Plate Cultures
Plate cultures usually contain agar as a solidifying agent. Some agars, such as PDA, are
enriched with nutrients and can be purchased as “ready-made,” whereas minimal agars
require addition of relevant nutrients such as a carbon and nitrogen source. In addition to
nutrients essential for growth, various other components can be introduced into the agar
media. For example, antibiotics may be added for the selection of fungal transformants or to
prohibit bacterial contamination. The surfactant Triton-X100 is typically applied to restrict
the growth of fungal colonies on the plates.
Liquid Cultures
Liquid cultures are typically carried out in conical (Erlenmeyer) flasks placed on a shaker.
This type of culturing is usually performed with a view of testing particular properties of the
fungal strains of interest such as production of an enzyme or a metabolite. Fungi are excellent
protein secretors; thus, high amounts of proteins can be found in the culture supernatants.
Composition of the growth medium and cultivation conditions depend on the goal of the
experiment. The procedure involves choosing the carbon or the nitrogen source and setting
the pH and the shaker speed. A fungal shake culture in a laboratory is typically carried out in
50 mL of medium placed in 250 mL Erlenmeyer flasks, but cultivations can also be carried
out on a smaller or a larger scale such as 15 mL test tubes, 11 mL “Duetz System” deep well
plates, and laboratory fermenters ranging from 0.5 to 20 L.
Methods
Carry out all procedures at room temperature unless otherwise specified. Take care for not
dusting the environment with agar or other easily spreadable medium components.
Autoclaving is carried out at 121 °C for 20 min if not stated otherwise. Cultivation media can
also be sterilized in a pressure cooker for 30 min at 121 °C (at 15 psi).
Fungal Sporulation