Li Jinting, Han Xueping, Wang Can, Qi Wanzhen, Zhang Weiyu, Tang Li, Zhao Xiting
College of Life Sciences, Henan Normal UniversityXinxiang, China.
Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude DrugsXinxiang, China.
Front Plant Sci. 2017 May 16;8:776. doi: 10.3389/fpls.2017.00776. eCollection 2017.
Real-time quantitative polymerase chain reaction (RT-qPCR) is a sensitive technique for gene expression studies. However, choosing the appropriate reference gene is essential to obtain reliable results for RT-qPCR assays. In the present work, the expression of eight candidate reference genes, (elongation factor 1-), (glyceraldehyde 3-phosphate dehydrogenase), (ubiquitin-conjugating enzyme), (polyubiquitin), (actin), (-tubulin), (adenine phosphoribosyltransferase 1), and (18S ribosomal RNA), was evaluated in samples using two algorithms, geNorm and NormFinder. The samples were classified into groups according to developmental stages, various tissues, stresses (cold, heat, drought, NaCl), and hormone treatments (MeJA, IBA, SA). Suitable combination of reference genes for RT-qPCR normalization should be applied according to different experimental conditions. In this study, , , and genes were verified as the suitable reference genes across all tested samples. To validate the suitability of the reference genes, we evaluated the relative expression of , which is a gene that may be involved in phytosterol synthesis. Our results provide the foundation for gene expression analysis in and other species of Amaranthaceae.
实时定量聚合酶链反应（RT-qPCR）是一种用于基因表达研究的灵敏技术。然而，选择合适的内参基因对于获得可靠的RT-qPCR检测结果至关重要。在本研究中，使用geNorm和NormFinder两种算法评估了8个候选内参基因（延伸因子1-、甘油醛-3-磷酸脱氢酶、泛素结合酶、多聚泛素、肌动蛋白、β-微管蛋白、腺嘌呤磷酸核糖转移酶1和18S核糖体RNA）在苋属植物样本中的表达情况。样本根据发育阶段、不同组织、胁迫（冷、热、干旱、NaCl）和激素处理（茉莉酸甲酯、吲哚丁酸、水杨酸）进行分组。应根据不同的实验条件应用适合RT-qPCR标准化的内参基因组合。在本研究中，延伸因子1-、甘油醛-3-磷酸脱氢酶和肌动蛋白基因被验证为所有测试样本中的合适内参基因。为了验证内参基因的适用性，我们评估了可能参与植物甾醇合成的基因的相对表达。我们的结果为苋属植物及其他苋科物种的基因表达分析奠定了基础。
