• Journal of Environmental Science International
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• v.18 no.12
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• pp.1331-1338
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• 2009

Marine dinoflagellate Alexandrium minutum producing paralytic shellfish toxins is responsible for paralytic shellfish poisoning (PSP). To investigate its temporal distributions in Chinhae Bay where PSP occurs annually, SYBR Green I based A. minutum-specific real-time PCR probe was developed on the LSU rDNA region. Assay specificity and sensitivity were tested against related species, and its specificity was further confirmed by sequencing of field-derived samples. Ten months field survey in 2008 (a total 100 surface water samples) by using the real-time PCR probe showed that A. minutum was detected at very low densities of 1-4 cells $L^{-1}$ in May and June being spring in Chinhae Bay, Korea.

• 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.

• 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.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.132-133
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• 2003

The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

• ALGAE
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• v.39 no.1
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• pp.1-16
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• 2024

Many dinoflagellates produce bioluminescence. To estimate the intensity of bioluminescence produced by populations of the heterotrophic dinoflagellates Noctiluca scintillans and Polykrikos kofoidii and autotrophic dinoflagellate Alexandrium mediterraneum in Korean waters, we measured cellular bioluminescence intensity as a function of water temperature and calculated population bioluminescence intensity with cell abundances and water temperature. The mean 200-second-integrated bioluminescence intensity per cell (BL_cell) of N. scintillans satiated with the chlorophyte Dunaliella salina decreased continuously with increasing water temperature from 5 to 25℃. However, the BL_cell of P. kofoidii satiated with the mixotrophic dinoflagellate Alexandrium minutum continuously increased from 5 to 15℃ but decreased at temperatures exceeding this (to 30℃). Similarly, the BL_cell of A. mediterraneum continuously increased from 10 to 20℃ but decreased between 20 and 30℃. The difference between highest and lowest BL_cell of N. scintillans, P. kofoidii, and A. mediterraneum at the tested water temperatures was 3.5, 11.8, and 21.0 times, respectively, indicating that water temperature clearly affected BL_cell. The highest estimated population bioluminescence intensity (BL_popul) of N. scintillans in Korean waters in 1998-2022 was 4.22 × 10¹³ relative light unit per liter (RLU L^-1), which was 1,850 and 554,000 times greater than that of P. kofoidii and A. mediterraneum, respectively. This indicates that N. scintillans populations produced much brighter bioluminescence in Korean waters than the populations of P. kofoidii or A. mediterraneum.

• ALGAE
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• v.31 no.4
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• pp.391-401
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• 2016

To develop an easy and rapid method of quantifying lipid contents of marine dinoflagellates, we quantified lipid contents of common dinoflagellate species using a colorimetric method based on the sulpho-phospho-vanillin reaction. In this method, the optical density measured using a spectrophotometer was significantly positively correlated with the known lipid content of a standard oil (Canola oil). When using this method, the lipid content of each of the dinoflagellates Alexandrium minutum, Prorocentrum micans, P. minimum, and Lingulodinium polyedrum was also significantly positively correlated with the optical density and equivalent intensity of color. Thus, when comparing the color intensity or the optical density of a sample of a microalgal species with known color intensities or optical density, the lipid content of the target species could be rapidly quantified. Furthermore, the results of the sensitivity tests showed that only $1-3{\times}10^5cells$ of P. minimum and A. minutum, $10^4cells$ of P. micans, and $10^3cells$ of L. polyedrum (approximately 1-5 mL of dense cultures) were needed to determine the lipid content per cell. When the lipid content per cell of 9 dinoflagellates, a diatom, and a chlorophyte was analyzed using this method, the lipid content per cell of these microalgae, with the exception of the diatom, were significantly positively correlated with cell size, however, volume specific lipid content per cell was negatively correlated with cell size. Thus, this sulpho-phospho-vanillin method is an easy and rapid method of quantifying the lipid content of autotrophic, mixotrophic, and heterotrophic dinoflagellate species.