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Strains, media, and growth conditions

HR Huomiao Ran
WY Wen-Bing Yin

Last updated date: May 10, 2022 Views: 616 Forks: 0

original An abbreviated version of this protocol was published in Science Advances in Apr, 2022
Fungal-fungal cocultivation leads to widespread secondary metabolite alteration requiring the partial loss-of-function VeA1 protein
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Co-cultivation of fungi in liquid media

 

Detailed protocol:

  1. Inoculate Aspergillus strains on GMM media with appropriate supplements corresponding to the auxotrophic markers, 37°C for 3 days.
  2. Inoculate Epicoccum dendrobii on PDA media, 28°C for 5 days.
  3. Collect Aspergillus conidia by scraping with 0.1% (v/v) Tween-80.
  4. Transfer E. dendrobii mycelia and Aspergillus conidia together into 250-ml flask containing 80 ml of PDB liquid medium. Incubated on a rotary shaker at 180 rpm and 28°C for 6 days.
  5. Extract with 80 ml of a solvent mixture (ethyl acetate:methanol:acetic acid, 89:10:1) for secondary metabolite analysis.


Media:

  1. GMM (glucose minimum medium): 1.0% (w/v) glucose, salt solution (50 ml/liter), trace element solution (1 ml/liter) and 1.6% (w/v) agar, autoclave.
  2. PDA medium: 39g Difco™ Potato Dextrose Agar to 1L of water, autoclave.
  3. PDB liquid medium: 24g Difco™ Potato Dextrose Broth to 1L of water, autoclave.


Solutions:

 

20 × Nitrate Salts (w/v)1 L
12% NaNO3 120 g
1.04% KCl10.4 g
1.04% MgSO4·7H2O10.4 g
3.04% KH2PO430.4 g
Add ddH2O to 1 L, autoclave, and stored at room temperature

 

1000 x Trace Elements (w/v)100 mL
5% Na4EDTA5 g
2.2% ZnSO4∙7H2O2.2 g
1.1% H3BO31.1 g
0.5% MnCl2∙4H2O0.5 g
0.16% FeSO4∙7H2O0.16 g
0.16% CoCl2∙5H2O0.16 g
0.16% CuSO4∙5H2O0.16 g
0.11% (NH4)6Mo7O24∙4H2O0.11 g
Adjust pH to 6.5 using KOH, add  ddH2O to 100 mL, autoclave, store at room temperature

 

Copyright: Content may be subjected to copyright.
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Ran, H and Yin, W(2022). Strains, media, and growth conditions. Bio-protocol Preprint. bio-protocol.org/prep1666.
  2. Wang, G., Ran, H., Fan, J., Keller, N. P., Liu, Z., Wu, F. and Yin, W.(2022). Fungal-fungal cocultivation leads to widespread secondary metabolite alteration requiring the partial loss-of-function VeA1 protein. Science Advances 8(17). DOI: 10.1126/sciadv.abo6094

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