General Process of Sapling Production from Tissue-Cultured Plantlets

a. Receipt of Plantlets

• Plantlets are supplied from laboratories in small polycups, test tubes, or culture bottles.
• They are delicate and accustomed to aseptic, controlled laboratory conditions.

b. Primary Hardening

• Plantlets are washed to remove agar media and transferred to sterilized potting media (cocopeat, vermiculite, perlite, or soilrite).
• Maintained in a greenhouse or shade house with controlled humidity (80–90 percent), temperature (25–28°C), and partial shade.
• Gradual exposure to external environment develops photosynthetic ability and stress tolerance.

c. Secondary Hardening

• Hardened plantlets are shifted to polybags containing soil, sand, and farmyard manure (2:1:1).
• Maintained under shade net or greenhouse for 30–45 days with regular irrigation and nutrient sprays.
• Plantlets attain robust root systems and 4–6 healthy leaves, suitable for field planting.

d. Nursery Management Practices

• Regular monitoring for pests and diseases.
• Application of water-soluble fertilizers and bio fertilizers.
• Maintaining optimum spacing to avoid etiolation.