How to Successfully Deflask and Acclimate Tissue Culture Plants

Tissue culture plants offer an exciting way to cultivate rare and exotic species, but the process of deflasking and acclimating them can be delicate and requires meticulous care. By following the right steps, you can ensure that these plants thrive in their new environment. This guide will walk you through the process of deflasking, planting, and acclimating tissue culture plants using sterile equipment and a humidity dome kit.

Step 1: Prepare Your Equipment and Environment

Before you begin, it’s crucial to ensure that all your equipment and workspace are sterile to prevent contamination. Here’s what you need:

• Purified or Reverse Osmosis (RO) water
• Sterilized 50% peat moss/50% perlite mix
• Sterilized containers for the planting mix
• Rubber-tipped tweezers
• Humidity dome kit
• Sterilizing solution (e.g., 70% isopropyl alcohol)

Sterilization Process:

1. Clean Your Work Area: Wipe down all surfaces with a sterilizing solution.
2. Sterilize Equipment: Dip the rubber-tipped tweezers in isopropyl alcohol and let them dry. Do the same with any other tools you’ll be using.
3. Prepare the Planting Mix: Mix the peat moss and perlite in a sterilized container.

Step 2: Deflasking the Tissue Culture Plants

1. Remove the Plants from the Flask: Gently remove the tissue culture plants from their flask using the sterilized rubber-tipped tweezers. Be careful not to damage the delicate roots.
2. Rinse in Purified Water: Place the plants in a bowl of purified or RO water to wash off the gel medium. This step is crucial to remove any remaining nutrients and chemicals that could harm the plants.
3. Inspect the Roots: Check for any signs of damage or disease. Healthy roots are essential for successful acclimation.

Step 3: Planting in the Sterilized Mix

1. Fill the Pots: Fill small, sterilized pots with the 50% peat moss/50% perlite mix.
2. Plant the Seedlings: Using the tweezers, carefully plant the seedlings in the mix, ensuring that the roots are adequately covered and the plant is secure.
3. Watering: Lightly water the plants with purified water to settle the mix around the roots.

Step 4: Using the Humidity Dome Kit

1. Place Pots in the Humidity Dome: Arrange the pots inside the humidity dome. The dome will help maintain high humidity levels, which are crucial for the initial stages of acclimation.
2. Close the Dome: Secure the lid of the dome to keep the humidity high.

Step 5: Acclimating the Plants

1. Initial Care: Keep the humidity dome in a location with bright, indirect light. Avoid direct sunlight, as it can overheat the plants.
2. Gradual Acclimation: Over the course of a few weeks, gradually increase the airflow by opening the vents or slightly lifting the dome. This process helps the plants adjust to lower humidity levels.
   • Week 1: Open the vents or lift the dome for 1-2 hours daily.
   • Week 2: Increase the exposure to 3-4 hours daily.
   • Week 3: Open the dome halfway for 5-6 hours daily.
   • Week 4: Completely remove the dome for a few hours each day, gradually extending the duration.

Step 6: Transition to Open Environment

1. Final Transition: Once the plants can handle full days outside of the humidity dome, you can move them to their permanent location.
2. Monitor and Adjust: Continue to monitor the plants closely for any signs of stress or dehydration. Adjust watering and humidity levels as needed.

Importance of Sterile Conditions

Maintaining a sterile environment throughout this process cannot be overstated. Contaminants can easily cause diseases that may kill the delicate tissue culture plants. Sterilize all tools, containers, and the workspace before and during the deflasking and planting process to ensure the best chances of success.

By following these steps and emphasizing cleanliness, patience, and gradual acclimation, you can successfully transition tissue culture plants from their flasks to a thriving state in your collection. Happy growing!