RFLP analysis of rice organelle DNA in regenerated plant

再生植株水稻细胞器 DNA 的 RFLP 分析

• 批准号: 03660002
• 负责人: ABE Toshinori
• 金额: $ 1.54万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03660002/
• 关键词: Chloroplast DNA; Mitochondria DNA; RFLP; Rice callus; Plant regeneration; Mutation; Insertion; Deletion; 分子育種; 再分化植物; オルガネラDNA; サザンハイブリダイゼ-ション

RFLPs of rice organelle genomes were analysed by Southern hybridization technique. Mutation which occurred in organelle genomes through rice callus cultures were detected by this method and gained RFLPs.1. Generally, RFLPs were seen in organelle genomes (chloroplast and mitochondoria DNAs) in rice calli subcultured after 4 months. Mutation ratio of calli which showed RFLPs was higher after long term subculture (7 passages) than short term subculture (4 passages). The ratio of organelle DNAs after long term subcultures (33 passages) were high in chloroplast, pst 2 probe (12-22%), and in mitochondoria, atp 6 probe (26-37%).2. Some RFLP patterns of regenerated plants were the same as those of calli which initiated the plantlets. Mutation ratio of organelle DNAs in regenerated plants was below 10% and lower than those of calli.3. Specific fragments other than normal ones were frequently observed, indicating that specific fragments were exclusively polymerized and detected by hybridization. Dletion and insertion which meke other types of RFLPs were also observed. Deletion of psa A and psa B region in chloroplast DNA and insertion of rrn 26 region in mitochondria DNA in rice were clearly observed.It is required to elucidate the relationship between mutation in RFLP level and protein translation and/or agronomic characteristics.

利用Southern杂交技术对水稻细胞器基因组进行了RFLP分析。利用该方法对水稻愈伤组织培养过程中细胞器基因组发生的突变进行检测，获得RFLPs1。继代培养4个月后，水稻愈伤组织的细胞器基因组(叶绿体和线粒体DNA)中普遍存在RFLPs。长期继代(7代)的愈伤组织突变率高于短期继代(4代)。长期传代(33代)后，细胞器DNA在叶绿体、PST-2探针(12-22%)和线粒体DNA(26-37%)中的比例较高。再生植株的某些RFLP图谱与启动植株的愈伤组织的RFLP图谱相同。再生植株细胞器DNA突变率低于10%，且低于愈伤组织。经常观察到正常片段以外的特定片段，表明特定片段被专一性聚合并通过杂交检测到。还观察到了与其他类型RFLP类似的缺失和插入现象。在水稻叶绿体DNA中发现了PSA A和PSA B区的缺失，在线粒体DNA中插入了rrn 26区，这需要阐明RFLP水平上的突变与蛋白质翻译和/或农艺性状的关系。

项目成果

Chowdhury, M. K. U. et al.: "Molecular analysis of organelle DNA of different subspecies of rice and the genomic stability of mtDNA in tussue cultured cells of rice." Theor. Appl. Genet.76. 533-539 (1987)

Chowdhury, M.K.U. 等人：“不同水稻亚种的细胞器 DNA 的分子分析以及水稻 tussue 培养细胞中 mtDNA 的基因组稳定性。”

阿部 利徳・笹原 健夫,平井 篤志: "イネミトコンドリアDNAのRFLP分析 II.種間差異" 育種学雑誌. 42(別2). 244-245 (1992)

Toshinori Abe、Takeo Sasahara、Atsushi Hirai：“水稻线粒体 DNA 的 RFLP 分析 II。种间差异”《育种科学》42（第 2 部分）。