• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.134-135
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• 2003

The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

• Journal of the Korean Society for Marine Environment & Energy
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• v.15 no.2
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• pp.133-141
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• 2012

Effects of temperature and salinity on the growth of the toxic dinoflagellate $Alexandrium$ $tamarense$ and $A.$ $catenella$ isolated from the southern coast of Korea were examined in the laboratory. Growth experiment was conducted under the following combinations of temperature and salinity; 10, 15, 20, 25 and $30^{\circ}C$, 10, 15, 20, 25, 30 and 35 psu at a constant irradiance of 300 ${\mu}mol$ photons $m^{-2}s^{-1}$. Temperature and salinity conditions for maximum growth rate were indicated as follows: temperature $15^{\circ}C$ and salinity 30 psu (0.31 $d^{-1}$) in $A.$ $tamarense$, temperature $25^{\circ}C$ and salinity 30 psu (0.36 $d^{-1}$) in A. catenella. Temperature and salinity ranges for optimum growth condition of two species were $10{\sim}20^{\circ}C$, 25~35 psu and $120{\sim}30^{\circ}C$, 25~35 psu, respectively. The result of two-factor ANOVA indicated significant effects (P<0.001) of temperature and salinity on the growth rate, and two species were more effected by a temperature than a salinity on the growth. In addition, prediction equations were obtained through the multiple regressions of the specific growth rates as ${\mu}=0.04+0.0193T-0.0339S- 0.0005T^2+0.0021S^2+0.00073TS-0.000022T^3-0.000038S^3+0.00000086TS^2-0.0000255T^2S$ in $A.$ $tamarense$ and ${\mu}=1.01-0.1288T-0.0778S+0.0067T^2+0.0038S^2+0.00204TS-0.0001T^3-0.000059S^3-0.0000131TS^2-0.0000392T^2S$ in $A.$ $catenella$. Correlation coefficient between experimental values and simulated values was highly indicated. These results seem to provide information for understanding the spreading mechanism of $A.$ $tamarense$ and $A.$ $catenella$.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.128-128
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• 2003

The annually outbreak of paralytic shellfish poisoning (PSP) were caused by toxic dinolagellate A. tamarense and A. catenella in Korea. The purpose of this study were to investigate the distribution of PSP-causative organisms, A. tamarense and A. catenella and their species classification. Sediment (Saemangeum, the south open sea) and water samples (southeastern coast) were sampled to establish clonal isolates in 2003. After isolation and purification, strains were cultured under $17^{\circ}C$, f/2 media, 14:10=L:D cycle. PST analysis and species identification were performed by HPLC-FD method and specific DNA probe, respectively. Thirty-ons strains were isolated from the Saemangeum reclamation, southeastern coast including Jinhae Bay and south open sea. PSTs were detected in all cultured strains. In eight strains from south offshore, major toxin components are GTX5, C1/2 and minors are GTX3/4, dcGTX3, neoSTX. Sixteen strains from south coastal area have GTX1/4, neoSTX, C1/2 as major toxin components and GTX2/3 as minors. Seven strains from the Saemangeum reclamation have GTX5, C1/2 as major toxin components and GTX1/2/3/4 as minors. Thus, among eight south offshore isolates, four A. tamarense have more toxic (38.31~l19.16 fmol.$cell^{-1}$) than A. catenella (3.78~13.13 fmol.$cell^{-1}$). With the previous results of different toxin composition, toxin components and toxin contents, .it is toxin profile that could used to diagnosis of regional toxic population and geographical distribution of both A. tamarense and A. catenella and their toxigenic strains.

• Journal of Life Science
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• v.12 no.6
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• pp.821-834
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• 2002

In order to measure the inter- and intraspecific genetic divergences within the genus Alexandrium, the variations within the internal transcribed spacer (ITS1 and ITS2) regions and 5.85 ribosomal RNA gene of eight Alexandrium species were examined for 33 strains from diverse geographical locations by direct sequencing. Five isolates of A. tamarense (AT-2, AT-6, AT-10, AT-A and AT-B) from Jinhae Bay, Korea were found to be completely identical to a Japanese strain OFX151-A. The length of the amplified ITSI-5.85-ITS2 region varied from 481 nucleotides (in A. margalefi) to 528 nucleotides (in A. affine CU1-1). ITS1 and ITS2 nucleotide lengths were negatively correlated, whereas a positive correlation was found between their G＋C content. The degree of sequence divergence ranged from 0.3% (1 bp) to a maximum of 53% (305 Up). Pairwise sequence comparisons revealed a small degree of divergence between A. tamarense and A. Pundyense isolates (1.2 - 2.3% ＝ 6-12 bp), but a high degree of divergence between A. tamarense and A. catenella (19.8% ＝ 102 bp), and between A. catenella and A. Pundyense (19.7%). Although most nodes were weakly supported by bootstrap values, some types tend to form independent molecular groups. A. catenella isolates also formed an independent molecular sub-group, with relaticula strong bootstrap values (94% or 85% and 79% or 98%, respectively in PAUP and NJ trees). Interestingly, A. cohorticula and A. frateculus always clustered within the same sub-group, this result being supported by strong bootstrap values. Our results indicate that the ITS regions provide useful informations on hierarchical population genetic structure and a high phylogenetic resolution in intraspecific and interspecific Alexandrium population.

