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Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions
Published on: August 15, 2019
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Cultivation of ammonia-oxidising archaea on solid medium
Timothy Klein1, Lianna Poghosyan1, J Elaine Barclay2
1School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, UK.
FEMS Microbiology Letters
|March 24, 2022
Summary
Growing ammonia-oxidising archaea (AOA) on solid media is challenging. A new Liquid-Solid (LS) method using Phytagel™ enabled colony formation for two AOA strains, paving the way for genetic studies.
Area of Science:
- Microbiology
- Environmental Science
- Biogeochemistry
Background:
- Ammonia-oxidising archaea (AOA) are crucial for nitrogen cycling.
- Current cultivation methods for AOA are limited to liquid cultures.
- Solid medium cultivation is needed for AOA purification and genetic manipulation.
Purpose of the Study:
- To develop a reliable method for growing AOA colonies on solid medium.
- To assess the efficacy of different gelling agents for AOA growth.
- To establish a foundation for genetic studies of AOA.
Main Methods:
- Tested three distinct AOA strains: 'Ca. N. franklandus C13', N. viennensis EN76, and 'Ca. N. sinensis Nd2'.
- Evaluated agar, Bacto-agar, and Phytagel™ as gelling agents.
- Developed and applied a Liquid-Solid (LS) method using Phytagel™ with liquid overlay.
Main Results:
- Agar and Bacto-agar inhibited growth of all tested AOA strains.
- 'Ca. N. franklandus C13' and N. viennensis EN76 tolerated Phytagel™.
- 'Ca. N. sinensis Nd2' was inhibited by Phytagel™.
- The LS method successfully produced visible colonies for 'Ca. N. franklandus C13' and N. viennensis EN76.
Conclusions:
- Phytagel™ is a suitable gelling agent for specific AOA strains.
- The developed LS method facilitates AOA cultivation on solid medium.
- This method supports future genetic manipulation of AOA.
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