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.

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.

History

6 Advantages of Tissue Culture1. 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 Culture4. 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 One1. 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.

Tissue Culture Process – Stage One

Tissue Culture Process – Stage Two1. 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.

Tissue Culture Process – Stage Two4. 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 Three1. Once the plantlets are developed they

are ready to begin forming roots.2. The shoots need to be transplanted

into another medium containing auxins that induce the growth of roots.

3. The plantlets are given higher light intensity.

Tissue Culture Process – Stage Four1. 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.

Popular Plants to CultureAfrican Violet Ferns Orchid

Bamboo Plumeria Rose

Carnivorous Hosta Banana

Cactus Hibiscus PawPaw

Palm/Cycad Arabidopsis Daylily

DemonstrationCleaning the hoodUsing sterile technique while pouring plates

Basically – spray EVERYTHING with ethanol, and don’t touch anything!

DemoStorage of clean agar plates

In Plastic bags

Review Important VocabularyAGARCALLUSCYTOKININEXPLANTSHORMONESLAMINAR FLOW HOODPARENT PLANTPLANTLETSSTERILE TECHNIQUETISSUE CULTURE