Emily Johnson
Cloning mushrooms involves taking a piece of tissue from an existing fungus and creating a genetically identical copy. This technique is used by mycologists to faithfully reproduce the characteristics of a specific specimen and maximise crop yields. It is also a way to guarantee that the genetics of the cloned mushroom will be the same as the original. The best mushrooms to clone are saprophytic mushrooms that grow on dead and decaying organic material in the wild, such as oyster mushrooms, shiitake and lion's mane. Cloning mushrooms is easier than you may think and can be done at home.
| Characteristics | Values |
|---|---|
| Purpose | To create an identical copy of a mushroom with desirable traits like yield, speed of growth, colour, substrate suitability, and disease resistance. |
| Mushroom types | Saprophytic, parasitic, and mycorrhizal mushrooms. Of these, only saprophytic mushrooms are easy to clone and cultivate. Oyster, shiitake, and lion's mane mushrooms are some of the easiest to grow and clone. |
| Mushroom characteristics | Choose a mushroom that is relatively large, healthy, and free from signs of disease or contamination. |
| Equipment | Agar plates (petri dishes with a sterile nutrient-rich agar solution), scalpel, alcohol-soaked cloth, magnetic stir plate or glass, syringe, needle. |
| Process | Clean the mushroom with an alcohol-soaked cloth. Tear the mushroom in half and use a needle to extract a tissue sample. Place the tissue sample on agar. Seal the petri dish and place it in a temperature-controlled environment (21-24°C). Monitor the growth of the mycelium and transfer healthy parts to new media for purification. |
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What You'll Learn
Choosing the right mushroom
Type of Mushroom
The best mushrooms for cloning are saprophytic mushrooms, which grow on dead and decaying organic matter in the wild. Saprophytic mushrooms are easy to clone and cultivate, and some varieties are more beginner-friendly than others. Oyster mushrooms, shiitake, and lion's mane mushrooms are known for their hardiness, fast growth, and ease of cultivation. Shiitake, king oyster, and cremini mushrooms are also good choices due to their thick, fleshy consistency, making them easier to work with.
Age and Development Stage
It is recommended to use young mushrooms still in their button stage for cloning. Younger mushrooms tend to colonize faster and can double in size daily. However, it is also suggested to select a relatively large fruiting body, as it can be more challenging to obtain clean tissue samples from smaller or thin-fleshed mushrooms.
Desirable Traits
When selecting a mushroom to clone, choose one that exhibits the traits you wish to cultivate. This could include characteristics such as rapid growth, large fruiting bodies, or high potency, depending on your specific goals. Cloning from a high-quality specimen increases the likelihood of replicating those desirable traits in future generations.
Health and Contamination
Always choose a healthy and fully-developed mushroom, free from any signs of contamination or disease. Issues like contamination or disease may persist in the cloned mycelium, affecting the success of the cloning process. Fresh mushrooms are generally preferred for cloning, as the chances of success are higher, and the process is less prone to contamination.
Source of Mushroom
While you can clone mushrooms you've grown yourself or store-bought mushrooms, it is important to note that growing mushrooms from spores can be unpredictable. Cloning wild mushrooms allows you to cultivate and preserve unique strains found in nature, contributing to the maintenance of genetic diversity among cultivated mushrooms.
Remember, the key is to select a mushroom with the traits you want to replicate and ensure it is healthy, young, and free from any contamination. By choosing the right mushroom and following the appropriate cloning techniques, you can successfully create identical copies of your desired mushroom.
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Preparing the work area
To prepare your work area for mushroom cloning, you'll need to gather the necessary supplies and create a sterile environment to reduce the chances of contamination. Here are the steps to follow:
First, clean your work surface thoroughly. This step is crucial to minimise the risk of introducing contaminants into your mushroom culture. Use a disinfectant suitable for the surface you're working on.
Next, gather your tools and equipment, such as scalpel(s), needle(s), petri dishes, and agar. Sterilise your tools by passing them over a flame to avoid introducing contaminants. If you're using agar plates, you can buy pre-made ones online, or you can make your own agar growing medium by purchasing agar in powdered form. Agar is a jelly-like substance derived from red algae, providing the nutrients needed for mycelium growth in a sterile environment.
Now, select your mushroom. It is recommended to use young mushrooms still in their button stage, as they grow faster and can double in size daily. Look for a relatively large fruit body, as smaller specimens can be challenging to work with. Clean the outside of the mushroom by wiping it down with an alcohol-soaked cloth. This step will damage the edibility of the mushroom, so be prepared to sacrifice it for the clone. The alcohol wipe will significantly reduce the potential for contamination from the mushroom's exposure to air.
Once your work area and equipment are sterile, and you have selected and cleaned your mushroom, you are ready to begin the cloning process.
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Tissue sampling
Sterilization and Preparation:
- Ensure your workspace is sterile by using a laminar flow hood or a Still Air Box (SAB) to minimize airborne contamination.
- Prepare a flame sterilizer, such as an alcohol lamp or burner, to sterilize your tools.
