Studies on Electrofusim of Plant Protoplasts Using High Speed Microscopy

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Actin is an important component of plant cytoskeleton that not only involved in contributing to the cytoarchitecture of cells but also in cell motility. Cell fusion is an important process for studying the development of the cell and for the subsequent cell division. The study is to report on three dimensional structures and dynamic aspects of plant protoplasts during early process of electrofusion. 1) New technology for recording the time-resolved phenomena has developed using a streak camera and a polarization microscope. We obtained that the technique is very useful in the study of muscle contraction and cell fusion. 2) We obtained sub microsecond and microsecond rotational motions with in the contractile myofibrils by analyzing the time-resolved birefringence. 3) We analyzed the location and movement of actin filaments after the application with AC field and DC pulse. 4) We observed that bundles of actin filaments are forming cables and networks 5) Three dimensional observations of protoplasts were also made by a high voltage electron microscope (1 MeV) equipped a revolving stage to get a pair of stereo-micrographs. Ring structure of actin filaments appeared at the contact areas between fused cells. We found the joint particle connecting between actin filaments. 6) A soft X-ray microscope with zone plate was installed at UVSOR.The X-ray micrographs of dry and wet protophasts were taken. X-ray micrographs of protoplasts were compared with those of optical and electron micrographs.