• 한국양식학회:학술대회논문집
• /
• 한국양식학회 2003년도 추계학술발표대회 논문요약집
• /
• pp.134-135
• /
• 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.

• 한국해양환경ㆍ에너지학회지
• /
• 제15권2호
• /
• pp.133-141
• /
• 2012

한국 남해연안해역에서 분리한 유독와편모조류 $Alexandrium$ $tamarense$와 $Alexandrium$ $catenella$의 성장에 영향을 미치는 수온과 염분의 특성을 실내 배양실험을 통해 살펴보았다. 수온은 10, 15, 20, 25, $30^{\circ}C$의 5단계, 염분은 10, 15, 20, 25, 30, 35 psu의 6단계를 조합한 총 30단계의 조건에서 성장속도 변화를 관찰하였다. 최대성장속도를 보이는 수온과 염분은 $A.$ $tamarense$에서 $15^{\circ}C$와 30 psu, $A.$ $catenella$에서 $25^{\circ}C$와 30 psu로 나타났다. 또한 최적성장조건은 $A.$ $tamarense$에서 $10-20^{\circ}C$, 25-35 psu이고, $A.$ $catenella$에서 $20-30^{\circ}C$, 25-35 psu였다. 얻어진 수온과 염분의 조건을 이용하여 중회귀분석에 의해 예측 모델식을 계산한 결과, $A.$ $tamarense$는 ${\mu}=0.04+0.0193T-0.0339S-0.0005T^2+0.0021S^2+0.00073TS-0.000022T^3-0.000038S^3+0.00000086TS^2-0.0000255T^2S$이며, $A.$ $catenella$는 ${\mu}=1.01-0.1288T-0.0778S+0.0067T^2+0.0038S^2+0.00204TS-0.0001T^3-0.000059S^3-0.0000131TS^2-0.0000392T^2S$로 나타났으며, 예측값과 실험값 사이에는 높은 상관관계를 보였다. 본 연구결과는 수온과 염분의 현장관측 값을 이용하여 성장속도를 추정할 수 있기 때문에, $A.$ $tamarense$와 $A.$ $catenella$의 개체군 확대를 이해하는데 도움이 될것으로 기대된다.

• 한국양식학회:학술대회논문집
• /
• 한국양식학회 2003년도 추계학술발표대회 논문요약집
• /
• pp.128-128
• /
• 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.

• 생명과학회지
• /
• 제12권6호
• /
• pp.821-834
• /
• 2002

알렉산드륨 적조생물의 리보소옴 알엔에이 유전자의 ITS1, 2 및 5.8S부위를 대상으로 종간 혹은 종내의 유전적 다양도를 조사하기 위하여 지리적으로 격리된 33 스트레인 유전자의 염기서열를 비교했다. 진해만에서 분리된 AT-2, AT-6, AT-10, AT-A, AT-B 5클론은 일본종 OFX151-A과 동일한 유전자임을 발견했다. ITS부위에서 가장 짧은 종은 A. margalefi로 481 bp이며 가장 긴 종은 A. affine으로 528 bp로 나타났다. ITS1과 ITS2 염기서열에 대한 상호관계는 역으로 나타낸 반면에, G＋C 함량에 대한 상호관계는 플러스로 나타났다. 유전적 변이율은 0.3% (1 bp)에서 53% (305 bp)였다. A. tamarense과 가장 적게 유전적 변이율을 보인 종은 A. fundyense(1.2-2.3% ＝ 6-12 bp)인 반면에, A. catenella와는 큰 변이율 (19.8% ＝ 102 bp)을 보였고, A. catenella와 A. fundyense은 19.7% 상이하였다. 알렉산드륨 적조생물의 bootstrap은 약하게 지지되는 데도 불구하고, A. catenella 분리종은 독립적인 그룹으로 형성하여 상호간에는 강력한 bootstrap 값은 PAUP과 NJ 분석에서 보였다. A. cohorticula와 A. frateculus 적조생물은 항상 sub-group 내에서 높은 bootstrap을 가졌다. 결론적으로 ITS부위의 염기서열 분석은 알렉산드륨 적조생물의 집단내 혹은 집단간의 계통분류을 밝히는데 유용한 것으로 보였다.

