Plant Tissue

Culture
CAPE BIO Workshops 2022
What is Plant Tissue Culture?
• Plant tissue culture is a collection of
techniques used to maintain or grow
plant cells, tissues or organs under
sterile conditions.

• The plant cells, tissues or organs are

grown on a nutrient culture medium.

• It is widely used to produce clones of a

plant in a method known as
micropropagation.
What is Micropropagation?

• Production of a
large number of
individual plants
from a small piece
of plant tissue
cultured in a
nutrient medium.
 Fundamental Principles of PTC
1. Totipotency

• The ability of cells such as an

embryonic stem cell to
differentiate into any type of body
cell.

• Plant cells are also totipotent

Fundamental Principles of PTC Cont’d

2. Plasticity

• the ability of living

organisms to change their
'state' in response to any
stimuli and applying the
most appropriate, adaptive
response.

• Plants can alter metabolism,

growth and development to
best suit their environment.
 What is needed for PTC
• Appropriate tissue

• A suitable growth medium

• Aseptic (sterile) conditions

• Growth regulators (e.g. auxins and cytokinins)

 PTC
• Stage 0 - Selection and preparation of the mother plant
• Appropriate plant cells or tissue is selected
• Sterilization of plant tissue
• Stage 1- Initiation of culture
• explant placed into growth media
• Stage 2- Multiplication
• Sub-culturing
• Stage 3- Rooting
• plantlets transferred to root media (hardening)
• Stage 4- Transfer to soil
• Plantlets transferred to soil
Selection of explant & Sterilization
• The parent plant material is cut into
many small pieces called explants.

• These could be tiny pieces of leaf,

stem, root or bud.

• The meristematic region is used

(region of mitotically dividing cells)
Selection of explant & Sterilization
• All materials, instruments, medium, plant material etc. used in PTC
must be free from microbes.

• The explants are sterilised with bleach and or alcohol to kill bacteria
and fungi.

• Flame sterilization, autoclave, alcohol, bleach, surface sterilization, washing of

hands etc.
• Steps 1-4 that follow are carried out under aseptic conditions
1. Initiation
• The sterilised explants are placed on a
growth medium

• Agar jelly containing nutrients such as

glucose, amino acids and phosphates, and
also high concentrations of the PGRs (auxin
and cytokinin).

• This causes the cells of each explant to

divide by mitosis to form a callus. A callus is
a mass of undifferentiated, totipotent cells.
2. Multiplication
• Multiplication is
done through sub-
culturing

• Each callus can then

be subdivided, to
increase the final
number of plants
that will be made.
3. Rooting
• The tiny plantlets are transferred to a
potting mixture and is slowly
acclimatised to normal growing
conditions.

• The process is sometimes called

hardening.

• The plant is eventually transferred to a

green house and is further cared for
prior to transfer to the field
4. Transfer to Soil
• The tiny plantlets are
transferred to soil under
natural conditions.
 Advantages of PTC
• Quick method for producing new plants compared to growing plants
from seed.

• Disease-free (no viruses).

• The plants chosen for cloning will be high-yielding and have other
desirable features such as maybe being resistant to a common pest or
disease of that crop.
 Advantages of PTC Cont’d
• The new plants all have the same phenotype. The uniformity makes
growing and harvesting easier.

• Infertile plants such as triploids can be reproduced, e.g. commercially

grown bananas.

• Plants that are hard to grow from seed can be reproduced, e.g.
orchids for the horticulture industry.
 Disadvantages of PTC
• an expensive process because it is labour intensive.

• The tissue culture process can fail due to microbial contamination.

• cloned offspring are susceptible to the same pest or disease. #

• There is no or very little genetic variation. The only source of variation

in a clone is mutation.
 Factors affecting PTC
• Plant species used

• Source of cultured tissue

• Age and health of donor plant

• Nutrient medium
 Applications of PTC
• Conservation of rare and endangered plant species

• Commercial production of plants used by florist

• To screen cells rather than plants for advantageous properties such as

herbicidal resistance/tolerance.