• Journal of Life Science
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• v.17 no.11
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• pp.1453-1463
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• 2007

This study presents a molecular phylogenetic analysis of the harmful dinoflagellate Cochlodinium polykrikoides, by use of partial sequence of small subunit (SSU) rRNA gene from most of the major taxa(24 species) in dinoflagellates. The class Dinophyceae clade formed a strong monophyletic relationship with C. polykrikoides and several taxa. On the basis of deeper nodes, the phylogenetic relationships placed C. polykrikoides closer to the order Prorocentrales rather than to the order Gymnodiniales, which was supported by a strong bootstrap value (100%) in the analyses of Neighbor-Joining and Parsimony methods. There is strong support for C. polykrikoides being placed in the same branch as Gymnodiniaceae and being connected in a clade with Prororcentrum micans among Prorocentrales. Morphological data show that C. polykrikoides is well associated with the genus Gyrodinium; however, this species is genetically closer to Gymnodinium than to Gyrodinium. The placement of C. polykrikoides always formed an independent branch separated from other dinoflagellates. In conclusion, planktonic P. micans plays an important role as an ancestor of Gymnodinium, whereas C. polykrikoides appears to be used an intermediate position between P. micans and Gymnodinium based on evolution.

• Fisheries and Aquatic Sciences
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• v.7 no.3
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• pp.122-129
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• 2004

Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

• ALGAE
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• v.32 no.3
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• pp.189-197
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• 2017

To quantify the abundance of the harmful dinoflagellate Cochlodinium polykrikoides in natural seawaters, we developed the innovative procedure using a droplet digital PCR (ddPCR) with C. polykrikoides-specific primers targeting the internal transcription sequence (ITS). The abundance of C. polykrikoides was estimated by the specific copy number of target ITS DNA segments per cell in cultures and natural water samples. The copy number per C. polykrikoides cell as acquired by ddPCR was $157{\pm}16$, which was evaluated against known cell numbers through a simplified protocol preparing DNAs. The abundances of C. polykrikoides in the waters of different locations estimated by ddPCR agreed with the number of cells visually counted under a microscope. This protocol was used to measure the abundance of C. polykrikoides close to and further off the southern coast of Korea in August of 2016 and 2017. The practical application showed that this method can reduce time for analysis and increase accuracy.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.171-179
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• 2022

The present study investigated the survival rate, respiration rate, plasma stress index, and histological changes according to exposure time of cultured red seabream (Pagrus major) and olive flounder (Paralichthys olivaceus) exposed to Cochlodinium polykrikoides red tide. Fish cultured in natural seawater were used as the control group. Cochlodinium polykrikoides density was set to 5,500±200 cells·ml^-1 in the experimental groups. All red seabreams died within 1 hour of exposure to red tide, whereas all olive flounders died within 5 hours of exposure. Analysis of physiological response revealed that in red seabream, plasma glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), catalase (CAT), and glutathione peroxidase (GPx) concentrations were increased; plasma glucose and superoxide dismutase (SOD) concentration were decreased. Meanwhile, in olive flounders, plasma cortisol, GOT, and GPT concentrations were increased; plasma glucose concentrations were increased during the first hour of exposure, followed by decrease after 5 hours; and plasma SOD, CAT, and GPx concentrations decreased during the first hour of exposure. Histological analysis revealed structural damage to the gills of both red seabream and olive flounder. In conclusion, the exposure of red seabream and olive flounder to Cochlodinium polykrikoides red tide at the density of 5,500 cells·ml^-1 induces oxidative stress, which activates antioxidant defense mechanisms, ultimately leading to liver and gill damage.

• Journal of Environmental Science International
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• v.19 no.12
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• pp.1385-1395
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• 2010

Harmful dinoflagellate Cochlodinium polykrikoides blooms have been frequently occurred in coastal areas of the East Sea since 1995. We compared the oceanic conditions in years 1995, 2001 and 2003 when the C. polykrikoides bloom was strong, and in years 1998 and 2004 when the C. polykrikoides bloom was not appeared. We studied temporal and spatial variation of upwelling and geostrophic currents on the western channel of Korean Strait, an entrance of the East Sea. The period and occurrence area of C. polykrikoides bloom was depended on variation of upwelling in summer. In the distributions of geostrophic current, southward current was dominant near the coast in August, 1998 and 2000. Whereas northward current was dominant near and off the coast in August, 1995 and 2003 which the C. polykrikoides bloom was strong. When compared dominant phytoplankton of the coastal areas in each year, Kuroshio indicator species Proboscia alata and Chaetoceros affine were dominant, respectively, in 2001 and 2003 at every stations. However, the dominant species was variable at each coastal area in 1998 and 2000. In 2003, the abundance of Sagitta elegans which is known as the cold water indicator was low, but the abundance of S. enflata, warm water indicator, was very high in Gangneung compared to Sokcho. It seemed that the distribution of S. elegans is restricted by strong warm water current. In conclusion, it was estimated that the distribution of C. polykrikoides bloom in the coastal area of the East Sea was closely related with the strength of East Korea Warm Current and upwelling.

