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564

Silicon transporter genes of Fragilariopsis cylindrus (Bacillariophyceae) are differentially expressed during the progression of cell cycle synchronized by Si or light

Oh, Han Sang, Lee, Sung-eun, Han, Chae-seong, Kim, Joon, Nam, Onyou, Seo, Seungbeom, Chang, Kwang Suk, Jin, EonSeon, Hwang, Yong-sic

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.2 2018 pp.191-203

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Fragilariopsis cylindrus is one of the most successful psychrophiles in the Southern Ocean. To investigate the molecular mechanism of biomineralization in this species, we attempted to synchronize F. cylindrus growth, since new cell wall formation is tightly coupled to the cell division process. Nutrient limitation analysis showed that F. cylindrus cultures rapidly stopped growing when deprived of silicate or light, while growth continued to a certain extent in the absence of nitrate. Flow cytometry analysis indicated that deprivation of either silicate or light could effectively arrest the cell cycle of this diatom species at the G1 phase, suggesting that synchrony can be established using either factor. Fluorescence labeling of new cell walls was faintly detectable as early as approximately 6 h after silicon repletion or light irradiation, and labeling was markedly intensified by 18 h. It is revealed that the synthesis of girdle bands begins before valve synthesis in this species, with active valve synthesis occurring during the G2 / M phase. Expression profiling revealed that selective member(s) of the F. cylindrus SIT genes (FcSIT) respond to silicate and light, with a different set of genes being responsive to each factor. The Si / light double depletion experiments demonstrated that expression of one FcSIT gene is possibly correlated to transition to G2 / M phase of the cell cycle, when the valve is actively formed.

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563

Calcium-related genes associated with intracellular calcification of Emiliania huxleyi (Haptophyta) CCMP 371

Nam, Onyou, Shiraiwa, Yoshihiro, Jin, EonSeon

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.2 2018 pp.181-189

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Emiliania huxleyi (a haptophyte) is the most abundant coccolithophore species that produces delicate calcite scales called coccoliths. In this study, we identified several candidate genes associated with coccolith production by comparing the transcriptomes of the calcifying (CCMP 371) and non-calcifying (CCMP 2090) strains of E. huxleyi. Among the candidates, genes highly expressed in CCMP 371 were identified. To confirm whether these genes are associated with calcification, we modulated coccolith production in CCMP 371 by culturing it at different calcium concentrations. At an ambient (10 mM) concentration of calcium in the growth medium, CCMP 371 sustained its calcifying ability. However, at a low (0.1 mM) concentration or absence of calcium, there was no calcite formation, demonstrating that calcium-limiting conditions negatively affect calcification. We also evaluated the expression patterns of the putative genes in cells grown at different calcium concentrations by quantitative reverse transcription polymerase chain reaction. In addition, we showed that the growth rate of cells cultured under calcium-limiting conditions does not differ from that under ambient conditions. Further studies are required to investigate the roles of the putative calcification-associated genes at the molecular level.

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562

Next-generation sequencing reveals the diversity of benthic diatoms in tidal flats

An, Sung Min, Choi, Dong Han, Lee, Howon, Lee, Jung Ho, Noh, Jae Hoon

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.2 2018 pp.167-180

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Benthic diatoms are ubiquitous in tidal flats and play major roles in maintaining coastal ecosystems. Spatio-temporal variations in diatom diversity have not been well-studied, mainly because of difficulties in morphological identification and the lack of appropriate genetic tools. To overcome these problems, we used the gene encoding the ribulose bisphosphate carboxylase large-subunit (rbcL) as a molecular marker, and sequenced these genes with the aid of the MiSeq platform. In this manner, we explored the genetic diversity of benthic diatoms in tidal flats of Guenso Bay on the west coast of Korea; differences in the spatial distributions of benthic diatoms were evident. The diatom communities were dominated by Nitzschia, Navicula, and Amphora; their relative distributions were affected by the sand proportion, grain size, and air exposure time. Our results suggest that meta-barcoding of the rbcL gene and next-generation sequencing can be used to explore the diversity of benthic diatoms.

