4.3 B. cinerea inoculation and disease evaluation
B. cinerea (isolate BcI16) was cultured on Potato Dextrose Agar
(PDA) in Petri dishes incubated at 22°C. Conidia were harvested from 10-
to 14-day-old cultures by agitating 1 cm2 of agar
bearing mycelium and conidia in a glass tube with tap water. The
suspension was then filtered through cheesecloth. The concentration of
conidia was determined using a haemocytometer under a light microscope,
and adjusted to 106 cells mL-1.
0.1% glucose and 0.1% K2HPO4 were
added to the final conidial suspension. Whole plants were inoculated with
this conidial suspension. The severity of the resulting necrotic lesions
was determined as the percentage of necrotic area, according to the
0–100% scale described previously (Meller Harel et al., 2014). The
level of disease was evaluated every 2–3 days for a period of 10 days.
Alternatively, 3 mm diameter discs of 3 day-old PDA cultured B.
cinerea were placed on each of 4 leaflets of leaf number 5 and rot area
was measured 3 to 5 days after inoculation.