Studies on the Induction of Cytoplasts from the Protoplasts of CMS(Cytoplasmic Male Sterility) Line of Nicotiana and the Fusion of the Cytoplast and the another Protoplasts

담배 CMS line의 원형질체로부터 cytoplast의 유도 및 이와 타품종 원형질체와의 융합에 관한 연구

This study was investigated as a step for the purpose of successful introduction of cytoplasmic inherited characters between the different plants. Cytoplasts were separated from the protoplasts of CMS(cytoplasmic male sterility) line such as MS Burley 21 which carried from Nicotiana megalosiphon. The cytoplasts were fused to protoplasts derived from Nicotiana tabacum Br 64 with PEG(polyethylene g1yco1). The cytoplasts were separated by density gradient centrifugation. Efficient separation of cytoplasts depended on the difference of specific density of gradient solution. However, the iso-osmolality of gradient solution was not important to separate the cytoplasts. The cells for a cybrid were fused with 50% concentration of PEG.

세포질 유전형질의 도입을 성공적으로 수행코자 세질 웅성 불임(eMS; cytoplasmic male sterili­t ty) 계통의 담배 MS(male sterility) Burley 21의 callus tissue에서 cytoplast를 분리시키고 이와 Ni­c cotiana tabacum Burley 64의 mesophyll protoplast 를 PEG방법에 의한 세포융합으로 cybrid cell올 유도할 수 있었다. cytoplast의 분라는 density gradient solution의 osmolarity를 일정하게 하는 것보다 gradient간의 차이가 더 효과적이였으며 분리된 cytoplast의 양은 재료로 이용된 protoplast의 20-30% 정도를 나타 내었다. 또한 cytoplast와 protoplast의 융합은 P PEG농도 50%에셔 효과적이었다.