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. Author manuscript; available in PMC: 2011 Nov 6.
Published in final edited form as: Plant J. 2010 May 6;63(3):417–429. doi: 10.1111/j.1365-313X.2010.04248.x

Figure 4. Transcription activation activity, subcellular localization and gene expression of ABO3.

Figure 4

(a) ABO3 promoter-driven GUS expression in different tissues: i, a whole seedling; ii, root; iii, stem; iv, leaf; v, flower; vi, total RNA was extracted from 10-day-old seedlings and reverse transcribed to cDNA. RT-PCR analysis of ABO3 transcript levels in the different tissues under internal control Actin was also displayed. The size of the amplified fragment is 180 bp for ABO3 and 220 bp for Actin.

(b) ABO3 promoter-driven GUS expression in the early development of seedlings, trichomes and guard cells. GUS expression in a seedling after seed germination for 24 (i), 48 (ii) or 72 h (iii). iv, GUS expression in trichomes. v, GUS expression in guard cells before and after 100 μM ABA treatment for 4 h. Scale bar: 10 μM.

(c) Subcellular localization of ABO3. An ABO3-GFP translational fusion construct was introduced into onion epidermal cells by particle bombardment: i, fluorescence image of ABO3-GFP; ii, the corresponding DAPI staining for the nucleus; iii, merged fluorescence and DAPI image; iv, the corresponding bright field; v, fluorescence image of GFP control; vi, the corresponding DAPI staining for the nucleus; vii, merged fluorescence and DAPI image; viii, the corresponding bright field. Scale bars: 50 μm.

(d) GAL4 DNA binding domain-ABO3 fusion analysis for transactivation activity and β-galactosidase quantitative assay. i, GAL4 DNA binding domain was fused with different parts of ABO3 and transformed into yeast strain AH109 containing the His3 and LacZ reporter genes. Three independent experiments were performed and each showed similar patterns. ii, Quantitative analysis of β-galactosidase activity of the relative yeast strains in liquid culture. Values are the means of data taken from three independent experiments. Error bars indicate the standard deviation. The labeling of the x-axis indicates the relative fold change of β-galactosidase activity between samples and the empty vector.