- Have a constant flame going on your worktop to sterilize your scalpel or needle between each tissue transfer.
- Put on disposable gloves to maintain sterility during the process.
- Clean the exterior of the mushroom with a sterilizing agent, such as isopropyl alcohol or a bleach solution, to reduce the risk of contamination.
Tissue Removal:
- Choose a fresh, healthy mushroom with desirable traits for cloning.
- Tear or pull the mushroom in half to expose the inner flesh, which is naturally sterile and contains clean, fertile mushroom mycelium cells. Tearing is preferred over cutting to avoid pushing contaminants from the outside into the center.
- Using a flame-sterilized scalpel or needle, remove a small piece of tissue from the innermost and fleshiest part of the mushroom, such as the interior of the cap or the stem base. Avoid using gill tissue as it may be contaminated with spores.
- If using a needle, you will have a core sample of tissue stuck on the tip, which can be injected directly into a jar of nutrient media.
- If using a scalpel, ensure the blade is cooled before removing the tissue to avoid damaging it with heat. You can dip the blade into the agar solution to cool it down.
Storing the Tissue Sample:
- Place the tissue sample onto a nutrient-rich agar plate, which provides the necessary environment for the mycelium to grow.
- Seal the agar plate with parafilm or masking tape to prevent contamination while still allowing the plate to breathe.
- Label the plate with the species, inoculation date, and mushroom strain.
- Store the sealed agar plate in a dark, temperature-controlled environment, typically around 70-75°F (21-24°C).
- Monitor the plate for mycelium growth, which can take 2-3 days to several weeks, depending on the strain.
- Be vigilant for any signs of contamination, such as off-color growth or unpleasant odors. If contamination occurs, you may need to transfer a clean sample of mycelium to a new, uncontaminated plate.
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Incubation and monitoring
Incubation
Seal the agar plates with tape to prevent contamination. Place the sealed plates in a temperature-controlled environment, maintaining a temperature range of 21-24°C. This temperature range is optimal for mycelium growth.
Monitoring
Regularly monitor the growth of the mycelium. Look for healthy parts of the mycelium and transfer them to new media for purification. This process is essential to ensure the successful growth of the cloned mushroom culture.
Isolation and Pure Culture
If the mycelium grows without contamination, you will have a pure culture ready for further expansion. Transfer this pure culture to a substrate like grains or sawdust, where it can continue to grow and develop into mature mushrooms.
Storage and Longevity
Agar plates are excellent for storing mushroom mycelium for extended periods. They provide a sterile environment that reduces the chances of contamination. Proper storage can help maintain the viability of the cloned mushroom culture over time.
Maintenance
To maintain the health and viability of your cloned mushroom culture, periodically shake the agar plates or use a magnetic stir plate to prepare the liquid culture with a magnetic stir bar. Alternatively, you can use broken glass pieces to break up the mycelium during shaking.
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Isolation and pure culture
To achieve isolation and pure culture, start by selecting a mushroom strain that is well-suited to your growing environment and conditions. Wild mushrooms can be cloned to cultivate and preserve unique strains, contributing to the maintenance of genetic diversity. Oyster mushrooms, for example, can be cloned from the basidiocarp onto a prepared medium, allowing the fungal mycelium to grow and be used as the mother culture inoculum.
The next step is to create a sterile environment to reduce the chances of contamination. Clean your work surface and tools, and disinfect the mushroom to be cloned. Using a sterilized scalpel, tear open the mushroom to access the sterile internal tissues. Take a small piece of flesh, approximately 2-3 mm², and place it on an agar plate or another sterile medium. Seal the plate with parafilm or masking tape to prevent contaminants from entering, and label it with the date and mushroom strain.
Store the agar plate in a dark, temperature-controlled environment, typically around 70-75°F (21-24°C). Monitor the plate regularly for mycelium growth, which can take several days to a few weeks, depending on the strain. During this incubation period, keep a close eye on any signs of contamination, such as off-color growth or unpleasant odors. If contamination occurs, you may be able to salvage a clean sample of mycelium and transfer it to a new, uncontaminated plate.
Once the agar plate is fully colonized with mycelium, you have achieved a pure culture. At this stage, you can store the plate in the refrigerator for several months, ensuring a continuous supply of mycelium for future cultivation projects. Additionally, you can transfer the pure culture to a mycelium on grains or sawdust substrate for further growth and cultivation.
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Frequently asked questions
The best mushrooms to clone are saprophytic mushrooms, which grow on dead and decaying organic matter in the wild. Some examples of saprophytic mushrooms that are easy to clone include oyster mushrooms, shiitake, and lion's mane.
Cloning mushrooms provides consistent and reproducible results, whereas growing from spores can be unpredictable. Cloning also reduces your reliance on spores, which can be expensive and have low germination rates.
You will need a sterile work environment, such as a still air box, and a sterile scalpel or needle to take a tissue sample from the mushroom. You will also need petri dishes or jars containing a sterile nutrient-rich agar solution for the tissue sample to grow in.