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561

A molecular investigation of Saccharina sessilis from the Aleutian Islands reveals a species complex, necessitating the new combination Saccharina subsessilis

Starko, Samuel, Boo, Ga Hun, Martone, Patrick T., Lindstrom, Sandra C.

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.2 2018 pp.157-166

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Cryptic species complexes are increasingly recognized in phycological research, obscuring taxonomy and raising questions about factors influencing speciation. A recent exploration of kelp genetic diversity on Haida Gwaii, British Columbia revealed the existence of a new species, Saccharina druehlii, which is cryptic with Saccharina sessilis. This suggests that molecular investigations further north may be required to elucidate the taxonomy and evolutionary history of this lineage. Although, for several decades, S. sessilis was considered a single highly variable species, its taxonomy has been far from straightforward. In particular, Hedophyllum subsessile (Areschoug) Setchell is now recognized as a synonym of S. sessilis in North America, but as a growth form of Saccharina bongardiana in Far East Russia. To resolve this taxonomic confusion, we sequenced mitochondrial (CO1-5P) and nuclear (internal transcribed spacer) markers of S. sessilis populations from the Aleutian Islands, Alaska, USA. Interestingly, none of our sequences matched S. sessilis sensu stricto. Instead, CO1-5P sequences from populations in the central and eastern Aleutians matched exactly S. druehlii with increasing sequence divergence occurring westward. Samples from Attu, the western-most island, composed a genetic group that clearly represents Kjellman's concept of Hafgygia bongardiana f. subsessilis and is distinct enough from S. druehlii and S. sessilis to potentially constitute a distinct species. Therefore, Saccharina subsessilis comb. nov. is proposed for this entity. Our results suggest the existence of a species complex at the crown node of S. sessilis and thus further investigation of Saccharina in Alaskan waters should be conducted to reconstruct the evolutionary history of this fascinating lineage.

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560

Morphological characterization and molecular phylogenetic analysis of Dolichospermum hangangense (Nostocales, Cyanobacteria) sp. nov. from Han River, Korea

Choi, Hye Jeong, Joo, Jae-Hyoung, Kim, Joo-Hwan, Wang, Pengbin, Ki, Jang-Seu, Han, Myung-Soo

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.2 2018 pp.143-156

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Dolichospermum is a filamentous and heterocytous cyanobacterium that is one of the commonly occurring phytoplanktons in the Han River of Korea. Morphological observations led to the identification of D. planctonicum-like filaments in seasonal water samples. In the present study, we successfully isolated these filaments using culture methods, and examined its morphology using light and scanning electron microscopy. The morphology of the D. planctonicumlike species differed from that of typical D. planctonicum; it had thin cylindrical-shaped akinetes, which were narrower towards the ends than at the center. This morphology is firstly described in the genus Dolichospermum. In addition, the akinetes in the filament developed solitarily and were distant from the heterocytes. Phylogenetic analysis of the 16S rRNA sequences showed that our Dolichospermum clustered with D. planctonicum and D. circinale, which have coiled trichome. However, phylogenetic analysis of the gene encoding rivulose-1,5-bisphosphate carboxylase (rbcLX) clearly separated our species from other Dolichospermum, forming a unique clade. Additionally, structures of D. planctonicum and D. hangangense strains were different type in Box-B and V3 region. These results demonstrated that the new Dolichospermum species was unique in morphology and molecular traits. Therefore, we propose this to be a new species belonging to genus Dolichospermum with the name Dolichospermum hangangense sp. nov.

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559

Insertional mutations exhibiting high cell-culture density HCD phenotypes are enriched through continuous subcultures in Chlamydomonas reinhardtii

Thung, Leena, He, Jing, Zhu, Qingling, Xu, Zhenyu, Liu, Jianhua, Chow, Yvonne

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.127-141

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Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

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558

Intensive land-based production of red and green macroalgae for human consumption in the Pacific Northwest: an evaluation of seasonal growth, yield, nutritional composition, and contaminant levels

Gadberry, Bradley A., Colt, John, Maynard, Desmond, Boratyn, Diane C., Webb, Ken, Johnson, Ronald B., Saunders, Gary W., Boyer, Richard H.

