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Development of genetic diagnosis method of toxic microalgae

Research Project

Project/Area Number 13556033
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Fisheries chemistry
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SAKO Yoshihiko  Kyoto University, Applied Biosciences, Professor, 農学研究科, 教授 (60153970)

Co-Investigator(Kenkyū-buntansha) CHRISTOPHER Scholin  米国カリフォルニア州モントレー湾, 水族館研究所, グループリーダー
KOTANI Yuichi  National Res.Inst.Fish.Sci.Yokohama Main St., Group leader, 企画調査室, 課長
UCHIDA Aritsune  Kyoto University, Applied Biosciences, Professor, 農学研究科, 教授 (50027190)
SCHOLIN Christopher  Monterey Bay Aquarium Res.Inst., Group leader
小谷 祐一  瀬戸内海区水産研究所, 赤潮環境部, 室長
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2002: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥8,000,000 (Direct Cost: ¥8,000,000)
Keywordsparalytic shellfish poison / dinoflagellates / Alexandrium / molecular phylogenetic analysis / rRNA gene / bivale poisoning / poison / molecular identification / アレキサンドリウム属
Research Abstract

Recent paralytic shellfish poisoning caused by toxic dinoflagellates which have very similar morphological features are serious problems in aquatic farms and public health. The purpose of this study are the establishment of molecular identification of toxic dinoflagellates Alexandrium spp. and development of genetic diagnosis method of toxic dinoflagellates
(1) Phylogenetic analysis based on 18S rDNA sequences and the D1/D2 region in 28S rDNA were examined among the many isolates of toxic A.tamarense, A.catenella, A.tamiyavanichii, A.ostenferdii and nontoxic A.affine, A.fraterculus, A.insuetum, A.psedogonyaulax.
(2) A rapid and precise identification method was established using a fluorescent, rRNA-targeted, oligonucleotide probes for toxic A.tamiyavanichii which are recently observed in Harima-Nada and the nontoxic A.affine and A.frateruculus. Each probe was species specific when applied using fluorescence in situ hybridization (FISH).
(3) Real-time PCR assay was designed, evaluated and established for rapid detection and quantification of the vegetative cells and cysts (hypnozygotes) of toxic dinoflagellates A.tamarense and A.catenella.
(4) The screening methods of the specific genes expressed in differential stages, such as cyst formation and germination of A.tamarense and A.catenella, were developed by use of differential display and subtractive hybridization methods. As a result, some genes specific for the sexual fusion and germination stages were detected.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] T.Yoshida et al.: "Geographic differences in paralytic shellfish poisoning toxin profiles among Japanese populations of Alexandrium tamarense and A.catrenella (Dinophyceae)"Phycological Research. 49. 13-21 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Sako et al.: "Purification and characterization of a sulfotransferase specific to N-21 of saxitoxin and gonyautoxin 2+3 from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae)"J.Phycol.. 37・6. 1044-1051 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Yoshida et al.: "Purification and characterization of sulfotransferase specific to O-22 of 11-α,β-hydroxy STX from the toxic dinoflaggelate Gymnodinium catenatum (Dinophyceae)"68. 634-642 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Tanabe et al.: "Simple and rapid detection of the toxic marine dinoflagellates Alexandrium tamarense and A.catenella with fluorescence in situ hybridization (FISH) using rRNA-targeted probes"Fisheries Sciences. 68 Supl I. 515-518 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Sako, S.Hosoi-Tanabe, A.Uchida: "Fluorescence in situ hybridization using rRNA-targeted probes for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella"J.Phycol.. 40(In press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hosoi-Tanabe, Y.Sako: "Rapid detection of natural cells of Alexandrium tamarense and A.catenella (Dinophyceae) by fluorescence in situ hybridization"Harmful Algae. 3(In press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Sako: "Molecular identification of harmful microalgae in "Red Tide""TERA Pub. & Kluwer. 439 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshida, T., Sako, Y., Uchida, A.: "Geographic differences in paralytic shellfish poisoning toxin profiles among Japanese populations of Alexandrium tamarense and A.catenella (Dinophyceae)"Phycol.Res.. 49. 13-21 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sako, Y., Yoshida, T., Uchida A., Arakawa, O., T.Noguchi, Y.Ishida: "Purification and characterization of a sulfotransferase specific to N-21 of saxitoxin and gonyautoxin 2+3 from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae)"J.Phycol.. 37. 1044-10551 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshida, T., Sako, Y., Uchida A., Kakutani, T., Arakawa O., Noguchi, T., Ishida, Y: "Purification and characterization of sulfo-transferase specific to O-22 of 11-hydroxy saxitoxin from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae)"Fish.Sci.. 68. 634-642 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tanabe, S., Sako, Y., Baba, T., Uchida, A.: "Simple and rapid detection of the toxic marine dinoflagellates Alexandrium tamarense and A.catenella with fluorescence in situ hybridization (FISH) using rRNA-targeted probes."Fish.Sci.Sup.II. 68. 515-518 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sako, Y.: "Molecular identification of harmful microalgae"Red Tide (Ed.Okaichi, T.) (TERA Pub.& Kluwer). 84-94 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sako, Y., S.Hosoi-Tanabe, A.Uchida: "Fluorescence in situ hybridization using rRNA-targeted probes for simple and rapid identification of the toxic dinoflagellate Alexandrium tamarense and Alexandrium catenella"J.Phycol.. 40(in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hosoi-Tanabe, Y.Sako: "Rapid detection of natural cells of Alexandrium tamarense and A.catenella (Dinophyceae) by fluorescence in situ hybridization"Harmful Algae. 3(in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Sako et al.: "Fluorescence in situ hybridization using rRNA-targeted probes for simple and rapid identification of the toxic dinoflagellates"Journal of Phycology. 40(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] C.-J.Kim et al.: "PSP toxin analysis of the genus Alexandrium (Dinophyceae) occurring in Korean coastal waters"Fisheries Science. 70(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y.Sako, T.Okaichi ed.: "Red Tides"Terra Scientific Pub. Com. and Kluwer Academic Pub.. 439 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Toshida et al.: "Purification and characterization of sulfotransferase specific to O-22 of 11-hydroxy saxitoxin from the toxic dinoflagellate Gymnodinium catenatum (dinophyceae)"Fisheries Science. 68・3. 634-642 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Tanabe et al.: "Simple and rapid detection of the toxic marine dinoflagellates Alexandrium tamarense and A.catonella with fluorescence in situ hybridization (FISH) using rRNA-targeted probes"Fisheries Science. 68 Supl I. 515-518 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 田辺(細井)祥子, 左子芳彦: "日本藻類学会創立50周年記念出版 21世紀初頭の藻学の現況"日本藻類学会. 153 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Yoshida et al.: "Geographic differences in paralytic shellfish poisoning toxin profiles among Japanese populations of Alexandrium tamarense and A.catenella (Dinophyceae)"Phycological Research. 49. 13-21 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Sako et al.: "Purification and characterization of a sulfotransferase specific to N-21 of saxitoxin and gonyautoxin 2+3 from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae)"J. Phycol.. 37・6. 1044-1051 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Yoshida et al.: "Purification and characterization of sulfotransferase specific to O-22 of 11-α, β-hydroxy STX from the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae)"Fish.Sci.. 68(in press). (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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