Tissue Culture

Tissue Culture Specifications

Aseptic (Sterile) Conditions

• All procedures must be carried out in a contamination-free environment
• Use of laminar airflow cabinet, sterilized tools, and disinfected explants

Suitable Explant

• A small piece of plant tissue (leaf, stem, root, meristem, etc.)
• Must be healthy, disease-free, and actively growing

Nutrient Culture Medium

• Provides essential nutrients for growth
• Contains:
  • Macronutrients (N, P, K, Ca, Mg)
  • Micronutrients (Fe, Mn, Zn, etc.)
  • Carbon source (usually sucrose)
  • Vitamins and amino acids
  • Plant growth regulators (auxins, cytokinins)
  • Solidifying agent (agar, if solid medium)

Controlled Environmental Conditions

• Temperature: usually 20–27°C
• Light: controlled photoperiod (light/dark cycle)
• Humidity: maintained for proper growth
• pH of medium: around 5.6–5.8

Growth Regulators

• Hormones regulate development:
  • Auxins → root formation
  • Cytokinins → shoot formation
• Proper balance is essential for desired growth

Subculturing

• Transfer of growing tissue to fresh medium at regular intervals
• Prevents nutrient depletion and maintains growth

Hardening (Acclimatization)

• Gradual adaptation of plantlets to external environment
• Transferred from lab to soil under controlled conditions

Equipment Required

• Laminar airflow chamber
• Autoclave (for sterilization)
• Culture bottles or tubes
• Growth room or incubator