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.109-125

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Turkish towel (Chondracanthus exasperatus), Pacific dulse (Palmaria mollis, also known as Red ribbon seaweed), and sea lettuce (Ulva spp.) were cultivated in a land-based intensive culture system at the Manchester Research Station, USA from August 2013 to September 2014. Macroalgae were grown in tumble-aerated tanks, harvested bimonthly for seasonal growth calculations, and analyzed for protein, lipid, ash, and amino acid content. Growth rate of all three species exhibited a similar pattern, with the highest specific growth rates occurring during the summer months (Turkish towel: 7.8%, Pacific dulse: 8.2%, and sea lettuce: 6.2%). Growth of all three species was lowest around winter solstice; with negative growth only observed in sea lettuce. On a dry weight basis significant differences in protein content existed between the three species with highest values for sea lettuce ($29.5{\pm}1.4%$). Lipid content varied between species (0.95-2.78%) with significantly higher lipid observed in sea lettuce (0.58-4.82%). No significant differences were detected on a seasonal basis among each species. Essential amino acids accounted for $43{\pm}0.9$ to $47{\pm}1.2%$ of total amino acids with Turkish towel having the highest value. Turkish towel had a significantly higher taurine level ($0.82{\pm}0.27$) than the other macroalgae. The levels of persistent organic pollutants and heavy metals were low. The estimated annual product of the three species ranged from 50- to $70-mt\;dry\;weight\;ha^{-1}\;y^{-1}$, significantly higher than conventional crops. Land-based culture of these species can produce year-round harvest, consistent product quality, and low contaminant levels.

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557

Comparison of the cultivation performance between Korean (Sugwawon No. 301) and Chinese strains (Huangguan No. 1) of kelp Saccharina japonica in an aquaculture farm in Korea

Hwang, Eun Kyoung, Liu, Fuli, Lee, Ki Hyun, Ha, Dong Su, Park, Chan Sun

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.101-108

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Saccharina japonica was introduced to both Korea and China from Hokkaido, Japan, and it has become an economically important species in both nations. After a long period of cultivation, several varieties of S. japonica have been developed in Korea and China. In this study, we conducted aquacultural research on the persistence of thalli from two kelp cultivars, one from China (Huangguan No. 1) and one from Korea (Sugwawon No. 301), between December 2015 and November 2016 in Haenam, Korea. The maximum length was $247.8{\pm}13.0$ and $227.5{\pm}42.0cm$, respectively, which were significantly longer in Sugwawon No. 301 than in Huangguan No. 1. The maximum width was $29.9{\pm}5.4$ and $23.2{\pm}1.9cm$, respectively, which were significantly wider in Huangguan No. 1 than in Sugwawon No. 301. The mean biomass obtained from the culture ropes was for Sugwawon No. 301 was $3.5{\pm}0.3kg\;wet\;wt\;m^{-1}$ and for while Huangguan No. 1 was $3.1{\pm}1.0kg\;wet\;wt\;m^{-1}$ of culture rope. After August, the persistence of the thalli of Sugwawon No. 301 was two months longer than that of Huangguan No. 1. We found that the Sugwawon No. 301 performed as well as the Huangguan No. 1 in Korean waters possibly due to increased flexibility as a result of the different cell arrangements of the two cultivars. Overall, the use of the Sugwawon No. 301 cultivar rather than the Huangguan No. 1 cultivar of S. japonica appears the best alternative to help to ensure a stable year round algal feed supply for the Korean abalone industry.

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556

Spatial and temporal dynamics of the abundance of crustose calcareous algae on the southernmost coral reefs of the western Atlantic (Abrolhos Bank, Brazil)

Amado-Filho, Gilberto M., Bahia, Ricardo G., Mariath, Rodrigo, Jesionek, Michel B., Moura, Rodrigo Leao, Bastos, Alex C., Pereira-Filho, Guilherme Henrique, Francini-Filho, Ronaldo Bastos

