Plant Tissue Culture - General Procedure & Definitions

Plant Tissue Culture (PTC) is the technique of growing plant cells, tissues, or organs in a sterile, nutrient-rich medium under controlled conditions.
It is used for micropropagation, genetic modification, conservation, and medicinal compound production.

Selection of Explant
- A small plant part (leaf, stem, root, meristem, embryo) is chosen for culture.
Sterilization (Aseptic Conditions)
- Explants are cleaned and sterilized using ethanol, mercuric chloride (HgCl₂), or sodium hypochlorite (NaOCl) to remove microbes.
- This process is performed in a laminar airflow chamber to prevent contamination.
Preparation of Culture Media
- Murashige and Skoog (MS) medium is commonly used, containing macronutrients, micronutrients, vitamins, sucrose, and agar.
- Growth hormones:
  - Auxins (IAA, NAA, 2,4-D) → Root and callus formation.
  - Cytokinins (BAP, Kinetin) → Shoot formation.
Inoculation
- Explants are placed onto the sterile medium in Petri dishes or test tubes.
Incubation
- Cultures are maintained at:
  - Temperature: 22-27°C
  - Light cycle: 16-hour light/8-hour dark
  - Humidity: 50-60
  - Cells grow into callus, shoots, or roots.
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Subculturing
- Tissues are transferred to fresh media every 2-4 weeks to maintain growth.
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Regeneration and Rooting
- Shoots and roots develop using cytokinins (for shoot formation) and auxins (for root formation).
Acclimatization (Hardening)
- Plantlets are moved to a greenhouse or soil for adaptation to external conditions.
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Transfer to Field
- Fully grown plants are transplanted into the natural environment.