Transcript Plant Tissue Culture

What is Tissue Culture
• A very technical method of
asexual propagation
• The growing of plantlets from
small pieces of plant tissue from a
parent plant.
• Uses an artificial medium under
sterile conditions.
• There are several advantages
to tissue culture.
History
• In 1902 it was proposed by
Haberlandt that single plant
cells could be cultured all
attempts failed.
• 1930’s plant growth regulators were
discovered as well as the importance
of vitamins for shoot and root growth.
• 1958 orchids and dahlias are cultured
providing disease free plants.
History
1962 Tissue Culture recipe
for medium is published.
1970’s Tissue Culture is promoted
for commercial plant production.
Today Tissue Culture is used all
over to world to produce genetically
identical disease free plants.
6 Advantages of Tissue Culture
1. Many plants can be produced
from a single plant in small space
and short period of time.
2. Diseases can be eliminated by
quickly dividing cells.
3. Produce plants with identical
flower color for the cut flower
industry.
6 Advantages of Tissue Culture
4. Excellent basal branching of
foliage plants.
5. Horticultural cultivars can be
improved by selecting plants
based on desirable traits.
6. Promote the growth of
genetically engineered
plant cells.
Tissue Culture- why so clean?
1. Tissue Culture must be done
under sterile conditions.
2. Anything that enters the hood
must be sprayed with ethanol.
3. Media, tools and containers are
autoclaved to kill bacteria and
fungi.
Tissue Culture- why so clean?
4.
Tissue Culture is done under
laminar air-flow hoods.
5. Flow hoods filter bacteria and fungal
spores from the air.
Tissue Culture Process
 The tissue culture propagation
process has 4 main stages.
 The first three stages must take
place under sterile conditions.
Tissue Culture Process – Stage One
Small pieces of plant material called
explants are removed from parent plant.
2. Explants are cleaned of bacteria or fungus
using 70% alcohol, 10% bleach solution
and sterile water.
3. Explants are placed on an agar media in a
glass bottle or test tube.
4. The agar media is a gel that contains water,
sugars, nutrients and plant hormones.
1.
Tissue Culture Process – Stage One
Tissue Culture Process – Stage Two
Cells of the explants begin to multiply.
2. They may form a callus, which is a
group of cell with no particular
function.
3. With the right hormones
the
callus cells can develop
into a normal plant.
1.
Tissue Culture Process – Stage Two
4. The other possibility is the rapid
multiplication of plantlets.
5. Cytokinins placed in the media
encourage an increase in the number
of buds on the explants, usually 6-8
per shoot.
6. Branching occurs as the buds
develop into shoots or plantlets.
Tissue Culture Process – Stage Three
1.
2.
3.
Once the plantlets are developed they are ready
to begin forming roots.
The shoots need to be transplanted into
another medium containing auxins that
induce the growth of roots.
The plantlets are given higher light intensity.
Tissue Culture Process – Stage Four
Plantlets are removed from the glass
container.
2. They are divided, planted in sterile
growing medium and placed in a
greenhouse.
3. The plants must be acclimated to their
new environment.
1.
Popular Plants to Culture
African Violet
Ferns
Orchid
Bamboo
Plumeria
Rose
Carnivorous
Hosta
Banana
Cactus
Hibiscus
PawPaw
Palm/Cycad
Arabidopsis
Daylily
Demonstration
 Cleaning the hood
 Using sterile technique while pouring plates
 Basically – spray EVERYTHING with ethanol, and don’t
touch anything!
 Demo
 Storage of clean agar plates
 In Plastic bags
Review Important Vocabulary
 AGAR
 CALLUS
 CYTOKININ
 EXPLANTS
 HORMONES
 LAMINAR FLOW HOOD
 PARENT PLANT
 PLANTLETS
 STERILE TECHNIQUE
 TISSUE CULTURE