• Journal of the Korean Society of Marine Environment & Safety
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• v.26 no.1
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• pp.84-92
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• 2020

To supply a stable amount of standard material to detect paralytic shellfish toxin (PST), we examined possible increases in toxic content in Alexandrium catenella and A. pacificum using a light emitting diode (LED), which is one of the most eco-friendly and economical lighting method. When comparing the growth rates of organisms and wavelengths of light used, the half saturation constants (Ks) of red wavelength were higher than those of other wavelengths. In contrast, the Ks of blue wavelength were lower than those of other wavelengths. Moreover, when comparing the toxic contents and wavelengths of light used, red wavelength produced approximately 8 times more toxic content in A. catenella and approximately 3.2 times more toxic content in A. pacificum than other wavelengths. Thus, the toxic content present in the organism might be closely related to the Ks of light. The optimum light source to be used to ensure economically ef ective and productive growth in an Alexandrium culture system (photo-bioreactor) would likely consist of a two-phase culture, wherein a blue LED is used during the lag and exponential phases to increase growth rates, followed by the use of a red LED during late exponential and stationary phases to achieve increased PST yields.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.136-137
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• 2003

The marine dinoflagellate genus Alexandrium has been recognized as the most representative toxic phytoplankton on account of production of paralytic shellfish poisoning (PSP) throughout the world. PSP producers, generally A, tamarense and A. catenella, within the genus Alexandrium have caused high level intoxicauon of fisheries products and even death of human. In addition, more recent increasing of geographical range of this deleterious species has given rise to alarming tension. The study presented here aimed construction of the molecular phylogenetic relationships through sequences-determination from 16 morphotypic species (containing newly sequenced 3 morphotypic species, A. tamiyavainchii, A. fraterculus and A. pseudogonyaulax) in LSU rDNA D1-D2 and 12 morphotypic species (containing newly sequenced 6 - morphotypic species, A. catenella, A. tamiyavanichii, A. fraterculus, A. affine, A. insuetum and A. pseudogonyaulax) in SSU rDNA region, and the sequences were subjected to comparative-analysis in respect to regional population using functionally expressed rDNA genus and pseudogenes. And we discussed on genetic differentiation between A. tamarense and A. catenella together with putative PSP divegence of the genus Alexandrium. The results of phylogenetic analysis showed the robust monophyletic 14 distinct classes of A. tamarense, A. excavatum, A. catenella, Tasmanian A. tamarense, A. affine (and/or A. concavum), Thai A. tamarense, A. tamiyavanichii, A. fraterculus, A. margalefii, A. andersonii, A. ostenfeldii, A. minutum (and/or A. lusitanicum), A. insuetum, and A, pseudogonyaulx clade. A. fraterculus and A. tamiyavanichii were sister relationship and they were positioned independently between A, affine cluster and those of A. margalefi, A. andersonii, A. ostenfeldii, A. minutum and A. insuetum. A. pseudogonyaulax appeared to be an ancestral taxon among Alexandrium.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.252-254
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• 2001

A great deal of effort has been put into the identification of Alexandrium tamarense/fundyense/catenella complex by understanding correlation between morphological and subcellular characteristics. To date, the most promising tool for the study of these species is sequence analyses of rRNA genes that have been useful for various organisms' taxonomy and phylogeny, and its application such as in situ hybridization. (omitted)

• Journal of the korean society of oceanography
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• v.34 no.3
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• pp.167-171
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• 1999

