I got friable embryogenic callus form hypocotyl explant of cotton on MSB5 medium supplemented with 2,4-D and kinetin and got well proliferation with TDZ+IBA. After proliferation, i transferred the callus to PGR free medium. Did subculture at regular intervals (every 2 weeks). Maintaining the callus for 3 months. Now i dont know how to get somatic embryo from this callus. Does anyone have idea about this?
You can augment the osmotic potential into the medium with more contents of sucrose (6-9%) for example, combined with ABA (concentration you need determined). Usually the process of induction of somatic embryos is based in physiological stress where the induced desiccation to generate the induction embryos.
my experience with ziziphus , to get embryos from embryogenic callus is to transfer the embryogenic callus to MS medium free of hormones.
Literature indicates strong genotypic response dependent. That means some genotypes might be easier than others to achieve embriogenic response... You may check this in the literature...
get the embryo and culture in ms media or B5 and use different combination of auxin (IAA,IBA) and kinetine