Plant tissue culture
• Broadly refers to technique of growing plant cells, tissues,
Plant tissue culture
a.k.a. micropropagation
Why do plant tissue culture?
• Fast commercial propagation of new cultivars
– Cost of culture (orchid price decrease)
• Agriculture
– Fast selection for crop improvement – nutritional value,
pest control, hardiness
– Cultivation virus free plants
• Pharmaceuticals – ginseng and taxol
• Cloning of rare and endangered plants
• Plant cultures in approved media are easier and safer to
export
Further progress
• 1958-60 Morel cultured orchids and dahlias freed them
from a viral disease
• 1962 Murashige and Skoog published recipe for M&S
medium
• 60’s and 70’s Murashige cloned plants in vitro
– raised haploid plants from pollen grains
– used protoplast fusion to hybridize 2 species of tobacco
into one plant contained 4N
• 70’s and 80’S beginning of genetic engineering
organs, seeds or other plant parts in a sterile environment
on a nutrient medium
• The first commercial use of plant propagation on artificial
media was in the germination and growth of orchid plants
in the 1920’s
• It was only after the development
of a reliable artificial medium by
Murashige & Skoog in 1962 that
plant tissue culture really took off
commercially
History
• First attempted by Haberlandt (1902) - grew palisade cells
from leaves of various plants but they did not divide
• 1934 - White generated continuously growing culture of
meristematic cells of tomato on medium containing salts,
yeast extract and sucrose and 3 vit B (pyridoxine,
thiamine, nicotinic acid) – established the importance of
additives
• 1953 Miller and Skoog, University of Wisconsin – Madison
discovered kinetin, a cytokine that plays an active role in
organogenesis
Basis for the cell culture
• Plant cells are totipotent
• Have the ability to develop into whole plants or plant
organs in vitro when given the correct conditions
• Not all plant cells are totipotent. However, there are a
sufficient number of totipotent cells in the plant (e.g. in
the pith)
• Differentiated cells have to be dedifferentiated into callus
and redifferentiated back to somatic embryo that will
regenerate the entire plant
1

Callus
• Is a natural response of the plant tissue to wounding
• A mass of actively dividing undifferentiated cells produced
by plant tissue explant
• Cells are totipotent
• Callus can be
– Resuspended in liquid media to create a susupension
culture of single totipotent cells
– Or differentiated into plant with the appropriate
manipulations of culture conditions
Culturing (micropropagating) plant
tissue - the steps
• Selection of the plant tissue (explant) from a healthy
vigorous ‘mother plant’ - this is often the apical bud, but
can be other tissue
• Establishment of the explant in a culture medium - the
medium sustains plant cells growth and encourages cell
division
– Each plant species (and sometimes the variety within a
species) has particular medium requirements that must
be established by trial and error
• Multiplication - the explant gives rise to a callus
• Differentiation and organogenesis

What is needed? Plant tissue culture media

• Explant (some tissues culture better than others) • Mineralals (17 essential elements)
• A suitable growth medium • Energy and carbon source - sucrose is preferred
• Aseptic conditions, as microorganisms grow much more • Growth regulators – auxins and cytokinins
quickly than plant and animal tissue (phytohormones)
• Growth regulators - in plants, both auxins & cytokinins • Vitamins
• Frequent subculturing to ensure adequate nutrition and to • Organic compounds
avoid the build up of waste metabolites • Water

• Usually 5.0-5.7

Vitamins Growth regulators

• Thiamine Auxins and cytokinins
• Pyridoxin • Auxins stimulate cell division in explants
• Nicotinic acid – Used for callus induction and growth
• Biotin • High concentration suppresses the organized growth and
• Citric acid promotes formation of meristem-like cells
• Ascorbic acid • IAA, indole-3-acetic acid a light sensitive, natural auxin
• Inositol
• Cytokinins
• Promote cell division and regeneration possibly through
the effects on protein synthesis
• Zeatin – natural cytokine that induces morphogenesis

2
 Homework
• In your notebook write a detailed content of a selected
plant culture medium. It has to have concentrations of all
ingredients!
• Make sure you understand what is in it and why. Again,
there will be a question about it on the first exam.
• Find a name of the plant that is grown on that medium