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Cho DH, Ramanan R, Kim BH, et al. Novel approach for the development of axenic microalgal cultures from environmental samples. J Phycol. 2013;49(4):802-10doi: 10.1111/jpy.12091.
Cho, D. H., Ramanan, R., Kim, B. H., Lee, J., Kim, S., Yoo, C., Choi, G. G., Oh, H. M., & Kim, H. S. (2013). Novel approach for the development of axenic microalgal cultures from environmental samples. Journal of phycology, 49(4), 802-10. https://doi.org/10.1111/jpy.12091
Cho, Dae-Hyun, et al. "Novel approach for the development of axenic microalgal cultures from environmental samples." Journal of phycology vol. 49,4 (2013): 802-10. doi: https://doi.org/10.1111/jpy.12091
Cho DH, Ramanan R, Kim BH, Lee J, Kim S, Yoo C, Choi GG, Oh HM, Kim HS. Novel approach for the development of axenic microalgal cultures from environmental samples. J Phycol. 2013 Aug;49(4):802-10. doi: 10.1111/jpy.12091. Epub 2013 Jul 09. PMID: 27007211.
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J Phycol. 2013 Aug;49(4):802-10. doi: 10.1111/jpy.12091. Epub 2013 Jul 09.

Novel approach for the development of axenic microalgal cultures from environmental samples.

Journal of phycology

Dae-Hyun Cho, Rishiram Ramanan, Byung-Hyuk Kim, Jimin Lee, Sora Kim, Chan Yoo, Gang-Guk Choi, Hee-Mock Oh, Hee-Sik Kim

Affiliations

  1. Environmental Biotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806, Korea.
  2. Green Chemistry and Environmental Biotechnology, University of Science and Technology, 113 Gwahang-ro, Yuseong-gu, Daejeon, 305-333, Korea.
  3. Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 305-701, Korea.

PMID: 27007211 DOI: 10.1111/jpy.12091

Abstract

We demonstrated a comprehensive approach for development of axenic cultures of microalgae from environmental samples. A combination of ultrasonication, fluorescence-activated cell sorting (FACS), and micropicking was used to isolate axenic cultures of Chlorella vulgaris Beyerinck (Beijerinck) and Chlorella sorokiniana Shihira & R.W. Krauss from swine wastewater, and Scenedesmus sp. YC001 from an open pond. Ultrasonication dispersed microorganisms attached to microalgae and reduced the bacterial population by 70%, and when followed by cell sorting yielded 99.5% pure microalgal strains. The strains were rendered axenic by the novel method of micropicking and were tested for purity in both solid and liquid media under different trophic states. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene confirmed the absence of unculturable bacteria, whereas fluorescence microscopy and scanning electron microscopy (SEM) further confirmed the axenicity. This is the most comprehensive approach developed to date for obtaining axenic microalgal strains without the use of antibiotics and repetitive subculturing.

© 2013 Phycological Society of America.

Keywords: Axenic culture; Chlorella sp.; FACS; Microalgae; Micropicking; Scenedesmus sp.; Ultrasonication

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