Plant cells are totipotent

Totipotency:
ability of a cell or tissue or organ to grow and develop into a fully
differentiated organism.
Ex-plant: any living tissue: leaf, root, zygotic embryos

Culture Media:
1. Minerals
2. Carbon source
3. Plant Growth Regulators

Callus Unorganized, growing mass of cells

organogenesis Somatic embryogenesis

Plant regeneration
 Organogenesis
The formation of organs (such as leaves, shoots,
roots) on a plant organ, usually of a different
kind.
 Organogenesis
• Auxin/cytokinin 10:1-100:1 induces roots.

• 1:10-1:100 induces shoots

• Intermediate ratios around 1:1 favor callus growth.

Somatic Embryogenesis
• The production of
embryos from
somatic or “non-
germ” cells.
• Usually involves a
callus intermediate
stage which can
result in variation
among seedlings
 Somatic embryos

Somatic embryos are bipolar structures with both apical

and basal meristematic regions, which are capable of
forming shoot and root, respectively.

A plant derived from a somatic embryo is sometimes referred

to as an “embling”.
 Somatic Embryogenesis and
Organogenesis
• Both of these technologies can be used as
methods of micro propagation.

• Not always desirable because they may not

always result in populations of identical
plants.

• The most beneficial use of somatic

embryogenesis and organogenesis is in the
production of whole plants from a single cell
(or a few cells).
 Induction of somatic embryogenesis

♦ Auxin is usually applied for induction of somatic

embryogenesis.

♦ Nitrogen and carbohydrate balance are also important.

♦ Embryos usually develop, once they are transferred to a

medium with no auxin.
Direct somatic embryogenesis
Direct embryo formation from an ex-plant
 Indirect Somatic Embryogenesis
Explant → Callus Embryogenic → Maturation →
Germination

1. Callus induction
2. Callus embryogenic development
3. Maturation
4. Germination
 Development
 Auxin must be removed for embryo development
 Continued use of auxin inhibits embryogenesis
 Stages are similar to those of zygotic embryogenesis
– Globular
– Heart
– Torpedo
– Cotyledonary
– Germination (conversion)
 Maturation
• Require complete maturation with apical
meristem, radicle, and cotyledons

• Often require ABA for complete

maturation

• ABA often required for normal embryo

morphology
 Germination
• May only obtain 3-5% germination

• Sucrose (10%), mannitol (4%) may be

required
 Somatic embryogenesis as a
means of propagation is seldom
used
 High probability of mutations
 The method is usually rather difficult.
 Losing regenerative capacity become greater with
repeated subculture
 Induction of embryogenesis is very difficult with many
plant species.
 A deep dormancy often occurs with somatic
embryogenesis
Plant tissue culture systems used for
micropropagation
somatic embryo culture
 Somatic Embryogenesis
l Parthenocarpy

l Apomixis

l In vitro somatic embryogenesis

 Somatic Embryogenesis
• Somatic embryogenesis, encapsulation of the somatic embryos and
their storage are very significant steps towards conservation of genetic
resources.

• These somatic embryos can be used for mass propagation also.

• Somatic embryogenesis is a process by which embryo-like structures
develop from the callus.

• The process of somatic embryogenesis takes place in two stages: the

induction of proembryonic cell masses or proembryos and the
development of proembryos to somatic embryos.

• Induction of proembryogenic masses takes place under high auxin

content and development of somatic embryos under low auxin content.
 Somatic Embryogenesis
♦ Somatic embryogenesis is best achieved in suspension culture.

♦ Somatic embryos are encapsulated in sodium alginate.

♦ The encapsulated embryos are placed in calcium for 20 minutes to form

complex, rinsed in water and stored in closed containers.

♦ The coating material is packed with nutrient hormones and bio-

fertilizers, if desired.
 Factors that Influence SE

• Genotype
• Growth regulators
• Carbon source
• Nitrogen
 Somatic Embryos

• Bipolar
• Not connected to ex-plant or callus cells
by vascular tissue
 Various terms for non-
zygotic embryos
 Adventious embryos
Somatic embryos arising directly from other organs or
embryos.
 Parthenogenetic embryos (apomixis)
Somatic embryos are formed by the unfertilized egg.
 Androgenetic embryos
Somatic embryos are formed by the male gametophyte.
 Somatic embryogenesis differs from
organogenesis
• Bipolar structure with a closed radicular end rather than a monopolar
structure.
• The embryo arises from a single cell and has no vascular connection
with the mother tissue.
 Techniques of Micropropagation

• Somatic Embryogenesis – Developing embryos from

vegetative cells instead of egg (gametes) cells.
– Can take place from explants or cell suspension of callus
culture
– Relies on totipotency and plasticity of cells
– Direct method – plant is produced from cell(s) without a callus
– Indirect method – Callus (suspension or is made from explant,
then differentiated into a large number of whole new plants
– Relies on plant regeneration through organogenesis
 Techniques of Micropropagation

• Somatic Embryogenesis – Developing embryos from

vegetative cells instead of egg (gametes) cells.
– Organogenesis can lead (plant and condition dependent) to
genetic variation – somaclonal variation
– Phenomenon mostly associated with callus culture rather than a
direct culture method
– Mutants may arise spontaneously or induced with chemical DNA-
altering mutagenic agents
– Results in stable genetically inheritable (some) epigenetic
changes.
– Allows for the generation of new strains to be screened for
attractive plant characteristics
• Herbicide resistance, stress tolerance, disease resistance
 Somatic Embryogenesis
Under the right conditions, plant tissues regenerate into
whole plants via two distinct processes; “somatic
embryogenesis” and “shoot morphogenesis”.
For somatic embryogenesis, embryos form, which can
develop and then germinate into a whole plant.
Somatic embryos are complete entities, similar to seeds.
They possess a shoot and root tip and have a
complete vascular system. They form under specific
conditions that are different from shoot
morphogenesis.
 Control of organogenesis
Cytokinin
Leaf strip

Adventitious
Shoot

Root

Callus Auxin

07 March 2002 Dr. Michael Parkinson 28