Institute of Plant Genetics, Breeding and Biotechnology, University of Life Sciences in Lublin, Akademicka 13, 20-950, Lublin, Poland.
Sci Rep. 2022 Aug 26;12(1):14601. doi: 10.1038/s41598-022-18746-z.
In this study we evaluated eleven candidate reference genes in Avena sativa during compatible and incompatible interactions with two different pathotypes of Puccinia coronata f. sp. avenae in six time points post-inoculation. The identification of genes with high expression stability was performed by four algorithms (geNorm, NormFinder, BestKeeper and ΔCt method). The results obtained confirmed that the combination of two genes would be sufficient for reliable normalization of the expression data. In general, the most stable in the tested plant-pathogen system were HNR (heterogeneous nuclear ribonucleoprotein 27C) and EF1A (elongation factor 1-alpha). ARF (ADP-ribosylation factor) and EIF4A (eukaryotic initiation factor 4A-3) could also be considered as exhibiting high expression stability. CYP (cyclophilin) was shown by all assessment methods to be the worst candidate for normalization in this dataset. To date, this is the first report of reference genes selection in A. sativa-P. coronata interaction system. Identified reference genes enable reliable and comprehensive RT-qPCR analysis of oat gene expression in response to crown rust infection. Understanding the molecular mechanisms involved in the host-pathogen interactions may expand knowledge of durable resistance strategies beneficial to modern oat breeding.
在这项研究中，我们评估了 11 个候选参考基因在与两种不同致病型的雀麦生棒黑粉菌在六个时间点相互作用过程中的表达稳定性。通过四种算法（geNorm、NormFinder、BestKeeper 和 ΔCt 方法）对高表达稳定性的基因进行了鉴定。结果证实，两个基因的组合足以可靠地对表达数据进行归一化。总的来说，在测试的植物-病原体系统中，最稳定的是 HNR（异质核核糖核蛋白 27C）和 EF1A（延伸因子 1-α）。ARF（ADP-核糖基化因子）和 EIF4A（真核起始因子 4A-3）也可以被认为具有高表达稳定性。CYP（亲环蛋白）在所有评估方法中均被证明是该数据集归一化的最差候选基因。迄今为止，这是首次在雀麦-生棒黑粉菌互作系统中报道参考基因的选择。鉴定出的参考基因可用于可靠和全面的 RT-qPCR 分析燕麦基因对冠锈病感染的反应。了解宿主-病原体相互作用中涉及的分子机制可能会扩展对现代燕麦育种有益的持久抗性策略的知识。