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.85-99

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Crustose calcareous algae (CCA) constitute one of the main reef builders on the Abrolhos Bank, Brazil. Once CCA taxonomy is locally understood, differences in growth-forms may be useful for the delimitation of taxa using characteristics such as the presence or absence of surface protuberances. Here, growth-forms were used to identify and quantify the most common CCA taxa on the shallow reefs (3-10 m) of the Abrolhos Bank to determine possible changes in the CCA community over a period of 10 years, and the ecological significance of CCA to local reefs was interpreted. The CCA assemblages were surveyed from 2006-2015 by using fixed photoquadrats at four sites in the inner (10-20 km from the mainland) and mid-shelf reefs (40-75 km from the mainland). The five most common CCA taxa were Pneophyllum conicum, the Lithophyllum kaiserii / Lithophyllum sp. complex, Melyvonnea erubescens, the Hydrolithon boergesenii / Porolithon onkodes complex and Peyssonelia sp. The overall mean CCA cover on the reefs was 20%. A comparison with a previous monitoring study in the same region indicated that the CCA cover nearly doubled from 2003-2008 to 2006-2015. This study reveals that the coral-killing species P. conicum dominated CCA flora on the shallow Abrolhos reefs in the last decade, and the local specific abundance of CCA slightly fluctuated over time and was species- and site-specific. The information obtained in this study contributes to the understanding of the ecology of the key calcifying components of the Abrolhos reefs and provides a useful baseline for exploring the responses of CCA to future environmental changes.

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555

Taxonomic revision of the genus Herposiphonia (Rhodomelaceae, Rhodophyta) from Korea, with the description of three new species

Koh, Young Ho, Kim, Myung Sook

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.69-84

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We examined the species diversity of Herposiphonia on Korean coasts, based on a combination of morphology and molecular analyses of the mitochondrial COI-5P DNA barcode marker and plastid rbcL gene. We report the presence of eight species including three novel species: H. donghaensis sp. nov., H. jejuinsula sp. nov., H. sparsa sp. nov., H. caespitosa, H. fissidentoides, H. insidiosa, H. parca, and H. subdisticha. Specimens were separated into eight clades in both the COI-5P and rbcL gene analyses, with 1.3-19.6 and 6.6-15% interspecific sequence divergence, respectively. These eight species are also distinguishable by several morphological characteristics such as: branching pattern (d/i pattern in H. donghaensis sp. nov. and H. sparsa sp. nov.; d/d/d/i pattern in others), shape of determinate branch (ligulate in H. fissidentoides; terete in others), number of vegetative trichoblasts (1-2 in H. insidiosa and H. sparsa sp. nov.; 3-4 in H. caespitosa; absent in others), and number of segments and pericentral cells in determinate branches. About three novel species revealed by our analyses, H. donghaensis sp. nov. is newly discovered, and H. jejuinsula sp. nov. and H. sparsa sp. nov. were previously reported in Korea as H. nuda and H. secunda, respectively. Our results show that DNA barcoding and rbcL analyses are useful for delimiting species boundaries and discovering cryptic species diversity in the genus Herposiphonia.

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554

Biogeographic pattern of four endemic Pyropia from the east coast of Korea, including a new species, Pyropia retorta (Bangiaceae, Rhodophyta)

Kim, Sun-Mi, Choi, Han-Gu, Hwang, Mi-Sook, Kim, Hyung-Seop

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.55-68

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Foliose species of the Bangiaceae (Porphyra s. l.) are very important in Korean fisheries, and their taxonomy and ecophysiology have received much attention because of the potential for developing or improving aquaculture techniques. Although 20 species of foliose Bangiales have been listed from the Korean coast, some of them remain uncertain and need further comparative morphological studies with molecular comparison. In this study, we confirm the distribution of four Pyropia species from the east coast of Korea, Pyropia kinositae, P. moriensis, P. onoi, and P. retorta sp. nov., based on morphology and rbcL sequence data. Although P. onoi was listed in North Korea in old floral works, its occurrence on the east coast of South Korea is first revealed in this study based on molecular data. P. kinositae and P. moriensis, which were originally described from Hokkaido, Japan, are first reported on the east coast of Korea in this study. Pyropia retorta sp. nov. and P. yezonesis share a similar thallus color and narrow spermatangial patches in the upper portion of the frond, and they have a sympatric distribution. However, P. retorta can be distinguished by the curled or twisted thalli and by molecular data. The biogeographic pattern of the two native species, P. kinositae and P. retorta, suggests that the east coast of Korea may have been a place of refugia during the Last Glacial Maximum (LGM), and then recolonized to the northern part of Japan through the restored East Korean Warm Current after the LGM.

