Microtubule Orientation During Stomatal Differentiation in Grasses

Abstract

The changing orientation of microtubules (MTs) during formation of the stomatal complex in grasses was observed by immunofluorescence microscopy, beginning with the asymmetrical division of the cell that gives rise to the guard cell mother cell, i.e. the guard cell grandmother cell. The asymmetrically placed preprophase band (PPB) of guard cell grandmother cells and hair cell mother cells is always laid down parallel to the distal end wall even when this wall is oblique to the long axis of the cell. The first step in formation of the PPB of a subsidiary cell mother cell appears to be establishment of an incomplete band of MTs. Whereas the mature PPB forms a curved line in a face view of a subsidiary cell mother cell, in this early stage MTs form fan-shaped arrays that focus on two points along the edge of the subsidiary cell mother cell. Replacement of the transversely oriented interphase microtubule band of the guard cell mother cell with the longitudinally oriented PPB involves several distinctive stages: (1) appearance of MTs directed toward the centre of the periclinal surface along the entire length of the lateral walls. (2) Appearance of another set of MTs along the entire width of both end walls, likewise focused toward the centre of the periclinal surface. Together these two groups of MTs form a cross with broadened tips in face view of the leaf. (3) Disappearance of the first set of MTs, and formation of an increasingly narrow band from the latter at the site of future cytokinesis. Although the anaphase spindles of guard cell grandmother cells, hair cell mother cells and guard cell mother cells are usually diagonally oriented relative to the site occupied previously by the PPB, the line connecting the centres of the spindle poles that are established at prophase is perpendicular to the persisting PPBs. Unlike the situation in certain other hair cells, MTs in leaf hair cells are transversely oriented even when the cells are highly elongated.