Plant tissue culture is:
A tool in biotechnology, used in
micropropagation, secondary metabolic
production(taxol), hybridisation etc.
It refers to the growth of plant
parts(organs, tissues etc.) in a sterile
environment in a nutrient medium.
Why Plant T.C.??
Saves time & labour in comparison to
conventional hybridization techniques.
Useful when other techniques are not
Eg.: Apical meristems which are free of
viruses(present in shoot tips) can be
harvested & nurtured to grow into virus-free
plants in times of viral attack.
• To save species from extinction
• To isolate disease from plants
• To produce plants with enhanced
stress or pest resistance
• To create new plant varieties
• To make money
Plant tissue culture is based on
Plant cells have the ability to produce the
whole plant from single cells. This is called
Plants have the ability to reproduce
asexually through a type of cell division-
• Daughter cells of identical chromosome
numbers are produced.
Beginning of T.C.
In 1902, a German physiologist, Gottlieb
Haberlandt proposed that single plant
cells could be cultured.
He isolated single fully differentiated
individual plant cells from different plant
species and was first to culture them in
Knop’s salt solution enriched with glucose.
In the 1930s, importance of vitamins was
determined for shoot and root culturing
Indole-Acetic Acid(IAA), a natural auxin,
and the most important, was discovered
during the 1930s.
Many more synthetic auxins were
discovered. Eg.: 2,4d &
Miller and Skoog discovered kinetin, a
synthetic cytokinin, in the University of
Wisconsin – Madison.
Kinetin plays active role in organogenesis.
Steward developed somatic embryo from
Morel cultured orchids and dahlias & freed them
from a viral disease
• Murashige and Skoog published recipe for MS
Medium for T.C.
Functions: provide H2O, provide mineral &
vitamin nutritional needs.
H2O is usually distilled
minerals must provide 17 essential
energy source - sucrose is preferred
provide access to atmosphere for gas
serve as a dumping ground for plant
Macronutrients-eg: K, Ca, Mg,
N, P, O etc.
Micronutrients-eg: Fe, Mn, Mo,
Vitamins & Amino
nicotinic acid, biotin, citric
acid, ascorbic acid, Inositol,
Growth Regulators: auxins and
pH usually 5.0-5.7
1. Explant collection
2. Callus generation
3. Organogenesis (shoot and root
Micropropagation does not involve the
second step. There is direct
portion of plant removed
and used for T.C.
source - some tissues are
better than others
physiological age -
young portions of plant
are most successful
degree of contamination
external infestation - soak plant
in sodium hypochlorite solution
internal infection - isolate cell
that is not infected
roots - especially difficult
because of soil contact
easier to culture than woody
• The dedifferentiation of a plant
cell(explant) produces callus.
• Callus is then expanded into a larger
mass of undifferentiated cells.
• Callus is then activated, by selective
use of plant hormones to
redifferentiate to produce, shoots,
roots and ultimately, plantlets
Usually induced by changes in hormonal
Shooting: Higher cytokinin concentration
& lower auxin concentration(cytokinins
promote shoot growth).
Rooting: lower cytokinin concentration
and increase auxin(auxins promote shoot
Hardening involves the formation of the
waxy cuticle on the leaves of the plant.
Plants in T.C. do not have cuticle
usually done in greenhouse with high
gradually increase light intensity and
lower RH after rooting occurs
allows plants to harden and helps plants
The inventors of tissue culture
The internet’s information
Microsoft Office 2010