Figure 2

Physiological characterization of LeMYC2 transgenic lines. A and B, Accumulation of chlorophyll in 6-day-old seedlings grown in white light (WL: 30 μmol/m²/s) and blue light (BL: 30 μmol/m²/s), respectively. C, Accumulation of anthocyanin in 6-day-old seedlings grown in BL (30 μmol/m²/s). Asterisks indicate that Ri1 and Ri2 are significantly different from the vector control (*P < 0.05, Student’s t test; n = 5). D and E, Real time PCR analysis for LeCAB1 and LeRBCS3 transcript levels in 6-day-old seedlings of various LeMYC2 transgenic lines grown in BL (30 μmol/m²/s). For experimental detail, see Figure 1A. F and G, The abundance of LeRBCS3 and LeCAB1 transcripts, respectively, from different transgenic seedlings grown in darkness for 5 days and then transferred to BL (30 μmol/m²/s) for 2 h and 4 h was determined by quantitative real time PCR. For experimental detail, see Figure 1A. H, LeMYC2 interacts with G-box containing LeRBCS3A promoter. Upper panel, Diagrammatic representation of 127 bp long LeRBCS-3A promoter fragment containing G-box used in the electrophoretic mobility shift assays. Lower panel, Gel shift assays using the GST-LeMYC2 and 127 bp G-box containing LeRBCS-3A promoter as probe. No protein was added in lane 1, and approximately 500 ng of GST protein was added in lane 2. In lanes (3–7) about 350 ng of GST-LeMYC2 protein was added. Competition was performed with 50 (lane numbers 4 and 6) and 100 (lane numbers 5 and 7) molar excess of wild type or mutated versions of 80 bp DNA fragment of LeRBCS-3A promoter as shown by the triangles in the figure. The arrowheads indicate the DNA-protein complex formed. The plus (+) and minus (−) signs indicate the presence and absence, respectively.