Plant Tissue Culture and Cloning

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 What is it?
 Tissue culture had its origins at the beginning of the 20th century
with the work of Gottleib Haberlandt (plants) and Alexis Carrel
(animals)
 Growing cells artificially -Tissue culture is the term used for “the
process of growing cells artificially in the laboratory”
 Clones- Tissue culture produces clones, in which all product cells
have the same genotype (unless affected by mutation during
culture)
 Natural products- Cell and tissue cultures today are not only used
for the propagation of plants but also evaluated for the
commercial production of natural products.
 Genetic modification- A more recent advance is the use of
genetically modified plant and animal tissue culture

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 Principle of tissue culture
 Totipotency- many plant cells have the ability
to regenerate a whole plant.
 Phytohormone-The ratio of auxin to cytokinin
plays an important role in plant growth.
 When cytokinin and auxin are present in equal
levels, the parenchyma cells form an
undifferentiated callus.
 More cytokinin induces growth of shoot buds
 more auxin induces root formation

cytokinin and auxin are

More cytokinin more auxin
present in equal levels

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 What is needed?
 Appropriate tissue (some tissues culture better than others)
 A suitable growth medium containing energy sources and inorganic salts
to supply cell growth needs. This can be liquid or semisolid
 Aseptic (sterile) conditions, as microorganisms grow much more quickly
than plant and animal tissue and can over run a culture
 Growth regulators - . The two types of plant hormones used in this
experiment are cytokinins and auxins
H Cl
N
O O
O

HO HO
OH
N
H
indole-3-acetic acid 4-chloroindole-3-acetic acid 2-phenylacetic acid
HO
O
O NH2
N N
NH NH N
N N N N
N N H
NH
N N N N
H H
zeatin kinetin diphenyl urea 6-benzylaminopurine
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 How is Tissue Culture Done?
 Explant selection
plant cells can be removed from various parts of a
plant and placed on media in petriplates.
 Callus formation
The media does not contain the growth hormones,
the cells do not differentiate and instead form a
mass of cells called a callus
 Differentiation
Since plant cells are totipotent, growth hormones
can be added to the media triggering the callus cells
to develop roots, shoots and eventually entire
plants.
 Plants regenerated from tissue culture will be clones
genetically identical to the cell they originated from.
 The only animal cells that have this totipotent
characteristic are fertilized eggs.
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 Why do Plant Tissue Culture?
 A single explant can be multiplied into several thousand plants in less than
a year
 does not usually destroy the mother plant,
 rare and endangered plants can be cloned safely
 give a continuous supply of young plants
 virus free plants (In plants prone to virus diseases)
 Plant tissue can be stored in ‘tissue banks’ in frozen state then
regenerated through tissue culture
 easier to export (most current plant export is now done in this manner)
 crop improvement - explants are chosen from superior plants, then cloned
 Tissue culture clones are ‘true to type’ as compared with seedlings, which
show greater variability
 Preservation of medicinal plants
 Propagation of medicinal plants
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 Production of natural products
 Secondary metabolites such as alkaloids, terpenes, anthocyanins and
anthraquinones have been reported to be produced by plant cell cultures
 However, some well-known drawbacks are the often-low productivity and
instability of the productive cell lines
Strategies to enhance production of secondary metabolite
 Select highly productive cell line
 Precursor feeding
 Elicitor (compounds of pathogen origin)
 Suitable media that give easy isolation
 Permeation
 Adsorption
 Immobilization of plant cell (fixing the cell on solid support)
Bioreactor
 Gas sparged
 Stirred tank
 Mist bioreactor
 Roller drum bioreactor
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 Cell culture Vs plant
In optimize condition the cell culture give better yield plant part extract

Biotechnology Advances 20 (2002) 101-153

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 conclusion
 Plant Cell Culture An essential step in creating transgenic crops.
 Plant cell culture also provides us with an environmentally sound
and economically feasible option for obtaining naturally occurring
products with therapeutic value.
 Plant cell culture is also an important source of compounds used as
flavors, colors and aromas by the food-processing industry.
 Cloning technology allows us to generate a population of genetically
identical molecules, cells, plants or animals.
 Because cloning technology can be used to produce molecules,
cells, plants and some animals, its applications are extraordinarily
broad.