I want to preserve the fungal strains I have isolated for my work for further work.
If you are preserving sporulating cultures than lyophilization is good method.
But other methods like preserving under liquid nitrogen or under syerile water is less painful.
Please find the details of some the methods in
Deshmukh S.K. (2003). The Maintenance and Preservation of Keratinophilic Fungi and related Dermatophytes. Mycoses. 46 (5-6): 203-207.
The best method for preserving microorganisms is one which does not injure cells through freezing. The method which I routinely use if L-drying (liquid drying). This is where water is removed from cell suspensions under high vacuum without freezing the suspension. the methodology goes back to the 50's but this method has been validated many times are efficacious. I have cultures which are still viable upon revival after 35 years. Here is a reference to _drying:
Lapage, S.P.; Shelton, J.E.; Mitchell, T.G. and Mackenzie, A.R. (1970) Culture collections and the preservation of bacteria. In: “Methods in Microbiology Volume 3A”; J.R. Norris and D.W. Ribbons (eds.); Academic Press, London; pp. 135 - 228.
and here is another which reports the use of L-drying:
Tommerup, I.C. and Kidby, D.K. (1979) Preservation of spores of vesicular-arbuscular endophytes by L-drying. Applied and Environmental Microbiology 37: 831 - 835.
Clem
Dear mam, kindly sporulating the fungal in liquid culture broth and try the glycerol stock like storing of bacterial cell.80% glycerol and 20% desired broth.
You want to preserve the spores. Promote sporulation of your strain in whatever medium works best. Harvest the spores (trying to not take mycelium). Transfer spore suspension into a sterile "grainy" medium (Perlite, soil, you can even try filter paper discs, dry milk, etc) depending on how often you plan to use them I have had success with discs that do not even need to be freeze dried (dried in incubator inside Petri dish for continuous use, one disc into one shake flask) and soils that do get freeze dried. When using the freeze dryer just have a "control" of liquid that has 20% more volume than what your spore suspension had so that you can visually check that all the moisture has been removed. As long as you can keep the vials, tubes or whatever container moisture free (i.e. wrap in Parafilm) they will keep for many many years (20+ in my experience). Spores will withstand the freezer as long as they are dry during this time.
Here is a variety of methods for Neurospora
http://www.fgsc.net/neurosporaprotocols/HowtopreservestocksK%20final.pdf
Good luck, they will also keep at room temperature.
See this paper, by Bunse, T., and G. K. Steigleder. "The preservation of fungal cultures by lyophilization Die Konservierung von Pilzkulturcn mittels Gefriertrocknung." mycoses 3.l73: l76-l99l. to achieve your goals and ojectivities
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