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Analyses of plant cell division and differentiation using structure-visualized cells and image processing

使用结构可视化细胞和图像处理分析植物细胞分裂和分化

基本信息

批准号：
18370015
负责人：
HASEZAWA Seiichiro
金额：
$ 10.64万
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：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

I have analyzed the vacuolar and cytoskeletal dynamics during the cell cycle progression and environmental response using the structure-visualized cells and image processing, as follows..(Analyses of plant cell division using dual-structure-visualized cells) I have time-sequentially followed the dynamics of chromosome segregation and spindle elongation in tobacco BY-2 cells using histone-RFP and GFP-tubulin. The image processing revealed that anaphase B contributes to about 40% of the chromosome separation in distance in higher plant cells.(Analyses of actin microfilaments in vacuolar dynamics and cell plate development) I have observed the co-localization of the actin microfilaments (MFs) and vacuolar membrane (VM), as visualized by vital VM staining with FM4-64 in living tobacco BY-2 cells stably expressing GFP-fimbrinm and found that the MFs support the vacuolar dynamics and the act-myosin system plays an essential roles in vacuolar morphogenesis. Moreover, by the expression of GFP-fimbrinm and vital staining with FM4-64, dynamics of MFs and cell plate could be followed throughout plant cytokinesis in living cells. Live imaging analysis allowed us to quantify the MF's contribution to the cell plate expansion throughout cytokinesis, suggesting a role transition of MFs in cell plate formation and expansion via regulation of endomembrane dynamics.(Dynamic organizations of vacuoles and MFs in guard cells during stomatal closure and diurnal cycle) I have found that cytokinins and auxins inhibit ABA-induced stomatal closure through the modulation of ethylene biosynthesis and that the guard cell vacuolar structures become complicated during stomatal closure by 3-D reconstruction. Moreover, in Arabidopsis plants, I have found that the cortical MFs are organized in radial and random array, following the increase and decrease in the stomatal aperture respectively, suggesting that the MF array of guard cells may control diurnal stomatal movement.

我使用结构可视化细胞和图像处理技术分析了细胞周期进程和环境响应过程中的空泡和细胞骨架动力学，具体如下：(利用双重结构可视化细胞分析植物细胞分裂)我使用组蛋白-RFP和GFP-微管蛋白对烟草BY-2细胞染色体分离和纺锤体伸长的动态进行了时间顺序的跟踪。图像处理表明，在高等植物细胞中，后期B约占染色体距离分离的40%。(肌动蛋白微丝在液泡动力学和细胞板发育中的分析)我观察到了肌动蛋白微丝(MFS)和空泡膜(Vm)的共存，通过FM4-活VM染色观察到稳定表达gfp-fimbrinm的活烟草BY-2细胞中的MFS支持空泡动力学，ACT-肌球蛋白系统在液泡形态发生中起重要作用。此外，通过GFP-Fimbrinm的表达和FM4-的活染色，可以跟踪活细胞中MFS和细胞板的动态变化。通过实时成像分析，我们能够量化MF在细胞质分裂过程中对细胞板扩张的贡献，表明MFS通过调节内膜动力学在细胞板形成和扩张中的角色转换。(气孔关闭和昼夜周期中保卫细胞中空泡和MFS的动态组织)我发现细胞分裂素和生长素通过调节乙烯的生物合成来抑制ABA诱导的气孔关闭，并且在气孔关闭过程中通过三维重建使保卫细胞的空泡结构变得复杂。此外，在拟南芥植物中，我发现皮层MFS分别随着气孔开度的增加和减少而呈放射状和随机排列，这表明保卫细胞的MF阵列可能控制气孔的昼夜运动。