FIGURE 3.

Confirmation of the 24-nt V. dahliae rRNA-derived VdsRNA (VdrsR-1) loaded into host AGO1 protein. (A) The schematic diagram shows the complementarity between VdrsR-1 and MIR157d. The mature miR157d and VdrsR-1 sequences are shown in red and green letters, respectively. (B) Northern blotting results with three biological replicates showed that V. dahliae generated abundant rRNA-derived ∼24-nt and smaller sRNAs. U6 served as loading control. (C) VdrsR-1 was detected by RT-PCR in AGO1-IPed sRNA from V. dahliae-infected plants. Two known AGO1-loaded miR162 and miR168 were used as positive controls. A plant endogenous 24-nt siRNA1003 and a V. dahliae miRNA-like VdmilR-1 that were not present in AGO1-IPed sequencing data were also examined and not detected. Anti-α-myc antibody was used to detect the amount of AGO1-IPed from mock and infected plants. The results of three independent replicates are shown.