Maintenance of in plant tissue culture

• Maintenance of in plant tissue culture ensures healthy growth, prevents contamination, and supports long-term research use.
• Maintenance of in plant tissue culture preserves explants under controlled light, temperature, and nutrient conditions.

1. Inoculation and Aseptic Handling

   • Work in a laminar flow cabinet to maintain sterility.
   • Use sterile tools and techniques to transfer explants or subculture tissues.

2. Subculturing

   • Transfer tissues to fresh medium periodically (every 4–6 weeks for many species).
   • Prevents nutrient depletion and accumulation of toxic metabolites.
   • Maintains vigor of callus/suspension/shoot cultures.

3. Monitoring and Contamination Control

   • Inspect cultures regularly for signs of bacterial or fungal
   • Discard contaminated cultures promptly to prevent spread.

4. Acclimatization (Hardening Off)

   • Gradual exposure of regenerated plantlets to lower humidity and non-sterile conditions.
   • Transfer plantlets to soil or potting mix in a controlled environment (greenhouse) for a few weeks.
   • Ensures a high survival rate when moved outdoors.

5. Storage / Cryopreservation

   • For long-term preservation of germplasm, explants or shoot tips can be cryopreserved in liquid nitrogen.
   • Slows metabolic processes, maintaining viability for extended periods.

6. Record-Keeping

   • Essential to track media compositions, explant sources, dates, and culture observations.
   • Ensures reproducibility and helps troubleshoot problems.

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