1. MICROPROPAGATION

2. Introduction In nature plants propagate either
Sexually (seeds generation) results heterogeneity Or
Asexually (vegetative multiplication) produce genetically identical plants.
Multiplication of genetically identical copies of a cultivar by asexual reproduction is called clonal propagation.
Via tissue culture called micropropagation,

3. What is Micropropagation?
“… the asexual or vegetative propagation (multiplication) of plants in vitro “
Implies - regeneration
- multiplication
- uniformity ??

4. Basic in vitro propagation ...

5. Micropropagation (contin)
Positives and negatives of micropropagation
positives
rapid multiplication rates
low space requirement
negatives
labor costs
high overhead (equipment, facilities, supplies)
loss by contamination
danger of variation

6. STAGES 2. Establish aseptic culture 3. Multiplication
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization

7. 2. Establish aseptic culture 3. Multiplication 4. Shoot elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization

8. Starting material for micropropagation
Tip bud
Starting material for micropropagation
Leaf
Axillary bud
Internode
Root

10. Selection of plant material ...
Part of plant
Genotype
Physiological condition
Season
Position on plant
Size of explant

11. Physiological state - of stock plant
Vegetative / Floral
Juvenile / Mature
Dormant / Active
Carbohydrates
Nutrients
Hormones

12. 2. Establish aseptic culture 3. Multiplication 4. Shoot elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization

14. Disinfestation Stock plant preparation Washing in water
Disinfecting solution
Internal contaminants
Screening

15. Organic ‘growth factors’ Growth regulators Gelling agent
The medium
Minerals
Sugar
Organic ‘growth factors’
Growth regulators
Gelling agent
Other additives

16. Physical Environment
Temperature
Moisture
Light

17. 2. Establish aseptic culture 3. Multiplication 4. Shoot elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization

18. Lateral / Axillary buds Adventitious (de novo, re-differentiation)
Origins of new shoots ...
Terminal extension
Lateral / Axillary buds
Adventitious (de novo, re-differentiation)
Callus differentiation

19. Role of growth regulators ...
Cell division
Differentiation
Cell expansion
Apical dominance
auxins
abscisic acid
cytokinins
gibberelic acid
ethylene

20. 2. Establish aseptic culture 3. Multiplication 4. Shoot elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization

21. Carry-over of hormones
Shoot elongation ...
Basal ‘hormone free’ medium
Gibberellins
Carry-over of hormones

22. 2. Establish aseptic culture 3. Multiplication 4. Elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Elongation
5. Root induction / formation
6. Acclimatization

23. Juvenility / rejuvenation Genotype
Root initiation ...
Auxins
Co-factors
C : N ratio
Light / darkness
Initiation vs growth
Juvenility / rejuvenation
Genotype

24. 2. Establish aseptic culture 3. Multiplication 4. Elongation
STAGES
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Elongation
5. Root induction / formation
6. Acclimatization

25. - survival of the new plant when removed from the in vitro environment
Acclimatization (hardening)
- survival of the new plant when removed from the in vitro environment

26. Micropropagation of almost all the fruit crops and vegetables is possible
Some examples: dwarfing sweet cherry, Shade trees, Ornamental shrubs, Roses, Clematis, Lilacs, Saskatoon berries, Nutraceutical Plants, Rhododendron, Azalea, mustard, corn, soybeans, wheat, rice, cotton, tomato, potato, citrus, turf, legumes

27. Advantages of Micropropagation
economical in time and space
greater output -can produce millions of uniformly flowering and yielding plants
African Biotechnologies - fruit crops banana and indoor pot flowers- 6 million pieces per year
disease free
elite plants with exceptional characteristics

28. Advantages Cont’d
facilitates safer movements of germplasm across nations - In vitro germplasm assures the exchange of pest and disease free material
great for
vegetatively reproduced crops
crops which produce few seeds or highly heterozygous seeds.

29. Uses of Micropropagation
Used to create transgenic, first generation plants
Used in horticulture to produce orchids, African Violets, lilies, and ferns
Used in nurseries to grow fruit trees, evergreens, roses, and shade trees

30. Benefits of Micropropagation
Many genetically identical plants can be created from one parent plant
Because plants are clones, the uniformity assures quality
Allows many plants to grow in a small place in a short time
In some species this method will produce healthier plants

31. Thank
You