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Trebouxia Medium



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Trebouxia Medium

Modification of Ahmadjian's medium for the phycobionts of lichens. Suitable for most axenic cultures of Trebouxia, Pseudotrebouxia, Trentepohlia and Cephaleuros.

Directions

For 1 L Total
  1. Prepare 1 L of Bristol Medium. Transfer approximately 825 mL to a 2-L Erlenmeyer flask. Save the remaining Solution.
  2. To approximately 825 mL of Bristol Medium, add each of the components in the order specified while stirring continuously.
  3. Bring total volume to 1 L with the Bristol Medium saved from step 1.
    • For 1.5% (w/v) agar medium: add 15 g of agar into the flask; do not mix.
  4. Cover and autoclave medium.
  5. Store at refrigerator temperature.

# Component Amount Stock Solution Concentration Final Concentration
1 Bristol Medium 825 mL/L
2 Soilwater: GR+ Medium 140 mL/L
3 Proteose Peptone
(BD 211684)
10 g/L
4 Glucose
(BACTO-dextrose or Sigma D 9634)
20 g/L 0.11 M

Important Notes To Consider:

  • The quality of water, including natural seawater, is important. The term 'dH2O' generally refers to distilled, deionized, distilled/deionized water, Milli-Q water (Millipore Corp.), etc. Natural seawater (and natural freshwater) should be obtained from a non-polluted source.
  • When dissolving chemicals, wait for the first component to dissolve before adding the second. Stirring, and sometimes the addition of heat, is often required to dissolve the chemicals efficiently. Preparation of stock solutions for frequently made algal culture media make preparing media convenient but also necessary to avoid errors from weighing very tiny amounts.
  • Attention should be given to the pH of the final medium. In most cases, if pH adjustment is required, this occurs before sterilization. Please note that autoclaving removes carbon dioxide from media lacking carbonate stabilizing buffers. This would make the medium very alkaline soon after removal from the autoclave. In these cases, you should wait approximately 24 hours for gaseous equilibrium before inoculating the medium.