Manipulation of mitochondria and chloroplast genome organization by artificially-synthesized chimera

我的目标是：(1)通过嵌合体的形成和维持，明确从正常线粒体到CMS线粒体的异质性变异和转换。(2)为此，必须明确细胞器的基因组结构，特别是有叶绿体的线粒体。(3)在(1)和(2)的基础上，探讨嵌合体合成诱导CMS的调控机制。结果表明，用已知的线粒体和叶绿体探针探测到的RFLP模式与含有orf138指示剂的〓萝卜CMS几乎一致。利用orf138指示剂，可以鉴定出3个具有英雄质粒的序列。从orf138向3kb 3’方向延伸的序列中，99%与CMS萝卜（Maccolof）相同。然而，叶绿体碱基的序列尚未确定。完整的地图或序列尚未确定。然而，在没有分子间重组的情况下，两种细胞器从正常基因组分子转变为CMS类型的总体情况非常清楚。这些结果与萝卜和卷心菜的属间细胞融合杂交结果相同。从我们的比较结果来看，将有可能通过组织培养、嵌合体合成等方法诱导和控制CMS的定量和定性基因组构建。进一步开展机理研究，建立新的CMS和杂交育种体系

My objective was to (1) make clear the heteroplasmic variation and shuft from normal mitochondria to CMS one by chimera formation and maintainance. (2) For that, genome structure of organelle must be nade clear, especially for mitcondria with chloroplast. (3) Base on the results of (1) and (2), controlling mechanism of the induction of CMS by chimera synthesis.As the results, RFLP patterns probed with known mitochodra and chloroplast probes were almost corresponded with that 〓radish CMS that has indicator orf138. By using the indicator orf138, three has heroplasimic sequnenses could be identified. Totally 99% of those sequences that were extended 3kb 3' direction from orf 138 were the same as that of CMS radish (by Maccolof).However, chloroplast bases based on sequence is not identitifoed. Full map or sequence could not be identified, yet. However, general ssituation that has been shited from normal genome molecule(s) to CMS types in the both organella is very clear without inter-molecular recombination.Those results were just the same as in intergeneric cell fusion hybrid between radish and cabbage. From our comparable results, it will be possible to induce and control CMS quantitative and qualitative genome constraction by tissue culture, chimera synthesis etc. Further the mechanism research should originate new CMS and hybrid breeding system