Figure 2. Pum1 orchestrates meiotic progression and basidial maturation.

(A) Violin plot analysis shows that disruption of PUM1 cascade members led to a decrease of high BMS basidial population during both unisexual and bisexual development (n = 150 for each strain). (B) Morphotype-specific enrichment of Dha1, Fas1, and Fad1. >50 cells in each morphotype were examined for mCherry-labelled proteins expression. (C) Dynamics of Fad1-mCherry expression during unisexual development. Fad1-mCherry shows a remarkably biased expression in the basidium structure and displays different localization patterns. Scale bar: 5 μm. (D) Cells were placed onto a V8 plate at 25°C in the dark for unisexual induction, and incubated for 7 days. For each BMS range, >20 basidia expressing Fad1-mCherry were examined. The right panel highlights the dynamic enrichment of patterns I, II and IV at various stages during basidial maturation. (E) Predominant Fad1 protein exhibited a subcellular localization identical to pattern IV in post-meiotic basidia (sporulated basidia) during unisexual reproduction. Thirty-seven sporulated basidia expressing Fad1-mCherry were measured. ND = Not Detected. (F) Sporulation phenotypes for wild-type XL280α, the fad1Δ deletion mutant (unisexual reproduction), a wild-type cross between XL280α and XL280a, and the fad1Δ bilateral mutant cross. Scale bar: 10 μm (upper and middle panels), 5 μm (bottom panels).

Figure 2—figure supplement 1. Morphotype-specific expression patterns of Dha1, Fas1 and Fad1, which are fused by mCherry at their C-terminus.

XL280α cells harboring genes encoding different mCherry-fused proteins were incubated on V8 agar for 7 days. Different morphotypes expressing mCherry-fused extracellular proteins were visualized. Representative images of n > 5 experiments. Scale bar: 5 μm.