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Lesson 6
Propagating Plants by Tissue
Culture
Next Generation Science/Common Core Standards Addressed!
 HS‐LS1‐1. Construct an explanation based on evidence for how the
structure of DNA determines the structure of proteins which carry out the
essential functions of life through systems of specialized cells. [Assessment
Boundary: Assessment does not include identification of specific cell or
tissue types, whole body systems, specific protein structures and functions,
or the biochemistry of protein synthesis.]
 HS‐LS2‐5. Develop a model to illustrate the role of photosynthesis and
cellular respiration in the cycling of carbon among the biosphere,
atmosphere, hydrosphere, and geosphere. [Clarification Statement:
Examples of models could include simulations and mathematical models.]
[Assessment Boundary: Assessment does not include the specific chemical
steps of photosynthesis and respiration.]
WHST.9‐12.7 Conduct short as well as more sustained research projects to
answer a question (including a self generated question) or solve a problem;
narrow or broaden the inquiry when appropriate; synthesize multiple sources
on the subject, demonstrating understanding of the subject under investigation.
(HS‐LS1‐3)
Agriculture, Food, and Natural
Resource Standards Addressed
 PS.03.01.03.a. Summarize optimal conditions for
asexual propagation and demonstrate techniques used
to propagate plants by cuttings, division, separation,
layering, budding and grafting.
Bell Work!
1. Discuss the importance of tissue culture
2. Discuss a tissue culture method of
propagation used in the greenhouse industry
3. Where can tissue culture be applied in the
commercial field of agriculture?
4. Discuss what type of plants are best suited
for tissue culture propagation
Terms
Auxins
Callus
Cytokinins
Explants
Plantlet
Sterile agar
medium
Sterile technique
Tissue culture
Cultivar
Resistance
Explants
Interest Approach
 We’ve talked about how you can
produce African violets through
leaf cuttings.
 But, what if you had to produce
1,000 of them that are identical to
each other in a short period of
time?
 What problems/challenges would
this produce for the grower?
 How can tissue culture help solve
this problem?
What Is Tissue Culture and Why
Is It Important?
Tissue culture is the practice of growing
plant cells on artificial media
It involves the culture or growing of small
pieces of plant tissue
It is performed on artificial medium under
sterile conditions
Foliage plants, pot plants and cut flowers are
propagated by this method
Advantages of Tissue Culture
1. Large numbers of
plants can be produced
from a single plant in
relatively small space
in a short period of time
This reduces growing
space, labor and plant
maintenance
requirements
Advantages Continued
2. Viruses and other systemic diseases are
eliminated by propagating the quickly dividing cells
of the shoot tip
3. The grower is able to produce plants with
identical flowers
4. Horticultural cultivars can be improved by
selecting plants, which vary slightly from the mother
plant
Examples are leaf shape, disease resistance,
growth habit and flower color
5. The growth of identically engineered plant cells
What Process Is Used For Tissue
Culture Propagation?
The tissue culture propagation process can
be defined in four main stages:
A. First stage - small pieces of plant
material, called explants, are carefully
removed form the parent plant
Explants are obtained from the actively
growing part (shoot tips, sections of leaves,
stems and roots, embryos, etc) of a desired
plant
The explants are cleaned and
placed on sterile agar medium in
glass bottles or test tubes
The sterile agar medium is a gel
that contains water, sugars,
nutrients, and plant hormones to
support and promote plant
growth
Tiny leaves, stems and roots
make tissue culture possible
Removing explants
B. Stage two - the cells of the explants
multiply in one of two ways:
1. The cells may form a callus, which is a
group of cells with no particular function
Supplied with the correct hormones in the
medium, these callus cells can develop into a
normal plant
2. The explant may produce many new
explants if cytokinins, hormones responsible
for cell division and differentiation, are placed
in the medium
Stage Two: Explant Multiplication
Cytokinins encourage the increase in the
number of buds on the explants to six or more
per shoot
Each bud is capable of becoming a plant and
producing more buds
Branching occurs as these buds develop into
plant shoots, or plantlets
These plantlets are divided and transferred to
new containers
In this way, a single explant can produce
millions of plantlets in a year
D. Stage four - the plantlets are
removed form the glass container
C. Stage three - the plantlets
have developed and are ready for
root formation
Shoots are transplanted to
another medium containing
auxins, a hormone that induces
the growth of roots
The plantlets are also given
higher light intensity in
preparation for stage four
They are divided, planted in a
sterile medium, and placed in
a greenhouse
Care must be taken during
this transition to acclimatize
the plant to their new
environment
Tissue Culture Process
Continued
One of the most important aspects of tissue
culture is sterile technique
Sterile technique is the maintenance of an
environment that is free of bacteria, fungi and
viruses
Sterilization of the agar media is essential
In addition, the slightest air movement can
stir spores of bacteria and fungi
Special sterile work
stations, called
laminar hoods, are
used when possible
Cleaning of the plant before removal of the
explant is usually accomplished by a brief
soaking in a bleach solution, followed by a
rinse in sterile water
The tissue culture agar medium and other
materials used to prepare and place the
explant must be sterilized
This is usually done by an autoclave
The autoclave uses
pressurized steam
to sterilize medium,
glassware, and
instruments
Cultures are transferred from one container
to another at various stages in their
development
This transfer must occur under sterile
conditions to prevent contamination by
microorganisms
Sterilized equipment must be used for each
transfer
Summary
Why is tissue culture important in the
horticulture industry?
What parts of a plant can be used in tissue
culture?
Define explant.
Give two advantages for using tissue culture.
What is a sterile agar medium?
What is the first stage in the tissue culture
propagation method?
Summary Continued
What is a callus?
What must be added to a callus in order for
it to continue to develop?
What is a plantlet?
How do auxins help an explant?
What are some practices of sterile
technique?
The End!