Isolation, Culture and Electroporation of Rice Protoplasts

벼 원형질체의 분리, 배양 및 Electroporation에 관한 연구

Culture of embryogenic callus and suspension were induced from rice seeds in MS2.5 medium. In hormone free N6 medium, whole plantlets were regenerated from embryogenic callus. We observed cell division and reformation of embryogenic callus on culture of protoplast isolated from embryogenic cell suspensions. In addition, we studied the influencing factors on viability of protoplast treated with electroporation. Viability was decreased according to the increase of voltage and capacitance during electroporation. An optimal level of viability was obtained after treatment with $200-300\;V/1180\;\mu\textrm{F}$ in HEM buffer at $4^{\circ}C$..

벼의 종자로부터 embryogenic callus를 유도하고 이로부터 개체로의 재분화를 유도하였으며, embroyogenic cell suspension culture를 통하여, 이로부터 원형질체를 분리, 배양하여 embryogenic callus를 형성하였다. 또한, 분리된 원형질체를 electroporation하였을 때 생존율(viability)에 미치는 여러 요인들을 조사하였다. 원형질체의 생존율은 voltage와 capactiance가 증가할수록 감소를 보였으며, HBM buffer에서 $4^{\circ}C$로 electroporation 하였을 때 생존율에 보다 효과적이었다.