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

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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.