Spores from 10-d-old cultures grown
at 20–25 ◦C were collected in sterile water containing 0.1% (v/v)Tween 20 The suspension was filtered through a double layer of
lens cleaning tissue (Whatman 105) to remove mycelial fragments
and then centrifuged at 11,000 × g (2 min, 4 ◦C). The supernatant
was decanted and the spore pellet re-suspended in 0.01% (v/v)
Tween 20 to remove nutrients from the medium. This procedure
was repeated twice. Spores were re-suspended in sterile water and
their concentration was adjusted by dilution to104 fungal spores
per milliliter (Neubauer chamber)