Induction System | Biotech Method | Undergrad Thesis | Enzyme Blend | FusaBlaze™
UNDERGRADUATE THESIS PROPOSAL
Title:
Advanced Biotechnological Induction of Agarwood Using Selected Microbial Consortia and Enzymatic Triggers in Aquilaria spp.
1. Introduction
Agarwood is one of the most valuable aromatic natural products globally, formed as a result of a complex biochemical defense response in Aquilaria species following injury and microbial infection. Natural agarwood formation may take decades and occurs unpredictably, leading to supply shortages and unsustainable harvesting practices.
Recent advances in biotechnology and microbial ecology suggest that controlled inoculation using specific resin-inducing microbes and their enzymes can stimulate agarwood resin biosynthesis in a shorter and more sustainable timeframe. These advanced methods aim to replicate natural biochemical pathways while maintaining chemical authenticity.
This study proposes to investigate an advanced biotech induction method using selected fungal and bacterial strains and key enzymes to induce agarwood formation and evaluate resin quality using chemical analysis.
2. Statement of the Problem
Conventional agarwood induction methods often rely on mechanical wounding or chemical inducers, which may result in:
- Inconsistent resin quality
- Lower chromone content
- Tree stress or mortality
- Reduced market acceptance
There is limited undergraduate-level research on microbe-enzyme-based induction systems that integrate biological specificity with biochemical control.
3. Objectives
General Objective
To evaluate the effectiveness of selected microbes and their enzymes in inducing high-quality agarwood resin in Aquilariaspp. using an advanced biotechnological approach.
Specific Objectives
- To identify microbial strains capable of stimulating agarwood resin biosynthesis
- To assess the role of selected enzymes in enhancing resin formation
- To compare resin yield and quality among different induction treatments
- To analyze chemical profiles of induced resin using GC-MS
- To compare induced resin profiles with reference natural agarwood markers
4. Significance of the Study
- Contributes to sustainable agarwood production
- Supports scientific validation of biological induction
- Provides data for quality grading and traceability
- Benefits agarwood farmers, researchers, and the perfumery industry
- Aligns with eco-friendly and ethical sourcing initiatives
5. Scope and Limitations
Scope
- Conducted on young to mature Aquilaria spp. trees
- Focus on controlled inoculation using microbes and enzymes
- Chemical analysis limited to GC-MS profiling
Limitations
- Time-bound resin development
- Limited number of microbial strains
- Environmental variability
6. Review of Related Literature (Summary)
Studies have demonstrated that fungal pathogens such as Fusarium oxysporum and Lasiodiplodia theobromae induce sesquiterpene and chromone production. Enzymes such as cellulases, laccases, and peroxidases facilitate tissue penetration and oxidative stress, triggering secondary metabolite pathways.
However, few studies integrate microbial consortia with enzyme supplementation to optimize resin induction.
7. Methodology
7.1 Research Design
Experimental, Completely Randomized Design (CRD)
7.2 Materials
- Aquilaria spp. trees
- Selected microbes:
- Fusarium oxysporum
- Lasiodiplodia theobromae
- Beneficial bacteria (Bacillus spp.)
- Enzymes:
- Cellulase
- Laccase
- Peroxidase
- Sterile inoculation equipment
- GC-MS instrumentation
7.3 Treatments
| Treatment | Description |
|---|---|
| T1 | Mechanical wound (Control) |
| T2 | Microbial inoculation only |
| T3 | Enzyme application only |
| T4 | Microbial + enzyme induction |
| T5 | Untreated control |
7.4 Inoculation Procedure
- Drilling standardized holes
- Inoculating microbial cultures
- Applying enzyme solutions
- Sealing inoculation points
- Monitoring over 6–12 months
7.5 Data Collection
- Visual resin formation
- Resin yield
- Tree health indicators
- Chemical profile (GC-MS)
- Sesquiterpene and chromone content
8. Data Analysis
- ANOVA for treatment comparison
- GC-MS chromatogram interpretation
- Principal Component Analysis (optional)
- Comparative chemical fingerprinting
9. Expected Results
- Microbial-enzyme treatments will produce higher resin yield
- Increased chromone diversity
- Chemical profiles closer to natural agarwood
- Improved consistency and sustainability
10. Ethical & Biosafety Considerations
- Use of non-invasive microbial strains
- Controlled inoculation protocols
- Environmental containment
- Institutional biosafety approval
11. Proposed Timeline
| Activity | Duration |
|---|---|
| Literature review | 1 month |
| Culture preparation | 1 month |
| Inoculation | 1 month |
| Resin development | 6–9 months |
| Chemical analysis | 1 month |
| Thesis writing | 2 months |
12. Expected Output
- Undergraduate thesis manuscript
- GC-MS chemical fingerprint database
- Recommendations for sustainable induction
13. Conclusion
This study seeks to bridge traditional agarwood knowledge with modern biotechnology by developing a scientifically controlled, microbe-enzyme-based induction method that enhances resin quality while promoting sustainability.