The Cytoplasmic pH Influences Hyphal Tip Growth and Cytoskeleton-Related Organization

The regulatory role of protons in hyphal tip growth was investigated by using membrane-permeant weak acids to acidify cytoplasm of the oomycete Saprolegnia ferax. Acetic acid decreased cytoplasmic pH from approximately pH 7.2 to 6.8, as shown by SNARF-1 measurements of cytoplasmic pH. Inhibition of growth in a dose-dependent manner by acetic, propionic, and isobutyric acid was accompanied by changes in positioning and morphology of mitochondria and nuclei, condensation of chromatin, disruptions in peripheral actin, and increases in hyphal diameter. These cellular alterations were fully reversible, and during recovery, major cytoplasmic movements and extensive apical vacuolations were observed. The results are consistent with proton regulation of the cytoskeleton, nuclear matrix, and/or chromosomes. However, a macroscopic cytoplasmic gradient of H+ in hyphae was not revealed by SNARF-1, indicating that if such a H+ gradient were required, it must occur at a finer level than we detected.