Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Application of Species-specific DNA Probe to Field Samples of Alexandrium tamarense (Lebour) Balech

자연 시료로부터 Alexandrium tamarense을 위한 종 특이적 DNA탐침의 응용

  • Cho, Eun-Seob (Harmful Algal Research Division, National Fisheries Research and Development Institute) ;
  • Kim, Gi-Young (Department of Microbiology, Pusan National University) ;
  • Park, Hyung-Sik (Department of Microbiology, Pusan National University) ;
  • Kim, Hak-Gyoon (Harmful Algal Research Division, National Fisheries Research and Development Institute) ;
  • Moon, Sung-Ki (Department of Biology, College of Science, Kyungsung University) ;
  • Lee, Jae-Dong (Department of Microbiology, Pusan National University)
  • 조은섭 (국립수산진흥원 적조생물과) ;
  • 김기영 (부산대학교 미생물학과) ;
  • 박형식 (부산대학교 미생물학과) ;
  • 김학균 (국립수산진흥원 적조생물과) ;
  • 문성기 (경성대학교 생물학과) ;
  • 이재동 (부산대학교 미생물학과)
  • Published : 2002.06.01
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Abstract

Fluorescent species-specific DNA probe (AT1) of toxic dinoflagellate Arexandrium tamarense was tested on several other species, on comparison of binding activity at different preservatives for fixation of the cells, at different culture age and estimation of cell density by light microscope or epifluorescent microscope using whole cell hybridization. Th AT1 probe specifically bound to Alexandrium tamarense, whereas it did not bind to other phytoplankton, in particular Alexandrium catenella, morphologically similar to Alexandrium tamarense, could not react to AT1 probe. When cells were fixed with all three preservatives, labeling cells of Alexandrium tamarense emitted strong fluorescent signal intensity. In addition, regardless culture days, binding activity with AT1 probe was strong. The tell densities estimated by epifluorescent microscope were than those estimated by light microscope. The enumeration and identifying of Arexandriurn tamarense using DNA probe method will be contributed to a new biotoxin monitoring and prediction system in field.

독성이 있는 특이한 편모충인 Alexandrium tamarense의 구별하기 위한 종특이적인 형광 DNA탐침 AT1이 서로 다른 3가지의 고정액에서, 그리고 배양기간의 차이와 whole cell hybridization을 사용하는 광학 현미경 또는 형광 현미경에 의한 세포밀도 측정에 있어서 hiding activity를 비교하는 방법으로 여러 다른 종의 실험에 사용되어졌고, 그의 결과를 보고하는 바이다. 형광 DNA탐침 AT1은 특이적으로, A. tamarense에 결합했지만, 형태학적으로 유사한 다른 편모충에는 결합하지 않았다. 특히 형태적으로 A. tamarense에 유사한 A. catenella는 형광 DNN탐침 AT1에 결합하지 않았다. A. tamarense은 다른 세가지 고정액을 처리하였을 때, 고정액과 상관없이 강한 형광신호를 발산하였다. 추가해서 말하면, 배양기간에 관계없이 형광 DNA탐침 AT1의 biding activity는 강했다. 광학 현미경에 의해서 측정 되어지기 보다는 형광현미경에 의해서 세포밀도가 측정되었다. 형광 DNA탐침 AT1을 이용한 A. tamarense의 동정과 계산은 이 분야에서 새로운 생물독성 감시와 예측 시스템에 기여 할 것이다.

