Magnolia champaca

Here is a comprehensive tissue culture protocol for Magnolia champaca (Golden Champaca), a fragrant flowering tree valued for its essential oils and ornamental use. Like many woody plants, it presents challenges due to high phenolic content and slow in vitro response—but tissue culture offers a sustainable way to propagate elite lines.

???? Tissue Culture Protocol for Magnolia champaca

1. Explant Selection:

  • Preferred: Nodal segments, shoot tips, or immature zygotic embryos
  • Source: Seedlings or juvenile plants (less phenolic exudation)

2. Surface Sterilization:

  • Wash explants under running water for 20–30 minutes
  • Dip in 70% ethanol for 30 seconds
  • Sterilize in:
  • 0.1% HgCl₂ for 5–8 min or
  • 1.0% sodium hypochlorite + 1 drop Tween-20 for 10–15 min
  • Rinse thoroughly with sterile distilled water (3–5 times)

3. Initiation Medium:

  • Basal medium: MS (Murashige & Skoog)
  • Supplements:
  • BAP (1.0–2.0 mg/L) – shoot induction
  • NAA (0.1–0.5 mg/L) – promotes callus or shoot base swelling
  • Ascorbic acid (100 mg/L) or citric acid – to minimize browning
  • Activated charcoal (0.1–0.2%) may help in reducing phenolics

4. Shoot Multiplication:

  • Subculture to MS medium with:
  • BAP (2.0 mg/L) + Kinetin (0.5–1.0 mg/L)
  • Optional: GA₃ (0.1 mg/L) to elongate shoots
  • Repeat subculturing every 4 weeks
  • Maintain under 25 ± 2°C, 16-hour photoperiod (cool white fluorescent light)

5. Rooting:

  • Use ½-strength MS or WPM (Woody Plant Medium)
  • Auxins:
  • IBA (1.0–2.0 mg/L) or NAA (0.5–1.0 mg/L)
  • Add 0.2% activated charcoal to enhance rooting
  • Rooting can take 3–5 weeks

6. Acclimatization:

  • Transfer rooted plantlets to trays with sterilized cocopeat + vermiculite/sand
  • High humidity under dome or mist chamber for first 2–4 weeks
  • Gradually reduce humidity and increase light exposure over another 2–4 weeks

✅ Important Tips:

  • Use young explants to reduce phenolic browning.
  • Activated charcoal is useful but may adsorb growth regulators—balance carefully.
  • Subculture regularly to prevent vitrification and tissue necrosis.

Would you like a flowchart diagram or a combined PDF protocol sheet for M. champaca, M. fragrans, and C. verum?

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