Journal Article

Dynamic Organization of Vacuolar and Microtubule Structures during Cell Cycle Progression in Synchronized Tobacco BY-2 Cells

Natsumaro Kutsuna and Seiichiro Hasezawa

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 9, pages 965-973
Published in print September 2002 | ISSN: 0032-0781
Published online September 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf138
Dynamic Organization of Vacuolar and Microtubule Structures during Cell Cycle Progression in Synchronized Tobacco BY-2 Cells

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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In higher plant cells, vacuoles show considerable diversity in their shapes and functions. The roles of vacuoles in the storage, osmoregulation, digestion and secretory pathway are well established; however, their functions in cell morphogenesis and cell division are still unclear. To observe the dynamic changes of vacuoles in living plant cells, we attempted to visualize the vacuolar membrane (VM) by pulse-labeling tobacco BY-2 cells with a styryl fluorescent dye, FM4-64. By time-sequence observations using confocal laser scanning microscopy (CLSM), we could follow the dynamics of vacuolar structures throughout the cell cycle in living higher plant cells. We also confirmed the dynamic changes of VM structures by the observation using transgenic BY-2 cells expressing GFP-AtVam3p fusion protein (BY-GV). Furthermore, by using transgenic BY-2 cells that stably express a GFP-tubulin fusion protein [BY-GT16, Kumagai et al. (2001) Plant Cell Physiol. 42: 723], we could study the relationship between the dynamics of vacuoles and microtubules. From these observations, we identified, for the first time, some remarkable events: (1) at the late G2 phase, tubular structures of the vacuolar membrane developed in the central region of the cell, probably in the premitotic cytoplasmic band (phragmosome), surrounding the mitotic apparatus; (2) from anaphase to telophase, these tubular structures invaded the region of the phragmoplast within which the cell plate was being formed; (3) at the early G1 phase, some of the tubular structures expanded rapidly between the cell plate and daughter nuclei, and subsequently developed into large vacuoles at interphase.

Keywords: Keywords: AtVam3p — Cell cycle — FM4-64 — Microtubule— Tobacco BY-2 cells — Vacuole; Abbreviations: CLSM, confocal laser scanning microscopy; ER, endoplasmic reticulum; GFP, green fluorescent protein; LS, Linsmaier and Skoog; MT, microtubule; PM, plasma membrane; PPB, preprophase band; PSV, protein storage vacuole; TVM, tubular structure of vacuolar membrane; VM, vacuolar membrane.

Journal Article.  5726 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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