HRT4803
PLANT
MICROPROPAGATION
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Course Instructor
Dr. Mohd Hakiman Mansor
Department of Crop Science
Office
Block D, 2nd Floor, Room 19
Faculty of Agriculture, UPM
Contact
Tel. : 03-8947 4903
H/P : 016-222 1070
Email : [email protected]
[email protected]
2
Course Assessment
No. Subject Percent
1 Course work 60
- Test 1 and Test 2 20 + 20
- Culture 20
2 Final Examination 40
Total 100
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Last week lecture
What is tissue culture?
- art and science of plant multiplication and
manipulation in vitro
Characteristics of PTC
- Micro scale, optimized environment, sterile
4
Last week lecture (cont.)
History of tissue culture
Justification – when PTC needed?
- When seed; not uniform, differs to original,
slow growth, difficult to handle, not
produce
- Cutting; slow growth, low survival rate,
high maintenance, easily infected by
disease, limited supply
5
Chapter 2
Basis, Objectives and
Applications of
Plant Micropropagation /
Plant Tissue Culture
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Learning Outcome
Scope:
1. What is the basis of tissue culture?
2. Why do some farmers choose tissue
culture and some are not?
3. Is tissue culture for all?
Learning Outcome:
>>> to learn the of basis, objectives,
advantages and limitations tissue culture
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Botanical Basis for Tissue
Plant cell
Culture
Endoplasmic
recticulum
Nucleolus Chloroplast
Nucleus Cell Wall
Ribosome
Vacuole
Plasma membrane Mitochondria
• totipotent capable of independent deviation
~ complete individual similar original (mother)
plant
• basis of in vitro propagation
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Botanical Basis for Tissue Culture (cont.)
Theoretically,
>>> any part / organs of plant can be tissue
cultured
- stem, bud, root, leaf
leaf petiole leaf bud
leaf section leaf blade
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Objectives of Plant Tissue Culture
To a plant propagator:
1. Elimination of viruses
2. Rapid multiplication of clones
3. Vegetative propagation of difficult to
propagate plant species
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Objectives (cont.)
4. All year round propagation of clones
5. Rapid multiplication of seedlings (in case
where seed is difficult to get)
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Micropropagation Applications
Micropropagation;
>>> rapid build up of new, superior variety
• Production of pathogen free plants through
elimination of endophytic diseases
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Micropropagation Applications (cont)
Crop improvement, plant breeding through
special techniques:
> haploid culture
> in vitro fertilization
> protoplast hybridization
> genetic engineering
• Production of enzymes, flavour, dyes,
fragrances.
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Micropropagation Applications (cont)
Germplasm storage
~ Cryopreservation
In vitro secondary products
Scientific purpose, basic research
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Similarities, Differences, Advantages and
Disadvantages of In vitro and Conventional Systems
Similarities :
1. Both promote axillary growth >> induced
by cytokinin
2. Both promote rooting >> induced by
auxin
3. Both systems conducted in high humidity
environment
4. Resulting plantlets require acclimatisation
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Differences in vitro conventional
Place Container Greenhouse
Sterility Required Not required
Skill Highly Required
both systems require required
knowledge : botany,
physiology, genetic, Medium Nutrient Sand,
and technical skill medium vermiculite
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Advantages of In Vitro System
• Rapid and efficient propagation system
>>> rapid and high multiplication rate
(exponential multiplication)
• Conducted in sterile environment;
free from microorganisms
• Virus free plantlet obtainable through
meristem culture 17
Advantages of In Vitro System (cont.)
Long-term germplasm storage ~ reduce
stock plant space
Does not require management during
growth
Explant used : small plantlets; small
space requirement
Propagate difficult species
e.g.: oil palm, coconut
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Advantages of In Vitro System (cont.)
International exchange made easier
Environment easy to monitor / control
growth in ideal environment
Hybrid production of incompatible species
Can be conducted year round
Genetically identical
New variants
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Advantages of Conventional Propagation
Equipment cost minimal
Experience can substitute for technical
expertise
Cost per unit are minimal
Potential for disease-free plantlets
Specialized techniques for growth control
(grafting onto dwarfing rootstocks)
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Disadvantages of In Vitro System
1. Higher initial cost
2. Require technical skill
3. Require specific procedures
4. Higher cost of finished product
5. Mutation / somaclonal variations possible
6. Acclimatisation and transfer in stages
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Micropropagation Limitations
Equipment / facility intensive operation
Technical expertise in management
positions
Protocols not optimized for all species
May not fit industry standard
Plantlets may be too expensive
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THE END
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