TC Acclimatization Guide

Initial Acclimatization (Days 0-2)

Environmental Conditions:

• Temperature: Maintain at 68-78 °F.
• Humidity: Keep at 85%. Remove humidity domes briefly twice a day at a minimum to exchange air.
• Irrigation pH: Adjust water pH to 5.7-6.5. Depending on your selected media requirements. 
• EC: Maintain between 0.5-1.7. If you are unsure, always elect a lower EC.
• Light Level: Set light intensity between 80-150 PPFD. Start low and increase as needed.
• Water Temperature: Ensure water temperature is 65-70 °F.
• Misting: Mist the plantlets with plain water 2-3 times daily to maintain high humidity.

Gradual Acclimation (Days 3-10)

Gradual Acclimatization (Days 3-10)

Final Acclimatization (Days 10+)

Remove Humidity Covers:

• After 10 days, remove humidity domes or covers completely.

Monitor Environmental Conditions:

• Continue to monitor and adjust temperature, humidity, and light levels to match the vegetative growing environment.

Nutrient Management:

• Begin a regular feeding schedule with a balanced fertilizer, starting at low concentrations and increasing as the plantlets show vigor and health.

Check for Stress:

• Regularly check plantlets for signs of stress (wilting, yellowing) and make necessary adjustments to the environment.

Now What?

• At this point, your plantlets are acclimatized to your environment. They can now be transplanted and continue with your vegetative growth protocol.

Notes:

• Timing: The duration of each stage may vary depending on plant species, environmental conditions, and initial health of tissue culture plants.
• Temperature: Ensure plants are not exposed to extreme temperatures during the hardening process, as they may still be sensitive.
• Nutrition: Consider fertilization needs based on plant species and growth stage; avoid excessive nitrogen during the hardening process to promote root development.

Tips:

If you notice any signs of stress in your plants during the first 10 days, we recommend reducing the light intensity. The ideal light levels for plantlets range from 80 to 400 PPFD. If you’re unsure, it’s better to start with lower light levels. Particularly for first-time growers using tissue culture plantlets, prioritize plant health over rapid growth. Trying to accelerate growth with excessive light can lead to significant losses and an uneven canopy, rather than achieving a 100% success rate with uniform plants.

Acclimatization Page Version 1

Objective:

To gradually acclimatize tissue culture plants from controlled laboratory conditions to external environmental conditions, ensuring their successful adaptation and growth.

Materials Needed:

1. Tissue culture plants
2. Propagation trays or flats (similar to those used in traditional cloning)
3. Humidity domes  (similar to those used in traditional cloning)
4. 2”-4” pots or containers that will fit in the propagation trays (seed flats)
5. Fresh filtered water ranging from 65°-70°F in temperature. 
6. Suitable light ranging between 80-150 PPFD 
7. Fertilizer: Any commercially available base nutrient product that can be measured to achieve EC of 0.5-1.7
8. Growing media. Any media you prefer. rockwool cubes, coco, or commercially available soilless seed starting mix.
9. Ph meter w/ temperature reading capabilities.
10. A fine misting spray bottle. 

Procedure:

1. Preparation:
   • Ensure all materials and equipment are clean and sterilized to prevent contamination. Use a <10% bleach solution to sanitize any material or tool used in this procedure.
   • Select an appropriate location for hardening, preferably a partially shaded area with good air circulation. Generally, any area used for traditional cloning or seed starting will work great. No direct sunlight or intense HID lights.  If possible measure and record the light at the level of the plantlet canopy. The proper amount is in the range of 80-150 PPFD. Make certain you have the capability to adjust this on the fly in case your plantlets exhibit stress. 
   • Mix a feed solution of your preferred fertilizer at a EC of 0.5-1.7
   • Adjust the PH in accordance with your media requirements. Generally:
     Coco Coir: pH 5.5-6.2
     Rockwool: pH 5.5-6.0
     Soilless Media: pH 5.8-6.5
   • Saturate your media with your nutrient solution and let drain until no runoff is observed. (Image)
   • Fill the 2”-4” pots or containers that will fit into your propagation tray fitted with your humidity domes. These are the same used in traditional cloning. (image)

