Edible fungus mycelium grows rapidly, is prone to mutation, and is challenging to maintain over long periods. To help preserve fungal strains effectively, here are four practical and easy-to-implement methods that have proven successful in laboratory and small-scale settings.
1. **Liquid Paraffin Preservation Method**
This method involves using sterile liquid paraffin to create a protective layer around the mycelium. First, sterilize the paraffin by autoclaving or placing it in a desiccator for several days. Then, using a sterile pipette or syringe, carefully inject about 1 cm of paraffin into each test tube containing the fungal strain, ensuring it covers the top of the culture. Seal the tube with a cotton plug and wax. Store the tubes upright in a cool, dark place. This method can keep the strain viable for up to five years.
2. **Bran Preservation Method**
Start by mixing dry bran with water or a nutrient solution until it's moist but not soggy. Fill test tubes halfway with this mixture, then sterilize them. Once cooled, inoculate with the mycelium and incubate at an appropriate temperature until the mycelium fully colonizes the bran. Afterward, dry the test tubes at room temperature. Once completely dry, store them in a cool, dry place below 20°C. This technique can preserve the strain for 3 to 5 years.
3. **Salt Preservation Method**
In this approach, the fungal strain is first grown in potato broth. Prepare a flask with 250 ml of broth and add 60 ml of the culture medium. Shake the flask 5 to 10 times daily for 5 to 7 days until mycelial balls form. Transfer these balls into test tubes containing 5 ml of sterile saline. Place 4 to 5 pieces of mycelium into each tube, seal with a cotton plug, and wax the top. This method allows storage at low temperatures for up to one year.
4. **Wheat Preservation Method**
Use high-quality wheat grains that have been washed and soaked in water at 20°C for 5 hours. Allow them to air-dry slightly before filling test tubes to about 1/4 to 1/3 of their length. Sterilize the grains, cool them, and inoculate with the mycelium. Incubate until most of the grains are covered with sparse hyphae. Once ready, store the test tubes in a dry place below 25°C. This method can preserve the strain for 1 to 2 years.
These preservation techniques are simple, cost-effective, and widely used in both research and cultivation settings. Choosing the right method depends on the specific fungus and available resources. Regular monitoring and proper storage conditions are key to maintaining viable cultures over time.
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