ALGAE

The Korean Society of Phycology (한국조류학회(藻類))
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An advanced tool, droplet digital PCR (ddPCR), for absolute quantification of the red-tide dinoflagellate, Cochlodinium polykrikoides Margalef (Dinophyceae)

  • Lee, Hyun-Gwan (Department of Oceanography, Chonnam National University) ;
  • Kim, Hye Mi (Department of Oceanography, Chonnam National University) ;
  • Min, Juhee (Department of Oceanography, Chonnam National University) ;
  • Kim, Keunyong (Department of Oceanography, Chonnam National University) ;
  • Park, Myung Gil (Department of Oceanography, Chonnam National University) ;
  • Jeong, Hae Jin (School of Earth and Environmental Sciences, College of Natural Sciences, Seoul National University) ;
  • Kim, Kwang Young (Department of Oceanography, Chonnam National University)
  • Received : 2017.04.03
  • Accepted : 2017.09.10
  • Published : 2017.09.30
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Abstract

To quantify the abundance of the harmful dinoflagellate Cochlodinium polykrikoides in natural seawaters, we developed the innovative procedure using a droplet digital PCR (ddPCR) with C. polykrikoides-specific primers targeting the internal transcription sequence (ITS). The abundance of C. polykrikoides was estimated by the specific copy number of target ITS DNA segments per cell in cultures and natural water samples. The copy number per C. polykrikoides cell as acquired by ddPCR was $157{\pm}16$, which was evaluated against known cell numbers through a simplified protocol preparing DNAs. The abundances of C. polykrikoides in the waters of different locations estimated by ddPCR agreed with the number of cells visually counted under a microscope. This protocol was used to measure the abundance of C. polykrikoides close to and further off the southern coast of Korea in August of 2016 and 2017. The practical application showed that this method can reduce time for analysis and increase accuracy.

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References

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