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Saintpaulia ionantha
African Violet

Difficulty: ⭐ BeginnerSuccess Rate: 80-90%Explant: Leaf Petiole

African violets are the gold standard for beginner tissue culture. They respond excellently to standard protocols, have high success rates, and provide clear visual feedback on technique quality.

The protocol leverages the natural regenerative capacity of Saintpaulia leaf tissue. Callus forms readily at the petiole base, followed by organized shoot development with minimal hormone requirements.

This species demonstrates exceptional acclimatization characteristics. Rooted plantlets transfer easily to soil-based substrate with minimal shock or mortality, making it ideal for learning proper techniques.

Why Start with African Violets?

African violets are highly recommended for beginners because they:

  • High Success Rates: Respond excellently to standard protocols with 80-90% success
  • Clear Feedback: Visual indicators show technique quality immediately
  • Forgiving Nature: Tolerate minor protocol variations without failure
  • Fast Response: Shoots appear within 2-3 weeks, providing quick results
  • Easy Acclimatization: Transfer to soil with minimal shock or mortality

Materials Needed

  • Healthy, non-flowering African violet plant
  • 70% ethanol for pre-treatment
  • 15% bleach solution (0.75% sodium hypochlorite)
  • Surfactant (1-2 drops liquid soap per 100 mL bleach solution)
  • MS medium with appropriate hormones
  • Sterile distilled water
  • Sterile tools (forceps, scalpel)
  • Still Air Box or laminar flow hood

Step 1: Explant Selection & Preparation

Choose Optimal Explants:

  • Select healthy, mature leaves with petioles attached
  • Avoid flowering plants (reduces success rates)
  • Use leaves from disease-free, vigorous plants
  • Morning harvest provides best results

Initial Cleaning:

  • Rinse explants under running tap water
  • Remove any visible debris or damaged tissue
  • Pat gently dry with paper towels
  • Prepare for surface sterilization immediately

Step 2: Surface Sterilization Protocol

  1. Alcohol Pre-treatment
    Immerse explants in 70% ethanol for . Agitate gently to ensure complete coverage. This breaks surface tension for better bleach penetration.
  2. Primary Sterilization
    Transfer to 15% bleach solution with surfactant. Duration: . Ensure complete submersion and agitate every 5 minutes for uniform treatment.
  3. Sterile Rinses
    Rinse 3 times minimum in sterile distilled water: First rinse , second rinse , third rinse . Additional rinses if bleach odor persists.

Step 3: Culture Initiation

Explant Preparation:

  • Work within sterile environment ( SAB or laminar flow)
  • Cut petiole to 2-3 cm length using sterile scalpel
  • Make fresh cuts on leaf base to expose Meristematic tissue tissue
  • Handle gently to avoid tissue damage

Initiation Medium Formula:

  • MS basal medium
  • Sucrose 30 g/L
  • BAP ( Cytokinin ) 1.0 mg/L
  • IAA ( Auxin ) 0.1 mg/L
  • Agar 8 g/L
  • pH pH 5.6-pH 5.8

Culture Conditions:

  • Temperature: 22°C-25°C
  • 16-hour photoperiod with moderate light intensity
  • Sealed culture vessels to maintain high humidity

Step 4: Multiplication Phase (4-6 weeks)

Growth Monitoring:

  • Shoots typically appear after 2-3 weeks
  • Multiple shoots develop from petiole base
  • Healthy shoots show bright green coloration
  • Watch for contamination signs daily

Subculturing Process:

  • Transfer to fresh multiplication medium every 4-6 weeks
  • Carefully separate individual shoots
  • Each shoot can be cultured as independent Explant
  • Maintain sterile technique during transfers

Step 5: Rooting (2-4 weeks)

Natural Rooting:

African violets often develop roots spontaneously on multiplication medium. If rooting occurs, proceed directly to Acclimatization .

Dedicated Rooting Medium (if needed):

  • Half-strength MS medium (2.22 g/L)
  • Sucrose 20 g/L (reduced from multiplication)
  • Hormone-free or very low hormone concentration
  • Agar 8 g/L
  • pH pH 5.6-pH 5.8

Transfer well-developed shoots to rooting medium. Roots typically develop within 2-4 weeks.

Step 6: Acclimatization (2-3 weeks)

Root Cleaning

Gently remove all agar from roots under running water. Avoid damaging delicate root hairs. Pat dry gently with paper towel.

Transplanting

Use sterile potting mix (peat/perlite/vermiculite blend). Small pots (2-3 inches) for initial transplant. Plant at same depth as in culture.

Humidity Management

Cover with clear plastic bag or container. Maintain high humidity initially (90-95%). Gradually increase ventilation over 2-3 weeks. Monitor for wilting and adjust accordingly.

Success Indicators Timeline

Timeline Expected Results
Week 1-2 No contamination present, explants remain green and healthy
Week 3-4 Shoot primordia visible at petiole base, small green bumps forming
Week 5-8 Multiple shoots developing, bright green coloration, 1-2 cm height
Week 9-12 Rooted plantlets ready for acclimatization, well-developed root systems

Troubleshooting Common Problems

Problem Solution
No Shoot Formation Increase BAP concentration to 1.5-2.0 mg/L. Ensure fresh medium and proper light conditions. Check explant quality.
Excessive Callus Reduce IAA concentration or increase BAP ratio. Transfer to fresh medium with adjusted hormones.
Poor Rooting Try IBA at 0.5-1.0 mg/L in rooting medium. Ensure shoots are well-developed before transfer. Check temperature and light.
Acclimatization Loss Maintain higher humidity longer (95%+ initially). Gradual reduction over 3-4 weeks. Ensure proper root development before transfer.

Key Success Factors

Critical Success Points:

  • Use healthy, non-flowering donor plants
  • Maintain strict sterile technique throughout
  • Monitor cultures daily for contamination
  • Provide consistent environmental conditions
  • Be patient—shoots take 2-3 weeks to appear

Why This Protocol Works:

  • Balanced hormone ratios promote organized development
  • Proper sterilization ensures clean cultures
  • Gradual acclimatization prevents shock
  • Species-specific conditions optimize growth
  • Clear timeline helps track progress