Figure 4
qRT-PCR validation of nitrate-responsive genes identified in light and dark conditions. Total RNA was isolated from control (H2O) and nitrate-treated leaves of 10 days old light-grown and etiolated rice seedlings. Relative transcript abundance was calculated by comparative Ct method using actin as a reference gene for data normalization. The data represent the mean ± SE of three technical replicates. Final relative fold change values (qPCR) of DEGs and their corresponding values obtained in microarray experiment were plotted for parallel comparison of DEG trends in qPCR and microarray experiments. To avoid additional bars and better clarity of the results, control value (H2O) obtained in qPCR were not included in the image. However, the statistical analyses were performed between control (H2O) and nitrate treated value (comparative Ct method) obtained by qPCR. Statistical unpaired T test analyses [control (H2O) vs. nitrate] were performed in the GraphPad Prism 6 software using technical replicates (**p value < 0.01, ***p value < 0.001). The experiments were performed with three independent biological replicates. MYB family transcription factors (BGIOSGA006030, BGIOSGA018651); universal stress protein ___domain containing protein (BGIOSGA005763); calcium-dependent protein kinase (BGIOSGA026567); efflux transporter of nicotianamine 1 (BGIOSGA034485); bHLH120 (BGIOSGA030896).
