FIGURE 4.

Preferential targeting of miR396 isoforms against OsGRFs. (A) Multiple alignment of miR396d target sites in OsGRF’s. The target genes were classified into four categories (Reporter 1, Reporter 2, Reporter 3, and Reporter 4) based on sequence homology of target site. (B) Confocal images show that miR396 isoforms differentially suppresses the protein accumulation of different OsGRF’s reporter genes. The indicated YFP-based reporter constructs were transiently expressed alone or co-expressed with miR396 isoforms in Nicotiana benthamiana. Bars = 25 μm. (C) RNA samples were collected from infiltrated leaves and the transcriptional regulation of OsGRF’s homologs by miR396 isoforms were studied using RT-qPCR. Error bars indicate SD from three technical replicates. The letters above the bars indicate significant differences at a P-value < 0.01, as determined by a one-way ANOVA followed by post hoc Tukey HSD analysis. This experiment was repeated at least three times with similar results.