Sozoniuk Magdalena, Czapla Karolina, Sugier Danuta, Kowalczyk Krzysztof
Institute of Plant Genetics, Breeding and Biotechnology, University of Life Sciences in Lublin, Akademicka Street 15, 20-950, Lublin, Poland.
Department of Biochemistry and Molecular Biology, Medical University of Lublin, Chodźki 1, 20-093, Lublin, Poland.
Sci Rep. 2024 Dec 30;14(1):32049. doi: 10.1038/s41598-024-83709-5.
A better understanding of secondary metabolites biosynthesis requires comprehensive research at the molecular level. Although the medicinal importance of secondary metabolites extracted from Arnica spp. has been well documented, the very plants themselves have been poorly studied. Characterization of biosynthetic pathways and their complex regulatory mechanisms may be enhanced via transcriptomic approach. To ensure the reliability of data, RT-qPCR based gene expression studies should be preceded by the reference genes selection step. Here, ten candidate reference genes were analyzed in terms of their expression stability in developing flowers of two arnica species, Arnica montana and Arnica chamissonis. After evaluating their expression stability with four distinct algorithms (geNorm, NormFinder, BestKeeper and delta Ct method), we found that SKIP16 and F-box were the most stable reference genes in A. montana, whereas SAND and F-box were best-performing genes in A. chamissonis. On the other hand, ACT1 in combination with F-box could be used as internal control for cross-species investigations. This study will aid in further research on molecular mechanisms underlying the processes of flowering and secondary metabolites production in medicinal plants.
更好地理解次生代谢产物的生物合成需要在分子水平上进行全面研究。尽管从山金车属植物中提取的次生代谢产物的药用价值已有充分记载，但对这些植物本身的研究却很少。通过转录组学方法可以加强对生物合成途径及其复杂调控机制的表征。为确保数据的可靠性，基于RT-qPCR的基因表达研究应先进行参考基因选择步骤。在此，对10个候选参考基因在两种山金车属植物——高山山金车和山金车花发育中的花朵中的表达稳定性进行了分析。在用四种不同算法（geNorm、NormFinder、BestKeeper和delta Ct法）评估它们的表达稳定性后，我们发现SKIP16和F-box是高山山金车中最稳定的参考基因，而SAND和F-box是山金车花中表现最佳的基因。另一方面，ACT1与F-box结合可用作跨物种研究的内参。这项研究将有助于进一步研究药用植物开花和次生代谢产物产生过程的分子机制。
