Plant and Cell Physiology Advance Access published online on August 18, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm109
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Elicitor-induced cytoskeletal rearrangement relates to vacuolar dynamics and execution of cell death: in vivo imaging of hypersensitive cell death in tobacco BY-2 cells
1 Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha Kashiwa, Chiba 277-8562, Japan
2 Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan
3Genome & Drug Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan
4Research Center for Health Science of Nanoparticles, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan
5These authors equally contributed to this work.
*Corresponding author: Professor Dr. Kazuyuki Kuchitsu. Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan. Tel: +81-4-7122-9404, Fax: +81-4-7123-9767, E-mail: kuchitsu{at}rs.noda.tus.ac.jp
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Abstract |
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Disintegration of vacuolar membrane has been proposed to be a crucial event in various types of programmed cell death in plants. However, its regulatory mechanisms are mostly unknown. To obtain new insights on the regulation of vacuolar membrane disintegration during hypersensitive cell death, we investigated structural dynamics and permeability of the vacuolar membrane, as well as cytoskeletal reorganization during programmed cell death in tobacco BY-2 cells induced by a proteinaceous elicitor, cryptogein. From sequential observations, we have identified the following remarkable events during the programmed cell death: (stage 1) bulb-like vacuolar membrane structures appear within the vacuolar lumen and the cortical microtubules are disrupted, while the cortical actin microfilaments are bundled. Simultaneously, transvacuolar strands including endoplasmic microtubules and actin microfilaments are gradually disrupted and the nucleus moves from the center to the periphery of the cell; (stage 2) cortical actin microfilament-bundles and complex bulb-like vacuolar membrane structures disappear. The structure of the large central vacuole becomes simpler, and small spherical vacuoles appear; (stage 3) vacuolar membrane is disintegrated and a fluorescent dye, BCECF leaks out of the vacuoles just prior to the programmed cell death. Application of an actin polymerization inhibitor facilitates both the disappearance of bulb-like vacuolar membrane structures and induction of the cell death. These results suggest that the elicitor-induced reorganization of actin microfilaments is involved in the regulation of hypersensitive cell death via modification of the vacuolar structure to induce vacuolar membrane disintegration.
Keywords: Actin microfilament - Green fluorescent protein - Hypersensitive cell death - Tobacco BY-2 cells - Transvacuolar strand - Vacuol
6Present address: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
7Present address: RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Kanagawa 230-0045, Japan.
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