Li Xi, Tang Dongqin, Shi Yimin
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
Heliyon. 2018 Jul 9;4(7):e00686. doi: 10.1016/j.heliyon.2018.e00686. eCollection 2018 Jul.
Quantitative real-time PCR (qRT-PCR) has been a widely used accurate technique for gene expression analysis in various species. However, its results require data normalization by reliable reference genes. Despite the horticultural importance of , and genome sequence has become available for the species, no gene expression study based on the stability of reference genes in qRT-PCR has been conducted. To boost the use of qRT-PCR in , we uncovered eight commonly used candidate reference genes for their stability. The expression levels of the eight genes were detected for the normalization in five different organs (bulbs, scapes, leaves, perianths and coronas) of three cultivars ('Marieke', 'Pinza' and 'Slim Whitman') by qRT-PCR. Subsequently, three commonly used computational programs were applied for evaluating the stability of the candidate reference gene's expressions. It turned out that for all the samples and most subgroups, and were the most suitable reference genes for normalization. However, the best reference genes were found not always the same one across diverse samples by different computational programs. Our study was the first reference gene evaluation in and will promote future studies on gene expression levels of .
定量实时聚合酶链反应（qRT-PCR）是一种广泛应用于各种物种基因表达分析的精确技术。然而，其结果需要通过可靠的内参基因进行数据标准化。尽管[物种名称]具有园艺重要性且其基因组序列已可获取，但尚未开展基于qRT-PCR中内参基因稳定性的基因表达研究。为了促进qRT-PCR在[物种名称]中的应用，我们筛选了八个常用的候选内参基因以评估其稳定性。通过qRT-PCR检测了这八个基因在三个[品种名称]（‘玛丽克’、‘平扎’和‘苗条惠特曼’）的五个不同器官（鳞茎、花葶、叶片、花被和副冠）中的表达水平，用于标准化分析。随后，应用三个常用的计算程序评估候选内参基因表达的稳定性。结果表明，对于所有样本和大多数亚组，[基因名称1]和[基因名称2]是最适合用于标准化的内参基因。然而，不同的计算程序显示，在不同样本中，最佳内参基因并非总是相同的。我们的研究是首次对[物种名称]进行内参基因评估，将推动未来关于[物种名称]基因表达水平的研究。
