Zhang M F, Liu Q, Jia G X
Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, National Engineering Research Center for Floriculture and College of Landscape Architecture, Beijing Forestry University, Beijing, China.
Genet Mol Res. 2016 Apr 29;15(2):gmr7982. doi: 10.4238/gmr.15027982.
Quantitative real-time polymerase chain reaction (qRT-PCR) is an important technology used to analyze gene-expression levels. Reference genes, which are assumed to be expressed consistently across various developmental stages and in different tissues, were selected for expression level analysis. Using digital gene expression technology, we selected nine reference genes (18S, EF, CYCOL, SAND, GAPDH, ACTIN, BHLH, TIP, and Clathrin) as candidate reference genes for further study. Using three different analysis methods (GeNorm, NormFinder, and BestKeeper), a total of 144 lily (Lilium x formolongi "Raizan 3") samples were analyzed. The samples were collected from four different tissues under various developmental stages. In addition, leaves treated with different plant hormones were collected and analyzed. The data showed that the stability of the nine reference genes differed among samples, but TIP, EF, Clathrin, and BHLH could be identified as the most stable genes overall. In addition, the relative expression level of LfFT in different lily tissues with the competence to flower was also analyzed to verify the selected reference genes. This study constitutes an important source for selecting reference genes when analyzing the expression patterns of flowering time and floral development regulation genes in lily cultivars.
定量实时聚合酶链反应（qRT-PCR）是一种用于分析基因表达水平的重要技术。选择假定在各个发育阶段和不同组织中均持续表达的内参基因进行表达水平分析。利用数字基因表达技术，我们选择了9个内参基因（18S、EF、CYCOL、SAND、GAPDH、ACTIN、BHLH、TIP和网格蛋白）作为进一步研究的候选内参基因。使用三种不同的分析方法（GeNorm、NormFinder和BestKeeper），对总共144个麝香百合（Lilium x formolongi "Raizan 3"）样本进行了分析。这些样本取自不同发育阶段的四种不同组织。此外，还收集并分析了用不同植物激素处理过的叶片。数据表明，9个内参基因的稳定性在不同样本间存在差异，但总体而言TIP、EF、网格蛋白和BHLH可被确定为最稳定的基因。此外，还分析了具有成花能力的不同麝香百合组织中LfFT的相对表达水平，以验证所选择的内参基因。本研究为分析百合品种开花时间和花发育调控基因的表达模式时选择内参基因提供了重要来源。
