Relationship between preprophase band organization, F-actin and the division site in Allium: Fluorescence and morphometric studies on cytochalasin-treated cells

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Abstract
The preprophase band (PPB) of microtubules (Mts), which appears in the G2 phase of the cell cycle in higher plants but disappears well before the end of karyokinesis, is implicated in the determination of the division plane because its location marks the site at which the phragmoplast/cell plate will fuse with the parental plasmalemma during cytokinesis. The PPB first appears as a rather wide array, which progressively narrows before or during prophase. Actin-containing microfilaments (Mfs) have recently been reported in the PPB, but the role of these elements in PPB organization and/or function remains unclear. The present study employed fluorescence and pharmacological methods in symmetrically and asymmetrically dividing epidermal cells of Allium to probe this problem. Our results show that PPBs in cells treated with 2–200μM cytochalasin D (CD) are still transversely aligned but remain two to three times wider than mature bands in control cells. Treatment for 0.5 h at 20 μM is sufficient to make the PPBs abnormally widel Premitotic nuclear migration in asymmetrically dividing cells is also inhibited by CD, as is the positioning of the mitotic apparatus and the new cell plate. The plate is still transverse, however. Band-like arrays of cortical Mfs become evident in most interphase cells by prophase. The band remains quite wide compared to the final dimensions of the Mt PPB, and clearly encompasses it. Levels of CD as high as 200μM decrease the number of cells with transverse actin bands, although a majority still retain them. Other F-actin arrays are disrupted by the drug. Thus, while CD does not inhibit the formation of an initial, broad, transverse PPB in most cells, it does prevent the narrowing process that defines the precise division site. The role of actin in this effect is discussed.