Development of Transformation System for Plant Mitochondria

植物线粒体转化系统的开发

• 批准号: 06556002
• 负责人: HIRAI Atsushi
• 金额: $ 9.34万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06556002/
• 关键词: Mitochondrial genome; Plasmid-like DNA; Rice; Transformation; アンチマイシン; エディティング

Mitochondria are the sites of energy transduction in eukaryotic cells which carry out aerobic metabolism. Energy transduction is conducted by several kinds of peotein complexes in the mitochondrial membrane. Genetic information of these complexes are stored in the nuclear and mitochondrial genomes. Thus, mitochondrial as well as nuclear genome should be transformed in order to improve mitochondrial activity.Plasmid DNA was constructed to transform rice mitochondrial DNA.Mitichondrial coded cob gene was mutated to antimycin resistant form by the site directed mutagenesis. The mutated cob gene, together with promoter region of atpl and 3' non-translated region of atp6 were cloned into pBSKS+ plasmid. Plasmid like DNA,B4, in rice mitochondria was also connected to the plasmid in order to accelerate the replication of the plasmid DNA in rice mitochondria. This plasmid DNA for mitochondria transformation was introduced into rice calli by a particle gun. However, transformed cells were not obtained so far. Construction of the improved plasmid is in progress.

线粒体是真核细胞中进行有氧代谢的能量传递部位。能量转导是通过线粒体膜上的几种人蛋白复合体进行的。这些复合体的遗传信息存储在核和线粒体基因组中。因此，为了提高线粒体的活性，必须同时对线粒体基因组和核基因组进行转化。构建了转化水稻线粒体DNA的质粒DNA，通过定点突变将线粒体编码的cob基因突变为抗霉素抗性的形式。将突变的cob基因与atp1启动子和atp6的3‘非翻译区一起克隆到pBSKS+载体中。为了加速水稻线粒体中的复制，还将水稻线粒体中的B4等质粒连接到该质粒上。用粒子枪将线粒体转化质粒DNA导入水稻愈伤组织。然而，到目前为止还没有获得转化细胞。改进后的质粒的构建正在进行中。

项目成果

Nishiwaki, S. et al.: "Structure of a gene for subunit 9 of NADH dehydrogenase (nad9) in rice mitochondria and RNA editing of its transcript." Plant Cell Physiol.36. 1135-1138 (1995)

Nishiwaki, S. 等人：“水稻线粒体中 NADH 脱氢酶 (nad9) 亚基 9 的基因结构及其转录物的 RNA 编辑。”

Miyata,S.et al.: "Polymorphic distribution and molecular diversification of mitochondrial plasmid-like DNAs in the genus Oryza." Jpn.J.Genet.70. 601-614 (1995)

Miyata,S.et al.：“稻属线粒体质粒样 DNA 的多态性分布和分子多样化。”

Kadowaki,K.et al.: "Targeting presequence acquisiton after mitochondrial gene transfer to the nucleus occurs by duplication of existing targeting signal." EMBOJ.15. 6652-6661 (1996)

Kadowaki,K.等人：“线粒体基因转移到细胞核后，通过复制现有的靶向信号来获取靶向前序列。”

Nakazono, M.et al.: "Homologous recombination mediated by two palindromic repeated sequences in the mitochondrial genome of Oryza" Theor. Appl. Genet.91. 1-8 (1995)

Nakazono, M.等人：“Oryza 线粒体基因组中两个回文重复序列介导的同源重组”理论。

Nakazono,M.et al.: "The CRTA motif is present in the promoters of mitochondrial genes of rice." Plant.Sci.(in press.).

Nakazono,M.等人：“CRTA 基序存在于水稻线粒体基因的启动子中。”