The possibility of nitrogen fixation by genetic improving the rice chioroplast

通过基因改良水稻叶绿体固氮的可能性

• 批准号: 12460001
• 负责人: TSUTSUMI Nobuhiro
• 金额: $ 2.88万
• 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 (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460001/
• 关键词: stromule; ribosomal protein; GFP; Kaede; protein transport; mitochondria; 葉緑体; rpl36; トランジットペプチド; 水平伝播; rpl16

In order to visualize the shapes of plastids in a living cell, a fusion gene of GFP with the transit peptide of rice chloroplast ribosomal protein L12-1 (cpTP-GFP) was introduced into rice epidermal cells by the particle bombardment method. Confocal laser scanning microscopy showed that the GFP fluorescence was localized in the plastids.The cpTP-GFP signals were matched specially with the signals of chlorophyll autofluorescence. In addition, there were some cells that had plastids with various lengths of protuberances without chlorophyll, i.e., "stromules". In a few cells, stromules connected almost all the plastids in a cell, appearing as a pan-cytoplasmic network. One plausible function of stromule connections is to exchange plastid components such as proteins. This would unify or coordinate all of the plastids in a cell and, by lessening the demands on the nucleus, contribute to the productivity of the cell.To determine whether fusion of plastids mediated by stromules occurs, we labeled plastids in onion epidermal cells with a plastid-targeted fluorescent protein (Kaede) and then altered the fluorescence of some of the plastids within a cell from green to red. No transition to a uniform color (yellow) indicating the fusion of plastids was observed, even two days after the photo-conversion.

为了观察活细胞中质体的形态，采用基因枪法将绿色荧光蛋白（GFP）与水稻叶绿体核糖体蛋白L12-1转运肽（cpTP-GFP）融合基因导入水稻表皮细胞。激光共聚焦扫描显微镜显示GFP荧光定位在质体中，cpTP-GFP信号与叶绿素自发荧光信号特异性匹配。此外，还有一些细胞的质体具有不同长度的突起，没有叶绿素，即，“stromules”。在少数细胞中，几乎所有的质体都由基质连接起来，形成泛细胞质网络。子实体连接的一个可能的功能是交换质体成分，如蛋白质。这将统一或协调所有的质体在一个细胞中，通过减少对细胞核的需求，有助于生产力的cell.To确定是否发生融合的基质介导的质体，我们标记的质体在洋葱表皮细胞与质体靶向荧光蛋白（Kaede），然后改变荧光的细胞内的一些质体从绿色到红色。即使在光转化两天后，也没有观察到指示质体融合的均匀颜色（黄色）的转变。