The organization of tip-growth-related organelles and microtubules revealed by quantitative analysis of freeze-substituted oomycete hyphae

Abstract

The distribution of organelles and microtubules in hyphal tips of the oomycete, Saprolegnia ferax, were quantitatively determined at high resolution from serial-section electron microscopy of freeze-substituted cells. All the organelles and the microtubules were non-uniformly distributed, each showing a characteristic longitudinal gradient starting at a different point behind the tip. In addition, when the cytoplasmic cross-sectional area was divided into radial regions, all organelles occurred preferentially in either the central (mitochondria and Golgi bodies) or the peripheral (microtubules, wall vesicles and spherical vesicles) region. The nuclei were so large as to span both regions but were always oriented with their centrioles facing the plasmalemma. Microtubules occurred in the extreme tips, became more abundant sub-apically, were predominantly short but increased in mean length with distance from the tip. The correlated patterns of organelle and cytoskeleton organization from this and previous work show that neither the microtubules nor the detected arrays of actin are sufficient to account for most organelle arrangements. However, on the basis of the distribution and orientation of the predominantly elongated wall vesicles, we suggest that the wall vesicles travel radially from their origin at the centrally located Golgi bodies to the cell periphery where they are transported longitudinally to the hyphal tip in conjunction with the plasmalemma-associated actin cables. Our data also suggest that the hyphae contain a cortical ectoplasm with which the nuclei interact, at least in part, via their centrioles and centriole-associated microtubules, and whose mechanical integrity is increased by both the peripheral actin cables and a high density of microtubules. We suggest that the endoplasm is less strong and has physiological properties that enhance the differentiation of endoplasmic reticulum and nuclear envelope into Golgi body production.