Tissue culture is an advanced propagation technique used to produce genetically uniform, disease-free, and fast-growing Aquilaria malaccensis plants. This method ensures a sustainable supply of high-quality agarwood-producing trees while conserving wild populations.
Step-by-Step Tissue Culture Process for Aquilaria malaccensis
1. Selection of Plant Material (Explant Preparation)
✔ Source: Young, healthy shoots, nodes, or immature seeds from a mature Aquilaria malaccensis tree.
✔ Disinfection:
- Wash explants under running water for 15–30 minutes.
- Soak in 0.1% Mercuric Chloride (HgCl₂) or Sodium Hypochlorite (NaOCl) for 10–15 minutes.
- Rinse 3–5 times with sterile distilled water to remove any chemical residue.
2. Establishment of Explants in Culture Media
✔ Media Selection: Murashige and Skoog (MS) medium is commonly used.
✔ Growth Regulators:
- For Shoot Induction: Cytokinins (BAP or Kinetin) at 0.5–2.0 mg/L.
- For Root Induction: Auxins (NAA or IBA) at 0.1–1.0 mg/L.
✔ pH Adjustment: Adjust medium pH to 5.7–5.8 before autoclaving.
✔ Sterilization: Autoclave media at 121°C for 15–20 minutes.
Explants should be transferred under sterile conditions in a laminar airflow chamber to prevent contamination.
3. Shoot Multiplication (Proliferation Phase)
✔ Subculturing: Transfer growing shoots to fresh MS medium every 3–4 weeks.
✔ Growth Enhancers: Adding 0.5–1.0 mg/L BAP enhances shoot proliferation.
✔ Light & Temperature Conditions: Maintain at 25°C ± 2°C, with 16-hour photoperiod (light:dark cycle).
Multiple shoots will develop from a single explant, increasing the propagation rate!
4. Root Induction & Plantlet Development
✔ Transfer well-developed shoots to MS medium with auxins (IBA or NAA at 0.5–1.0 mg/L) for root formation.
✔ Keep cultures in dark conditions for the first 5–7 days to stimulate root growth.
✔ After root emergence, shift to normal light conditions (16-hour photoperiod).
5. Hardening & Acclimatization
✔ Once plantlets develop strong roots, transfer them to sterile soil mix (peat moss + vermiculite + perlite in a 1:1:1 ratio).
✔ Place in a high-humidity chamber or greenhouse for 2–4 weeks to adjust to external conditions.
✔ Gradually expose plants to normal environmental conditions before transferring to the field.
Key Takeaways
✔ Tissue culture allows rapid multiplication of Aquilaria malaccensis for large-scale agarwood plantations.
✔ Proper sterilization and growth regulator balance are critical for success.
✔ Acclimatization is essential to prevent transplant shock and ensure survival.