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-inco mpatibility of common buckwheat. Therefore, experiments were conducted to i solate viable protoplasts from egg cells for use in vitro fertilization. Pr otoplasts 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 digeste d and the protoplasts of the egg cells came out of the micropylar or chalaz al 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 t he synergids.