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Aquaculture International

, Volume 19, Issue 4, pp 693–704 | Cite as

Development of primers and a procedure for specific identification of the diatom Thalassiosira weissflogii

  • Saengchan Senapin
  • Kornsunee Phiwsaiya
  • Pauline Kiatmetha
  • Boonsirm Withyachumnarnkul
Article

Abstract

A recent study showed Thalassiosira weissflogii to be a diatom containing suitable nutrition for larviculture of the black tiger shrimp, Penaeus monodon. Accurate and practical identification of this diatom species is therefore important for commercial hatcheries. The purpose of this study was to establish a DNA-based method of identification to supplement morphological examination, avoiding confusion with other Thalassiosira sp. Primers, 18SF/28SR1, specific for ribosomal DNA genes (3′-end of 18S rDNA through 5′-end of 28S rDNA, covering two internal transcribed spacers), were employed as a first-step polymerase chain reaction, followed by a second nested amplification using specifically designed primers, ITS1-F-D/ITS1-R-D. The nested-PCR result revealed specificity in the detection, distinguishing T. weissflogii from T. pseudonana, Cyclotella meneghiniana, and Chaetoceros sp., and the PCR fragment of the amplified region had a sequence that was 99% identical to the T. weissflogii sequence held by GenBank.

Keywords

Diatom ITS Larviculture PCR rDNA Shrimp Thalassiosira 

Notes

Acknowledgments

This study was funded by the Thailand Research Fund, grant No. DBG4980008.

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Saengchan Senapin
    • 1
    • 2
  • Kornsunee Phiwsaiya
    • 1
    • 2
  • Pauline Kiatmetha
    • 3
    • 4
  • Boonsirm Withyachumnarnkul
    • 1
    • 4
    • 5
  1. 1.Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp)BangkokThailand
  2. 2.National Center for Genetic Engineering and Biotechnology, National Science and Technology Development AgencyPathumthaniThailand
  3. 3.Department of Biochemistry, Faculty of SciencePrince of Songkla UniversitySongkhlaThailand
  4. 4.Shrimp Genetic Improvement CenterSurat ThaniThailand
  5. 5.Department of Anatomy, Faculty of ScienceMahidol UniversityBangkokThailand

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