SOMATIC EMBRYOGENESIS

SUDHANSHU SHEKHAR M.Tech (Biotech) IIIrd Sem A7110709009

‡ Embryogenesis occurs naturally as a result of sexual fertilization and the formation of the zygotic embryos. ‡ Formation of the embryo begins with the division of the fertilized eggs or zygote within the embryo sac of the ovule (Zygotic variation). ‡ The embryo along with other cells from the mother plant develops into the seed or the next generation, which, after germination, grows into a new plant.

In the sexual cycle: New plants arise after fusion of the parental gametes, and, generally, each seed represents a new combination of genes. After germination they represent, a bipolar structure having both a shoot and root pole.

Somatic embryogenesis is an asexual form of plant regeneration in which it is formed from somatic tissues in nature and develops into a whole new plant in a way analogous to zygotic embryos which mimics many of the events of sexual reproduction. During the maturation stage somatic embryos Somatic embryos accumulate storage products that exhibit the same characteristics as those of the zygotic embryos.

Somatic embryogenesis in plant tissue cultures has been considered as a model system for the understanding of plant embryo development. Embryogenic cultures are an attractive target for gene transformation and crop improvement. Exogenously plant growth regulators (PGR) are essential to the process of somatic embryogenesis.

WHY«There is a need/approach of Somatic Embryogenesis«if seed propagation is easy?

‡Nature circumvents to produce hybrid vigour in between two interspecific species because there is some barrier to fertilize. So, in this way somatic embryogenesis is a tool of Plant Biotechnology in which we can obtain such a plant of two different genetic properties.
‡Barriers (Ex-Psidium guajava)

Wilt disease caused by Fusarium oxysporium psidii Stress sensitivity Low fruit growth Short shelf life High seed content

‡ Somatic embryogenesis could be induced from callus derived from various explants. ‡ Explants of Somatic Embryogenesis: Roots Stems Flower buds Petioles Immature leaf Immature hypocotyls

Figure: Schematic representation of various genes identified influencing somatic embryogenesis in higher plants.

‡ Embryogeneic cultures are compact dry, amorphous, and white in colour as compared to non embryogenic cultures which are watery dirty white to light brown in colour and soft in nature.

Embryonic Cultures

Non-Embryonic Cultures

SOMATIC EMBRYOGENESIS Direct Somatic Embryogensis: In direct embryogenesis the embryo is formed directly from a cell or group of cells without the production of an intervening callus. Indirect Somatic Embryogeneisis: When embryo is formed via intervening callus phase i.e. involvement of callus, it is called indirect Somatic embryogenesis.

Figure: Somatic Embryogenesis Pathway

Characters of Somatic Embryo 
The origin is single cell  Bipolar i.e., they have both shoot and root primordia  Vascular connection between the somatic embryos and the explant is absent  The somatic embryos are easily separated from the explant tissue

Distinct stages in development of Somatic Embryogenesis
1. 2. 3. 4. 5. Single cells Group of cells Globular stage Heart shaped embryo Torpedo stage embryo

Stages of Somatic Embryogenesis
1. Embryo initiation: In the initial stage, a high concentration of 2,4 -D is used. 2. Embryo production: In the second stage, embryos are produced in a medium with no or very low level of 2,4D.

* In many systems it has been found that somatic
embryogenesis is improved by supplying a source of reduced nitrogen, such as specific amino acids e.g glutamine or casein hydrolysate.

Induction of Somatic Embryo¶s Formation
Induction of somatic embryogenesis requires a change in the fate of a vegetative (somatic) cell. In most cases, an inductive treatment is required to initiate cell division and establish a new polarity in the somatic cell. In alfalfa, 2,4-D is most commonly used as a inductive treatment, but other auxins such as 2,4,5-T are effective.
Plant Growth Regulators (PGRs) Auxin Cytokinin Abcisic acid Induction Role Initiator of somatic embryos Used for secondary embryogenesis Regulate carbohydrate metabolism, Seed formation, stress response Elongation of internodes, Inhibit shoot formation and adventitious root Examples 2,4-D, NAA, IAA, IBA BAP, Kinetin, Zeatin, 2-iP ABA

Gibberellins

GA3, GA1, GA20

Differences / comparison between organogenesis and embryogenesis
Characteristics Origin Organogenesis Many cells , usually superficial Unipolar Present; vascular strands connected with those present in callus / explant Embryogenesis Single cell, usually superficial Bipolar Absent; there is no vascular connection with callus/ explant

Polarity Vascular connection with callus / explant

Separation from callus/ explant

Not easity separated unless cut off

Easily separated since the radicular end is cutinized

1.

2. 3. 4. 5.

Importance of somatic embryogenesis Clonal propagation: - the mass production of adventitious embryos in cell culture is one of the ways of clonal propagation. For genetic transformation Raising somaclonal variation in tree species with the help of mutation Synthesis of artificial seeds Somatic embryogenesis provides potential plantlets in the form of somatic seeds.

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