In situ identification of symbiotic dinoflagellates, the genus Symbiodinium with fluorescence-labeled rRNA-targeted oligonucleotide probes

Hiroko Yokouchi, Haruko Takeyama, Hideaki Miyashita, Tadashi Maruyama, Tadashi Matsunaga*

*この研究の対応する著者

研究成果: Article査読

18 被引用数 (Scopus)

抄録

Fluorescence in situ hybridization has been used for the identification and analysis of populations of the dinoflagellate Symbiodinium that lives symbiotically in marine invertebrates. Conditions for in situ hybridization of Symbiodinium were optimized and used to identify the clade to which the isolate belongs using specific probes. The optimized in situ hybridization procedure used a combination of chlorophyll removal and permeabilization with hot ethanol. Incubation of the cells in 50% ethanol at 80°C for 20 min rendered the cell wall permeable to Cy3-labeled probes. Symbiodinium clade-specific probes were designed based on 18S rRNA sequences. Symbiodinium A, B and C were distinguished by in situ hybridization with the specific probes SymA, SymB and SymC, respectively. The hybridization results using clade-specific probes corresponded with results obtained using restriction fragment length polymorphism (RFLP) analysis. Symbiodinium isolated from jellyfish Cassiopea sp. and sea anemone Aiptasia sp. were classified as belonging to clades A and B using the FISH procedure established in this study.

本文言語English
ページ(範囲)327-334
ページ数8
ジャーナルJournal of Microbiological Methods
53
3
DOI
出版ステータスPublished - 2003 6月 1
外部発表はい

ASJC Scopus subject areas

  • 微生物学
  • 分子生物学
  • 微生物学(医療)

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