Abstract

<p>(A) Sexual developments of a wild type, <i>mkkB</i>Δ, <i>mkkB</i> complementation (comp+), <i>steC</i>Δ [lacking AnSte11] strains, which were point inoculated (1×10<sup>4</sup>) and grown on minimal medium in dark conditions (5 days at 37°C). Small squares are the close-up stereomicroscopic images of the strains. Red arrows indicate the mature black fruiting bodies of the wild type and complementation strains and yellow arrows denote the premature nests produced by <i>mkkB</i><b>Δ</b> and <i>steC</i><b>Δ</b> strains. (B) Raster electron microscopy (REM) image of the strains from (A). The wild type fruiting body (cleistothecium: Cl) is surrounded by the globose Hülle cells. <i>steC</i> and <i>mkkB</i> mutants produce only dispersed groups of Hülle cells (yellow arrows) instead of mature fruiting bodies. (C) Monitoring hyphal fusions and heterokaryons via fluorescence microscopy. Strains bearing either cytoplasmic synthetic green fluorescent protein (sGFP) or nuclear red fluorescent protein fused histone 2A (mRFP) were used in different combinations. Only two wild types form green and red fluorescent combinations through hyphal fusions. (D) Isolated protoplasts from two wild types (yellow and green), <i>steC</i>Δ (green), <i>mkkB</i>Δ (yellow or green) were used for protoplast fusions as shown in combinations and plated on selective medium. <i>wt</i>/<i>wt</i>, <i>wt</i>/<i>steC</i>Δ, <i>wt</i>/<i>mkkB</i>Δ combinations produce fruiting bodies after 7–8 days, whereas <i>mkkB</i><b>Δ</b>/<i>mkkB</i><b>Δ</b> combination only produces nests.</p

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