Figure 2.

Lowering ΔG free energy of a miRNA-target interaction can improve silencing efficacy. A, Nucleotide and amino acid changes introduced to generate mMYB81, MYB81-33, and MYB81-m33 compared to MYB81. The miR159 binding sequences are marked in red. Amino acid changes are marked in green, and nucleotide changes in blue. B, The number of primary transformants of MYB33, MYB81, mMYB81, MYB81-33, and MYB81-m33 falling into each phenotypic category (none, mild, or severe), as a percentage of the total number of transformants generated for each construct (n). C, Transcript levels of MYB81 and CP1 in primary transformants of MYB33, MYB81, mMYB81, MYB81-33, and MYB81-m33 measured by qRT-PCR. RNA was extracted from pools of rosette tissue of 10 to 15 randomly selected, 3- to 4-week-old primary transformants. Col-0 was used as wild-type control and mRNA levels were normalized to CYCLOPHILIN. Measurements are the average of three technical replicates.