Conservation of medicinal plants deals with the
controlled utilization & official supervision in order to
preserve or protect them.
Acc to WHO, as many as 80% of the world’s population
depends on traditional herbal medicine for their primary
health care needs.
Today many medicinal plants face extinction or severe
Tissue culture is one of the many techniques in
biotechnology which can be used for the conservation of
such medicinal plants.
Gottlieb Haberlandt, pioneer of plant tissue culture.
Murashige & Skoog medium, an important plant growth
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WHAT DO WE MEAN BY TISSUE
Plant tissue culture is a collection of techniques
used to maintain or grow plant cells, tissues or
organs under sterile conditions on a nutrient culture
medium of known composition.
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It is widely used to produce clones of a plant in
a method known as Micropropagation.
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MURASHIGE & SKOOG MEDIUM
Murashige & Skoog medium(MSO/MS0) is a plant
growth medium used in laboratories for the cultivation
of plant cell culture.
Invented by Plant scientists Toshio Murashige & Folke
K.Skoog in 1962 during Murashige’s search for new
plant growth regulator.
A number behind the letters MS is used to indicate the
sucrose concentration of the medium.
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MILE STONES IN PLANT TISSUE
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1902 Haberlandt proposed concept of invitro cell culture
1922 Kolte & Robbins successfully cultured root & stem tips respectively
1926 Went discovered first plant growth hormone- Indole acetic acid
1941 Overbeek was first to add coconut milk for cell division in Datura
Skoog & Miller discovered Kinetin as cell division hormone
Skoog & Miller gave concept of hormonal control of organ formation
Kanta & Maheswari developed test tube fertilization technique
Murashige & Skoog developed MS medium with higher salt concentration
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1974 Reinhard introduced biotransformation in plant tissue cultures
Chilton et al. successfully integrated Ti plasmid DNA from Agrobacterium
tumefaciens in plants
Melchers et al. carried out somatic hybridization of tomato & potato
resulting in Pomato
1981 Larkin & Scowcroft introduced the term somaclonal variation.
2005 Rice genome sequenced under International Rice Genome Sequencing
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It is labour intensive & expensive process.
All plants cannot be successfully tissue cultured.
It is usually because the medium of growth is not known.
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TYPES OF TISSUE CULTURE
Plant tissue culture includes two major methods
A. Type of in vitro growth- Callus & Suspension
B. Type of Explant-
Single cell culture
Shoot & root culture
Somatic embryo culture
Anther culture & haploid production
Protoplast culture & somatic hybridization
Embryo culture, Ovule culture, Ovary culture
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CHOICE OF EXPLANT
The tissue obtained from a plant to be cultured is called
In a totipotent, explant can be collected from any part of
In many plants, explants of various organs vary in their
rate of growth & regeneration.
The choice of explant material also determines if the
plantlets developed via tissue culture are haploid/diploid.
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STERILIZATION OF EXPLANTS
EXPLANTS ARE PLACED OVER SOLID/LIQUID MEDIUM
PROFOUND EFFECT ON THE MORPHOLOGY OF TISSUES
PIECES ARE TYPICALLY SLICED OFF &TRANSFERRED TO NEW MEDIA
SHOOTS EMERGE FROM CULTURE
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PERFORMED UNDER ASEPTIC CONDITIONS UNDER HEPA
FILTERED AIR PROVIDED BY A LAMINAR FLOW CABINET
MAY BE SLICED OFF
MATURED ONE ARE TRANSFERRED TO POTTING SOIL FOR
FURTHER GROWTH IN THE GREEN HOUSE AS NORMAL PLANTS
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The specific differences in the regeneration potential
*Differences in the stage of the cells in the cell cycle.
*Availability or ability to transport endogenous growth
*Metabolic capabilities of the cells
The most commonly used tissue explants are the
meristematic ends of the plants like the stem tip, auxillary
bud tip & root tip.
