Sarah Brown
Cloning wild mushrooms is a valuable skill for mycologists, enabling them to faithfully reproduce characteristics and maximise yields. The process involves scraping a small piece of tissue from a mushroom under sterile conditions and placing it on a sterile medium to grow. This simple technique can be performed by amateurs and experts alike and is a great way to preserve rare strains and discover new varieties. It's important to choose the right mushroom to clone, opting for young, fresh, healthy specimens with desirable traits. With the correct tools and environment, anyone can attempt to clone mushrooms at home.
| Characteristics | Values |
|---|---|
| Process | Harvest a piece of tissue from a mushroom fruitbody, place it on agar, and allow the mycelium to grow out until you have a pure culture. |
| Equipment | A laminar flow hood or still air box (SAB), scalpel or X-Acto knife, isopropyl alcohol, bleach solution, gloves, parafilm or masking tape, jars of grain spawn, cardboard, pressure cooker, Bunsen burner. |
| Mushroom Type | Wild, store-bought, or cultivated mushrooms can all be cloned. Saprophytic mushrooms that grow on dead and decaying organic material in the wild are the easiest to clone. |
| Mushroom Characteristics | Choose a mushroom that is healthy, fully developed, and free from contamination, disease, or discoloration. The fresher the mushroom, the higher the chances of cloning success. |
| Yield | Cloning can produce numerous generations but cannot be sustained indefinitely due to genetic drift and senescence. |
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What You'll Learn
Choosing the right mushroom to clone
When it comes to choosing the right wild mushroom to clone, there are several factors to consider. Firstly, it is crucial to select a mushroom that is healthy and fully developed, avoiding any specimens with signs of contamination or disease. These issues may persist in the cloned mycelium, so it is important to choose a fresh, healthy mushroom fruiting body. The fresher the specimen, the higher the chances of a successful cloning process.
Secondly, you should identify the traits you want to cultivate and replicate. For example, you may prioritise characteristics such as rapid growth, large fruiting bodies, high potency, yield, colour, substrate suitability, or disease resistance. By selecting a mushroom that exhibits these desirable traits, you increase the likelihood of successfully replicating those traits in future generations.
Thirdly, consider the type of mushroom and its ease of cloning. Saprotrophic mushrooms, which grow on dead and decaying organic material in the wild, are generally easier to clone and cultivate than parasitic or mycorrhizal mushrooms. Some specific types of mushrooms that are commonly suggested to be easy to clone include oyster mushrooms, shiitake, and lion's mane. Additionally, thick and fleshy mushrooms like shiitake, king oysters, and cremini mushrooms are reportedly easier to work with when cloning.
Lastly, it is recommended to choose young mushrooms that are still in their button stage. At this growth stage, mushrooms tend to grow faster and can even double in size daily. This rapid cell division is advantageous when transferring tissue to agar, as it allows for quicker growth and reproduction.
Remember, cloning wild mushrooms allows you to cultivate and preserve unique strains found in nature, contributing to the maintenance of genetic diversity among cultivated mushrooms and the discovery of new strains. By considering the health, traits, type, and growth stage of the mushroom, you can make an informed decision when choosing the right wild mushroom to clone.
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Preparing your work area and tools
Choose a Suitable Work Area
Select a workspace that can be easily sanitised and controlled to minimise contamination. A laminar flow hood or a still air box (SAB) is ideal for maintaining a sterile environment. If neither of these is available, a kitchen near a gas cooker or a workspace with a Bunsen burner can also help limit contaminants.
Gather the Necessary Tools and Materials
You will need a variety of tools and materials for the cloning process:
- Sterile scalpel or knife: Use a sharp and sterilisable instrument like a scalpel or X-Acto knife for precise tissue removal and transfer. Sterilise the blade with a flame before each use.
- Sterilising agents: Isopropyl alcohol, bleach solution, or other sanitising agents are essential for cleaning surfaces, tools, and even the exterior of the mushroom to prevent contamination.
