B. cinerea was isolated from infected grape berries, stored
at −80 ◦C on silica gel and cultured on potato dextrose agar
(PDA) for 2 weeks at 25±1 ◦C. A spore suspension was created
by flooding plates with a small volume of sterile distilled
water containing 0.05% (v/v) Tween-80 (Sigma Chemical Co.,
St. Louis, U.S.A.), and spores were removed by gently scraping
with a glass rod. The resulting spore suspension was filtered
through four layers of cheesecloth to remove mycelial fragments
and diluted with sterile water to obtain an absorbance of
0.25 at 425 nm as determined by a BioMate5 Union ultraviolet
spectrophotometer (UNI, Hamburg, Germany). The density was
about 1.2×106 spores/mL and was diluted with sterile water to
obtain the desired spore concentrations. A volume of 50mL
of inoculum per 1200 berries was applied with an air-brush
sprayer.