FIG 2.

Absence of HvPssA or HvPssD leads to defects in growth, cell morphology, and motility. (A) Wild-type (strain H53) and ΔartA, ΔhvpssA, and ΔhvpssD mutant cells were grown with shaking in 96-well plates with a total volume of 200 μl of liquid semidefined CA medium, and the growth of six biological replicates was monitored at the OD₆₀₀, with recordings taken every 30 min. For complementation analysis, artA, hvpssA, or hvpssD was expressed from pTA963 under the tryptophan-inducible p.tna promoter. The wild-type and deletion strains were transformed with an empty pTA963 plasmid as a control. (B) The wild-type (strain H53) and artA, hvpssA, or hvpssD deletion and complementation strains from individual colony on solid agar plates were individually stab inoculated with a toothpick into semisolid 0.35% agar in CA medium supplemented with tryptophan, followed by incubation at 45°C. (C) Phase-contrast images were taken from wild-type and mutant cells during mid-exponential-growth phase (OD₆₀₀, 0.3) and immobilized under 0.5% agarose pads. (D) Violin distributions of aspect ratio measurements from single cells. Rodlike (aspect ratio, >2) cells were prevalent to disklike (aspect ratio, <2) cells in the mutant strains compared to the wild type. Biological replicates were collected on three different days, and data were analyzed from >1,000 cells under each condition by automated image segmentation. Scale bars = 5 μm.