- Gloves: Wear disposable gloves to maintain sterility and prevent the introduction of unwanted microbes from your hands.
- Agar plates: Agar is the most common medium for cloning mushrooms. You can prepare your own agar plates using a basic malted agar recipe or purchase pre-poured plates online.
- Alternative growing media: Besides agar, you can use sterilised grain (such as rye, wheat, or rice) or damp cardboard as a growing medium. However, the risk of contamination is higher with these alternatives.
- Mushroom specimen: Choose a fresh, healthy mushroom with desirable traits to clone. Young mushrooms in their button stage are ideal as they grow faster and can double in size daily.
Prepare the Work Area and Tools
Once you have selected your workspace and gathered the necessary tools and materials, it's time to prepare:
- Sanitize your work area: Use sterilising agents to wipe down all surfaces, including your laminar flow hood or still air box. Ensure all supplies are within reach.
- Prepare the mushroom specimen: Clean the exterior of the mushroom with a sterilising agent like isopropyl alcohol or a diluted bleach solution. If using wild mushrooms, consider immersing them in hydrogen peroxide for 30 seconds to deactivate potential contaminants.
- Sterilise your tools: Ensure your scalpel or knife is sterile by flaming or using another suitable method. Put on your disposable gloves to maintain a sterile environment during the cloning process.
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Tissue extraction and transfer
Choosing the Mushroom
Select a fresh, healthy mushroom with desirable traits, such as rapid growth or large fruiting bodies. Wild mushrooms, store-bought fruits, or cultivated species can be cloned. However, it is important to ensure the mushroom is properly identified before proceeding.
Sterilization and Workspace Preparation
Prepare a sterile workspace by using a laminar flow hood or a still air box (SAB) to minimize contamination. Put on disposable gloves to maintain sterility and clean the mushroom's exterior with a sterilizing agent like isopropyl alcohol or a bleach solution.
Tissue Extraction
Use a flame-sterilized scalpel or X-Acto knife to extract a small piece of tissue from the mushroom. The ideal location for tissue extraction is the thickest, fleshiest part of the mushroom, such as the interior of the cap or the stem, as these areas are less likely to be contaminated. Avoid using gill tissue as it may be contaminated with spores.
To prevent contamination, tear the mushroom fruit rather than cutting it to avoid pushing contaminants from the outside into the center. Ensure the scalpel is cool before extracting the tissue, as heat may kill the tissue. You can cool the scalpel by dipping it into the agar solution or allowing it to cool in the stream of the flow hood.
Tissue Transfer
Place the extracted tissue on a nutrient-rich growing medium, such as agar or sterilized grain. Agar is the most common medium, and you can use alternative methods such as transferring the tissue into a jar of sterilized grain or between damp, sterilized cardboard. Seal the agar plate with parafilm or masking tape to minimize contamination and store it in a controlled environment, typically around 70-75°F (21-24°C).
Monitor the plate for mycelium growth, which may take several days to a few weeks. If contamination occurs, perform culture transfers by removing clean mycelium to a new dish. This process may need to be repeated several times to obtain a clean culture.
Dried Mushrooms
Although fresh mushrooms are ideal, it is possible to clone dried mushrooms. Rehydrate the dried mushroom tissue in sterile distilled water for up to a day, then transfer it to a sterile growing medium. However, cloning from dried mushrooms is less reliable as the drying process may have damaged the tissue.
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Using agar plates
To clone wild mushrooms using agar plates, you will need the following supplies:
- Agar plates: Pre-poured or homemade agar plates that provide a nutrient-rich medium for mycelium growth. Agar plates are petri dishes that contain a sterile, nutrient-rich agar solution. Agar is a jelly-like organic substance produced from red algae.
- A fresh, healthy mushroom fruiting body: Choose a specimen with desirable traits to clone. Thick, fleshy mushrooms like king oyster, shiitake, gold top, and button mushrooms are easiest to work with.
- A sterile workspace: Use a laminar flow hood or still air box (SAB) to minimize contamination.
