What is somatic embryogenesis?
Oxford dictionary describes “somatic” as “relating to the body, especially as distinct from the mind.”, however adding that in biology it means “relating to the soma”. The soma is all parts of an organism except for reproductive cells. In other words: all cells that are differentiated and part of an organ or tissue type etc. can be called somatic cells. Non-somatic cells are for example stem cells.
Embryogenesis describes the formation and development of an embryo. It is important to know that not only animals have embryos but plants have those as well.
The term totipotency describes the ability of (most) plants to form a complete organism from somatic cells given the right conditions. These conditions can be produced in vitro by offering culture media containing plants hormones, also called plant growth regulators (PGRs). Many plant species have the potential to form organs (roots / shoots) and thereupon to regenerate complete plants out of single plant cells or tissue fragments using a developmental pathway called organogenesis. In somatic embryogenesis however, a somatic cell is induced in vitro to adopt embryogenic properties. This artificial process is the only way to propagate most conifer species efficiently.
The generation of a cell culture with embryogenic competence is called initiation. The resulting embryogenic cell culture (clone) contains early embryos and can be propagated on media. The maintenance and multiplication of cultures is called proliferation. The fresh mass must periodically be placed on new nutrition medium; alternatively, it is possible to store the clone in liquid nitrogen. The next step in somatic embryogenesis is called maturation. During maturation the early embryos grow and develop into mature, cotyledonary embryos. To produce emblings (i.e. somatic seedlings), mature embryos can be placed on germination media under grow lights. Germination is the last in vitro step and still performed under sterile conditions. During acclimatization, seedlings are placed in soil and slowly accustomed to ex vitro conditions.
Somatic embryogenesis is an important tool in plant genetics and forestry, as it is possible to generate large numbers of embryos and thus plants from a single cell. This is particularly useful for plants where naturally obtained seeds are limited. Furthermore, this system allows us to closely monitor early plant development and long term studies on the same genetic material, as plants derived from the same initiation explant and position are considered clones.
Initiation of Embryogenic Cultures in Conifer Species
Initiation of cell cultures for woody plants is a little more complicated than placing any cell on culture media. For conifers, an immature zygotic embryo is prepared from healthy seeds and placed on culture media containing a combination of PGRs (auxin and cytokinin).
We’d love to establish a protocol for conifers to induce embryogenic cell cultures from adult trees in the future.

Maturation of Embryogenic Cultures of Conifer Species
For the maturation of conifer somatic embryos, a different combination of PGRs is needed. In this step, the immature somatic embryos grow from the P0 phase via a globular to a mature cotyledonary somatic embryo. The developmental stages of somatic embryos are similar to zygotic embryos in seeds, both morphologically and physiologically.

Germination of Somatic Embryos in Conifer Species
Somatic embryos, that show normal early development regarding their morphology, are stimulated to form roots on germination medium with activated charcoal (germination or conversion). Germination is the last step of somatic embryogenesis, which is carried out in vitro under sterile conditions. Under grow lights, the formerly pale embryos develop into seedling with green needles.


Germination of somatic embryos on black culture medium (supplemented with activated charcoal) into somatic emblings (corresponding to seedlings) of Larix x eurolepis.
Cryopreservation of Embryogenic Cultures
For cryopreservation, water needs to be removed from the embryogenic cell culture on osmotically active medium. Using antifreezing substances (e.g. dimethyl sulphoxide, DMSO) and by slowly lowering the temperature, the formation of ice crystals is reduced to protect the cells. Afterwards, the culture can be stored long-term in liquid nitrogen. At any point, the embryogenic culture can be thawed and cultivated again, ensuring the conservation of the clone.



