Cyclic somatic embryogenesis in indonesian elite Theobroma cacao L. clones

To date, in vitro somatic embryogenesis is the only option for the mass vegetative propagation of cocoa. The somatic embryogenesis of Indonesian cocoa clones SUL1 and SUL2 was investigated, focusing on primary and cyclic secondary embryogenesis. The study showed that staminode explants were more effective than petal explants in generating primary somatic embryos (SEs), especially when cultured in liquid medium containing 2 mg/L 2,4-D and 0.25 mg/L kinetin, with the staminodes of SUL2 producing a significant number of globular SEs. In contrast, SUL1 showed limited SE production. The study also demonstrated that fragmenting cotyledons and hypocotyls of the SEs of SUL2 and culturing them on an induction medium supplemented with 2,4,5-T and proline resulted in a high yield of secondary SEs. This cyclic embryogenesis process, in which the SEs remained attached to the maternal tissue, facilitated continuous SE production and development. The addition of proline was found to improve the quality of SEs, leading to higher production of well-organized, milky SEs with a better-defined meristematic structure. These results suggest a promising protocol to produce SEs from cocoa, with implications for plant transformation and gene editing applications.