Fig. 4

Silence of the MaGT gene in transgenic petals of grape hyacinth. Schematic representation of RNAi expression vectors, pFGC-MaGT1 RNAi, constructed for transformation (a). Silencing of MaGT in transgenic flower petals resulted in a clear phenotypic change in coloration (b). Shown are non-transformed controls (Control) and three MaGT gene-silenced lines. Scale bar in d: 100 μm. PCR amplification of a 281 bp DNA fragment of the Basta gene from the transgenic and non-transgenic lines (c). Total RNA was extracted from young petals prior to full expansion. Then, mRNA accumulation of the MaGT gene was assessed by semi-quantitative and quantitative real-time PCR. Actin was used as a reference gene (d, e). Anthocyanin analysis of non-transformed and transformed flower petals (f). Each bar represents means ± standard deviations from three dependent replicates. The symbols ‘**’ and ‘***’ above bars indicate statistically significant differences at P ≤ 0.01 and P ≤ 0.001, respectively, by the student’s t-test. Control: non-transgenic petals, GT RNAi1-3: transgenic petals silencing GUS gene expression