Jurczyk Barbara, Pociecha Ewa, Janeczko Anna, Paczyński Robert, Rapacz Marcin
University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland.
University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland.
J Plant Physiol. 2014 Oct 15;171(16):1541-4. doi: 10.1016/j.jplph.2014.07.008. Epub 2014 Jul 25.
Quantitative PCR studies need proper reference genes with expression stability exclusively validated under certain experimental conditions. The expression stability of several genes commonly used as references was tested under 24-epibrassinolide (EBR) and temperature treatment. Different statistical approaches (qBase(PLUS), BestKeeper, NormFinder) were used to prepare rankings of expression stability in two species of an economic importance: common wheat (Triticum aestivum) and perennial ryegrass (Lolium perenne). Candidate reference genes were shown to be regulated differentially in these two plant species. The maximum stability values indicated that the expression stability was higher in T. aestivum. Taking into account of all ranks it seems that TBP-1 and UBI in ryegrass and ACT, ADP and EF1A in wheat should be used as reference genes in the brassinosteroids and temperature involving studies.
定量PCR研究需要在特定实验条件下专门验证过表达稳定性的合适的内参基因。在24-表油菜素内酯（EBR）和温度处理下，测试了几种常用作内参的基因的表达稳定性。使用不同的统计方法（qBase(PLUS)、BestKeeper、NormFinder）对两种具有经济重要性的物种：普通小麦（Triticum aestivum）和多年生黑麦草（Lolium perenne）的表达稳定性进行排序。候选内参基因在这两种植物物种中表现出不同的调控方式。最大稳定性值表明普通小麦中的表达稳定性更高。综合所有排名来看，在涉及油菜素内酯和温度的研究中，黑麦草中的TBP-1和UBI以及小麦中的ACT、ADP和EF1A应作为内参基因。
