Here’s a professional, course-ready section on MS, WPM, and Customized Agarwood Media for COPI’s Biotechnological Propagation Course, including rationale, composition, and applications in Aquilaria tissue culture.
Culture Media for Agarwood (Aquilaria spp.) Tissue Culture
1. Overview
The choice of culture medium is critical for explant survival, growth, and organogenesis. Media provide:
- Essential macro- and micronutrients
- Vitamins, amino acids, and growth regulators
- A physiological environment conducive to regeneration
Common media in Aquilaria tissue culture:
- Murashige and Skoog (MS) medium
- Woody Plant Medium (WPM)
- Customized Agarwood Media (CAM) – optimized for Aquilaria species
2. Murashige and Skoog (MS) Medium
2.1 Overview
- Widely used general-purpose plant tissue culture medium
- High salt concentration suitable for fast-growing, herbaceous tissues
2.2 Composition Highlights
| Component | Typical Concentration |
|---|---|
| Nitrate (NO₃⁻) | 60 mM |
| Ammonium (NH₄⁺) | 20 mM |
| Potassium (K⁺) | 20 mM |
| Phosphate (PO₄³⁻) | 1.25 mM |
| Micronutrients | Fe, Mn, Zn, Cu, Mo, B |
- Vitamins: Thiamine, Nicotinic acid, Pyridoxine
- Carbon source: 3% sucrose
2.3 Applications in Aquilaria
- Nodal and shoot tip culture
- Organogenesis experiments
- Seedling culture in early propagation
2.4 Advantages
- Well-characterized, widely available
- Supports vigorous growth and shoot multiplication
2.5 Limitations
- High salt may cause hyperhydricity in woody species like Aquilaria
3. Woody Plant Medium (WPM)
3.1 Overview
- Developed by Lloyd & McCown (1981) for woody plant species
- Lower salt concentration than MS
- Optimized for slow-growing woody tissues
3.2 Key Features
- Lower total ionic strength → reduced stress on woody explants
- Adjusted micronutrients for woody plant physiology
- Supports direct organogenesis and callus induction
3.3 Applications in Aquilaria
- Nodal culture of mature trees
- Callus induction from leaf or hypocotyl explants
- Reduced tissue necrosis in long-term cultures
4. Customized Agarwood Media (CAM)
4.1 Rationale
Aquilaria species have species-specific nutrient requirements for:
- Optimized shoot multiplication
- Healthy rooting and acclimatization
- Improved resin biosynthesis potential
4.2 Example Customization Parameters
| Parameter | Adjustment |
|---|---|
| Macronutrients | Modified N:K ratio to reduce hyperhydricity |
| Vitamins | Addition of glycine, casein hydrolysate, or coconut water |
| Growth regulators | Tailored auxin:cytokinin ratios for direct/indirect organogenesis |
| Carbon source | Sucrose 2–3%, optional glucose or fructose |
| pH | Adjusted to 5.6–5.8 for Aquilaria tissues |
4.3 Applications
- Shoot multiplication of elite mother plant clones
- Callus induction for somatic embryogenesis research
- Pilot-scale propagation for plantation deployment
4.4 Advantages
- Species-specific optimization improves survival and regeneration
- Enhances resin-yield potential in propagated plantlets
- Reduces tissue necrosis and contamination
5. Practical Tips for COPI Laboratories
- Always prepare media fresh or store properly to prevent contamination
- Autoclave media at 121°C for 15–20 min
- Add heat-sensitive PGRs after autoclaving via sterile filtration
- Monitor pH and osmotic conditions to suit Aquilaria explants
- Maintain a media log for traceability
6. Course-Ready Key Statement
The choice of culture medium—MS, WPM, or Customized Agarwood Media—directly influences the success of Aquilaria tissue culture. Customized media tailored to woody plant physiology, optimal nutrient balance, and species-specific growth regulators maximize regeneration, maintain genetic fidelity, and enhance resin production potential.
I can next:
- Create a visual media comparison chart (MS vs WPM vs CAM) for the course
- Include PGR recommendations per explant type
- Make a lab SOP poster for media preparation
Do you want me to create the infographic version next?