A novel method for separation of somatic embryos from embryogenic suspension cultures by cold treatment

This study reports a novel method for embryo separation by cold treatment of heterogeneous suspension cultures which contain embryogenic single cells, cell clusters and embryos at various stages of development. The method was applied to embryo suspension cultures of pepper (Capsicum annuum) and sugarcane (Saccharum officinarum). In both plants, single cells lost their viability dramatically over a few days while the viability of embryos remained above 95% for 25-30 days when kept at 5 degreesC. The effect of duration of cold treatment on embryo germination was also tested. The optimal duration of cold treatment was found to be 10 days for sugarcane and 21 days for pepper. After the cold treatment, the germination percentages were 90% and 96% for sugarcane and pepper, respectively.