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Studies on dynamic changes of molecular distribution in plant cells by immuno-electron microscopy

免疫电镜研究植物细胞内分子分布的动态变化

基本信息

批准号：
02304007
负责人：
KUROIWA Tsuneyoshi
金额：
$ 13.25万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

We have developed an immuno-light-electron microscopy and examined the dynamic changes of DNA and proteins in plant cells. As a result, we have obtained new data about these problems. Especially, Osafune and Ueda improved the immuno-gold-electron microscopy (IGEM) to obtain a high resolution. The newly developed techniques are applied to various fields of these investigators including the group. Kuroiwa developed a technique for the simultaneous observations of light microscopic field and electron microscopic field and applied the technique to analyzing the multiplication of cells in root meristems of Pelargonium zonale, Nicotiana tabacum and Arabidopsis thaliana. The results showed that the mtDNA content per mt-nucleus in the cells just above quiescent center, where the synthesis of mtDNA is active, corresponds to approximately 3000 kilobase pairs but, in the meristematic cells just below the elongation zone of the root it falls less than 170 kbp. Murakami showed using IGEM that the photosystem I is located in thylakoid membrane systems, while the photosystem II in stroma thylakoids. Osafune found by a high resolution IGEM that LHCPII apoprotein transferred to plastid through the Golgi apparatus. Ueda showed that the actin is located on the surface of lysosomes in Trebouxia. Tanaka, Yamaguchi and Miyakawa revealed the behavior of proteinase, the formation of virus-like particles and the existence of mitchondorial nuclear proteins in yeast cells, respectively.

我们建立了一种免疫光电子显微镜，并研究了植物细胞中DNA和蛋白质的动态变化。结果，我们获得了关于这些问题的新数据。特别是Osafune和上田改进了免疫金电子显微镜（IGEM），以获得高分辨率。新开发的技术适用于这些调查人员的各个领域，包括该集团。Kuroiwa开发了一种同时观察光镜场和电镜场的技术，并将该技术应用于分析天竺葵、烟草和拟南芥根分生组织中细胞的增殖。结果表明，在静止中心上方的细胞中，mtDNA的合成是活跃的，每个线粒体核中的mtDNA含量相当于大约3000个线粒体酶对，而在根伸长区下方的分生组织细胞中，它的含量福尔斯低于170 kbp。Murakami利用IGEM证明光系统I位于类囊体膜系统中，而光系统II位于基质类囊体中。Osafune通过高分辨IGEM发现LHCP Ⅱ脱辅基蛋白通过高尔基体转移到质体。上田等研究表明，肌动蛋白主要分布在Trebouxia的溶酶体表面。Tanaka，Yamaguchi和Miyakawa分别揭示了蛋白酶的行为，病毒样颗粒的形成和线粒体核蛋白在酵母细胞中的存在。