Figure 1.

Expression of ARF10, ARF16, and ARF17 in Plants, and Regulation by miR160.

(A) Alignment of partial mRNA sequences of ARF10, ARF16, ARF17, and miR160. Free energies of duplex structures were calculated using the Mfold method (Zuker, 2003). mARF10, mARF16, and mARF17 are the modified mRNAs that harbor synonymous nucleotide substitutions in miR160 binding sites.

(B) Expression of miR160, ARF10, ARF16, and ARF17 in Arabidopsis. Total RNAs from rosette leaf (L), stem (S), root (R), and inflorescence (I) were examined by RNA gel blot analysis with the radiolabeled probes indicated.

(C) Cleavage of ARF16 transcripts by miR160 in planta. Constructs harboring the wild-type or the mutated ARF16 driven by the 35S promoter were coagroinoculated with the Pro_35S:MIR160 construct in tobacco leaves. The empty vector of pKYLX71 (Vector) was used as a negative control. Total RNAs were extracted after a 3-d inoculation and examined by RNA gel blot analysis.

Arrows and asterisks indicate the full-length and 3′ cleaved products of each mRNA, respectively.