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

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反应过程中的液泡和细胞骨架动力学，如下图所示。（利用双结构可视化细胞分析植物细胞分裂）我利用组蛋白- rfp和gfp -微管蛋白对烟草BY-2细胞的染色体分离和纺锤体伸长进行了时间顺序的动态观察。图像处理显示，在高等植物细胞中，后期B约占染色体距离分离的40%。在稳定表达GFP-fimbrinm的活烟草by -2细胞中，我观察到了肌动蛋白微丝（MFs）和液泡膜（VM）的共定位，用FM4-64对其进行了活体VM染色，发现MFs支持液泡动力学，而act-myosin系统在液泡形态发生中起着重要作用。此外，通过gfp - fibrinm的表达和FM4-64的活体染色，可以在活细胞的整个细胞质分裂过程中跟踪MFs和细胞板的动态。实时成像分析使我们能够量化MF在整个细胞质分裂过程中对细胞板扩增的贡献，表明MF通过调节细胞膜动力学在细胞板形成和扩增中的作用转变。（保护细胞在气孔关闭和日循环过程中液泡和mf的动态组织）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，通过三维重建发现保护细胞在气孔关闭过程中液泡结构变得复杂。此外，在拟南芥植物中，我发现皮层MF排列呈径向和随机排列，分别随气孔孔径的增加和减少而变化，这表明保护细胞的MF排列可能控制着气孔的昼夜运动。

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.