FIGURE 2.

Effect of dsGFP on M. oryzae GFP. (A) Microscopic assessment of GFP intensity 3 dpt, among axenic cultures of M. oryzae GFP fed with and without dsGFP. While the culture that was not fed with any dsRNA (DEPC water- treated) was kept as the negative control, the culture that was fed with dsDES1 (non-specific for GFP) served as the positive control. (B) Time-dependent fluorescence trend of M. oryzaeGFP grown on 300 nM dsGFP-sprayed leaves with respect to the unsprayed set, across 0–5 days. (C) Fluorescence of M. oryzaeGFP on wounded (W) and intact (I) rice leaves sprayed with dsGFP, 24 h prior to infection. GFP fluorescence was observed under an FITC filter of AxioCam mRm 0 [4 h post treatment (hpt)], 4, and 7 dpt, and fluorescence emitted by M. oryzaeGFP on Tris EDTA (TE)-sprayed leaves were kept as mock. TE was sprayed 24 h prior to infection, as replacement for dsGFP. Comparisons were made between the sets, at similar time points, with observations made from at least 10 leaves for each set. The experiment was repeated thrice, and the results were compared independently. (D) Relative abundance of GFP transcripts in dsRNA-sprayed wounded and intact leaves, with respect to TE-sprayed leaves, at 4 dpt. OsACTIN was used as internal control for normalization. The experiments were repeated thrice, each time with three biological replicates. Error bars represent standard deviation (n = 3), where * and ** denote statistical significance at the P ≤ 0.05 and P ≤ 0.01 levels, respectively.