Aquilaria malaccensis – Crown Agroforestry Group

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.