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. 2011 Mar;11(2):223-32.
doi: 10.1111/j.1567-1364.2010.00712.x. Epub 2011 Jan 14.

Flo11p adhesin required for meiotic differentiation in Saccharomyces cerevisiae minicolonies grown on plastic surfaces

Melissa G White  1 , Sarah Piccirillo, Vladimir Dusevich, Douglas J Law, Tamas Kapros, Saul M Honigberg
Affiliations

Flo11p adhesin required for meiotic differentiation in Saccharomyces cerevisiae minicolonies grown on plastic surfaces

Melissa G White et al. FEMS Yeast Res. 2011 Mar.

Abstract

Saccharomyces cerevisiae grown on plastic surfaces formed organized structures, termed minicolonies, that consisted of a core of round (yeast-like) cells surrounded by chains of filamentous cells (pseudohyphae). Minicolonies had a much higher affinity for plastic than unstructured yeast communities growing on the same surface. Pseudohyphae at the surface of these colonies developed further into chains of asci. These structures suggest that pseudohyphal differentiation and sporulation are sequential processes in minicolonies. Consistent with this idea, minicolonies grown under conditions that stimulated pseudohyphal differentiation contained higher frequencies of asci. Furthermore, a flo11Δ mutant, which fails to form pseudohyphae, yielded normal sporulation in cultures, but was defective for minicolony sporulation. When minicolonies were dispersed in water and cells were then allowed to settle on the plastic surface, these cells sporulated very efficiently. Taken together, our results suggest that sporulation in minicolonies is stimulated by pseudohyphal differentiation because these pseudohyphae are dispersed from the core of the colony.

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Figures

Fig. 1
Fig. 1. Comparison of yeast strain backgrounds for minicolony sporulation and affinity to plastic
A) 6-day minicolonies of the indicated strain backgrounds before washing (left column of images) and after washing (right column). Strains used: W303 (SH3881), ©1278b (SH2533), S288C (SH2081), SK1 (SH4329). Scale bar = 200 μ. B) Growth rates of the same strains as in (A) on a plastic surfaces, growth measured as the total number of cells/well at the indicated times, data expressed as mean ± S.E.M., n=3. C) Affinity to plastic for the indicated strains after 6-day incubation, “sheared cells” are cells removed in the supernatant or in any of four sequential washes, “retained cells” are cells remaining after these washes. D) Affinity to plastic and sporulation of SK1 strain. At the indicated times, the fraction of cells retained on the plastic after washing (open triangles) and the fraction of asci in minicolonies (filled circles) were assayed.
Fig. 2
Fig. 2. Effect of nutrient concentration on pseudohyphae formation and cell-surface affinity
A) Morphology of SK1 (SH4329) minicolonies grown for 6 days in YPA-1 (low concentration nitrogen source), YPA-2 (intermediate concentration nitrogen source), and YPA-3 (high concentration nitrogen source). B) Growth rate of SK1 in the indicated media as in (A). C) Affinity to plastic for 6 day SK1 minicolonies grown in the indicated media as in (A), and assayed as in Fig. 1.
Fig. 3
Fig. 3. Yeast minicolony development
Scanning electron micrographs of minicolonies incubated for the indicated times. The left column of images are magnified 1000×, 10 μ bar. The right column are magnified 2000×, 5 μ bar. Arrows indicate representative chains of elongated cells. Arrowheads indicate representative sporulated cells.
Fig. 4
Fig. 4. Roles of FLO11 in minicolony development
A) Sporulation levels of wild type (WT, SH4329) and flo11Δ (SH4325) in suspension cultures and minicolonies. Strains were grown in suspension for 4 or 8 days in YPA-1 (n=3) or grown as minicolonies for 8, 10, or 12 days in the same medium (n=6), and the percentage asci in the population determined B) Growth rate of WT and flo11Δ minicolonies. C) Affinity of 6-day WT and flo11Δ minicolonies to well of microtiter plate assayed as in Fig. 1. D) Morphology of WT and flo11Δ minicolonies after 6 days incubation. Scale bar = 50 μ.
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