These tissues have high rates of cell division & produce
required growth regulating substances including auxins &
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Shoot culture : Performed in 4 stages for mass
production of plantlets through in vitro vegetative
Organogenesis : Common method of
Micropropagation that involves tissue regeneration of
adventitious organs/axillary buds directly or indirectly
from the explants.
Non-zygotic embryogenesis: Important pathway for
producing somaclonal variants, developing artificial
seeds & synthesizing metabolites.
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The commercial production of plants which uses
meristem & shoot culture to produce large numbers of
To conserve rare or endangered plant species.
A plant breeder may use tissue culture to screen cells
rather than plants for advantageous characters.
Large scale growth of plants in liquid culture in
bioreactors for the production of valuable compounds.
To cross distantly related species by protoplast fusion &
regeneration of the novel hybrid.
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To rapidly study the molecular basis for physiological,
biochemical & reproductive mechanisms in plants.
To cross pollinate distantly related species & then tissue
culture the resulting embryo which would otherwise
normally die (Embryo Rescue)
For chromosome doubling & induction of polyploidy.
As a tissue for transformation, followed by either short
term testing of genetic constructs or regeneration of
Certain techniques such as meristem tip culture can be
used to produce clean plant material from virused stock.
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HAIRY ROOT CULTURE
It is also called Transformed root culture.
It is used to study plant metabolic processes or to
produce valuable secondary metabolites or recombinant
proteins, often with plant genetic engineering.
A naturally occurring soil bacterium that contains root
inducing plasmids can infect plant roots & cause them
to produce a food source for the bacterium & to grow
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The abnormal roots are particularly easy to culture in
artificial media because hormones are not needed.
These roots will be having a high growth rate as well
as genetic & biochemical stability.
It is also used for regeneration of whole plants & for the
production of artificial seeds.
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RECOGNITION OF TISSUE CULTURE
The need for a certification programmes for the plant
tissue culture is imperative since inadvertent
Micropropagation of virus infected plants will not only
result in its poor performance, but also in undesired
spread of viruses wherever such plants are grown.
Also, failure to use prescribed standard protocols will
result in variation in the plants produced.
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The National certification system for tissue culture
raised plants(NCS-TCP) has been developed for the
first time to provide support to Plant Tissue Culture
Industry to facilitate production of quality planting
material through tissue culture/ Micropropagation.
The Department of Biotechnology, Ministry of Science
& Technology, Government of India as authorised
under section 8 of seeds act 1966,Vide Gazette
Notification dated 10th march 2006 is the Certification
Agency for the purpose for certification of the Tissue
culture raised propagules upto laboratory level & to
regulate its genetic fidelity as prescribed.
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Tissue culture production facilities can get their
material certified under this programme from
accredited Test laboratories as per prescribed criteria.
Only recognised tissue culture production facilities are
eligible to register for certification of their material.
The Project management unit(PMU) has been set up at
Biotech consortium India Ltd, New Delhi for the
implementation of this programme & in undertaking
and recognition of Tissue culture production facilities.
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Invitro propagation of Garlic by shoot
Invitro propagation of Potato (Solanum tuberosum . L),
G. Hussey, N.J. Stacey-Annals of Botany 1981
Invitro propagation and low temperature storage of
Saussurea lappa CB Clarke – An Endangered ,
Medicinal plant, R. Arora , S. S. Bhojwani - Plant Cell
Invitro propagation of Gymnema sylvestre-A
multipurpose medicinal plant, N.Komalavalli,M.V.Rao-
Plant cell, Tissue & organ culture 2000
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It is important for a researcher to be ethical while
performing Tissue Culture, as this technique comes
with great responsibility
Plant tissue Culture is meant to produce products that
are useful to the human kind or the ecosystem.
Plant tissue culture is our hope to end world hunger.
However when it comes to manipulating a living
organism many ethical issues will arise.
Hence, this technique must be performed with caution
to minimize the risks while capitalizing on the benefits.
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