- Scalpel or X-Acto knife: A sterilizable, sharp instrument for tissue removal and transfer.
- Sterilizing agents: Isopropyl alcohol, bleach solution, or another sanitizing agent for cleaning surfaces and tools.
- Gloves: Disposable gloves for maintaining sterility during the cloning process.
- Parafilm or masking tape: For sealing agar plates to minimize contamination.
Before you begin, clean and sanitise your workspace, including your laminar flow hood or still air box.
To begin the cloning process, first, wipe down the outer surface of the mushroom with alcohol on a paper towel. Then, carefully tear open the mushroom to expose the inner tissue. Using a sterilised scalpel, remove a sample of the mushroom fruiting body. Place the tissue sample onto the centre of the agar plate. Only remove the lid from the plate momentarily and replace it as quickly as possible. Seal the agar plate with parafilm or plastic wrap. Ensure that all plates are labelled with the species and date. Store the plates at room temperature, away from direct sunlight, and observe the growth. Contaminants will usually become visible within 3–5 days. Using parafilm ensures that the agar plates can breathe while maintaining a tight seal and preventing the development of contaminants.
After a few days, you should notice mycelium growing out from the mushroom tissue. After a week or two, you should observe decent growth. During this process, you may have picked up some contamination, especially if you are working with wild mushrooms, and performing multiple transfers is advisable. Choose the healthiest, fastest-growing mycelium and transfer each to multiple plates. After around three transfers, you should have clean mycelium that can be used for future cultivation projects.
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Storing your cloned mushrooms
When storing cloned mushrooms, it is crucial to choose specimens with desirable traits that are healthy and fully developed. Avoid any mushrooms with signs of contamination or disease, as these issues may persist in the cloned mycelium. Ideally, you should select fresh mushrooms as the fresher the specimen, the higher the chances of success. However, it is possible to clone dried mushrooms by hydrating them in sterilized water.
In terms of storage methods, there are several options. One approach is to use agar plates, which are the most common medium for cloning mushrooms. Once the agar plate is fully colonized with mycelium, you can store it in the refrigerator for several months, providing a continuous supply of mycelium for future projects. To ensure the agar plates remain uncontaminated, seal them with parafilm or masking tape and store them in a Ziploc bag on a shelf at room temperature, away from direct sunlight.
Another method for cloning mushrooms is to use sterilized jars of grain, also known as grain spawn. Various types of grain can be used, including rye, wheat, and wild bird seed. This technique involves transferring a small piece of internal mushroom tissue directly into the jar of sterilized grain, allowing the mycelium to grow throughout.
Additionally, some people use sterilized cardboard instead of agar or grain. This method has a higher chance of contamination and lower yield, but it is simple and low-tech. It involves pushing the mushroom tissue into the ribs of the prepared cardboard and then placing the inoculated cardboard into a sterilized jar.
Overall, storing your cloned mushrooms requires careful selection of specimens, attention to freshness, and the use of appropriate storage methods to minimize contamination.
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Frequently asked questions
The best mushrooms to clone are saprophytic mushrooms, which grow on dead and decaying organic material in the wild. Some species are easier to clone than others, including oyster mushrooms, shiitake, and lion's mane.
You will need a sterile workspace, such as a laminar flow hood or still air box. You will also need a scalpel or X-Acto knife, sterilizing agents, and disposable gloves. Agar plates or liquid culture can be purchased online and act as a nutrient-rich medium for the fungus to grow on.
First, choose a healthy, young mushroom with desirable traits. Clean the mushroom with a brush and water, and then immerse it in hydrogen peroxide to remove spores from potential contaminants. Next, harvest a small tissue sample from the mushroom and place it on an agar plate or alternative growing medium. The mycelium will grow out until you have a pure culture.
It is important to maintain a sterile environment when cloning mushrooms to avoid contamination with bacteria or mold. Clean and sanitize your workspace and tools with isopropyl alcohol or a bleach solution. Choose a mushroom that is healthy and free from mold or other contaminants, as these issues may persist in the cloned mycelium.
