Liquid culture

Normally spores are injected into a substrate. It takes time for the spores to germinate and to form mycelium. Mushroom growers find this a time consuming growing method. That’s why they created the liquid culture technique.

Liquid culture is a fluid containing water and essential nutrients to ensure mycelium growth (mostly consists of malt extract or honey). Spores are injected into the liquid. There they germinate and within a couple of days you’ll have a liquid culture full of mycelium.
This liquid culture is then used to inoculate new substrates. Using liquid cultures, the substrate is colonized much faster compared to spores.
I read Otto’s method and decided to make my own liquid culture so I could teach you how to do it as well. Quick side note: this method of developing mycelium should work with every mushroom species.

Materials needed:

• 1 clean glass jar (about 250ml)
• Jar lids
• Hammer and a nail
• Tin foil
• Water
• Honey
• Table spoon
• Cooking pot
• A tong or any kind of large tweezer
• Micropore tape
• Ethanol (rubbing alcohol)
• Spore print/ spore syringe/ piece of mycelium from agar plates or cardboard
• Scalpel or sharp knife

Instructions:

Step 1: take the lids of your jars and puncture a hole in the middle with a hammer and a nail. Smoothen the edge a little so you don’t cut yourself.
Clean the jars thoroughly, as well as the lids. Take some water and honey and proceed to step 2.

Step 2: Put the jars in a cooking pot along with the lids. Add some water and bring to a boil for about 30 minutes. This sterilizes the jars to avoid contaminations of bacteria and molds later on. Use a lid on your cooking pot to capture the steam.
After the 30 minutes are up, don’t do anything. First read step 3.

Step 3: Using the tong, take out the jars and place them on a clean working area whilst still hot. The water should still be hot or even boiling. Now pour about 250ml of the boiled water in the jars. Use a clean measuring cup if necessary.
Now add a tablespoon of honey to the sterilized water. Put the lids on the jars and go on to step 4.
Side note: the picture images a teaspoon, not a tablespoon.

Step 4: Cover the jars with tin foil. Make sure the tin foil is on there snugly, since this prevents water from entering the jar during sterilization in step 5.

Step 5: We’re going to repeat step 2. Place the jars in the pot and boil the water for 30 minutes, as pictures above. Use a lid on your cooking pot to capture the steam. Take the jars out after the 30 minute boil and leave them to cool over night.
WARNING : leave the tin foil ON the jars to cool. This prevents airborne contaminants to enter your jars.

Step 6: After the jars have cooled down over night, gently take away the tin foil. Using your ethanol and a tissue, thoroughly clean the lid of the jar. You’re now ready for inoculation.
To inoculate, you need a spore print, a live culture or a spore syringe.
A spore syringe/live culture: Rub your scalpel or knife using the ethanol. Gently rub about a knife tip worth of spores (you really don’t need much) off of the print. Unscrew the lid and insert the spores into the jar in one smooth motion. Make sure you close the lid as fast as possible. The same goes for live cultures. Cut out a piece of culture from a petri dish or cardboard and place it in the jar.
A spore syringe: Heat the needle of your syringe until it’s red hot. Wipe it down using ethanol and insert the needle in the hole in the jar lid in one smooth motion. Inject 1cc of spore solution. Eject your syringe.
Step 7: Cover the hole in the jar lid with micropore tape to keep out contaminants. Store somewhere for about one week at room temperature. Make sure you agitate the liquid every day by shaking it. You will start to see mycelium. Keep shaking it daily. This will prevent the mycelium to clump together. But be careful not to wet the micropore. Wet micropore is a open invention for contaminants.
All the little spots you see in the image above are clumps of mycelium. Suck these up using a sterile syringe for future use. You can keep these syringes for a couple of months in the fridge.