Orientational variability of parallel arrays of cortical microtubules under the outer cell wall of the Helianthus hypocotyl epidermis

The epidermis of Helianthus hypocotyl can be peeled off and, in the form of detached strips can be used as a model system to study the effect on cortical microtubule (cMTs) orientation of these factors, which are difficult to be manipulated in situ, such as apoplastic pH or applied stress. In the first step, however, the orientation and reorientation of cMTs in the epidermis in situ must be described. The cMTs under the epidermal wall in hypocotyl epidermis at different positions along the hypocotyl and on its opposite sides were studied by means of immunostaining, using epi-fluorescence microscopy. The angle λ that parallel array of cMTs makes with cell longitudinal axis was measured. The variation of λ in a population of cells was documented by λ-histogram (frequency of cells exhibiting a particular λ λ plotted against λ value). The histograms were of either transverse type (maximum at λ ~90o, denoted as type A) or oblique type (two maxima on both sides of the transverse direction, denoted as type B) in the apical part of the hypocotyl, and were either of B type or of longitudinal type (maximum at λ ~0o or 180o denoted as type C) in the basal part. The change from A or B to C basipetally may be considered as due to the developmental trend in cMT orientation. The occurrence of B above A in some hypocotyls in their apical part strengthens the hypothesis on the autonomous reorientation of cMTs. The intermingled occurrence of A and B reorientation in the upper part of hypocotyl is interpreted as a manifestation of a subtle control of cell growth in latitudinal direction. The majority of histograms were asymmetric showing predominance of cMT parallel arrays inclined as the middle part of the letter Z.