I store mycelium on sterile filter papers in the freezer. When I grow culture I put piece of sterile filter paper on the media in petri dish and when fungus spreads on the surface of the filter paper I take the paper out, put it in sterile coin envelope, let it air dry in a laminar hood for 12 hours and store in the freezer. This technique works pretty well in our lab, we have cultures 15 years old stored this way which still easy to regenerate just putting the piece of filter paper with dried frozen mycelium back on the media.

Dear Chao,

One way to store stock cultures of microbes is in cryogenic vials containing ~20-40% glycerol. For bacteria and yeast, the cells can be grown to a high cell density, then small volumes (~0.5-1.0 mL) added to cryogenic vials containing the same volume of sterile 40% glycerol. After gentle mixing, the vials can be frozen. A temperature of -80C is best; however, you can use -20C if you don't have an ultra-low freezer. Cells stored this way can be preserved for years.

With filamentous fungi, it may be a little more tricky to transfer the cells to the vials, since you won't have a uniform cell suspension. However, it can be done and it should work.

I hope this helps you.

Bill Colonna, Center for Crops Utilization Research, Dept. Food Science & Human Nutrition, Iowa State University, Ames, IA   USA   [email protected]

Hello Chao, 

I use cryotubes with glycerol 80%. Approximately, in cryotubes of 2 ml, with 600-800 microlitres of glycerol and them I put 4-5 portions of mycelia cut in square or circle immersed inside, stored at -80 ºC. When I need recuperate one of this I just have to take one of this little portion and culture in a rich medium like MEA or PDA. Therefore, you can use the cryotube more times. For short-term I do the same but I used 2 ml tubes (like eppendorf), distilled water instead of glycerol and stored at 4 ºC.

I hope that it be useful.

Hi: Also you can use silica gel method to preserve fungal for long time 7-10 years, its very simple and easy .

Hi Chao

Its advisable to store your fungal spores in 40-50% glycerol.

Dear Chao,

As you have mentioned that you are using only refrigerated temperature for storage it appears that you don't want to go for lyophilisation or deep freeze preservation techniques.

You may prepare a spore culture of a desired fungus and suspend the same in 50% aqueous glycerol to be stored in a test tube at refrigerated temperature. Before transferring the culture to a refrigerator add about 3-4 cm high column of liquid paraffine over the spore suspension in the test tube. Addition of liquid paraffin would further help to slow down the biological activity of the refrigerated culture; thereby extending the shelf life significantly.

Dear Chao,

As you have mentioned that you are using only refrigerated temperature for storage it appears that you don't want to go for lyophilisation or deep freeze preservation techniques.

You may prepare a spore culture of a desired fungus and suspend the same in 50% aqueous glycerol to be stored in a test tube at refrigerated temperature. Before transferring the culture to a refrigerator add about 3-4 cm high column of liquid paraffine over the spore suspension in the test tube. Addition of liquid paraffin would further help to slow down the biological activity of the refrigerated culture; thereby extending the shelf life significantly.

Dear Chao,

As you have mentioned that you are using only refrigerated temperature for storage it appears that you don't want to go for lyophilisation or deep freeze preservation techniques.

You may prepare a spore culture of a desired fungus and suspend the same in 50% aqueous glycerol to be stored in a test tube at refrigerated temperature. Before transferring the culture to a refrigerator add about 3-4 cm high column of liquid paraffine over the spore suspension in the test tube. Addition of liquid paraffin would further help to slow down the biological activity of the refrigerated culture; thereby extending the shelf life significantly.

Dear colleague, 

One good option is to grow the fungus on a poor medium for one or two weeks, to cut small fragments of the colony and put them inside cryotubes or hermetically closed glass tubes with sterilized water. Poor media enhance the production of resistance structures such as chlamydospores or (micro)sclerotia. 

I have preserved dematiaceous fungi and dermatophytes using this method for many years and it works very well. 

Success!

Hugo

From our experience we preserve yeast and filamentous fungi using cryotubes with beads from Mast diagnistics Company, UK. these are propably with 5-10% glycerol, and organisms stay viable over 12 years at -80C. Also some laboratories preserve filamentous fungi in sterilised water in a test tubes at room temperature.

I wish you all the best in your research

Saad J. Taj-Aldeen

I store mycelium on sterile filter papers in the freezer. When I grow culture I put piece of sterile filter paper on the media in petri dish and when fungus spreads on the surface of the filter paper I take the paper out, put it in sterile coin envelope, let it air dry in a laminar hood for 12 hours and store in the freezer. This technique works pretty well in our lab, we have cultures 15 years old stored this way which still easy to regenerate just putting the piece of filter paper with dried frozen mycelium back on the media.

The method suggested by Olga Kozhar appears to be the best.

In water at room temperature

From my experience, you could also try:

- keeping the fungal strains on growth medium slants covered by glycerol in a fridge,

- freeze-drying (echnically known as lyophilisation) of spores/hyphae, however this method requires a deep-freezing freezer in the lab,

- sealing the growth medium plates with parafilm (this way the medium does not dry out, and exogenic fungi/bacteria do not infect our colonies.

I also found this tiny article, it might be helpful (it's in the link).

https://blog.mycology.cornell.edu/2008/01/10/a-simple-way-to-preserve-fungal-cultures/

Dear Chao Yuan

you could also try for this:

1- Transferring mycelial plugs to the sterile water and then stored in the refrigerator

2-freeze-drying of spores/hyphae- sealing the growth medium plates with parafilm

3- Grow fungal isolates on the filter paper and then stored in the freezer (-15 to-20 ° C)

4-Freezing fungal spores (spore suspension) in to Water/Glycerol (20:80 vol.).

5- For Fusarium spp. you can use sterilized sand and kept in the refrigerator

One can add a plug of fungal mycelium in

1. double sterile distilled water

2. Milli Q water

3. Skimmed milk.

for long time preservation.

What is expected shelflife at ambient and refrigerated temperature?

In Copenhagen sol: ethanol 52 ml

Glycerol 6 ml and d.w. 42 ml