Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench.)

In vitro fertilization could be useful to overcome the problem of self-incompatibility of common buckwheat. Therefore, experiments were conducted to isolate viable protoplasts from egg cells for use in vitro fertilization. Protoplasts from viable egg cells of common buckwheat were isolated by enzyme treatment followed by mechanical isolation. Incubation of ovules for up to 4 h allowed the boundary wall of the egg cells to become partially digested and the protoplasts of the egg cells came out of the micropylar or chalazal end after squashing with a mean isolation frequency of approximately 30% (6 out of 20 ovules). The egg cell protoplasts were larger than those of the synergids. Key words: Common buckwheat, Fagopyrum esculentum Moench., protoplast, egg cell solation, embryo sac