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553

Intraspecific variation of gene structure in the mitochondrial large subunit ribosomal RNA and cytochrome c oxidase subunit 1 of Pyropia yezoensis (Bangiales, Rhodophyta)

Hwang, Il Ki, Kim, Seung-Oh, Hwang, Mi Sook, Park, Eun-Jeong, Ha, Dong-Soo, Lee, Sang-Rae

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.49-54

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Red algal mitochondrial genomes (mtDNAs) can provide useful information on species identification. mtDNAs of Pyropia / Porphyra (Bangiales, Rhodophyta) have shown diverse variation in their size and gene structure. In particular, the introns and intronic open reading frames found in the ribosomal RNA large subunit gene (rnl) and cytochrome c oxidase subunit 1 gene (cox1) significantly vary the mitochondrial genome size in Pyropia / Porphyra species. In this study, we examined the exon / intron structure of rnl and cox1 genes of Pyropia yezoensis at the intraspecific level. The combined data of rnl and cox1 genes exhibited 12 genotypes for 40 P. yezoensis strains, based on the existence of introns. These genotypes were more effective to identify P. yezoensis strains in comparison to the traditional DNA barcode cox1 marker (5 haplotypes). Therefore, the variation in gene structure of rnl and cox1 can be a novel molecular marker to discriminate the strains of Pyropia species.

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552

Morphological and molecular identification of Alaria paradisea (Phaeophyceae, Laminariales) from the Kurile Islands

Klimova, Anna V., Klochkova, Nina G., Klochkova, Tatyana A., Kim, Gwang Hoon

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.37-48

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Alaria is the second largest genus of the Laminariales, which is distributed far into the northern Pacific and Atlantic oceans. Due to its high morphological plasticity, over 100 specific and sub-specific names have been used in Alaria, this has been tailored down to the present 17 species through morphological revision and molecular phylogenetic analysis. Endemic species of Alaria from Russian Far East have not been thoroughly revised since their original description, and few of them were confirmed using molecular data until recently. We carried out morphological and molecular studies on A. paradisea which is an endemic species distributed on the Kurile Islands, first described by Miyabe and Nagai in 1932 as Pleuropterum paradiseum. The range of morphological variability and its distribution was re-evaluated using the type specimen as well as other specimens. Analyses of partial mitochondrial cytochrome c oxidase subunit 1 and nuclear-encoded internal transcribed spacer sequences showed that A. paradisea nested within the genus Alaria, but differs morphologically from any other Alaria species in having additional sporophylls with a central midrib (${\beta}-sporophylls$). Our results showed that A. paradisea clearly belongs to the genus Alaria based on DNA data, although the key morphological character that was used to include this species to the genus Pleuropterum, ${\beta}-sporophylls$, is stable and distinguishes it from other Alaria species.

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551

Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

Lee, Sung Yeon, Jeong, Hae Jin, You, Ji Hyun, Kim, So Jin

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.21-35

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The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

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550

Revisiting the Parvilucifera infectans / P. sinerae (Alveolata, Perkinsozoa) species complex, two parasitoids of dinoflagellates

