Southern blot and GFP fluorescence analyses of transgenic and NT plants for studying the promoter activity of <i>OsbZIP23</i> gene.

<p><b>(A)</b> Southern blot analysis of rice T₀ transformants of RuP and 20P lines developed with the <i>O</i>. <i>rufipogon</i> promoter-GFP construct (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150763#pone.0150763.s001" target="_blank">S1 Fig</a>) and <i>O</i>. <i>sativa</i> IR20 promoter-GFP construct (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150763#pone.0150763.s002" target="_blank">S2 Fig</a>), respectively. The genomic DNA of plant sample was digested with <i>Hind</i>III, and the CDS of GFP gene was used as hybridization probe. Lane NT- non-transgenic, Lane M- molecular weight marker. (<b>B</b>) GFP expression in finely cross-sectioned leaves of drought stressed single integration transgenic rice lines under the confocal microscope (Olympus FV1200). Scale bar 50 μm. (<b>C</b>) Real time PCR analysis showing relative expression level of <i>GFP</i> in leaf samples of RuP and 20P lines under drought stress condition, where rice polyubiquitin1 (<i>OsUbi1</i>) gene was taken as internal reference. Data bars represent the mean ±SD of triplicate measurement. Statistical analysis of Student’s t test indicated significant differences (**P<0.01).