In this comprehensive guide on how to store agar plates with mycelium, you will gain in-depth understanding of the appropriate storage procedures for these biology essentials. Given the sensitivity of mycelium in a laboratory setting, the proper storage of agar plates becomes critical for maintaining its viability. With a focus on temperature conditions, handling techniques, and duration of storage, this guide provides you with practical and tested methods to ensure optimal preservation of your agar plates with mycelium.
Understanding Agar Plates and Mycelium
As you venture into understanding fungi and their behavior, agar plates and mycelium are two fundamental concepts you will encounter. The study of these two elements serves as a foundation in understanding fungi’s growth, propagation, and storage.
Defining Agar Plates
Begin first by understanding what agar plates are. They are petri dishes filled with a gel-like substance primarily composed of polysaccharides derived from certain seaweed species. Agar plates serve as a nutrition source for microorganisms, giving them the necessary nutrients to grow and multiply in a controlled environment. They are typically used in scientific studies to culture microorganisms like bacteria and fungi.
Understanding Mycelium
Mycelium, on the other hand, can be likened to the root structure of a plant, but for fungi. It’s this white, fibrous structure you often observe in rich soils or decomposing logs, but it’s also what allows fungi to absorb nutrients, grow, and proliferate. This filamentous structure is vital because it’s where the magic of the fungi happens — and is a crucial area to focus on when growing fungi.
Relevance of Agar Plates in Mycelium Growth
Agar plates play a significant role in the growth of mycelium. The nutrients in the agar provide the necessary sustenance for the mycelium to grow. By creating a conducive environment for growth, agar plates provide an effective means to observe, study, and propagate mycelium in a controlled environment.
Preparing Agar Plates for Mycelium Storage
When it comes to mycelium growth, the preparation of agar plates is a critical process. This involves sterilization, pouring of agar medium, cooling and solidification, and inoculation of mycelium.
Sterilization of the Agar Plates
Your first step involves sterilizing the agar plates. As you are seeking to grow a very specific strand of fungi, it’s crucial that other, perhaps competing, microorganisms do not establish themselves on the plate. Typically, sterilization is achieved by heating the plates over a flame or in a high-pressure steam autoclave to kill any existing microorganisms.
Pouring Agar Medium
Next, you will pour a nutrient-rich agar medium into each sterilized plate. The agar, a seaweed extract, contains all the nutrients necessary for mycelium growth.
Cooling and Solidifying of Agar
After pouring the agar medium, allow it to cool and solidify. Keep the plates in a safe, flat surface to solidify without disturbance. Do ensure that conditions are sanitary to prevent contamination.
Inoculation of Mycelium
Once the agar is solidified, it’s time to introduce the mycelium. This process, called inoculation, involves adding a small sample of mycelium to the agar plate and then sealed to eliminate contamination risks.
Ideal Conditions for Mycelium Growth on Agar Plates
Different fungi species thrive under different conditions. However, some environmental factors like optimal temperature, lighting conditions, required humidity level, and adequate air circulation play a vital role in mycelium growth on agar plates.
Optimal Temperature
Mycelium growth rates can be sensitive to temperature variations. For maximum growth, the temperature should typically be maintained between 20°C to 24°C, depending on the specific strain.
Lighting Conditions
Light provides an important signal for mycelium growth. These organisms best thrive in indirect, low-level light. It’s notable that exposure to light can also stimulate mycelium to produce fungal body, or the mushrooms themselves.
Required Humidity Level
Maintaining the right humidity levels is crucial for mycelium growth on agar plates. This usually involves creating a moist environment, but not overly damp, as it can lead to contamination issues.
Air Circulation
Proper air circulation ensures that the mycelium has enough oxygen necessary for development.
Proper Handling and Management of Agar Plates
The way you handle and manage your agar plates can significantly influence mycelium growth success. Key areas to focus on include avoiding contamination, monitoring the growth, dealing with mycelium contamination, and transferring mycelium.
Avoiding Contamination
One of the most important aspects of managing your agar plates is avoiding contamination. Everything from the air, to your hands, to the tools you use can contaminate and compromise the mycelium growth.
Monitoring the Growth
Part of handling and managing agar plates involves observing how well the mycelium is growing over time. This can involve looking at aspects like how quickly it takes up the nutrients in the agar, the shape and branching form it takes, among other things.
Dealing with Mycelium Contamination
At some point, you might find your plate has been contaminated. In such scenarios, it’s wise to start afresh and discard the contaminated plate, given that it’s challenging to eradicate unwanted microbial growth once established.
Transferring Mycelium
There may come a time when it becomes necessary to transfer mycelium. This could be due to a need for further growth, because the agar nutrients have been completely consumed, or the potential of contamination has increased.
Short-Term Storage of Agar Plates
After the mycelium has fully colonized the agar plates, you may not immediately need to use the plates. This ushers in the need for storage. In the short term, this involves understanding the right placement, sealing and wrapping, monitoring, and deciding when to move them to longer-term storage.
