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The Three Pillars of Yield
BREEDING
Strategically breed plants
to create new, more robust
seeds that perform better –
and longer – in the field.
AGRONOMICS
Use precision ag, planting
density, plant health protection,
and conservation tillage to make
acres more productive.
BIOTECHNOLOGY
Supplement breeding
advancements by
adding special
beneficial genes to the
plant.
ALL THREE ARE CRITICAL IN
DELIVERING YIELD TODAY – AND
In Vitro Culture
The culture of organized living material cell
under artificial condition in tubes, glasses, dishes
 The process of growing organized living
materials artificially in the laboratory
 The culture and maintenance of organized living
materials, in sterile, nutritionally (synthetic
media) and environmentally (controlled)
supportive conditions

Organized living material
Homogeneous
(tissues)
Heterogeneous
(organs)
Tissue
Hooke (1665) and Leeuwenhoek (1677) using
microscope discovered tissues were made of
cells
Cells
The cells of a particular tissue had a common
structure.
Cell




Taken to its simplest
form
A plasma membrane…
Surrounding
cytoplasm…
Containing hereditary
material.
Level of complexity


Xavier Bichat (1771-1802):
An organ is composed of
different tissues
Several organs can be
grouped together as an
organ system (e.g. the
digestive system)
Organism
Organsystem
Organ
Tissue
Cell
Level of complexity

Purkinje (1835) Observed a fertilised hen's egg
(a single cell) could develop into an embryo
(many specialised cells in a compact mass)
 C19th botanists showed that plant tissues
consist of many different types of cells.
THE CELL THEORY

Schleiden (1838) & Schwann (1839)
“The cell is the basic unit of living tissue”
The cell is an autonomous unit (“a citizen”)
grouped together to form an organism (“the
society”).
« Omnis cellula e cellula »
Virchow (1858)
“all cells come
from pre-existing
cells”
Cell culture
The process by which prokaryotic,
eukaryotic or plant cells are grown under
controlled conditions.
Cell culture was first successfully undertaken
in animal cell by Ross Harrison (1907) and in
plant cells by Haberlandt (1902)
ORGANISMAL THEORY
Reichert (a morphologist):
An organism has a structured plan
Image Credit Frog embryo fate map
Plasmodesmata
Strasberger (a cytologist):
Cells are connected in an
organism sometimes by
cytoplasmic bridges
Acellular organisms
Some organisms do not
have cellular
compartments
Unicellular organisms


Some organisms only
consist of a single cell
But these do usually have
the cell components
(nucleus, membrane,
etc)
Tissue culture
In vitro cultivation of organs, tissues & cells at defined
temperature using an incubator & supplemented with a
medium containing cell nutrients & growth factors


Cells can be cultured
away from a body
But this often requires
elaborate support
systems (technology)
Organized living material
1.Animal Cells
2.Plant Cells
Plant Tissue Culture
Plant Tissue Culture
“the aseptic culture of plant protoplasts,
cells, tissues or organs under conditions
which lead to cell multiplication or
regeneration of organs or whole plants “
Totipotency
…. each living cell has a complete genetic
blueprint and therefore has the potential to
develop into an entire plant.
… cells differentiate
…
animal
cells,
plant cells can
… unlike
living plant
cells
canliving
re-differentiate
de-differentiate and then re-differentiate to
form different cell types
Early Plant Cell Culture
…Haberlandt .. early 1900’s
… proposed concept of totipotency
… cells cultured under right conditions
Callus cultured from tree cambium
Gautheret et al. (1930).
… cells kept alive but did not develop
Early Plant Tissue Culture
- dependent on discovery of
“growth regulators”
 Cell enlargement … role of auxins
 Cell division ... role of cytokinins
 Regeneration from tobacco pith ..
(Skoog and Miller) … interaction of auxin
and cytokinin gives differentiation.
Carrot plants from root cells
(Stewart, 1964)
Tobacco plants from single cells
(Vasil & Hilderbrandt, 1965)
Requirement



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
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 tissue and can over run a culture
Growth regulators, both auxins & cytokinins.
Frequent sub-culturing to ensure adequate nutrition and to avoid
the build up of waste metabolites
Course Outline
Introduction to in vitro culture
 Determining factors in plant tissue culture
 Micropropagation
 Pattern of development
 Micropropation enterprice
