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Development of primer sets for multiplex and qPCR assays targeting Skeletonema species and their application to field samples

  • Natsumi Enjoji
  • Toshiya KatanoEmail author
  • Yuki Yoshinaka
  • Fuka Furuoka
  • Yutaro Ando
  • Machiko Yamada
  • Tomomi Hamasaki
  • Emika Miyamura
  • Mayuko Otsubo
  • Katsuhide Yokoyama
Original Article
  • 54 Downloads

Abstract

Skeletonema is a dominant diatom, especially in coastal waters. However, species identification and quantification in the natural environment has not yet been established, as species identification can be nearly impossible with light microscopy. In the present study, we developed primer sets for multiplex and real-time PCR to identify and enumerate the 10 Skeletonema species, and applied this technique to samples obtained from the tidal zone of the Chikugo River and its adjacent waters of the Ariake Sea from spring to summer. In total, eight Skeletonema species were detected during the investigation. Among the eight, the S. marinoidohrnii complex was detected at all three stations. Skeletonema potamos was the second most frequently detected, mainly in the Chikugo River. The overall number of detected species was low at the upper tidal zone of the Chikugo River, where salinity was < 2. These results reveal the diverse Skeletonema community in the brackish environment, and the primer sets designed in the present study are useful for analyzing species composition of Skeletonema.

Keywords

Skeletonema Identification Multiplex PCR Real-time PCR 

Notes

Acknowledgements

This study was supported by Grants-in-Aid for Scientific Research (25450259, 16K07508) from JSPS, Kasen-kikin (Nos. 261211018, 285211023) from Kasen Zaidan, and the Cooperative Monitoring Program of the Ariake Sea (COMPAS) of Saga University.

Supplementary material

10872_2018_504_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (docx 15 kb)
10872_2018_504_MOESM2_ESM.docx (20 kb)
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Copyright information

© The Oceanographic Society of Japan and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Natsumi Enjoji
    • 1
    • 2
  • Toshiya Katano
    • 1
    Email author
  • Yuki Yoshinaka
    • 1
  • Fuka Furuoka
    • 1
  • Yutaro Ando
    • 1
  • Machiko Yamada
    • 2
  • Tomomi Hamasaki
    • 2
  • Emika Miyamura
    • 2
  • Mayuko Otsubo
    • 2
  • Katsuhide Yokoyama
    • 3
  1. 1.Graduate School of Marine Science and TechnologyTokyo University of Marine Science and TechnologyTokyoJapan
  2. 2.Fukuoka Women’s UniversityFukuokaJapan
  3. 3.Graduate School of Urban Environmental SciencesTokyo Metropolitan UniversityTokyoJapan

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