Induction System | Fungal Consortium | Master Thesis | MycoResin Harmonia™
Development and Evaluation of a Fungal Consortium Blended with Fusarium oxysporum for Enhanced Agarwood Resin Induction in Aquilaria malaccensis
A Thesis Submitted to the Graduate School
In Partial Fulfillment of the Requirements for the Degree of
Master of Science (MS)
in
Plant Pathology / Agroforestry / Biotechnology
Author: Manuelito R. Putong
Institution: [University Name]
Adviser: [Adviser Name, PhD]
Year: 2026
ABSTRACT
Agarwood is a highly valuable resinous wood produced by Aquilaria species as a defensive response to injury and microbial invasion. Natural agarwood formation is rare and unpredictable, creating a supply gap in response to increasing global demand. This study focused on the development and evaluation of a fungal consortium blended with Fusarium oxysporum to enhance agarwood resin induction in Aquilaria malaccensis under plantation conditions.
Selected fungi were isolated from naturally resinous agarwood and grouped based on functional traits such as pathogenicity, lignocellulolytic enzyme activity, and elicitation of host defense responses. The fungal consortium was evaluated alone and in combination with F. oxysporum using a controlled inoculation experiment. Resin formation was assessed through visual scoring, gravimetric analysis, and chemical profiling using gas chromatography–mass spectrometry (GC–MS).
Results showed that trees inoculated with the fungal consortium blended with F. oxysporum produced significantly higher resin yield and improved sesquiterpene diversity compared with single-strain and uninoculated controls. The findings demonstrate that synergistic fungal interactions enhance agarwood resin biosynthesis and provide a biologically based, scalable approach for sustainable agarwood production.
KEYWORDS
Agarwood, Aquilaria malaccensis, Fusarium oxysporum, fungal consortium, resin induction, plant–microbe interaction
CHAPTER 1 – INTRODUCTION
1.1 Background of the Study
Agarwood, also known as oud or gaharu, is a fragrant resinous heartwood formed in trees of the genus Aquilaria. The resin develops as part of the tree’s defense mechanism following wounding and microbial infection, leading to the accumulation of sesquiterpenes and chromone derivatives. Due to overexploitation of wild resources and the slow, uncertain nature of natural resin formation, agarwood-producing species have been listed under CITES Appendix II.
Commercial plantations have emerged as a sustainable alternative; however, plantation-grown Aquilaria trees require artificial induction to produce economically viable agarwood. Among biological induction methods, Fusarium oxysporumhas been widely used, yet resin yield and quality remain inconsistent. Evidence from naturally formed agarwood suggests that multiple fungal species are involved in resin formation, indicating potential benefits of a consortium-based approach.
1.2 Statement of the Problem
Single-fungal induction systems often result in uneven resin distribution, low resin density, and variable chemical composition. There is limited experimental research evaluating the use of fungal consortia for agarwood resin induction under plantation conditions.
1.3 Objectives of the Study
General Objective
To develop and evaluate a fungal consortium blended with Fusarium oxysporum for enhanced agarwood resin induction in Aquilaria malaccensis.
Specific Objectives
- To isolate and identify fungi associated with naturally formed agarwood.
- To formulate a compatible fungal consortium based on functional traits.
- To compare resin formation among consortium-based, single-strain, and control treatments.
- To analyze the chemical composition of induced agarwood resin.
- To assess tree health following inoculation.
1.4 Significance of the Study
The study provides scientific support for a multi-fungal induction strategy that can improve agarwood yield and quality. The findings are relevant to agarwood farmers, researchers, and conservation-oriented plantation developers.
1.5 Scope and Limitations
The research focused on Aquilaria malaccensis grown under plantation conditions. The study evaluated resin formation up to 18–24 months post-inoculation and did not include molecular-level gene expression analysis.
CHAPTER 2 – REVIEW OF RELATED LITERATURE
2.1 Biology of Aquilaria malaccensis
Aquilaria malaccensis is a tropical tree species native to Southeast Asia. Agarwood formation occurs within the xylem as resin accumulates in response to prolonged stress and microbial colonization.
2.2 Chemical Composition of Agarwood Resin
Agarwood resin primarily consists of sesquiterpenes and 2-(2-phenylethyl) chromones, which are responsible for its fragrance and commercial value.
2.3 Fungal Communities Associated with Agarwood
Studies have identified various fungi in agarwood tissues, including species of Fusarium, Aspergillus, Penicillium, Lasiodiplodia, and Cytospora. These fungi are believed to contribute to resin biosynthesis by inducing host defense pathways.
2.4 Fusarium oxysporum as an Agarwood Inducer
F. oxysporum induces agarwood formation by colonizing vascular tissues and triggering oxidative stress responses. However, excessive pathogenicity may negatively affect tree health.
2.5 Rationale for Fungal Consortium Use
A fungal consortium allows complementary interactions among different functional groups, potentially enhancing resin induction while moderating pathogenic effects.
CHAPTER 3 – MATERIALS AND METHODS
3.1 Study Site and Experimental Design
The study was conducted in a managed Aquilaria malaccensis plantation using a randomized complete block design with four treatments and replicates per treatment.
3.2 Fungal Isolation and Identification
Fungal isolates were obtained from naturally resinous agarwood samples and identified using morphological characteristics and ITS rDNA sequencing.
3.3 Consortium Formulation
Fungi were selected based on pathogenicity, enzyme activity, and compatibility with F. oxysporum. Compatibility testing was conducted prior to consortium formulation.
3.4 Inoculation Procedure
Trees were inoculated at breast height using a drill-and-inject technique. Spore suspensions were standardized at approximately 10⁶ spores mL⁻¹.
3.5 Assessment of Resin Formation
Resin formation was evaluated through visual scoring, wood core sampling, and gravimetric determination of resin content.
3.6 Chemical Analysis
GC–MS was used to identify and compare sesquiterpene profiles among treatments.
3.7 Statistical Analysis
Data were analyzed using analysis of variance (ANOVA), and mean comparisons were conducted at a 5% significance level.
CHAPTER 4 – RESULTS AND DISCUSSION
4.1 Resin Yield and Distribution
Trees inoculated with the fungal consortium blended with F. oxysporum exhibited significantly higher resin content and more uniform distribution compared with single-strain and control treatments.
4.2 Chemical Profile of Induced Resin
Consortium-treated samples showed increased diversity and abundance of commercially important sesquiterpenes.
4.3 Effects on Tree Health
No severe decline symptoms were observed, indicating that the consortium approach maintained a balance between induction efficacy and tree viability.
4.4 Comparison with Previous Studies
The findings align with previous reports on the role of multi-fungal interactions in natural agarwood formation and extend these observations to plantation systems.
CHAPTER 5 – CONCLUSIONS AND RECOMMENDATIONS
5.1 Conclusions
The fungal consortium blended with Fusarium oxysporum significantly enhanced agarwood resin induction in Aquilaria malaccensis compared with single-fungal inoculation. The approach offers a promising biological induction strategy for sustainable agarwood production.
5.2 Recommendations
Further research should evaluate longer induction periods, larger field trials, and integration with chemical or mechanical induction methods.
REFERENCES
[To be completed following APA or institutional format]
APPENDICES
A. Detailed Inoculation Protocol
B. GC–MS Chromatograms
C. Statistical Analysis Tables