Sterilized cellophane membrane (DINGGUO, Beijing, China), used to simulate plant root surface to induce formation of hyphopodium, was covered on MM medium (Zhao et al. 2016 (link)). Equal amounts of conidia collected from V. dahliae strains as indicated were incubated on the cellophane membrane and grown at 25 °C for 5 days. The cellophane membrane was then removed and observed under the Leica SP8 confocal laser scanning microscope system for determining hyphopodium formation. Further culture for an additional 3 days to observe whether there were hyphae grew on the underlying medium.
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