Application of real-time PCR assay for detection and quantification of bloom-forming diatom Chaetoceros tenuissimus Meunier

Kensuke Toyoda, Keizo Nagasaki, Yuji Tomaru

研究成果: Article査読

9 被引用数 (Scopus)

抄録

Viral infection of the bloom-forming diatom Chaetoceros tenuissimus was discovered in 2008, and is now assumed to have a significant influence on the dynamics of C. tenuissimus populations in natural environments. Enumeration of C. tenuissimus and its viruses is essential when examining the host-virus relationship in situ; however, the diatom species is so small in size that its identification and counting by optical microscope is almost impossible. To resolve this problem, we have developed a TaqMan-based real-time polymerase chain reaction (PCR) method for detection and quantification of C. tenuissimus. We designed primers and a TaqMan probe to target the D1 region of its 28S ribosomal RNA (rRNA) gene; the established real-time PCR was specific at the species level by testing 41 microalgal strains including C. tenuissimus. Tris-EDTA buffer-based boiling method was shown to be efficient for extracting DNA from filter-trapped C. tenuissimus cells in this study. The detection range of the established TaqMan-based real-time PCR method for C. tenuissimus was 101 to 106 cells collected on a filter; the method was applicable for C. tenuissimus cells accompanied with natural microorganisms.

本文言語English
ページ(範囲)56-61
ページ数6
ジャーナルPlankton and Benthos Research
5
2
DOI
出版ステータスPublished - 2010

ASJC Scopus subject areas

  • 海洋学
  • 生態、進化、行動および分類学
  • 水圏科学
  • 生態学

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