Plant Tissue Culture
Tissue Culture
A.K.A
•Micropropagation
•In vitro Culture
A rice plant
growing in
nutrient rich
agar
faperta.ugm.ac.id/newbie/download/pak_tar/Plantbiotechnology/TissuC
ulturebiotech2007.ppt
Plant Tissue Culture
 Learning Objectives
 Introduction to Tissue Culture
 Picture Dictation Exercise
Our Learning Objectives
in relation to Tissue Culture
 Reasons for propagating plants by tissue culture are
stated
 The culture process is described
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Describe some common tissue culture methods
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Including – meristem culture and embryo culture
Describe how the plant part is selected
Describe how the culture medium is made up
Explain the timing of plant selection
Introduction to Tissue Culture
 Tissue Culture (also known as Micropropagation or
In vitro culture) is:
The growing of plant cells, tissues, organs, seeds or
other plant parts in a sterile environment on a
nutrient medium.
Introduction to Tissue Culture
 Plant tissue culture is a bit like the equivalent of
‘Dolly the sheep’ but using plants.
Introduction to Tissue Culture
Cloning Animal Cells
Cloning Plant Cells
Both these processes use undifferentiated cells
Picture Dictation Exercise
 Take some meristematic cells from a plant.
 These cells are called an explant
 Place the explant on a sterile nutrient rich agar
 The explant grows into a ball of cells
 This ball of cells is called a callus
 The callus then develops roots, stem and leaves
 This is called a plantlet
 Transplant the plantlet into a traditional growing
media
Answer
Introduction to Tissue Culture
 Is …the technique of growing
plant cells, tissues, organs,
seeds or other plant parts in a
sterile environment on a
nutrient medium
 You might say … it is the plant
equivalent of dolly the sheep
Back to picture dictation
Back to Contents
What are undifferentiated cells
 What are undifferentiated cells
 In the human body we have lots of different types of cells
Blood cells
Muscle cells
Bones cells
Brain cells
An undifferentiated cell is a cell that does not yet have a
specialised job
Next
What are undifferentiated cells
 Undifferentiated cells in animals
Stem cells
 Undifferentiated cells in plants
?
You tube demonstrations
 http://www.youtube.com/watch?v=kje0YczE0Do
The Culture Medium (Media)
 The environment surrounding the developing
explants must provide the correct ‘necessities’ for
plant growth.
Neccessities
for plant
growth
The Culture Medium (Media)
 From this list of neccessaties – which ones are
provided by the agar?
Neccessities
for plant
growth
Hormones in the agar
 Two Hormones Affect Plant Differentiation:
 Auxin: Stimulates Root Development
 Cytokinin: Stimulates Shoot Development
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Generally, the ratio of these two hormones can
determine plant development:
 Auxin ↓Cytokinin = Root Development
  Cytokinin ↓Auxin = Shoot Development
 Auxin = Cytokinin = Callus Development
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What is Callus development
 A callus is a blob of tissue – (mostly undifferentiated
cells)
 A callus is naturally developed on a plant as a result
of a wound
 This callus can be left to develop or can be further
divided
 The culture medium contains a gel (agar) with
the proper mixture of nutrients, sugars, vitamins and
hormones (growth promotors),
 These causes the plant part to grow at very rapid
rates to produce new plantlets. It has been estimated
that one chrysanthemum apex placed in tissue
culture could produce up to 1,000,000 new plantlets
in one year.
 Tissue Culture Transfer Protocol
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 Dr. Dan Lineberger of Texas A&M University demonstrates the protocol to
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transfer African violets from tissue culture containers where they were grown into a
small �forest� of cloned plants (called multiplication culture tubes) to tissue
containers where the young clones will form new roots.� After the roots are
formed, they can be removed and potted into containers. This procedure must be
done in the sterile environment of a transfer hood.
Sterilize the surfaces of the transfer hood.
Sterilize all tools that touch the plants by first dipping in alcohol them flaming.
African violet clones in a shoot multiplication tube.
Remove the cluster of plants in the culture.
Insert the cluster of plants into the new culture container.
Break up the cluster of plants and spread them out.
Seal the container with paraffin film.
The culture before transfer (left) and after transfer (right).
Embry Culture
Plant Tissue Culture and G.E