1)Providing us a basic understanding of
physical and chemical requirements of
cell, tissue, organ culture, their growth
2)To learn and understand a procedure that
is often used to propagate many plants of
the same genetic background.
Banana belongs to Musaceae family, second largest food-fruit
crops of the world produced in the tropical and subtropical
several constraints including highly season dependent and
sometimes poor quality of planting material of banana become a
limiting factor for banana propogation. Therefore, tissue culture
technology was considered an appropriate option to overcome
Application of tissue culture technology to propagate banana
plants gives some advantages
resulting in disease free planting materials, more vigorous
growth, high yields, better quality of fruits, earlier fruiting and
more uniform crop since they are made from selected high
yielding mother plants.
The suckers are the major source of planting
material for banana and normally remain true-totype.
200 ml of distilled water is added to a 500 ml conical
15g of sugar
25ml of Macronutrients stock
2.5ml of Micronutrients stock
1ml of Vitamin stock
2.5 ml of Plant Regulator Growth stock
4g of agar is added
Distilled water is added to adjusted the volume to 500ml.
Natural regeneration of cultivated
bananas through suckers is very slow
due to hormone-mediated apical
dominance of the mother plant. So, as
an alternative, shoot tips from young
banana suckers with growing buds or
cut rhizomes called ‘bits’ and ‘peepers’
are most commonly used as explants
for accelerating the rate of in vitro
propogation of banana.
• The suckers of
banana tree is
trimmed by discarding
the outer layers of the
roots leaving only
shoot tips containing
• 70% bleach and
Tween-20 for (15
spray the surface
The half trimmed banana suckers then sterile immediately by put in
bottle contain 70% ethanol and shake it for 20 seconds and then 3
times of sterile distilled water.
WHY TISSUE CULTURE
• Multiplying plant in short time and in large
• Disease free plants
• Save space for early
plantation/germination thus reduce cost for
• Cultured specific/usefull part of plant
Sucker selection (sword sucker)
Growth media (MS media)
suitable nutrient composition (media)
duration of alcohol use in sterilization
Crowded work station
Working area/ method
•use shoot tips from young banana suckers
with growing buds or cut rhizomes called ‘bits’
and ‘peepers’ are accelerating the rate of in
vitro propogation of banana.
•Place the banana culture in complete darkness
first for 1 week.
•add antioxidant in media such as ascorbis
Importance of Tissue Culture
in Rice production
Rice is the most important crop in Malaysia because
it is staple food for Malaysian
In Malaysia, rice demand is higher than production
New technology must be developed to increase the
Tissue culture technology is one of the way to
produce high quality rice which is can produce much
rice grains, resistant to environmental stress and
shorter development stage ( cut time so that can
Callus can undergo mutation to achieve the
The following methods are used to induce
mutation in callus :
Physical mutagens (irradiation)
Chemical mutagens (carcinogens)
The application of somaclonal variation in
rice callus includes:
For crop improvement
To increase yield
Producing plants that resistant to
To increase plant fertility
Description : Contaminated
Factor : Technique of handling
Description : Contaminated
Factor : Contamination from media
Unorganized, undifferentiated, growing and dividing mass of
Form from the wounding part.
Rapid plant multiplication for quality seed material.
Performed in the dark as light can encourage differentiation of
Important in plant biotechnology.
Manipulation of the auxin to cytokinin ratio in the medium can
lead to the development of shoots, roots or somatic embryos
from which whole plants can subsequently be produced.
Callus Suspension culture
Place friable callus into a liquid medium and agitate
Single or small clumps of cells will be released into the medium,
where they keep growing and dividing to eventually a cell suspension.
The size of inoculum have to be sufficient , suspension can be build up
Too much cells and there is a risk that toxic products from damaged
cells could accumulate to a inhibiting level.
Cell suspension cultures should also be subcultured periodically.
It can be concluded dichlorophenoxyacetic
acid 2, 4- D is suitable for callus induction
of rice seed.
The role of 2,4-D in cell division
is to increase the rate of cell division and
attributes to the increased amount of
For callus induction MS medium
supplemented with different concentrations
DIFFERENT VARIETIES OF RICE.
ASD 16, ADT 43, Basmati 370, Pusa Basmati and