Altered patterns of tubulin polymerization in dividing leaf cells of Chlorophyton comosum after a hyperosmotic treatment

Microtubule organization and tubulin polymerization in meristematic leaf cells of Chlorophyton comosum treated with an aqueous solution of 1 M mannitol, inducing plasmolysis, were examined with immunofluorescence and transmission electron microscopy. Hyperosmotic treatment induced disintegration of the interphase microtubule systems. Free tubulin, either liberated from the depolymerized microtubules or preexisting as a nonassembled pool, was incorporated into a network of paracrystals. In most of the dividing cells, mitotic and cytokinetic microtubule systems were replaced by atypical spindle-like structures displaying bipolarity and atypical phragmoplasts, respectively These atypical mitotic and cytokinetic structures consisted of large densely packed bundles of macrotubules (32 nm diameter) or macrotubules and paracrystals. Tubulin paracrystals also occurred in ectopic positions in plasmolysed mitotic and cytokinetic cells. Dividing cells displaying paracrystals only did not form atypical mitotic and cytokinetic apparatuses. Short hyperosmotic stress causes disintegration of all microtubule arrays in dividing cells of C. comosum. Free tubulin is incorporated into macrotubules and tubulin paracrystals. The latter exhibit definite periodicity and characteristic fine structure.