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. 2014 Mar 13;10(3):e1004232. doi: 10.1371/journal.pgen.1004232

Figure 1. Artificial miR159a variants with up to two central mismatches are potent silencers of MYB33/MYB65.

Figure 1

(A) Structure of the MIR159a transgene used to generate miR159a variant constructs. Numbers indicating relative positions to the transcription start site. Black arrow: Transcription start site; Orange bar: Polyadenylation site. (B) The alignment between MYB33 and the miR159a variant sequences used to transform mir159ab. The percentage of free energy pairing with MYB33 compared to a perfect match is listed on the right. Green: original mismatches; Purple: corrected mismatches; Red: introduced mismatches. (C) Aerial view of rosettes from 30-day-old MIR159a0, MIR159a1 and MIR159a2 plants (mir159ab) grown side by side with Col-0 and mir159ab under long day conditions. Scale bar = 10 mm. (D) The mRNA levels of CP1 in the rosettes of mir159ab plants complemented by the MIR159 variants in comparison with wild type and mir159ab. The value for each variant is the average of three independent transgenic lines. All mRNA levels were normalized with CYCLOPHILIN 5. Error bars represent the SEM. (E) Mature miR159a levels in Col-0 and mir159ab, and mature miR159a variant levels in three independent lines of MIR159a0 (mir159ab), MIR159a1 (mir159ab) and MIR159a2 (mir159ab) transformants. All miR159a variant and miR159a levels were normalized with sno101 by comparative quantitation analysis. Error bars represent the SEM.