FIG. 4.

Northern blot analysis of PMR1 expression. (A) Expression of PMR1 in P. digitatum in the absence of triflumizole. Conidia from each strain were grown in the absence of triflumizole in PDB at 25°C for 1 day. Total RNA was prepared as described in Materials and Methods. Approximately 50 μg of total RNA from each strain (lanes: 1, sensitive strain PD5; 2, sensitive strain DF1; 3, sensitive strain U1; 4, resistant strain LC2; 5, resistant strain M1; 6, resistant strain I1; and 7, Δpmr1 mutant DIS33) was loaded into each lane. Ethidium bromide-stained rRNA bands are shown for comparison of the quantities of loaded RNAs. (B) Time course of expression of PMR1 in the presence of triflumizole. Approximately 50 μg of total RNA from DMI-sensitive strain PD5 (lanes 1 to 3) and DMI-resistant strain LC2 (lanes 4 to 6) after treatment with triflumizole (50 μg/ml) was loaded into each lane. Lanes: 1 and 4, 0 min of treatment; 2 and 5, 10 min of treatment; 3 and 6, 60 min of treatment.