| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Research Abstract |
In this study, I have developed the transgenic BY-2 cells and analyzed the dynamics of cytoskeleton and vacuole, as follows.
1) I introduced GFP-tubulin, -fimbrin and -AtVam3p into tobacco BY-2 cells and Arabidopsis Col-0 cells for visualizing microtubules (MT), actin microfilaments (MT) and vacuolar membrane (VM), respectively. Then, I succeeded in establishing the cell lines for the time-sequential observations.
2) I developed the new algorithm for three dimensional (3-D) reconstruction, using confocal images, named REANT (reconstructor and analyzer of 3-D structure).
3) I observed the developmental process of vacuoles in BY-2 mini-protoplasts, in which almost vacuoles were eliminated. The mini-protoplasts became to reorganize the large vacuoles through the tubular structures of vacuoles.
4) I performed time-sequential observations of MF dynamics during cell division, and revealed the appearance of two MF bands in metaphase. The cell plate formed exactly within the valley between the bands.
5) I succeeded in differentiating the Arabidopsis cells, expressing GFP-tubulin, into tracheary elements, and analyzed the roles of MTs in secondary wall thickening.
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