1. Stage 1: Transfer to Hardening Trays
   • Carefully remove tissue culture plants from the culture vessels using clean gloved hands. 
   • Place the plantlets in hardening trays filled with a pre-moistened potting mix. Make certain that the plantlet plug is surrounded by the moist media, touching it completely. Make sure the roots are completely covered in media. 
   • Water in the plantlets with a small amount of your nutrient solution. Drain off any excess water.
   • Lightly mist the plantlets before covering them with the humidity dome ensuring there is a tight fit to the propagation tray. . If your domes have vents, make sure they are completely closed. To maintain high humidity.
2. Stage 2: Observe
   • You will need to observe the plantlets often in the first 24 hours to see any possible stress. We recommend every 2 hours at a minimum. If there is a stresser in the environment i.e. low humidity, high temps etc. the results will be immediate and action must be taken. The first  24 hours are crucial. 
3. Stage 3: Acclimate
   • After the first 24 hours, open the vents on your humidity domes.
   • At least twice a day, remove the humidity domes to exchange the air. Take this opportunity to check the media moisture. You Likely won’t need to water the plantlets for the first few days because of the temperature and humidity levels.
   • Spot water gently as needed using a freshly mixed nutrient solution. Adjust the EC as needed to meet the demands of the plantlet. 
4. Stage 4: Monitoring and Adjustment
   • Monitor plant health and growth regularly throughout the hardening process.
   • Adjust watering frequency and light as needed. 

Initial Acclimation (Days 0-2)

 Environmental Conditions:

• Temperature: Maintain at 68-78 °F.
• Humidity: Keep at 85%. Remove humidity domes briefly twice a day at a minimum to exchange air.
• Irrigation pH: Adjust water pH to 5.7-6.5. Depending on your selected media requirements. 
• EC: Maintain between 0.5-1.7. If you are unsure, always elect a lower EC.
• Light Level: Set light intensity between 80-150 PPFD. Start low and increase as needed.
• Water Temperature: Ensure water temperature is 65-70 °F.

       Misting: Mist the plantlets with plain water 2-3 times daily to maintain high humidity.

4. Gradual Acclimation (Days 3-10)

Days 3-5:

• Temperature: Maintain at 68-84 °F.
• Humidity: Reduce to 75%-80%. Remove humidity domes briefly twice a day at a minimum to exchange air.
• Irrigation pH: Maintain water pH at 5.7-6.5.
• EC: Keep between 0.5-1.7
• Light Level: Increase to 100-200 PPFD.
• Water Temperature: Ensure water temperature remains 65-70 °F.
• Misting: Gradually reduce misting frequency to once daily.

Days 5-7:

• Temperature: Maintain at 68-84 °F.
• Humidity: Reduce to 70-75%. Remove humidity domes briefly twice a day at a minimum to exchange air.
• Irrigation pH: Keep water pH at 5.7-6.5.
• EC: Increase to 0.5-2.0.
• Light Level: Increase to 150-450 PPFD.
• Water Temperature: Ensure water temperature remains 65-70 °F.
• Misting: Continue misting as needed to maintain humidity.

Days 7-10:

• Temperature: Maintain at 68-84 °F.
• Humidity: Reduce to 60-75%. Remove humidity domes briefly twice a day at a minimum to exchange air.
• Irrigation pH: Keep water pH at 5.7-6.5.
• EC: Increase to 1.0-2.0.
• Light Level: Increase to 200-800 PPFD.
• Water Temperature: Ensure water temperature remains 65-70 °F.
• Misting: Discontinue misting and rely on environmental humidity control.

5. Final Acclimation (Days 10+)

Remove Humidity Covers:

• After 10 days, remove humidity domes or covers completely.

Monitor Environmental Conditions:

• Continue to monitor and adjust temperature, humidity, and light levels to match the vegetative growing environment.

Nutrient Management:

• Begin a regular feeding schedule with a balanced fertilizer, starting at low concentrations and increasing as the plantlets show vigor and health.

Check for Stress:

• Regularly check plantlets for signs of stress (wilting, yellowing) and make necessary adjustments to the environment.

Now what?

• At this point, your plantlets are acclimated to your environment. They can nowe be transplanted and continue with your vegetative growth protocolprotolol. 

Notes:

• Timing: The duration of each stage may vary depending on plant species, environmental conditions, and initial health of tissue culture plants.
• Temperature: Ensure plants are not exposed to extreme temperatures during the hardening process, as they may still be sensitive.
• Nutrition: Consider fertilization needs based on plant species and growth stage; avoid excessive nitrogen during the hardening process to promote root development.

Tips:

If you notice any signs of stress in your plants during the first 10 days, we recommend reducing the light intensity. The ideal light levels for plantlets range from 80 to 400 PPFD. If you’re unsure, it’s better to start with lower light levels. Particularly for first-time growers using tissue culture plantlets, prioritize plant health over rapid growth. Trying to accelerate growth with excessive light can lead to significant losses and an uneven canopy, rather than achieving a 100% success rate with uniform plants.