1. Plant Tissue Culture

2. 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.

3. 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.

4. 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.

5. 6 Advantages of Tissue Culture
Many plants can be produced from a single plant in small space and short period of time.
Diseases can be eliminated by quickly dividing cells.
Produce plants with identical flower color for the cut flower industry.

6. 6 Advantages of Tissue Culture
Excellent basal branching of foliage plants.
Horticultural cultivars can be improved by selecting plants based on desirable traits.
Promote the growth of genetically engineered plant cells.

7. 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.

8. 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.

9. Tissue Culture Process
The tissue culture propagation process has 4 main stages.
The first three stages must take place under sterile conditions.

10. Tissue Culture Process – Stage One
Small pieces of plant material called explants are removed from parent plant.
Explants are cleaned of bacteria or fungus using 70% alcohol, 10% bleach solution and sterile water.
Explants are placed on an agar media in a glass bottle or test tube.
The agar media is a gel that contains water, sugars, nutrients and plant hormones.

11. Tissue Culture Process – Stage One

12. Tissue Culture Process – Stage Two
Cells of the explants begin to multiply.
They may form a callus, which is a group of cell with no particular function.
With the right hormones the callus cells can develop into a normal plant.

13. 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.

14. Tissue Culture Process – Stage Three
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.

15. Tissue Culture Process – Stage Four
Plantlets are removed from the glass container.
They are divided, planted in sterile growing medium and placed in a greenhouse.
The plants must be acclimated to their new environment.

16. Popular Plants to Culture
African Violet
Ferns
Orchid
Bamboo
Plumeria
Rose
Carnivorous
Hosta
Banana
Cactus
Hibiscus
PawPaw
Palm/Cycad
Arabidopsis
Daylily

17. 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

18. Review Important Vocabulary
AGAR
CALLUS
CYTOKININ
EXPLANTS
HORMONES
LAMINAR FLOW HOOD
PARENT PLANT
PLANTLETS
STERILE TECHNIQUE
TISSUE CULTURE