SOMATIC EMBRYOGENESIS

 In somatic embryogenesis, the embryos regenerate from somatic cells, tissues or organs either de
novo or directly from the tissues (adventive origin), which is the opposite of zygotic or sexual
embryogenesis. Various forms of somatic embryos are reported viz. adventive embryos (somatic
embryos arising directly from other organs or embryos), parthenogenetic embryos (those formed
by the unfertilized egg), and androgenetic embryos (formed by the male gametophyte).
 In sexual embryogenesis, the act of fertilization triggers the egg cell to develop into an embryo.
However, it is not the monopoly of the egg to form an embryo. Any cell of the gametophytic
(embryosac) or sporophytic tissue around the embryo-sac may give rise to an embryo. Cells of the
nucellus or inner integument of members of Rutaceae family (e.g., Citrus) may develop into
embryos. The occurrence of asexual embryogenesis is generally restricted to intra ovular tissues.
 However, in genera, somatic embryos are those which are formed from the somatic tissues like
epidermis, parenchymatous cells of petioles or secondary root phloem in vitro. Somatic
embryogenesis differs from organogenesis as the embryo being a 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 maternal callus tissue or the cultured explant.
 The initiation and development of embryos from somatic tissues of Daucus carota was first
reported by Steward et al. (1958) and Reinert (1958, 1959). Embryos have also been obtained from
generative cells of Datura innoxia microspores (Guha and Maheshwari (1964). The list of species
from which somatic embryogenesis is reported is exhaustive.
 The members of Umbellifereae and Solanaceae, Leguminoseae and a range of other dicots along
with monocots families’ viz. Gramineae show the ability to generate somatic embryos. In this
chapter embryogenesis will be restricted to sporophytic tissue; the discusssion on androgenesis is
dealt separately. Sharp et al. (1980) described two routes to somatic embryogenesis.

Stages of somatic embryos

Globular, heart, torpedo and cotyledonaty states of somatic embryos
 Direct Embryogenesis
 The embryos initiate directly from the explant tissue without the intervening callus. This occurs
through ‘Pre-Embryogenic Determined Cells’ (PEDCs) where the cells are committed to develop
embryos and needs potentiation. Such PEDCs are found in embryonic tissues (e.g., scutellum of
cereals), and in certain tissues of young in vitro grown plantlets (e.g., hypocotyl in Daucus carota,
Ranunculusscleratus, Linum usitatissimum, Brassica napus), nucellus and embryo-sac (within ovules
of mature plants). Thormatie embryo development is triggered by transferring the PEDCs to a low
auxin containing medium or only to the basal medium.
 Subsequent conversion of somatic embryos into plantlets occurred mostly in hormone free media or
in specific plants on media supplemented with GA 3 , BAP and IBA in Psoralea corylifolia, Eryngium
foetidum and Aconitum carmichaeli (Chand and Sahrawat, 2002;Ignacimuthu et al., 1999; Hatano et
al., 1987).

Indirect Embryogenesis
 Indirect embryogenesis involves an intermittent callus formation process from which the somatic
embryos develop. The embryogenically determined cells in the callus form embryos and as these
are induced to do so, they are also called ‘Induced Embryogenic Determined Cells’ (IEDCs), e.g.,
secondary phloem of carrot, leaf tissues of coffee, Petunia, Asparagus, etc. Specific growth
regulator concentrations and/or cultural conditions are required for these stages in somatic
embryogenesis.
 Somatic embryos arise from single cells located within clusters of meristematic cells either in the
callus mass or in suspension. Such cells develop into proembryos, with polarity following a pattern
that tends to mimic the general pattern associated with the development of embryos in the ovule.
Proembryo initials may be single cells or multicellular groups.
 For some species any part of the plant body serves as an explant for embryogenesis (e.g. carrot),
whereas in some species only certain regions of the plant body may respond in culture e.g., Cereals.
Floral or reproductive tissue in general has proven to be an excellent source of embryogenic
material. Somatic embryos have been grown on a range of media from the dilute White’s medium
to very high salt MS medium. The addition of reduced nitrogen in the medium helps in both embryo
initiation and maturation. Of all the amino acids, L-glutamine seems to play a special role.
 Another factor is the chelated form of iron in the media. In the absence of iron, embryo
development fails to pass from the globularto the heart-shaped stage.
 Growth regulators in the medium, especially auxin or auxin in combination with cytokinin appear
essential for the onset of growth and the induction of embryogenesis. Of all the auxins, 2,4-D
followed by NAA has proven to be extremely useful. Effective concentration ranges are 0.5-27.6 µM
for 2, 4-D and 0.5-10.7 µM for NAA. Cytokinins have been used in the primary medium invariably
during embryogenesis of crop plants. The effective concentration range for kinetin is 0.5-5.0 µM.
Cytokinins are important in fostering somatic embryo maturation and especially cotyledon
development. Cytokinins are sometimes required for growth of embryos into plantlets. Gibberellins
are rarely incorporated in primary culture media. ABA, antiauxins (5-hydroxynitrobenzyl bromide)
and other growth inhibitors may serve to promote somatic embryo maturation by countering the
effects of growth promoters.

Limitation
High probability of mutation. ii) Methodological difficulty.
Loss of regenerative capacity with repeated subcultures.
Induction of embryogenesis is often very difficult or impossible with many recalcitrant plants.
Occurrence of somatic embryo dormancy.

Advantages
Somatic Embryogenesis Is the Preferred Method for In Vitro Propagation of Woody, crop and medicinal
Plants. Synthetic Seeds can be prepared with somatic embryos for cryopreservation and in vitro
cultivation for clonal propagation.

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