Since toxic Alexandrium catenella and non-toxic A. fraterculus are morphologically similar, they are difficult to discriminate under the light microscope. However, a novel technology, such as fluorescein isothiocyanate (FITC)-conjugated lectin probes enables easy and rapid differentiation. Toxic A. catenella bound seven different lectins, whereas the non-toxic A. fratercuzus did not bind Arachis hypogaea (PNA) lectin. In addition, Pseudo-nitrschia species in this study were also difficult to identify to species level with light microscope techniques, but it was possible to classify them using fluorescent lectins. Pseudo-nitzschia multistriata, P. subfraudulenta and P. pungens bound Canavalia ensiformis (ConA), whereas P. subpaclfica did not, and P. pungens also bound Ricinus communis (RCA). These results imply that lectin could be used as a critical tool in the differentiation of P. multistriata, P. subfraudulenta and P. pungens. However, P. subpacifica was not differentiated by the lectins tested. Therefore, it isconcluded that lectin probes are useful for discriminating toxic A. catenella from non-toxic A. fraterculus, and for the identification of some Pseudo-nitzschia species. In addition, this method has a great potential to speed and detection between non-toxic and toxic harmful algal blooms (HABs) in Korean biotoxin monitoring systems.

• Journal of the korean society of oceanography
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• v.39 no.3
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• pp.172-185
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• 2004

LSU rDNA D1-D2 and SSU rDNA genes of 23 strains in seven Alexandrium (Halim) species, A. tamarense (Lebour) Balech, A. catenella (Whedon et Kofoid), A. fraterculus (Balech) Balech, A. affine (Inoue et Fukuyo) Balech, A. insuetum Balech, A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo and A. tamiyavanichii Balech, were sequenced and the data were used for molecular phylogenetic analysis. The sequence data revealed 11 and 7 ribotypes in the LSU rDNA D1-D2 region and 4 and 17 ribotypes in the SSU rDNA region of A. catenella and A. tamarense, respectively. Other Alexandrium species had also 1 to 5 ribotypes in the two regions. With the exception of CMC2 and CMC3 of A. catenella, all A. tamarense and A. catenella strains had a common ribotype, a functionally expressed rRNA gene (here termed type A), in both gene regions. In addition to the functionally expressed gene, several pseudogenes were obtained that were found to be good tools to analyze the population designation of regional isolates by grouping them according to shared ribotypes. From the phylogenetic analysis of the sequence data determined in this study and retrieved from GenBank, the genus Alexandrium was divided into 14 groups: 1) A. tamarense, 2) A. excavatum, 3) A. catenella, 4) Tasmanian A. tamarense, 5) A. affine (and/or A. concavum), 6) Thai A. tamarense, 7) A. tamiyavanichii, 8) A. fraterculus, 9) A. margalefii, 10) A. andersonii, 11) A. ostenfeldii, 12) A. minutum (or A. lusitanicum), 13) A. insuetum, and 14) A. pseudogonyaulax. The SSU rDNA gene sequence of A. fundyense was so similar to those of A. tamarense used in this study that the two species were difficult to discriminate each other. A. tamiyavanichii was closest to the A. tamarense strain isolated in Thailand and close to the long chain-forming species of A. affine and A. fraterculus. The phylogenetic tree showed that A. margalefii, A. andersonii, A. ostenfeldii, A. minutum and A. insuetum constituted the basal relative complex, and that A. pseudogonyaulax is an ancestral taxon in the genus Alexandrium.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.3
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• pp.248-255
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• 2010

This study was carried out to assess the feasibility of the electron beam irradiation as a mean of red tide control in coastal water. Prorocentrum minimum, Prorocentrum micans, Cochlodinium polykrikoides, Heterosigma akashiwo, Alexnadrium catenella were selected and cultured for experiments, and red tide occurring in Tongyeong(2007. 8. 15) was also tested under the same conditions. The irradiation dose were 1 kGy, 2 kGy, 4 kGy and 8 kGy. The result showed 50~65% extinction in red tide cells was observed right after irradiation dose of 1 kGy and 86~97% within 1 day after irradiation, compared with control. Chlorophyll-a concentration of red tide was reduced by 50~64% immediately and it was drastically reduced up to 86~97% 1 day after irradiation. When the culture was irradiated at 1 kGy, 28~47% of s-protein was released immediately, and 77~138% was released 1day after irradiation. 77~212% of s-carbohydrate was excreted after 1 day while 16~45% of s-carbohydrate was excreted immediately. A transmission electron microscope(TEM) observation for the irradiated red tide revealed that the cell was destroyed and intracellular biopolymeric substance was leached out from the damaged cell as a result of electron beam irradiation. These results imply that electron beam irradiation is enable to control red tide by flocculation with extracellular biopolymer. The paralytic shellfish poisoning(PSP) toxin contents produced by Alexandrium catenella was decreased 48% by 1 kGy of electron beam irradiation compared with the unirradiated cells. As a result, electron beam irradiation was effective for detoxication as well as destruction of red tide.