Mechanism of the diletion of plastid DNA in pollen-derived albino and its application as a vector.

花粉源白化质体DNA的消耗机制及其作为载体的应用。

• 批准号: 03660001
• 负责人: HARADA Takeo
• 金额: $ 1.41万
• 依托单位: Hirosaki 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-03660001/
• 关键词: Rice; Anther culture; Albino; Chloroplast DNA; Plastid DNA; Linear DNA; Hairpin telomere; Vector; ベクター

Albino rice plants derived from pollen contain plastid genomes that have suffered large-scale deletions. From the roots of albino plants, we obtained several calli containing homogeneous plastid DNA differing in the size and position of the deletion. Southern blotting and pulsed field gel electrophoresis experiments revealed that the DNAs were linear molecules having a hairpin structure at both termini, existing as monomers (19 kb) or dimers, trimers and tetramers liked to form head-to-head and tail-to-tail multimers. This characteristic form is similar to that of the vaccinia virus, in which the replication origin is thought to lie at or near the hairpin termini. Farthermore, polymerase chain reaction experiments revealed complete loss of the ribosomal RNA genes of the plastid DNA. The results suggest that plant cells can grow without translation occurring in plastids. All of the deleted plastid DNAs commonly retained the region containing the tRNA^<Glu> gene(trnE), which is essential for biosynthesis of porphyrin. As porphyrin is the precursor of heme for mitochondria and other organelles, it is considered that trnE on the remnant plastid genome may be transcribed by an RNA polymerase encoded on nuclear DNA.

来自花粉的白化水稻植株含有遭受大规模缺失的叶绿体基因组。从白化植物的根中，我们获得了几个含有均一质粒DNA的愈伤组织，其缺失的大小和位置不同。Southern印迹和脉冲场凝胶电泳实验表明，DNA是线形分子，两端都有发夹结构，以单体(19kb)或二聚体、三聚体和四聚体形式存在，喜欢形成头到头和尾到尾的多聚体。这种特征形式与痘苗病毒相似，后者的复制起点被认为位于发夹末端或附近。此外，聚合酶链式反应实验显示，叶绿体DNA的核糖体RNA基因完全丢失。结果表明，植物细胞可以在没有翻译的情况下生长在叶绿体中。所有缺失的叶绿体DNA都保留了含有tRNA^&lt；Glu&gt；基因(TrnE)的区域，这是卟啉生物合成所必需的。由于卟啉是线粒体和其他细胞器的血红素的前体，因此认为残存的叶绿体基因组上的trnE可能是由核DNA上编码的RNA聚合酶转录的。

项目成果

原田 竹雄: "Large-scale deletions of plastid DNA in rice anther culture." Theor.Appl.Genet.81. 157-161 (1991)

Takeo Harada：“水稻花药培养物中质体 DNA 的大规模删除。”Theor.Appl.Genet.81（1991）。

原田 竹雄: "Pollen-derived rice calli that have large deletions in plastid DNA do not require protein synthesis in plastids for growth." Mol.Gen.Genet.233. 145-150 (1992)

Takeo Harada：“质体 DNA 中有大量缺失的花粉来源的水稻愈伤组织不需要质体中的蛋白质合成来生长。”Mol.Gen.Genet.233 (1992)。

原田 竹雄: "Large-scale deletions of plastid DNAin rice anther culture." Theor.Appl.Genet. 81. 157-161 (1991)

Takeo Harada：“水稻花药培养物中质体 DNA 的大规模删除。”Theor.Appl.Genet 81. 157-161 (1991)

原田 竹雄: "The conformation of largeーscale deleted plastid DNA in rice calli derived from pollen show no requirement for translation in plastid for plant cell growth to occur." Mol.Gen.Genet.

Takeo Harada：“来自花粉的水稻愈伤组织中大规模缺失的质体 DNA 的构象表明，植物细胞生长不需要质体翻译。”