Micropropagation

Micropropagation
• Micropropagation is the production
of whole plants from small sections
of plant such as a shoot tip, node,
meristem, embryo, or even a seed
• Plant tissue culture is basically the
same thing, except that it implies the
use of callus tissue generated from
plant cells cultured in-vitro.
 Micropropagation
• Why does micropropagation work?
– Plant cells have the ability to reproduce
the whole plant from single cells. This is
called totipotency.
– Totipotency is the ability of a single cell to
express the full genome in the cells to
which it gives rise by cell division.
 Micropropagation
• What part of the plant can be used for
micropropagation?
– Any part of a plant can be used.
– Meristems, shoot and root tips, leaf
tissue, anthers, embryos, flowers, all
parts of a plant can be used.
 Micropropagation
• The only limitation is that each plant is
propagated differently and not every
plant will respond the same way.
• Each genus, species and variety may
require a different tissue which will
obtain the best results.
Tip bud
Starting material for
micropropagation
Leaf

Axillary
bud

Internode

Root
 Micropropagation Methods
• Micro cuttings
• Small sections of plant
tissue are cut from the
mother plant
• Placed into media
• Grown out or
subdivided again to
produce more plants
 Micropropagation Methods
• Shoot Tip Culture (less 10 mm)
• Meristem culture (0.1-0.5 mm)
Surface sterilize, wash& culture.
Higher cytokinins to over come
apical dominance.
Adventitious bud production.
sub-culture frequently.
Basic in vitro propagation ...
 Micropropagation Stages

• The Basics of Micropropagation

• The Four stages
– Stage one – Explant establishment or initiation
– Stage two – Multiplication
– Stage three – Rooting
– Stage four – Acclimatization or hardening off
Benefits for Micropropagation ...

♣ Rapid multiplication of clones

♣ Difficult species ?
♣ Genetic uniformity ?
♣ Aseptic conditions
♣ Micro- stock plants
♣ Controlled environment
Other applications ...

♣ in vitro micro-grafting
♣ Genetic conservation
♣ Plant improvement
♣ Experimental system
 Problems arises during
micropropagation and their control
• Phenolic secretions:
Stress.
Phytotoxic causes plant death.
Antioxidant solution over night.
Activated charcoal.
Adenine sulphate.
Endogenous contamination
Sterilization (external contamination)
Viral, fungal or bacterial growth
(endogenous).
Check sample on different media (PDA)
antibiotics (rifamp,cefotax,tetracyc.)
 Vitrification:

Swelling/thickening of tissue just like

callus
Enclosed atmosphere CO2 and ethylene
and water vapors.
Causes morphological abnormalities.
Reduce temperature at bottom.
 Rooting and Acclimatization
• Proper rooting
• Specie to specie
• Critical growth regulators. NAA,IBA etc.
• In vitro plants are sensitive to
environment.
• Maintain the relatively high humidity.
• Never transplant the in vitro plants
directly to open fields.