Characterization of Fungal Vegetative Growth

For comparing vegetative growth, mycelial colony diameter was measured after growing the wild type and transformants on V8, oatmeal, and CM agar medium at 25°C without light for 6 days. The hydrophobicity of vegetative hyphae was tested by dropping sodium dodecyl sulfate (SDS) and water on 7-day-old mycelia. Conidiation was evaluated by counting conidia harvested in 5 ml distilled water from 7-day-old OMA medium. Lactophenol blue solution (Sigma-Aldrich, St. Louis, MO, USA) was used to distinguish conidiophores from mycelia (Kim et al., 2009 (link)). Conidial germination was performed by placing the conidial suspension (5 × 10⁴ /ml) on hydrophobic coverslips and counting germinated conidia 2 h later. Appressorium formation was conducted by incubating a conidial suspension on hydrophobic coverslips and hydrophilic slide glasses with or without the addition of 5 mM exogenous cAMP (Fu et al., 2018 (link)). Surface hydrophobicity was tested by dropping water or SDS solution onto 14-day-old mycelia bearing conidia and evaluated by comparing mycelial wettability. Experiments were repeated three times with three replicates for each repeat. All data were analyzed according to Duncan’s test at P < 0.05.