What is somaticembryogenesis?

ABT 451 - Commercial Plant Tissue Culture (0-+10) ROHITH s 2017021054

 Zygotie Embryogenesis Somatie Embryogenesis

Male Ganmetes Female Gametes Somatic or body cell (2n)

(n) (n)

-vitro.
ZygoticEmbryogenesis Epansion

ation
Geriation Callus

Q-0-0-0 Deication

Dormancy
and
Growth

Cety
Zygote (2n)
Somatic Embryogenesis
egeneratiom
Embryoids
of

whole plant
Embryo
Paet

New plant.
New plant.
Somatie Embryogenesis:

A where an embryo is derived from a single somatie cell or group of

process
somatic cells. Somatic embryos (SEs) are formed from plant cells that are not
normally involved in embryo formation.

Embryos formed by somaticembryogenesis are called Embryoids.

V The process was discovered for the first time in Daucas carota L. (carrot) by
Steward (1958),Reinert (1959).
 Cell Callus Induces Embryoids
Plant
(single) Nutrient (PGR) (Non- Zygoic )
medium
2n In-vitro

Explant

Explant in
Nutrient
NOTE:
media Differ
without membranous
Callusu Eg. Seed coat, endosperm

Pianlet But functional.

Embryold
Process offormation of somatic embryos:

Two main phases of somaticembryogenesis include:

1) Inductive phase: In this phase, the somatic cells acquire

embryonic competence and proliferate as embryonic clls

2) Expressive phase:
a) Development
b) Maturation
lobular heart torpedo cotyledonary
1) Inductive phase
* In this phase, the somatic cells acquire embryonic competence and proliferate as
embryonic cells

*An auxin, particularly 2,4-D, is generally necessary to induce embryogenesis.

Pro embryogenic masses are formed.

Once transferred to a medium with low or no auxin, these cells can start to
develop into mature embryos.

For callus induction MS medium supplemented with different concentration of

2,4-D(0, 1.0, 1.5, 2.5, 3.5 and 5 mg/l) was used in which 3.5, 5 mg/| 2,4-D
showed high callus induction percentage.
 Globular stage:
Embryo is small and round multi
2)Expressive phase cellular structure
Auxin must be
a) Development removed, continuous
use of auxin inhibits
Heart stage
- Globular structure embryogenesis
symmetry)
(bilateral

Globular shapechanges to heart

- Heart- shaped structure

Torpedo -shaped stage
Initialcells for shoot'root
- Torpedo-shaped structure meristem

Cotyledonary stage
Torpedo
b) Maturation Shape with more cotyledon
- Cotyledonary development

Embryo becomes
Germination cylindrical and germinated
 Online teaching ot

-Explant

-Medium Callus
A mature
Plant Direct
embryogenesis

Embryold
formation

Embryoids transfer to
semisolid medium
Regenerated
plantiets Cotyledonary Stage Torpedo Globular

Somatic embryos have a bipolar strueture, consisting of shoot apex and root apex, so
they are able to develop into shoot and root respectively in one step. usually without
any specific treatment
D m
 TYPES OF SOMATIC sOMATIC EMBRYOGENESis (SE)

EMBRYOGENESIS : Direct SE
Indirect SE

callus
No intervening callus

1. Direct SE
Globular stage
When embryos are fomed directly from explant tissue Globular stage

creating an identical clone without production of

intervening callus. Heart stage Heart stage

CAH

2. Indirect SE Torpedo stage CutinttOSIRE Torpedo stage

when explants produced undifferentiated mass of

cells(callus)which is maintained or differentiated into Cotyledonary stage Cotyledonary stage

embryo. Specific growth regulators and culture conditions

are required for callus formation. Embryo Embryo

PGR, medi,r,eplast,
arygrs, P,stress, tim, glt
 Examples of direct somatic Examples of indirect somatic
embryogenesis:
embryogenesis:
•Somatic embryogenesis has
been reported from many
•Secondary phloem
of carrot, leaf,
plants such as Coffea arabica
Alfalfa, Daucus carota tissues of coffee,
Ranunculus scleratus, Linum Petunia,
usitatissimum,Brassica
Asparagus Inetc.

napus,Arachis hypogea etc.

majority ofcases
•Leaves,scutellum,hypocotyl,nu
cellus and embryo-sac etc are embryogenesis is
used asexplants. through ndirect
Direct embryogenesis Indirectembryogenesis (in
method.
(in cassava) coffeu)
FACTORS AFFECTING SOMATIC EMBRYOGENESIS

1) Characters of explants:

O Even though a variety of explants can be utilized, the correct developmental

stage of the explants are also erucial for the initiation of embryogenic callus.

