Plant Tissue Culture

What is Plant Tissue Culture?

• 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.

General Procedure

1. Selection of Explant

   • A small plant part (leaf, stem, root, meristem, embryo) is chosen for culture.

2. 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.

3. 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.

4. Inoculation

   • Explants are placed onto the sterile medium in Petri dishes or test tubes.

5. 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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6. Subculturing

   • Tissues are transferred to fresh media every 2-4 weeks to maintain growth.
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7. Regeneration and Rooting

   • Shoots and roots develop using cytokinins (for shoot formation) and auxins (for root formation).

8. Acclimatization (Hardening)

   • Plantlets are moved to a greenhouse or soil for adaptation to external conditions.
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9. Transfer to Field

   • Fully grown plants are transplanted into the natural environment.

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