• 해양환경안전학회지
• /
• 제26권1호
• /
• pp.84-92
• /
• 2020

본 연구는 마비성 독소 표준물질의 안정적인 공급을 위해 친환경적이고 경제성이 우수한 발광다이오드 광원의 파장을 이용하여, PST의 원인종 Alexandrium catenella와 A. pacificum의 독 함량을 증대시킬 수 있는 가능성을 살펴보았다. 파장별 광량과 생장속도의 관계를 보면, 반포화상수(Ks)는 두 종 모두 적색파장에서 가장 높게 나타났고, 청색파장에서 가장 낮았다. 빛 파장에 따른 독함량의 경우, 적색 파장이 A. catenalla은 최고 약 8배 그리고 A. pacificum은 약 3.4배 다른 파장보다 높았다. 그리고 광량에 대한 친화성을 나타내는 Ks와 독농도와 밀접한 관련이 있는 것으로 나타났다. 따라서 세포당 독농도 증가를 위한 배양법은 초기에 세포수량의 확보 및 빠른 생장속도를 유도하기 위해서 청색파장을, 대수생장기 후기에는 적색파장을 주사하여, 세포내 독함유량을 증가시킬 수 있는 2단계 배양법을 제시할 수 있을 것으로 생각된다.

• 한국양식학회:학술대회논문집
• /
• 한국양식학회 2003년도 추계학술발표대회 논문요약집
• /
• pp.136-137
• /
• 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.

• 한국어업기술학회:학술대회논문집
• /
• 한국어업기술학회 2001년도 춘계 수산관련학회 공동학술대회발표요지집
• /
• pp.252-254
• /
• 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
• /
• 제34권3호
• /
• pp.167-171
• /
• 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
• /
• 제39권3호
• /
• pp.172-185
• /
• 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.

• 대한환경공학회지
• /
• 제32권3호
• /
• pp.248-255
• /
• 2010

본 연구는 연안해역에서 발생하는 적조의 현장 제어기술로 전자빔을 이용하고자 적조사멸에 대한 전자빔 조사의 효율성을 평가하였다. Prorocentrum minimum, Prorocentrum micans, Cochlodinium polykrikoides, Heterosigma akashiwo, Alexnadrium catenella의 5종을 실험대상 적조로 선정하여 배양하였으며, 실제 경남 통영(2007. 8. 15)에서 발생한 적조에 대해서도 동일한 실험을 수행하였다. 전자빔 조사선량은 1 kGy, 2 kGy, 4 kGy, 8 kGy로 1 kGy의 전자빔 조사로 50~65%의 적조사멸율을 보였으며, 1일 경과 시 86~97% 사멸하였다. Chlorophyll-a의 경우 조사 즉시 50~64% 감소하고, 1일 경과한 뒤 72~93%의 감소율을 보였다. 또한 전자빔 조사 즉시 S-protein은 28~47%, S-carbohydrate는 16~45% 가 증가하였으며, 1일의 시간 경과 후에는 S-protein이 77~138%, S-carbohydrate가 77~212%까지 증가하였다. 또한 투과 전자현미경을 통해 전자빔 조사 전후의 적조세포 관측을 통하여 세포의 파괴 및 세포내 구성물질의 용출을 확인할 수 있었다. 이렇듯 적조에 전자빔을 조사함으로써 Extracellular Biopolymer에 의한 Flocculation을 유도하여 적조를 침강 제어할 수 있다. 또한 전자빔 조사 전 후 Alexandrium catenella의 PSP(Paralytic Shellfish Poisoning) 패독 분석 결과 1 kGy의 전자빔을 조사함으로써 전자빔을 조사하지 않은 적조 대비 PSP 독성 함량이 48% 감소하여 전자빔조사가 유독적조의 사멸 뿐만 아니라 제독에도 효과가 있음을 알 수 있었다.