Plant tissue culture is a practice used to propagate plants under
sterile conditions, often to produce clones of a plant. Different
techniques in plant tissue culture may offer certain advantages over
traditional methods of propagation, including:
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The production
of exact copies of plants that produce particularly good
flowers, fruits, or have other desirable traits.
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To quickly
produce mature plants.
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The production
of multiples of plants in the absence of seeds or necessary
pollinators to produce seeds.
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The
regeneration of whole plants from plant cells that have been
genetically modified.
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The production
of plants in sterile containers that allows them to be moved
with greatly reduced chances of transmitting diseases, pests,
and pathogens.
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The production
of plants from seeds that otherwise have very low chances of
germinating and growing, i.e.: orchids and nepenthes.
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To clean
particular plant of viral and other infections and to quickly
multiply these plants as 'cleaned stock' for horticulture and
agriculture.
Plant tissue culture relies on the fact that many plant cells
have the ability to regenerate a whole plant (totipotent).
Single cells, plant cells without cell walls (protoplasts),
pieces of leaves, or (less commonly) roots can often be used to
generate a new plant on culture media given the required
nutrients and plant hormones.
Techniques
Modern plant tissue culture is performed under aseptic
conditions under filtered air. Living plant materials from the
environment are naturally contaminated on their surfaces (and
sometimes interiors) with microorganisms, so surface
sterilization of starting materials (explants) in chemical
solutions (usually alcohol or bleach) is required. Mercuric
chloride is seldom used as a plant sterilant today, as it is
dangerous to use, and is difficult to dispose of. Explants are
then usually placed on the surface of a solid culture medium,
but are sometimes placed directly into a liquid medium,
particularly when cell suspension cultures are desired. Solid
and liquid media are generally composed of inorganic salts plus
a few organic nutrients, vitamins and plant hormones. Solid
media are prepared from liquid media with the addition of a
gelling agent, usually purified agar.
Choice of explant
The tissue obtained from the plant to culture is called an
explant. Based on work with certain model systems, particularly
tobacco, it has often been claimed that a totipotent explant can
be grown from any part of the plant. However, this concept has
been vitiated in practice. In many species explants of various
organs vary in their rates of growth and regeneration, while
some do not grow at all. The choice of explant material also
determines if the plantlets developed via tissue culture are
haploid or diploid. Also the risk of microbial contamination is
increased with inappropriate explants. Thus it is very important
that an appropriate choice of explant be made prior to tissue
culture.
The specific differences in the regeneration potential of
different organs and explants have various explanations. The
significant factors include differences in the stage of the
cells in the cell cycle, the availability of or ability to
transport endogenous growth regulators, and the metabolic
capabilities of the cells. The most commonly used tissue
explants are the meristematic ends of the plants like the stem
tip, auxiliary bud tip and root tip. These tissues have high
rates of cell division and either concentrate or produce
required growth regulating substances including auxins and
cytoinins.
Some explants, like the root tip, are hard to isolate and are
contaminated with soil microflora that become problematic during
the tissue culture process. Certain soil microflora can form
tight associations with the root systems, or even grow within
the root. Soil particles bound to roots are difficult to remove
without injury to the roots that then allows microbial attack.
This associated micro flora will generally overgrow the tissue
culture medium before there is significant growth of plant
tissue.
Aerial (above soil) explants are also rich in undesirable micro
flora. However, they are more easily removed from the explant by
gentle rinsing, and the remainder usually can be killed by
surface sterilization. Most of the surface micro floras do not
form tight associations with the plant tissue. Such associations
can usually be found by visual inspection as a mosaic,
de-colorization or localized necrosis on the surface of the
explant.
An alternative for obtaining uncontaminated explants is to take
explants from seedlings which are aseptically grown from
surface-sterilized seeds. The hard surface of the seed is less
permeable to penetration of harsh surface sterilizing agents,
such as hypochlorite, so the acceptable conditions of
sterilization used for seeds can be much more stringent than for
vegetative tissues.
Applications
Plant tissue culture is used widely in plant science; it also
has a number of commercial applications. Applications include:
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Micro
propagation is widely used in forestry and in floriculture.
Micro propagation can also be used to conserve rare or
endangered plant species.
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A plant
breeder may use tissue culture to screen cells rather than
plants for advantageous characters, e.g. herbicide
resistance/tolerance.
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Large-scale
growth of plant cells in liquid culture inside bioreactors
as a source of secondary products, like recombinant proteins
used as biopharmaceuticals.
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To cross
distantly related species by protoplast fusion and
regeneration of the novel hybrid.
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To
cross-pollinate distantly related species and then tissue
culture the resulting embryo which would otherwise normally
die (Embryo Rescue).
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For
production of doubled monoploid (dihaploid) plants from
haploid cultures to achieve homozygous lines more rapidly in
breeding programmes, usually by treatment with colchicines
which causes doubling of the chromosome number.
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As a tissue
for transformation, followed by either short-term testing of
genetic constructs or regeneration of transgenic plants.
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Certain
techniques such as meristem tip culture may be employed that
can be used to produce clean plant material from viruses
stock, such as potatoes and many species of soft fruit.
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Micro
propagation using meristem and shoot culture to produce
large numbers of identical individuals
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Screening
programmes of cells, rather than plants for advantageous
characters
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Large-scale
growth of plant cells in liquid culture as a source of
secondary products
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Crossing
distantly related species by protoplast fusion and
regeneration of the novel hybrid .
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As a tissue
for transformation, followed by either short-term testing of
genetic constructs or regeneration of transgenic plants .
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Removal of
viruses by propagation from meristematic tissues.
Laboratories
Although some growers and nurseries have their own labs for
propagating plants via tissue culture, a number of independent
laboratories provide custom propagation services. The Plant
Tissue Culture Information Exchange lists many commercial tissue
culture labs. Since plant tissue culture is a very labour
intensive process, this would be an important factor in
determining which plants would be commercially viable to
propagate in a laboratory.
Organic farming is the form of agriculture that relies on
crop rotation, green manure, compost, biological pest control,
and mechanical cultivation etc... to maintain soil productivity
and control pests, excluding or strictly limiting the use of
synthetic fertilizers and synthetic pesticides, plant growth
regulators, livestock feed additives, and genetically modified
organisms. Since 1990, the market for organic products has grown
at a rapid pace, to reach $46 billion in 2007. This demand has
driven a similar increase in organically managed farmland.
Approximately 32.2 million hectares worldwide are now farmed
organically, representing approximately 0.8 percent of total
world farmland. In addition, as of 2007 organic wild products
are harvested on approximately 30 million hectares Organic
agricultural methods are internationally regulated and legally
enforced by many nations, based in large part on the standards
set by the International Federation of Organic Agriculture
Movements (IFOAM), an international umbrella organization for
organic organizations established in 1972. IFOAM defines the
overarching goal of organic farming as follows:
"Organic agriculture is a production system that sustains the
health of soils, ecosystems and people. It relies on ecological
processes, biodiversity and cycles adapted to local conditions,
rather than the use of inputs with adverse effects. Organic
agriculture combines tradition, innovation and science to
benefit the shared environment and promote fair relationships
and a good quality of life for all involved..
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