Jeon, Boo Seong, Nam, Seung Won, Kim, Sunju, Park, Myung Gil

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.33 No.1 2018 pp.1-19

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Members of the family Parviluciferaceae (Alveolata, Perkinsozoa) are the well-known dinoflagellate parasitoids along with Amoebophrya ceratii species complex and parasitic chytrid Dinomyces arenysensis and contain six species across three genera (i.e., Parvilucifera infectans, P. sinerae, P. rostrata, and P. corolla, Dinovorax pyriformis, and Snorkelia prorocentri) so far. Among Parvilucifera species, the two species, P. infectans and P. sinerae, are very similar or almost identical each other morphologically and genetically, thereby make it difficult to distinguish between the two. The only main difference between the two species known so far is the number of sporangium wall (i.e., 2 layers in P. infectans vs. 3 layers in P. sinerae). During sampling in Masan bay, Korea during the spring season of 2015, the dinoflagellate Akashiwo sanguinea cells infected by the parasite Parvilucifera were observed and this host-parasite system was established in culture. Using this culture, its morphological and ultrastructural features with special emphasis on the variation in the number of sporangium wall over developmental times, were investigated. In addition, the sequences of rDNA regions and ${\beta}-tubulin$ genes were determined. The result clearly demonstrated that the trophocyte at 36 h was covered with 4 layers, and then outer layer of the sporocyte gradually degraded over time, resulting in wall structure consisting of two layers, with even processes being detached from 7-day-old sporangium with smooth surface, indicating that the difference in the number of layers seems not to be an appropriate ultrastructural character for distinguishing P. infectans and P. sinerae. While pairwise comparison of the large subunit rDNA sequences showed 100% identity among P. infectans / P. sinerae species complex, genetic differences were found in the small subunit (SSU) rDNA sequences but the differences were relatively small (11-13 nucleotides) compared with those (190-272 nucleotides) found among the rest of Parvilucifera species (P. rostrata and P. corolla). Those small differences in SSU rDNA sequences of P. infectans / P. sinerae species complex may reflect the variations within inter- strains of the same species from different geographical areas. Taken together, all morphological, ultrastructural, and molecular data from the present study suggest that they are the same species.

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549

Protective effect of gallic acid derivatives from the freshwater green alga Spirogyra sp. against ultraviolet B-induced apoptosis through reactive oxygen species clearance in human keratinocytes and zebrafish

Wang, Lei, Ryu, BoMi, Kim, Won-Suk, Kim, Gwang Hoon, Jeon, You-Jin

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.32 No.4 2017 pp.379-388

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In the present study, we enhanced the phenolic content of 70% ethanol extracts of Spirogyra sp. (SPE, $260.47{\pm}5.21$ gallic acid equivalent $[GAE]mg\;g^{-1}$), 2.97 times to $774.24{\pm}2.61GAE\;mg\;g^{-1}$ in the ethyl acetate fraction of SPE (SPEE). SPEE was evaluated for its antiradical activity in online high-performance liquid chromatography-ABTS analysis, and the peaks with the highest antiradical activities were identified as gallic acid derivatives containing gallic acid, methyl gallate, and ethyl gallate. Isolation of ethyl gallate from Spirogyra sp. was performed for the first time in this study. In ultraviolet B (UVB)-irradiated keratinocytes (HaCaT cells), SPEE improved cell viability by 8.22%, and 23.33% and reduced accumulation of cells in the sub-$G_1$ phase by 20.53%, and 32.11% at the concentrations of 50 and $100{\mu}g\;mL^{-1}$, respectively. Furthermore, SPEE (50 and $100{\mu}g\;mL^{-1}$) reduced reactive oxygen species generation in UVB-irradiated zebrafish by 66.67% and 77.78%. This study suggests a protective activity of gallic acid and its derivatives from Spirogyra sp. against UVB-induced stress responses in both in vitro and in vivo models, suggesting a potential use of SPEE in photoprotection.

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548

Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis (Bangiales, Rhodophyta)

Du, Yu, Guan, Jian, Xu, Ruijun, Liu, Xin, Shen, Weijie, Ma, Yafeng, He, Yuan, Shen, Songdong

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.32 No.4 2017 pp.359-377

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Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

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547

Variations in carbon and nitrogen stable isotopes and in heavy metal contents of mariculture kelp Undaria pinnatifida in Gijang, southeastern Korea

