Visualization of actin cytoskeletal dynamics during the cell cycle in tobacco ( Nicotiana tabacum L. cv Bright Yellow) cells

Abstract

Background information . The actin cytoskeleton forms distinct actin arrays which fulfil their functions during cell cycle progression. Reorganization of the actin cytoskeleton occurs during transition from one actin array to another. Although actin arrays have been well described during cell cycle progression, the dynamic organization of the actin cytoskeleton during actin array transition remains to be dissected. Results . In the present study, a GFP (green fluorescent protein)‐mTalin (mouse talin) fusion gene was introduced into suspension‐cultured tobacco BY‐2 ( Nicotiana tabacum L. cv Bright Yellow) cells by a calli‐cocultivation transformation method to visualize the reorganization of the actin cytoskeleton in vivo during the progression of the cell cycle. Typical actin structures were indicated by GFP—mTalin, such as the pre‐prophase actin band, mitotic spindle actin filament cage and phragmoplast actin arrays. In addition, dynamic organization of actin filaments was observed during the progression of the cell from metaphase to anaphase. In late metaphase, spindle actin filaments gradually shrank to the equatorial plane along both the long and short axes. Soon after the separation of sister chromosomes, actin filaments aligned in parallel at the cell division plane, forming a cylinder‐like structure. During the formation of the cell plate, one cylinder‐like structure changed into two cylinder‐like structures: the typical actin arrays of the phragmoplast. However, the two actin arrays remained overlapping at the margin of the centrally growing cell plate, forming an actin wreath. When the cell plate matured further, an actin filament network attached to the cell plate was formed. Conclusions . Our results clearly describe the dynamic organization of the actin cytoskeleton during mitosis and cytokinesis of a plant cell. This demonstrates that GFP—mTalin‐transformed tobacco BY‐2 cells are a valuable tool to study actin cytoskeleton functions in the plant cell cycle.