Inocula of the four isolates of R. solani were prepared on
autoclaved-moist wheat grain. Wheat grain was soaked inwater for 12 h. After soaking, excess water was drained off
and water-soaked wheat grains were poured into 500-mL
Erlenmeyer flasks. The inoculum of each isolate was
prepared in a separate flask. Five-millimeter-diameter
mycelial discs were cut from the edge of 3-day-old PDA
cultures of the pathogen. Ten to 12 mycelial discs from
each isolate of the pathogen were added to the flasks
containing autoclaved wheat grains in the flasks and
incubated at 25 8C for 21 days. They were shaken by hand
at 2–3 days interval for even colonization. The colonized
wheat grain was air dried for 1 week and stored at 10 8C
before being used as an inoculum of the pathogen.