Shim, JeongHee, Kim, Jeong Bae, Hwang, Dong-Woon, Choi, Hee-Gu, Lee, Yoon

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.32 No.4 2017 pp.349-357

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Korean mariculture Undaria pinnatifida was collected during the months of January, February, March, and December of 2010, as well as from January of 2011 to investigate the changes in the carbon and nitrogen stable isotope ratios (${\delta}^{13}C$ and ${\delta}^{15}N$) and heavy metal with respect to it growth and to identify the factors that influence such changes. The blades of U. pinnatifida showed ${\delta}^{13}C$ and ${\delta}^{15}N$ in the range (mean) of -13.11 to -19.42‰ (-16.93‰) and 2.99 to 7.57‰ (4.71‰), respectively. Among samples with the same grow-out period, those that weighed more tended to have higher ${\delta}^{13}C$ suggesting a close association between the carbon isotope ratio and growth rate of U. pinnatifida. Indeed, we found a very high positive linear correlation between the monthly average ${\delta}^{13}C$ and the absolute growth rate in weight ($r^2=0.89$). Nitrogen isotope ratio tended to be relatively lower when nitrogen content in the blade was higher, probably due to the strengthening of isotope fractionation stemming from plenty of nitrogen in the surrounding environment. In fact, a negative linear correlation was observed with the nitrate concentration in the nearby seawaters ($r^2=0.83$). Concentrations of Cu, Cd, Pb, Cr, Hg, and Fe in the blades showed a rapid decrease in their concentration per unit weight in the more mature U. pinnatifida. Specifically, compared to adult samples, Cu, Hg, and Pb were concentrated by 30, 55, and 73 folds, respectively, in the young blades. Therefore, U. pinnatifida tissue ${\delta}^{13}C$ is as an indirect indicator of its growth rate, while ${\delta}^{15}N$ values and heavy metal concentrations serve as tracers that reflect the environmental characteristics.

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546

Growth, reproduction and recruitment of Silvetia siliquosa (Fucales, Phaeophyceae) transplants using polyethylene rope and natural rock methods

Gao, Xu, Choi, Han Gil, Park, Seo Kyoung, Lee, Jung Rok, Kim, Jeong Ha, Hu, Zi-Min, Nam, Ki Wan

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.32 No.4 2017 pp.337-347

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Silvetia siliquosa is an ecologically and commercially important brown alga that is harvested from its natural habitats, but its population has recently been diminishing along the Korean coast. To develop new techniques for algal population restoration, we tested two newly developed transplantation methods (using polyethylene ropes and natural rock pieces) at two study sites, Gwanmaedo and Yeongsando, on the southwest coast of Korea, from May to November 2014. The transplants on polyethylene ropes showed significantly greater survival, maturity, and growth than those on natural rocks at both study sites. Newly recruited juveniles (<3 cm) of S. siliquosa increased remarkably from May to December near the transplants on polyethylene ropes and natural rocks.Therefore, we suggest that transplantation using polyethylene ropes is more effective than using natural rocks to restore the population of S. siliquosa in Korea.

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545

Effects of disturbance timing on community recovery in an intertidal habitat of a Korean rocky shore

Kim, Hyun Hee, Ko, Young Wook, Yang, Kwon Mo, Sung, Gunhee, Kim, Jeong Ha

[Kisti 연계] 한국조류학회(藻類) ALGAE Vol.32 No.4 2017 pp.325-336

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Intertidal community recovery and resilience were investigated with quantitative and qualitative perspectives as a function of disturbance timing. The study was conducted in a lower intertidal rock bed of the southern coast of South Korea. Six replicates of artificial disturbance of a $50cm{\times}50cm$ area were made by clearing all visible organisms on the rocky substrate in four seasons. Each of the seasonally cleared plots was monitored until the percent cover data reached the control plot level. There was a significant difference among disturbance timing during the recovery process in terms of speed and community components. After disturbances occurred, Ulva pertusa selectively preoccupied empty spaces quickly (in 2-4 months) and strongly (50-90%) in all plots except for the summer plots where non-Ulva species dominated throughout the recovery period. U. pertusa acted as a very important biological variable that determined the quantitative and qualitative recovery capability of a community. The qualitative recovery of communities was rapid in summer plots where U. pertusa did not recruit and the community recovery rate was the lowest in winter plots where U. pertusa was highly recruited with a long duration of distribution. In this study, U. pertusa was a pioneer species while being a dominant species and acted as a clearly negative element in the process of qualitative recovery after disturbance. However, the negative effect of U. pertusa did not occur in summer plots, indicating that disturbance timing should be considered as a parameter in understanding intertidal community resilience in temperate regions with four distinct seasons.

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