Selection of reference genes for quantitative RT-PCR studies in Uncaria rhynchophylla

The real-time reverse transcription polymerase chain reaction (qRT-PCR) is a method for quantitative analysis and widely used for assessment of gene expression and regulation in medicinal plants such as uncaria (Uncaria rhynchophylla). During the practice, it is realized that selection of a reference gene has a great influence on data analysis. In this study, we identified the four candidate reference genes (GAPDH, alpha-tubulin, NDUFV and beta-actin) by analyzing the transcriptome and expression profiles from U rhynchophylla and evaluated the accuracy of these genes as a reference gene in qRT-PCR by Delta Ct, Bestkeeper, Normfinder and geNorm algorithms. The results showed that the performance of GAPDH and beta-actin was better than the other two genes while alpha-tubulin was the worst in the performance. Based on this study, we propose that GAPDH and beta-actin are the most reliable reference genes for the normalization of qRT-PCR data in quantitative analysis.