Quantification of Trehalose in Fungal Conidia

The amount of trehalose (α-d-glucopyranosyl α-d-glucopyranoside) in conidia was measured as previously described [28 (link),29 (link)]. Briefly, 10⁸ conidia were suspended in 200 μl of ddH₂O and incubated at 95 °C for 20 min. The supernatant was separated by centrifugation. Fifty microliters of supernatant were transferred to a new tube, mixed with an equal volume of 0.2 M sodium citrate (pH 5.5), and incubated with or without (as a negative control) trehalose (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C for 8 h to generate glucose. After incubation, the amount of glucose generated from the trehalose was assayed with a Glucose Assay Kit (Sigma-Aldrich, St. Louis, MO, USA).