O Young orjuvenile explants produced more somatic embryos than older explants

Various types of explants used in SE

[Link] zygotic embryos

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2. Plant growth regulators:

i) Auxins

*2,4-D has been the best synthetic auxin used for inducing SEs.

Continuous supply ofauxin causes embryogenic cells to divide (Proliferation medium) without
the appearanceofembryos.

Witherell(1971)have suggested that continuous supply of auxin induces endogenous ethylene

production which suppresses embryo development.

So,embryogenic cells after treatment with auxin must be transferred to auxin free medium that
constitute the embryo development medium.
i)Cytokinins:

Cyokinin produces globular embryo from initial embryos.

Zeatin is promotive when applied to embryogenic days 3-4 transfer from the
cells after

proliferation medium to ED medium whereas BAP and kinetin have inhibitory effect on
embryogenesis.

High ratio of eytokinin than auxin induces shoot formation and reverse ratio favours
rooting,

others include Gibberllins, inhibits SE.

ABA promote embryo maturation and prevent precocious germination and secondary
embryogenesis.

3) Nitrogen source: reduced form of nitrogen is the sole source of embryo formation.
4) Others:

* Light generally promotes embryogenesis

High temperature usually favorable for
embryogenesis

Parameters: sto

1) pH: 5.6 to 5.8

2) Temperature:26 °C to 28 °C
3) Humidity: 40 – 80%
4) a) light intensity: 5000 to 8000 lux
b) light duration: 16hrs light and 8hrs darkness
Advandages:

VCan produce artificial seeds by encapsulation of agarosegel or sodium alginate.

YGermplasm conservation.

VCryopreservation.
VTo study the biotechnological studies.

YHigh propagation rate.

Somaclonal variations.

VEliminationof diseases and viruses.

VSuitablefor suspension culture.

Disadvandages:

•Confined to few species only.

• Response tissue specific (Explants).
• Show very poor germination because of their physiological and biochemical
immaturity.

• Requires skilled labour.

•Time consuming.
Difference between zygotie and somatieembryo:

Variables Somatie embryo Zygotic embryo

Formed by Somatic cells Fertilized egg or
zygote

Covered by No covering seed coat

Output Only form embryo Seed

Nature of plantlet Weak Healthy

Alike Mother plant Not like mother plant

Propagation rate High Comparatively low

 Iduced cele
Call chumps
Gabuler tge
Heart tg

Cell
multiplication
Embryo
differentiaton Trgede t
Embryogenie
callus

Coffea
somatic
embryogenesis
process Pre-germination in
Leaf explant
temporary immersion
bioreactor (RITA)

Mother tree Ponet

Conversion in temporary
immersion bioreactor
CTwin Flask)
Direct Sowing and plant
acclimatization ex-vitro
Peanut somatie embryogenesis

Rubber tree from

somatic embryogenesis
References:

o Bhojwani S. S. and Razdan M. K. 1983. Plant Tissue Culture: Theory and Practice. Elsevier publications.
Pp: 236-250

$ Bajaj. Y. P.S. (1995). Somatic Embryogenesis and Its Applications for Crop Improvement. Somatic
Embryogenesis and Synthetic Sed 1. pp: 105-125.

$ Alemanno, Devic, M., Niemenak, N., Sanier, C., Guilleminot, J., Rio, M., et al. 2008. Characterization
L., of

leafy cotyledon I-likeduring embryogenesis in Theobrpma cacao L. Planta. pp: 227, 853-866

* Chalupa, V. 1985. Somatic embryogenesis and plantlet regeneration from cultured immature and mature
embryosof Picea abies (L)Karst. Commun. Inst. For Cech 14,pp: 57-63.

* ():
Joshi, R., Kumar, P, 2013, Regulation of Somatic Embryogenesis in Crops:A Review, Agri. Reviews, 34
1-21, 2013.

Waren, G.S, Fowler, M.W. 1981. Physiological interactions during the initial stages of embryogenesis in

cultures of Daucus carota L. New Phytol. pp: 481-486.