• Journal of the Korean Association of Geographic Information Studies
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• v.6 no.3
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• pp.139-150
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• 2003

There was a close relationship between spatio-temporal distribution of Cochlodinium polykrikoides red tide and meso-scale variation of oceanographic environment around the Korean waters. Oceanographic conditions of Narodo island, where red tide usually first occurred during summer seasons were formation of the thermohaline frontal zone from 1995 to 2001. Huge C. polykrikoides red tides were observed in every uneven year during the past 7 years (1995~2001) and quasi-biennial oscillation also occurred in the oceanographic variations of sea surface temperature and salinity in the northern part of the East China Sea during the same years. The distribution area and moving pattern of C. polykrikoides red tides were definitely depended on the temporal and spatial variation of upwelling cold water originated form the southeastern coast of the Korean peninsula in summer season.

• Journal of Environmental Science International
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• v.16 no.7
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• pp.799-804
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• 2007

Oil spill caused severe effects on the marine fauna and flora due to direct contact of organisms with the oil and even in regions not directly affected by the spill. This study was conducted to understand the effects of the oil spill accidents and the use of dispersant on the red tide of Cochlodinium polykrikoides. Crude oil produced in Kuwait, bunker-C, kerosene and diesel oil, and a chemical dispersant produced in Korea, were added with a series of 10 ppb to 100 ppm in the f/2-Si medium at $20^{\circ}C$ under a photon flux from cool white fluorescent tubes of $100\;mol\;m^{-2}\;s^{-1}$ in a 14: 10 h L:D cycle for the culture of C. polykrikoides. In low concentrations of ${\leq}$ 1 ppm of examined oils no impact on the growth of C. polykrikoides was recorded, while in high concentration of ${\geq}$ 10 ppm, cell density was significantly decreased with the range of 10 to 80% in comparison with the control. The growth of C. polykrikoides after the addition of the dispersant and the mixtures combined with oils and a dispersant of ${\geq}$ 10 ppm appeared to decrease, whereas the growth of C. polykrikoides exposed to ${\leq}$ 100 ppb showed little serious impact. However, almost all the C. polykrikoides cells were died regardless of a dispersant and combined mixtures within a few days after the addition of high concentrations.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.24 no.3
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• pp.422-428
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• 2019

Fish killing dinoflagellate Cochlodinium polykrikoides has been separated into four genetically differentiated subpopulations globally based on large-subunit (LSU) ribosomal RNA gene, and two subpopulations have been found in the South Sea, Korea. In this study, distributions of the East Asia and Philippines ribotypes were surveyed in the South Sea for 3 years (2014~2016) using quantitative real-time PCR (qPCR). The East Asia ribotype was detected in all sampling stations of the South Sea (Tongyeong~Wando) by 40~100% positives for 2014~2016, whereas the Philippines ribotype was detected in some areas of Tongyeong~Goheung by 1~2% positives for only 2016 when the Tsushima Warm Current (TWC) was particularly strengthened. These results indicate that the East Asia ribotype is the dominant subpopulation in the South Sea, also some of C. polykrikoides swimming cells might be transported from offshore to the South Sea via TWC.

• Journal of Life Science
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• v.14 no.2
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• pp.255-262
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• 2004

We applied nuclear DNA content stained with 4'-6'-diamidino-2-phenylindole (DAPI) and total protein concentration to predict the existence of Cochlodinium polykrikoides before huge blooms occurred, based on a short-term survey at sites in the South Sea. Fluctuations in environmental conditions and nutrients (nitrate, nitrite, and phosphate) were of a similar range, regardless of sampling sites or early and middle field observations. However, C. polykrikoides abundance was significantly different depending on the station, with a higher cell density of 34, 62, and 57 cells L$^{-1}$ at Stn C2, C5, and C6, respectively than what was found in early August, 2000. In mid August, 2000, the highest cell density of 547 cells L$^{-1}$ at Stn C3 was observed. The relationship between C. polykrikoides abundance, DAPI-stained DNA content, and total protein concentration was a positive correlation coefficient, in particular a higher positive correlation was exposed to even a smaller abundance of C. polykrikoides. These results suggest that DNA stained by DAPI and total protein concentration could play an important index in easily predicting the presence of C. polykrikoides before blooms.

• ALGAE
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• v.26 no.3
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• pp.229-235
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• 2011

Spliced leader (SL) trans-splicing is a mRNA processing mechanism in dinoflagellate nuclear genes. Although studies have identified a short, conserved dinoflagellate SL (dinoSL) sequence (22-nt) in their nuclear-encoded transcripts, whether the majority of nuclear-coded transcripts in dinoflagellates have the dinoSL sequence remains doubtful. In this study, we investigated dinoSL-containing gene transcripts using 454 pyrosequencing data (Cochlodinium polykrikoides, 93 K sequence reads, 31 Mb; Prorocentrum minimum, 773 K sequence reads, 291 Mb). After making comparisons and performing local BLAST searches, we identified dinoSL for one C. polykrikoides gene transcript and eight P. minimum gene transcripts. This showed transcripts containing the dinoSL sequence were markedly fewer in number than the total expressed sequence tag (EST) transcripts. In addition, we found no direct evidence to prove that most dinoflagellate nuclear-coded transcripts have this dinoSL sequence.