Hyperosmotic stress-induced actin filament reorganization in leaf cells of Chlorophyton comosum

Actin filament (AF) organization was studied during the plasmolytic cycle in leaf cells of Chlorophyton comosum Thunb. In most cells the hyperosmotic treatment induced convex or concave plasmolysis and intense reorganization of the AF cytoskeleton. Thin cortical AFs disappeared and numerous cortical, subcortical and endoplasmic AFs arranged in thick and well-organized bundles were formed. Plasmolysed cells displayed a significant increase in the overall AF content compared with the control cells. Cortical AF bundles were preferentially localized in the shrunken protoplast areas, lining the detached plasmalemma regions. The endoplasmic AF bundles were mainly found in the perinuclear cytoplasm and on the tonoplast surface. AFs also traversed some of the Hechtian strands. AF disorganization after cytochalasin B (CB) treatment induced dramatic changes in the pattern of plasmolysis, which lasted for a longer time and led to a greater decrease of the protoplast volume compared to the untreated cells. In many of the above cells the protoplasts assumed an 'amoeboid' form and were often subdivided into sub-protoplasts. Soon after the removal of the plasmolytic solution both CB-treated and untreated cells were deplasmolysed, while the AF cytoskeleton gradually reassumed the organization observed in the control cells. The findings of this study revealed for the first time in angiosperm cells that plasmolysis triggers an extensive reorganization of the AF cytoskeleton, which is involved in the regulation of protoplast shape and volume. The probable mechanism(s) leading to AF reorganization as well as the function(s) of the atypical AF arrays in plasmolysed cells are discussed.