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分析了细胞周期进程和环境响应过程中的液泡和细胞骨架动力学，如下。（使用双结构可视化细胞分析植物细胞分裂）我按时间顺序跟踪使用组蛋白-RFP 和 GFP-微管蛋白研究烟草 BY-2 细胞中染色体分离和纺锤体伸长的动态。图像处理显示，高等植物细胞中，后期B对染色体距离分离的贡献约为40%。（液泡动力学和细胞板发育中肌动蛋白微丝的分析）我观察到了肌动蛋白微丝（MF）的共定位和液泡膜（VM），通过在稳定表达 GFP-fimbrinm 的活烟草 BY-2 细胞中用 FM4-64 进行活体 VM 染色可见，并发现MF 支持液泡动力学，而肌动球蛋白系统在液泡形态发生中起着重要作用。此外，通过 GFP-fimbrinm 的表达和 FM4-64 的活体染色，可以在活细胞中的整个植物胞质分裂过程中跟踪 MF 和细胞板的动态。实时成像分析使我们能够量化整个胞质分裂过程中 MF 对细胞板扩张的贡献，表明 MF 通过调节内膜动力学在细胞板形成和扩张中的作用转变。（气孔关闭和保卫细胞中液泡和 MF 的动态组织）昼夜周期）我发现细胞分裂素和生长素通过调节乙烯生物合成抑制 ABA 诱导的气孔关闭，并且保卫细胞液泡结构变得通过 3-D 重建，气孔关闭期间变得复杂。此外，在拟南芥植物中，我发现皮质 MF 呈放射状和随机排列，分别跟随气孔孔径的增加和减小，这表明保卫细胞的 MF 阵列可能控制昼夜气孔运动。