Gu Chun-Sun, Liu Liang-qin, Xu Chen, Zhao Yan-hai, Zhu Xu-dong, Huang Su-Zhen
Institute of Botany, Jiangsu Province and Chinese Academy of Science, Nanjing, Jiangsu 210014, China.
College of Horticulture, Nanjing Agricultural University, Nanjing 210014, China.
ScientificWorldJournal. 2014;2014:532713. doi: 10.1155/2014/532713. Epub 2014 May 26.
Quantitative real time PCR (RT-qPCR) has emerged as an accurate and sensitive method to measure the gene expression. However, obtaining reliable result depends on the selection of reference genes which normalize differences among samples. In this study, we assessed the expression stability of seven reference genes, namely, ubiquitin-protein ligase UBC9 (UBC), tubulin alpha-5 (TUBLIN), eukaryotic translation initiation factor (EIF-5A), translation elongation factor EF1A (EF1 α ), translation elongation factor EF1B (EF1b), actin11 (ACTIN), and histone H3 (HIS), in Iris. lactea var. chinensis (I. lactea var. chinensis) root when the plants were subjected to cadmium (Cd), lead (Pb), and salt stress conditions. All seven reference genes showed a relatively wide range of threshold cycles (C t ) values in different samples. GeNorm and NormFinder algorithms were used to assess the suitable reference genes. The results from the two software units showed that EIF-5A and UBC were the most stable reference genes across all of the tested samples, while TUBLIN was unsuitable as internal controls. I. lactea var. chinensis is tolerant to Cd, Pb, and salt. Our results will benefit future research on gene expression in response to the three abiotic stresses.
定量实时聚合酶链反应（RT-qPCR）已成为一种测量基因表达的准确且灵敏的方法。然而，要获得可靠的结果取决于选择能使样本间差异标准化的内参基因。在本研究中，我们评估了七个内参基因在马蔺根中的表达稳定性，这七个内参基因分别是泛素蛋白连接酶UBC9（UBC）、微管蛋白α-5（TUBLIN）、真核翻译起始因子（EIF-5A）、翻译延伸因子EF1A（EF1α）、翻译延伸因子EF1B（EF1b）、肌动蛋白11（ACTIN）和组蛋白H3（HIS），实验条件为马蔺植株遭受镉（Cd）、铅（Pb）和盐胁迫。在不同样本中，所有七个内参基因均显示出相对较宽的阈值循环（Ct）值范围。使用GeNorm和NormFinder算法来评估合适的内参基因。两个软件单元的结果表明，在所有测试样本中，EIF-5A和UBC是最稳定的内参基因，而TUBLIN不适合作为内参。马蔺对镉、铅和盐具有耐受性。我们的结果将有助于未来关于基因表达对这三种非生物胁迫响应的研究。
