phase with spermatangia and tetrasporangia, or 5) bisexual/mixed‐phase plants with spermatangia, carpogonial branches, and tetrasporangia. About 70% of the plants regenerated from hybrid cells between male and female plants produced tetrasporangia that were always formed with spermatangia on a single cell. Some of those tetrasporangia released tetraspores, six of which gave rise to mature plants. The plants regenerated from hybrid cells between male and tetrasporangial plants developed into spermatangiate, tetrasporangiate, or mixed‐phase plants with spermatangia and tetrasporangia. The plants regenerated from hybrid cells between female and tetrasporangial plants developed into carpogonial, tetrasporangiate, or mixed‐phase plants with carpogonial branches and tetrasporangia. All types of reproductive structures we re functional.

Evidence for Taxonomic Status of Pachydictyon coriaceum (Holmes) Okamura (Dictyotales, Phaeophyceae) Based on Morphology and Plastid Protein Coding rbcL, psaA, and psbA Gene Sequences

The morphological and molecular characteristics of Pachydictyon coriaceum (Holmes) Okamura (1899) are described. Plants are collected from Korea all year round and have maximum height from August to September. The monthly variability of thallus growth is in the way with that of the seawater temperature. Two types of thallus structures, thick cortical layer tallus type and thin cortical cell layer type, are distinguished according to growing seasons. The habit of Korean plants is also classified into two thallus types, slender type and wide type, based on the length and the width of internodes, but this distinction between two types is not supported by either anatomical or molecular characteristics. P. coriaceum shares typical morphology in branching pattern and morphogenetic processes with the other species of Dictyota: 1) multi-cellular cortical and medullar layer in the partial of thallus, 2) same development of thallus from apical meristem cell, and 3) sub-lineage within Dictyota species lineage in rbcL, psaA and psbA gene sequences analyses. These characteristics lead to propose the new combination of Dictyota coriacea (Homes) I.K. Hwang, H.S. Kim et W.J. Lee, comb. nov.

The Occurrence of Griffithsia okiensis (Ceramiaceae, Rhodophyta) from Korea on the Basis of Morphology and Molecular Data

Despite continued studies on red algal flora in Korea, the taxonomy of the tiny ceramiaceous algae has received little attention. We report for the first time Griffithsia okiensis from Korea on the basis of morphology and molecular data. The species is small in thalli height (0.3-1.5 cm), and in diameter of vegetative cells (50-500 μm), and the ratio of cell length/breadth is 2-3 times. It has two carpogonial branches from the supporting cell of procarp. We generated psbA and rbcL sequences from ten specimens of G. okiensis isolated from Korea and Japan and from one G. japonica species isolated Japan. Eight specimens of G. okiensis from Korea were almost identical in both psbA and rbcL regions, nevertheless they differed from Japanese specimens by 4 ucleotides in psbA and 7 in rbcL. In all analyses of psbA, rbcL, and psbA + rbcL data sets, G. okiensis was determined to be a different species from G. japonica isolated from Japan, although both species showed a sister relationship. For all that extensive collection trips, we found no evidence for the occurrence of G. japonica in Korea.

Nuclear DNA Quantification of Some Ceramialean Algal Spermatia by Fluorescence Microscopic Image Processing and their Nuclear SSU rDNA Sequences

Nuclear DNA contents of spermatia from eight ceramiacean and four dasyacean algae (Ceramiales, Rhodophyta) and microspores from two land plants were estimated by fluorescence microscopic image processing and their nuclear SSU rDNA sequence data were analyzed. In frequency distribution patterns, the DAPI-stained nuclear volume (NV) of spermatia showed two peaks corresponding to 1C and 2C. Nuclear 2C DNA contents estimated from NV were 0.45-2.31 pg in ceramiacean and 0.40-0.57 pg in dasyacean algae and 8.42-9.51 pg in two land plants, Capsicum annuum and Nicotiana tabacum. By nuclear patterning of vegetative cells derived from an apical cell, 2C DNA contents of spermatia were 2.31 pg in an alga having uninucleate and non-polyploid nucleus (Aglaothamnion callophyllidicola), 0.45-1.94 pg in algae having uninucleate and polyploid nucleus (Antithamnion spp. and Pterothamnion yezoense), and 0.40-0.62 pg in algae having multinucleate and non-polyploid nuclei (Griffithsia japonica and dasyacean algae). Each mature spermatium and microspore (pollen grain) seemed to have a 2C nucleus, which may provide a genetic buffering system to protect the genetic content of a spermatium and microspore from potentially lethal mutations. Nuclear DNA content and SSU rDNA sequence of Antithamnion sparsum from Korea were reasonably different from those of Antithamnion densum from France. The data did not support the previous taxonomic studies that these two taxa could be conspecific.

Morphological Characteristics of Brown Alga Spatoglossum crassum Tanaka (Dictyotaceae, Dictyotales), New to Korea

Morphological and phonological characteristics of brown alga Spatoglossum crassum Tanaka new to Korea were described based on the field and the indoor cultured plants. The taxonomic characteristics of the plants were agreed to those from the type locality-submerged reproductive organs in cortex, anatomical features, and absence of phaeophycean hairs on the surface. But they have rudimentary midrib on lower portion of thallus. We can observe the young plants on November, adult ones in June, and senile ones in August. This species has an annual life-cycle in the field, starting with germ lings in early November. The differentiation of thallus is quite different from other species of genera in tribe Zonarieae, e.g. Zonaria and Homoeostrichus. Three different tissues, meristoderm, cortex and medulla are discerned. The outmost cortical one celled layer as a meristoderm produce cortex by unequal periclinal division. In the apical cell division, the primary inner cells are developed into 3-4 cell layered medulla of thallus. The distribution of this species extends from Korea to Shizuoka Peninsula (34°40`N) Japan, which is the type locality of this species.