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Somatic embryogenesis (SE) is the most important plant biotechnology process for plant regeneration, propagation, genetic transformation and genome editing of coffee,  Coffea arabica  L. Somatic embryo (SEs) conversion to plantlets is the principal bottleneck for basic and applied use of this process. In this study we focus on the maturation of SEs of  C. arabica  var. Typica. SEs conversion to plantlet up to 95.9% was achieved under osmotic stress, using 9 g/L gelrite, as compared with only 39.34% in non-osmotic stress. Mature SEs induced in osmotic stress developed shoot and root apical meristems, while untreated SEs were unable to do it.  C. arabica  regenerated plants from osmotic stress were robust, with higher leaf and root area and internode length. To understand a possible regulatory mechanism, gene expression of key genes of  C. arabica , homologous to sequences in the  Arabidopsis thaliana  genome, were analyzed. A set of two component system and cytokinin signaling-related coding genes ( AHK1, AHK3, AHP4  and  ARR1 ) which interact with  WUSCHEL  and  WOX5  homedomains and morphogenic genes,  BABY-BOOM, LEC1, FUS3  and  AGL15 , underwent significant changes during maturation of SEs of  C. arabica  var. Typica. This protocol is currently being applied in genetic transformation with high rate of success.

Osmotic stress-induced somatic embryo maturation of coffee Coffea arabica L., shoot and root apical meristems development and robustness