Placement of the Plates
For short-term storage, typically, agar plates are placed in a cool, dry area, shielded from direct sunlight to preserve the mycelium viability.
Sealing and Wrapping
To maintain sterility, it’s advisable to seal and wrap your agar plates using a high-quality laboratory film. This also helps to prevent dehydration of the agar.
Monitoring Stored Plates
Even in storage, it’s essential to regularly check the plates for signs of drying out or contamination.
Deciding When to Move to Longer-Term Storage
Depending on your plans for using the mycelium, you will need to decide when to move the plates to long-term storage, usually a refrigerator.
Long-Term Storage of Agar Plates
For long-term storage solutions, refrigeration and cryopreservation come in handy. You will also need to ensure sterility during long-term storage and determine the storage lifespan of your mycelium.
Refrigeration as a Method
Refrigeration can be an excellent option for long-term storage. It slows down the metabolic processes of the mycelium, giving it a longer viable period.
The Use of Cryopreservation
Cryopreservation is another option for long-term storage. It involves freezing the agar plates at very low temperatures. This method can effectively preserve mycelium for years.
Ensuring Sterility During Long-Term Storage
Irrespective of the storage method chosen, maintaining sterility is essential. This prevents contamination that may render the culture unviable.
Determining the Storage Life Span
The lifespan of stored mycelium varies considerably, depending on the type of fungi and the storage conditions.
Revival and Usage of Stored Agar Plates
When you’re ready to use your stored mycelium, certain steps support successful revival. These include warming, re-culturing, and transferring the mycelium back onto fresh agar plates.
Warming Process
After removing the plates from storage, you need to gradually warm them to room temperature. This gentle defrosting helps prevent shock that can damage the mycelium.
Re-culturing the Mycelium
Once the plates are back at room temperature, re-culturing the mycelium may be necessary before use. This could involve transferring a small sample from the old agar plate to a new one.
Transferring Mycelium Back onto Fresh Agar Plates
After re-culturing, you can transfer the mycelium onto fresh agar plates for continued growth or use.
Common Challenges in Storing Agar Plates with Mycelium
Despite the best efforts, you may encounter challenges in storing agar plates with mycelium. Common issues include potential contamination, agar drying, difficulty in mycelium revival, or dealing with fungal infections.
Potential Contaminations
As previously mentioned, one of the most common challenges is contamination. This can occur at any stage, and it’ll be necessary to discard the infected plates to maintain the integrity of your mycelium cultures.
Preventing Agar Drying
Agar drying is another common problem, which can occur due to storage conditions that are too dry or due to inadequate sealing.
Difficulty in Mycelium Revival
Reviving mycelium from long-term storage can be a difficult process if not done properly. It requires careful re-culturing, without which the mycelium might not survive the transition from storage to growth conditions.
Dealing with Fungal Infections
Fungal infection is another issue that can arise. It could manifest in different forms that make the mycelium unhealthy and unfavourable for your intended usage.
Profiling Specific Types of Mycelium and Their Storage Preferences
Different types of fungi have unique characteristics, and as such, understanding their unique conditions for growth and storage is critical. Here, we briefly profile some specific types of mycelium and their storage preferences.
Psilocybe cubensis
This species prefers moderately warm temperatures and high humidity for growth. For storage, it does well at cooler, refrigerated temperatures.
Shiitake (Lentinula edodes)
Shiitake thrives in cooler conditions with high humidity. For storage, refrigeration works best.
Oyster Mushroom (Pleurotus species)
Oyster mushrooms prefer cooler temperatures and high humidity. Upon storage, they should be kept in a refrigerator.
Lion’s Mane (Hericium erinaceus)
Lion’s Mane prefers cool to warm temperatures with a high level of humidity. For storage, a cool, refrigerated environment works best.
Safety Measures and Best Practices in Handling Agar Plates
In everything you do regarding agar plates and mycelium, safety comes first. This often entails proper hand hygiene, the use of personal protective equipment, safe disposal of used agar plates, and being aware of potential mycelium allergies and risks.
Proper Hand Hygiene
Hands are often significant contamination sources, and thus maintaining proper hand hygiene is key to successful mycelium growth and storage.
Use of Personal Protective Equipment
While working with agar plates and mycelium, using personal protective equipment like gloves, lab coats, and potentially a face mask can help mitigate contamination risks.
Safe Disposal of Used Agar Plates
Once done using your agar plates, the disposal process should be equally procedure-specific to prevent potential hazards from discarded fungal components.
Mycelium Allergies and Risks
Lastly, being aware of potential allergies to mycelium is vital. Everyone reacts differently, and in the case of suspected allergies, it’s advisable to consult a healthcare professional.
In conclusion, storing agar plates with mycelium requires basic understanding of mycology, meticulous preparation, appropriate storage conditions, and regular monitoring. This comprehensive guide sums up the processes you need to venture into the mycological world, ensure successful storage and revival of your mycelium cultures.