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

本研究的目的是：(1)通过嵌合体的形成和维持，明确正常线粒体向CMS线粒体的异质性变异和关闭。(2)细胞器的基因组结构必须清晰，特别是有叶绿体的有丝分裂。(3)在(1)和(2)结果的基础上，探讨了嵌合体诱导不育系的调控机制，结果表明，用已知的线粒体和叶绿体探针检测的RFLP图谱与指示基因orf138的〓萝卜不育系基本一致。通过使用指标orf138，可以识别出三个具有英雄气概的序列。从ORF 138向外延伸3kb 3‘方向的序列中，99%的序列与不育系萝卜(Maccolof)的序列一致，但基于序列的叶绿体碱基没有被鉴定。目前还无法确定完整的图谱或序列。然而，两种细胞器中的正常基因组分子(S)已分化为CMS类型的一般情况非常清楚，没有分子间重组，这一结果与萝卜与甘蓝属间细胞融合杂交的结果相同。从我们的可比性结果来看，通过组织培养、嵌合体合成等方法诱导和控制CMS数量和质量的基因组构建是可能的。进一步的机理研究将开创新的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