Aspergillus clavatus (DSM3410) was cultivated in Czapek-Dox medium according to DSM at 27°C and C. albicans (DSM3454) was cultivated in universal medium for yeasts (YM) according to DSM at 27°C.
In the vineyard, grape berries were placed in direct contact with plates containing Rose Bengal Chloramphenicol Agar (RBCA), a selective medium for yeasts and molds (15 g L−1 agar, 10 g L−1 glucose, 5 g L−1 papain-digested soybean meal, 1 g L−1 KH2PO4, 0.5 g L−1 MgSO4 × 7H2O, 0.05 g L−1 Rose Bengal, and 10 g L−1 chloramphenicol).
In this study a natural culture medium that mimics the synthetic yeast peptone glucose medium used for yeast fermentations was designed to screen and select yeasts capable of producing high levels of diacetyl and acetaldehyde.
The medium used for yeast strain maintenance was YPD containing 1% (w/v) yeast extract, 2% (w/v) peptone, and 2% (w/v) glucose.
The biofilms were carried out using sterile untreated 96-well polyethylene U-bottom plates (IPT) containing the specific medium (Sabouraud for yeast and BHI for bacteria) enriched with 2% sucrose.
It should be noted that yeast-extract-based rich media contains autolyzed S. cerevisiae extract and therefore may not be as optimal a growth medium for fission yeast as it is for budding yeast.
The medium for the yeast cultivations was the same as for the pre-culture, but with glucose, 1 mL/L trace elements solution (0.015 g/L FeCl3·6H2O, 9 mg/L ZnSO4·7H2O, 10.5 mg/L MnSO4·2H2O, and 2.4 mg/L CuSO4·5H2O), 1 mL/L vitamin solution (0.06 g/L myoinositol, 0.03 g/L Ca-pantothenate, 6 mg/L thiamine HCl, 1.5 mg/L pyridoxine HCl, and 0.03 mg/L biotin), and 200 µL/L antifoam agent.
Briefly, lectins were diluted in culture medium (SDB for yeasts and TSB for bacteria) to concentrations of 31.25 to 250 μg/mL, and 100 μL of each lectin was incubated with 100 μL of each microorganism (previously adjusted to 2 × 10 cells/mL).
