Wan Yingling, Hong Aiying, Zhang Yixuan, Liu Yan
1Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, National Engineering Research Center for Floriculture, Beijing Laboratory of Urban and Rural Ecological Environment, College of Landscape Architecture, Beijing Forestry University, Beijing, 100083 People's Republic of China.
Management Office of Caozhou Peony Garden, Heze, 274000 Shandong People's Republic of China.
Physiol Mol Biol Plants. 2019 Jul;25(4):1097-1105. doi: 10.1007/s12298-019-00684-2. Epub 2019 Jun 20.
The stem of will bend when it grows in greenhouse at a low light intensity. It is important to explore causes of morphological changes of peony to improve its quality. Gene expression can be evaluated by quantitative real-time PCR, based on reference gene. However, systematic selection of reference genes under weak lighting for herbaceous peony is lacking. To address this problem, we first selected 10 candidate reference genes based on a coefficient of variation of gene expression from peony stem transcriptome data. Then, geNorm, NormFinder and BestKeeper were applied to assess the stability of the genes, and RankAggreg was used to give a comprehensive ranking. The results show that there are some differences in optimal reference genes among samples from different organs and under the two lighting conditions, and the optimal number of suitable reference genes is distinct. Two selected suitable reference genes were then used to normalize target genes, and the results were compared with transcriptome data. Consistent gene expression trends were obtained, indicating the reliability of the method. To the best of our knowledge, this is the first time reference genes for herbaceous peony were selected in different organs, developmental stages and under two kinds of lighting conditions. The findings can provide a practical method for selecting reference genes for peony under these conditions and demonstrate a useful combination of reference genes.
芍药茎在弱光条件下于温室中生长时会弯曲。探究芍药形态变化的原因对于提高其品质很重要。基于内参基因，基因表达可通过定量实时聚合酶链式反应（qRT-PCR）进行评估。然而，目前缺乏对草本芍药在弱光条件下内参基因的系统筛选。为解决这一问题，我们首先基于芍药茎转录组数据中基因表达的变异系数，筛选出10个候选内参基因。然后，应用geNorm、NormFinder和BestKeeper评估这些基因的稳定性，并使用RankAggreg给出综合排名。结果表明，来自不同器官的样本以及在两种光照条件下，最佳内参基因存在一些差异，且合适的内参基因的最佳数量也不同。随后，使用两个选定的合适内参基因对靶基因进行标准化，并将结果与转录组数据进行比较。获得了一致的基因表达趋势，表明该方法的可靠性。据我们所知，这是首次在不同器官、发育阶段以及两种光照条件下筛选草本芍药的内参基因。这些发现可为在这些条件下筛选芍药内参基因提供一种实用方法，并展示了内参基因的有效组合。
