Microbial Enzyme | FusaBlaze™ | FusaBlaze™ Formulation | FusaBlaze™ Development | FusaBlaze™ Profile
BarIno™ FusaBlaze™ Microbial-Enzyme Synergy for Premium Agarwood Induction
I. OBJECTIVE
To formulate a live microbial inoculant utilizing Fusarium oxysporum (F. oxysporum) for inducing agarwood resin in Aquilaria trees by activating natural defense pathways through controlled fungal infection and enzymatic activity.
II. ACTIVE INGREDIENTS (Per Liter Batch)
| Component | Function | Amount |
|---|---|---|
| Fusarium oxysporum spores/culture | Primary microbial inducer | ≥10⁸ CFU/mL |
| Molasses or Glucose | Carbon source for fungal growth | 20 g |
| Soybean meal or Peptone | Protein/nitrogen source | 10 g |
| Yeast extract (optional) | Growth stimulant and vitamin source | 2 g |
| Laccase & Peroxidase Enzyme Mix | Lignin-degrading enzyme support | 1–2 mL or 100 U/mL |
| Tween 80 (emulsifier) | Enhances penetration and dispersion | 0.5–1.0 mL |
| Sterile distilled water | Solvent | To make 1,000 mL |
III. PREPARATION PROCEDURE
A. Culture Propagation (3–5 Days)
- Prepare Fungal Pre-Culture
- Inoculate F. oxysporum onto Potato Dextrose Agar (PDA) or Sabouraud Dextrose Agar (SDA) plate.
- Incubate at 25–28°C for 3–5 days until mycelial growth and sporulation.
- Prepare Liquid Culture Medium
- Mix 20 g molasses, 10 g soybean meal/peptone, and 2 g yeast extract in 1 liter of distilled water.
- Adjust pH to 5.8–6.2. Sterilize via autoclaving (121°C, 15 psi, 20 minutes).
- Inoculate Liquid Medium
- Transfer several agar plugs (6–8 mm) or a spore suspension (1 mL, 10⁷ spores/mL) into the sterile medium.
- Incubate at 28°C in a shaker (150 rpm) or static for 3–5 days.
B. Enzyme Addition & Formulation
- Filter or Decant Culture (Optional)
- Remove fungal biomass if a clarified enzyme-rich broth is desired.
- For full-spectrum bioactivity, leave mycelial content intact.
- Add Enzyme Blend
- Mix in purified laccase + peroxidase enzymes to enhance resin induction (1–2 mL/L or as per manufacturer activity unit).
- Add 0.5–1.0 mL Tween 80 as surfactant.
- Final Volume Adjustment
- Bring to 1,000 mL with sterile distilled water.
- Shake well before bottling.
IV. PACKAGING & STORAGE
| Parameter | Value |
|---|---|
| Container Type | Amber HDPE bottle, vented cap |
| Storage Temperature | 4–10°C (refrigerated), away from light |
| Shelf Life | 3–6 months (under refrigeration) |
| Label Details | Batch No., CFU count, expiry, usage |
V. APPLICATION GUIDELINES
| Target Tree | Aquilaria malaccensis, A. crassna, etc. |
|---|---|
| Minimum Age | 5 years (≥10 cm DBH) |
| Application Method | Drill-and-inject |
| Hole Dimensions | 6–8 mm wide, 4–6 cm deep |
| Spacing | 15–20 cm spiral pattern |
| Dose per Hole | 10–15 mL |
| Reinoculation | Optional after 6–9 months |
VI. SAFETY & HANDLING
- Use gloves and face mask while handling fungal cultures.
- Sterilize tools after use.
- Avoid inhaling spores or aerosolized liquid.
- Do not apply during rain or extreme heat.
VII. NOTES & VARIATIONS
- Optionally combine with FeCl₃ microdoses (1–2 g/L) for synergistic stress effect (as in BarIno™ Dual-Action protocols).
- For solid-state carriers (sawdust or corn grit), fungal culture can be adapted for field inoculation plugs.