Fig. 2

Expression and subcellular localization of GhYGL1d. a qRT-PCR analysis of GhYGL1d expression in different tissues. Total RNA was isolated from roots, stems, leaves, flowers and fibers. The GhUBQ7 gene was used as a reference gene for qRT-PCR. DPA, day post-anthesis. The values shown are means ± SE of three replicates. b GhYGL1d localizes to the chloroplast. Protoplasts from wild-type Arabidopsis were transiently transformed with a control GFP vector (designated 35S::GFP) or with a GhYGL1d-GFP vector. Fluorescence was observed by confocal microscopy of single protoplasts; green fluorescence = GFP; red = chlorophyll autofluorescence. Bars = 10 μm. c GhYGL1d localizes to the stroma and thylakoid fractions of chloroplasts. Total proteins extracted from the 35S::GhYGL1d-GFP transgenic line were used to confirm the specificity of the anti-GFP antibody. Intact chloroplasts were isolated from 35S::GhYGL1d-GFP transgenic seedlings and separated into the thylakoid and stroma fractions. Chloroplasts protein Cyt f and RbcL were used as a marker of thylakoid membrane and stroma fractions, respectively. The GFP antibody was used to detect the GhYGL1d-GFP fusion protein.