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

Sun Hee Woo¹., Taiji Adachi²., Seung Keun Jong³ and Clayton G. Campbell⁴

¹ Agriculture and Agri-Food Canada, Crop Diversification Centre, Morden, Manitoba, Canada R6M 1Y5; ² Applied Genetics and Biotechnology Division, Faculty of Agriculture, Miyazaki University, Gakuen Kibanadai Nishi 1-1, Miyazaki 889-2154, Japan; ³Dept. of Agronomy, College of Agriculture, Chungbuk National University, Cheongju 360-763, Korea; ⁴ Kade Research Ltd. Morden, Manitoba., Canada R6M 1E9.

Sun Hee Woo.,Taiji Adachi., Seung Keun Jong and Clayton G. Campbell. 1999. Isolation of protoplasts from viable egg cells of common buckwheat (Fagopyrum esculentum Moench). Can. J. Plant Sci. 79: 593-595.

Abstract: 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 four hours 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 isolation, embryo sac.