Mastering the Art of Tissue Culture: Proven Steps for Healthy and Vibrant Plants

Procedure Description Technical Details and Examples Numbers Products for Assistance
Deflasking Carefully remove plants from the tissue culture flask, ensuring minimal damage to roots. 🌿 - Sterilise equipment such as forceps and scissors. 🧼 - Rinse plants in a sterile solution (e.g., 70% ethanol) before transfer. 🚿 Sterilize equipment for 5 minutes ⏳. Rinse plants in a sterile solution for 1 minute ⏳. N/A
Plug Selection Choose appropriate plugs or small pots filled with a suitable growing medium for transplanting. 🪴 - Select plug size based on the plant's root system and growth stage. Example: 50-cell plug trays for small seedlings. 📏 Use 50-cell plug trays for small seedlings 🌱. N/A
Preparing Growing Medium Ensure the growing medium is well-draining and sterile to prevent contamination. 🏥 - Use a mix of peat moss, perlite, and vermiculite sterilized in an autoclave. 🌱 - Maintain a pH level of 5.5 to 6.5. 📐 Sterilize the growing medium at 121°C for 15 minutes 🔥. Maintain a pH level of 5.8 🧪. N/A
Transplanting Gently transfer tissue-cultured plants into plugs, ensuring proper placement and minimal stress. 🌱 - Handle plants with sterilized forceps. 🧤 - Ensure the plant's root collar is at the right depth in the plug. 🌱 Transplant with minimal root disturbance 🌱. Place the root collar 1 cm below the plug surface. 📏 N/A
Watering Provide sufficient water to settle the plants in plugs, but avoid overwatering to prevent root rot. 💧 - Use a misting system or gentle watering. 🚿 - Maintain moisture levels to avoid wilting. 🌿 Water the plugs until excess moisture drains from the bottom 💧. Maintain moisture at 70-80% 💧. Rooting hormone for enhanced root development 🌱. Antifungal solution for root rot prevention 🍄.
Humidity Control Maintain a high humidity environment around the plugs to reduce transplant shock. 🌧️ - Cover plugs with a clear plastic dome or use a misting system. ☁️ - Aim for 80-90% relative humidity. 🌡️ Maintain humidity at 85% for the first 7 days ☁️. Use a clear plastic dome with venting 🏡. Humidity trays with water reservoirs 💧.
Light Exposure Gradually introduce tissue-cultured plants to light to prevent sunburn or damage. 🌞 - Start with low-intensity light and increase gradually. 🌄 - Maintain a photoperiod suitable for the plant species. 🕒 Begin with 100 lux of light intensity for 2 hours daily ☀️. Maintain a 16-hour photoperiod 🕒. Grow lights for supplemental lighting 💡.
Temperature Control Keep the temperature stable and within the appropriate range for the plant species. 🌡️ - Example: Maintain a temperature of 22-25°C for most tropical plant species. 🌴 - Use heat mats for temperature control. 🔥 Maintain a constant temperature of 23°C 🌡️. Use heat mats to adjust temperature as needed 🔥. Heating mats for temperature regulation 🔥.
Fertilization Begin a light fertilization regimen to support plant growth and development. 🌾 - Start with a diluted, balanced fertilizer (e.g., 20-20-20). 🥤 - Adjust the fertilizer ratio based on plant requirements. 🌱 Use a 1/4 strength balanced fertiliser 🌱. Adjust based on weekly nutrient analysis 🧪. Liquid fertilizers for precise application 🥤.
Monitoring Regularly check for signs of stress, disease, or nutrient deficiencies and address them promptly. 👀 - Monitor plant growth, leaf color, and overall health. 👁️ - Conduct nutrient tests if needed. 🧪 Weekly visual assessments and monthly nutrient tests 👁️. Address issues within 48 hours 🕰️. Pest control products 🐜, pH meters 📏, and nutrient supplements 🌿.
Transplanting to Larger Containers When the plants have grown sufficiently, move them to larger pots or garden beds for further growth. 🪣 - Example: Transplant young tomato seedlings to individual pots when they have 2-4 true leaves. 🍅 Transplant to individual pots when plants reach a height of 10 cm with 2-4 true leaves 🪣. Potting mix 🌱 and larger containers 🪣.
Hardening Off If transplanting outdoors, gradually acclimatize plants to outdoor conditions before planting. 🌬️ - Reduce humidity and exposure to direct sunlight over several days or weeks. 🌤️ - Protect plants from strong winds. 🌪️ Over 7 days, reduce humidity by 10% daily ☁️. Gradually increase sunlight exposure 🌞. Protect from winds above 20 km/h 🌬️. Shade cloth ☂️, windbreaks 🌿, and plant supports 🌱.

 

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