RTG from starvation results in three distinct cell morphologies.

<p>(A) Representative morphologies of daughter cells following RTG from SPO throughout a meiotic time course. Top left panel: RTG at 0 hours, <i>i.e.</i>, after growth in BYTA, (RTG₀); top right panel: RTG at 3 hours (RTG₃), which is comparable to PH cells from solid nitrogen medium (bottom left panel). Bottom right panel: RTG at 6 hours (RTG₆). RTG₀ and RTG₃ cells were photographed after one complete cell cycle, corresponding to 160 minutes and 220 minutes after shift to rich medium, respectively (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002732#pgen.1002732.s001" target="_blank">Figure S1A</a>). RTG₆ cells were photographed three hours after shift to rich medium, at which point the majority of cells (>95%) had formed spores. Arrows indicate primary daughter cells upon RTG₀ and RTG₃. B) Quantification of axial ratio in wild-type (SAy821) cells upon RTG throughout a meiotic time course (red bars, left axis; n = 200 cells/time point) relative to percent of cells undergoing pre-meiotic DNA synthesis (<i>i.e.</i>, 4C cells) (blue diamonds, right axis, quantified by FACS, 3×10⁴ cells/time point) and percent cells undergoing meiotic divisions as assayed by DAPI staining (green triangles, right axis). Schematic at top defines axial ratio. The majority (>80%) of RTG₅, RTG₆, RTG₇, and RTG₉ cells either formed spores or remained unbudded three hours after shift to rich medium; axial ratio was therefore not quantified for these time points. C) <i>MAT </i><b>a</b>/α diploids (SAy821) (top left panel) or <i>MAT</i> α haploids (H224) (top right panel) were returned to growth in rich medium after meiotic induction. Arrows indicate primary buds. Bottom panels represent colony morphologies after growth on SLAD for 6 days. D) Distribution of axial ratios of primary daughter cells upon RTG₃ for strains in (C): wild-type diploid (SAy821 top panel), haploid <i>MAT</i> α (H224 bottom panel) (n = 200 cells/strain). RTG₀ is represented in red bars, RTG₃ in blue bars. E) Wild-type (SAy821), <i>flo11</i>Δ/Δ (SAy789) and <i>flo8</i>Δ/Δ (SAy905) daughter cell morphologies upon RTG₃ (top panels). Arrows indicate primary daughter cells. The same strains were photographed after growth on SLAD for 6 days (bottom panels). Axial ratios are quantified in (F) (n = 200 cells/strain).