Molecular and phenotypic analyses of transgenic lines expressing mutant variant 1 (Var1) of smiRNA(CHS) ECV. RNA was extracted from different transgenic lines expressing smiRNA(CHS) ECV Var1, and qRT-PCR experiments were performed to quantify the relative transcript levels (A) of the smiRNA(CHS) ECV Var1 precursor and (B) of the CHS mRNA target. Data were normalized to the highest values. wt, wild type seedlings; Line 11, smiRNA(CHS) line 11 as controls (compare with Figures 2 and 3). (C) Relative anthocyanin content of the same transgenic smiRNA(CHS) ECV Var1 lines. Wild type seedlings were grown either without (wt-) or with 4% sucrose (wt+). All other plants were grown with 4% sucrose. Control seedlings were measured for comparison: chs knockout (tt4) and smiRNA(CHS) line 4 (Line 4; compare with Figures 2 and 3). (D) smiRNA:target mRNA hybrid structures of all smiRNA(CHS) variants. Differences of Var1-4 to the original smiRNA(CHS) are indicated in red. Mfe, minimal free energy.