Microtubule organization during the cell cycle of cultured Vicia hajastana Grossh.

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Abstract
Microtubule (MT) organization was examined at each stage of the cell cycle in cell suspension cultures of Vicia hajastana Grossh. Simultaneous staining of MTs by immunofluorescence and DNA by Hoechst 33258, and microfluorimetric quantitation of DNA in interphase allowed direct correlation of MT configuration with mitotic stage and with G1, S, and G2 of interphase. The results indicated that in the majority of the cells the cortical MTs were disorganized in early G1, but then organized rapidly into parallel transverse arrays, remained ordered throughout S, and lost order in G2, probably near the onset of mitosis. A wide range of cell sizes were found in S, indicating that entry into S was loosely controlled. Preprophase bands, present in 55–80% of cells in prophase were observed in both disorganized cell clumps and regular cell files, indicating they were not exclusively associated with organized patterns of growth and division. In many cells, short MTs or MT clusters were observed in the cell cortex during all stages of mitosis. These MT remnants may serve as nucleating centres for new cortical MTs, which appear in late telophase as a disordered network. Ordering of MT occurs later in G1, indicating that MT nucleation and organization are two different processes.Key words: interphase, microfluorimetry, microtubules, mitosis, plant cell culture.
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