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Applications of Tissue Culture to Plant Improvement

Presentation · February 2017

DOI: 10.13140/RG.2.2.27290.24000

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Tarek Y S Kapiel
Cairo University
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Plant Tissue Culture
Tools for Plant Improvement

Plant Tissue Culture


Applications of Tissue Culture to Plant Improvement

Definition

Plant tissue culture can be defined as:

The culture of explants (seeds,

organs, tissues, cells, or protoplasts)
on nutrient media under
sterile conditions
 Applications of Tissue Culture to Plant Improvement
Introduction:
• Plant Tissue Culture starts through the sterile, in vitro
cultivation of plant parts such as organs, embryos,
seeds, and single cells on solidified or liquid media.
• Differentiated cells can be cultured to generate whole
plants, with the use of very little starting material.
• Meristematic tissue (growing cells) is used to grow
flowering plants, and is virus-free, which is important
for plant propagation.
 Applications of Tissue Culture to Plant Improvement
The basic steps of plant tissue culture are:
- Remove a piece of tissue from a plant, called an explant.
- Place the explant on a specific nutrient medium to force the
cells of the explant to become undifferentiated and form callus
tissue. This is called dedifferentiation.
- Callus tissue is transferred to another nutrient medium where it
is allowed to differentiate into plant tissue. This is called
redifferentiation.
- The plant is transferred to soil to complete plant growth.
- The ability of a plant cell to give rise to a whole plant through
dedifferentiation and redifferentiation is called “totipotency.”
 Applications of Tissue Culture to Plant Improvement
Characteristics of Plant Tissue Culture Techniques

1.Occur on a micro-scale.
2. Environmental conditions optimized (nutrition,
light, temperature).
3. All microbes (fungi, bacteria, viruses, and
nematodes are excluded).
 Applications of Tissue Culture to Plant Improvement
Characteristics of Plant Tissue Culture Techniques
4. The normal pattern of plant development
breaks down giving rise to callus, adventitious
roots, shoots, and embryos.
5. Ability to grow as single cells (protoplasts,
microspores, suspension cultures).
6. Plant cells are totipotent,
(able to regenerate a whole plant).
 Basis for Plant Tissue Culture
Growth Media
Minerals, Growth factors, Carbon source, Hormones
Environmental Factors
Light, Temperature, Photoperiod, Sterility, Media
Explant Source
Usually, the younger, less differentiated the explant, the
better for tissue culture
Genetics
Different species show differences in amenability to
tissue culture
In many cases, different genotypes within a species will
have variable responses to tissue culture; response to
somatic embryogenesis has been transferred
between melon cultivars through sexual hybridization
Tissue Culture Applications (1)

Callus Culture
 Factors Affecting Plant Tissue Culture
Two Hormones Affect Plant Differentiation:
Auxin: Stimulates Root Development
Cytokinin: Stimulates Shoot Development

Generally, the ratio of these two hormones can

determine plant development:
 Auxin ↓Cytokinin = Root Development
 Cytokinin ↓Auxin = Shoot Development
Auxin = Cytokinin = Callus Development
Callus, Root and Shoot formation in leaf disc and cotyledon explants
Callus Culture
Callus induction and Plant Regeneration in Lilium
 Tissue Culture Applications (2)

Micropropagation
 Micropropagation

• The art and science of plant multiplication

in vitro
• Usually derived from meristems
(or vegetative buds) without a callus stage
– Tends to reduce or eliminate somaclonal
variation, resulting in true clones
• Can be derived from other explants or
callus (but these are often problematic)
Micropropagation
 Steps of Micropropagation
Stage 0 – Selection & preparation of the mother plant
sterilization of the plant tissue takes place

Stage I - Initiation of culture

explant placed into growth media

Stage II - Multiplication
explant transferred to shoot media; shoots can be constantly
divided

Stage III - Rooting

explant transferred to root media

Stage IV - Transfer to soil

explant returned to soil; hardened off
Steps of Micropropagation
Steps of Micropropagation
 Steps of Micropropagation

• Stage 0 - Surface sterilization treatments

 Steps of Micropropagation

• Stage I - Initiation of culture & Explant selection

 Steps of Micropropagation

cv. Giza 21 cv. Giza 1

• Stage II – Multiplication
• Axillary Shoot Production and Shoot proliferation
from shoot tip explants of watermelons
 Steps of Micropropagation

• Stage III – Rooting of shoots

• explants transferred to root media
 Steps of Micropropagation

• Stage IV – Transplantation (Transfer to soil )

• explant returned to soil; hardened off
(Acclimatization)
 Steps of Micropropagation

Stage II
Stage 0 Stage I

Stage IV Stage III

 Features of Micropropagation
• Clonal reproduction
– Way of maintaining heterozygozity
• Multiplication Stage can be recycled many times
to produce an unlimited number of clones
– Routinely used commercially for many ornamental
species, some vegetatively propagated crops
• Easy to manipulate production cycles
– Not limited by field seasons/environmental influences
• Disease-free plants can be produced
– Has been used to eliminate viruses from donor plants
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