The improved inoculation method of Rhizoctonia solani AG-1 IA on Brachypodium distachyon. (a) Progression of infection and symptom appearance in the inoculation method. Leaves detached from 3 weeks old B. distachyon were placed in Petri dish with moist paper and 3 agar plugs prepared from R. solani-grown PDA medium were placed. The dishes were covered with lids and incubated at 25 °C. Photos were taken at the indicated hours after inoculation (hpi). (b) Inoculated leaves were sampled at the indicated timing and hyphae around leaves were removed before extraction of total RNAs. The relative abundance of fungal biomasses of the infected tissues was measured by qRT-PCR using cDNAs prepared from extracted RNAs with a specific primer for 18S rRNA gene.