Keywords

References

  1. Adachi, M., Y. Sako and Y. Ishida. 1993. Application of monoclonal antibodies to field samples of Alexandrium species. Nippon Suisan Gakkaishi 59, 1171-1175. https://doi.org/10.2331/suisan.59.1171
  2. Adachi, M., Y. Sako and Y. Ishida. 1996. Identification of the toxic dinoflagellates Alexandrium catenella and A. tamarense (Dinophyceae) using DNA probes and whole-cell hybridization. J. Phycol. 32, 1049-1052 https://doi.org/10.1111/j.0022-3646.1996.01049.x
  3. Bates, S. S., C. Leger, B. A. Keafer and D. M. Anderson. 1993. Discrimination between domoic acidproducing and non-toxic forms of the diatom Pseudonitzschia pungens using immunofluorescence. Mar. Ecol. Prog. Ser. 100, 185-195 https://doi.org/10.3354/meps100185
  4. Cho, E. S., J. G. Park, H. G. Kim, C. H. Kim, L. L. Rhodes and C. S. Chung. 1999. The rapid differeniation of toxic Alexandrium and Pseudonitzschia species using fluorescent lectin probes. J. Kor. Ocean. Soc. 34, 167-171
  5. Cho, E. S., G. M. Seo, S. G. Lee, H. G. Kim, S. J. Lee,L. L. Rhodes and Y. K. Hong. 1998. Application of FITC-conjugated lectin probes for the recognition and differentiation of some Korean coastal red tide microalgae. J. Fish. Sci. Tech. 1, 250-254
  6. Cho, E. S., S. G. Lee, I. S. Kim and M. Chinain. 2001. Molecular phylogeny of the genus, Alexandrium (Dinophyceae), based on internal transcribed spacers ITS1 and ITS2, including 5.8S nuclear rDNA sequences. Algae. (accepted)
  7. Costas, E., E. G. Chavarri, A. Aguilera, S. G. Gil, and V. L. Rodas. 1993. Use of lectins to recognize and differentiate unicellular algae. Bot. Mar. 36, 1-4. https://doi.org/10.1515/botm.1993.36.1.1
  8. Fukuyo, Y. 1985. Morphology of Protogonyaulax tamarensis (Lebour) Taylor and Protogonyaulax catenella (Whedon and Kofoid) Taylor from Japanese coastal waters. Bull. Mar. Sci. 37, 529-537.
  9. Guillard, R. R. L. and J. H. Ryther. 1962. Studies of marine planktonic diatoms 1. Cyclotela nana Hustedt, and Detonula confervacea (Cleve) Gran. Can. J. Microbiol. 8, 229-239. https://doi.org/10.1139/m62-029
  10. Lee, S. G. 1991. The taxonomy and distribution of marine toxic flagellates in Chinhae Bay. Ph. D. thesis. University of Kyungsung, Pusan, 125 pp. (in Korean)
  11. Miller, P. and C.A. Scholin. 1998. Identification and enumeration of cultured and wild Pseudo-nitzschia (Bacillariophyceae) using species-specific LSU rRNAtargeted fluorescent probes and filter-based whole cell hybridization. J. Phycol. 34, 371-382. https://doi.org/10.1046/j.1529-8817.1998.340371.x
  12. Sako, Y., C. H. Kim, H. Ninomiya, M. Adachi and Y. Ishida. 1990. Isozyme and cross analysis of mating populations in the Alexandrium catenella/tamarense species complex. pp. 320-323. In Graneli, E., Sundstrom, B., Edler, L. and Anderson, D. M. (eds.) Toxic Marine Phytoplankton, Elsevier, New York.
  13. Scholin, C. A., K. R. Buck, T. Britschgi, G. Cangelosi and E. P. Chavez. 1996. Identification of Pseudonitzschia austalis (Bacillariophyceae) using rRNA-targeted probes in whole cell and sandwich hybridization formats. Phycologia 35, 190-197. https://doi.org/10.2216/i0031-8884-35-3-190.1
  14. Scholin, C. A., P. Miller, K. R. Buck, E. P. Chavez, P. Harris, P. Haydock, J. Howard and G. Cangelosi. 1997. Detection and quantification of Pseudo-nitzschia australis in cultured and natural populations using LSU rRNA-targeted probes. Lim. Oceano. 42, 1265-1272. https://doi.org/10.4319/lo.1997.42.5_part_2.1265
  15. Seaby, D.A. 1996. Differentiation of Trichoderma taxa associated with mushroom production. Plant Patho. 45, 905-912. https://doi.org/10.1111/j.1365-3059.1996.tb02901.x
  16. Vrieling, E. G., A. Draaijer, W. Vanzeijl, L. Peperzak, W. W. C. Gieskes and M. Veenhuis. 1993. The effect of labelling intensity, estimated by real-time confocal laser scanning microscopy, on flow cytometric appearance and identification of immunochemically labelled marine dinoflagellates. J. Phycol. 29, 180-188. https://doi.org/10.1111/j.0022-3646.1993.00180.x

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