Hervé Etienne, Benoît Bertrand, Eveline Dechamp, Patrick Maurel, Frédéric Georget, Romain Guyot and Jean-Christophe Breitler
In plants, undifferentiated or totally differentiated cells can be easily in vitro cultured to generate undifferentiated embryogenic cells that can regenerate complete plants. This is the most spectacular expression of totipotency. Embryogenic cells represent a key material in plant biotechnologies as they are used in many processes aiming at asexual reproduction by somatic embryogenesis (SE), genetic transformation, protoplast regeneration and cryopreservation. Nevertheless, tissue culture systems that involve the acquisition of competence for totipotency and extensive cell division remain risky with respect to genome and epigenome instabilities. Particularly, the use of embryogenic cell suspensions has frequently been associated with an increased likelihood of genetic instability and somaclonal variation (SV) in the regenerated plants. SV is a major concern in all in vitro vegetative plant propagation systems because it leads to the loss of genetic fidelity. Since 20 years, embryogenic cell suspension based SE techniques have been applied to coffee for the large-scale dissemination of exceptional Arabica hybrids. Here, we present our production and research experience showing that SE is efficient and reliable for true-to-type propagation. Over 99% of coffee trees regenerated fully conform to the mother plant, both morphologically - they grow, flower and produce normally. Hence strong genetic and epigenetic changes in proliferating embryogenic cells are not a fatality. The results also demonstrated the importance of embryogenic culture age on SV and hence the non-random nature of this phenomenon. The genetic and epigenetic alterations are particularly limited during SE. The main change in most of phenotypic variants was aneuploidy showing that mitotic aberrations play a major role in SV in coffee. These results provide a proof of concept for the use of embryogenic cell suspensions with other plant species: a revolution in the world of plant micropropagation on an industrial scale.
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