title: Assessment of candidate reference genes for the expression studies with brassinosteroids in Lolium perenne and Triticum aestivum.
abstract: 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.
authors.affiliation: University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland. Electronic address: b.jurczyk@ur.krakow.pl.
authors.forename: Barbara
authors.initials: B
authors.lastname: Jurczyk
authors.affiliation: University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Pociecha
authors.affiliation: Institute of Plant Physiology, Polish Academy of Sciences, Niezapominajek 21, 30-239 Kraków, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Janeczko
authors.affiliation: University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland.
authors.forename: Robert
authors.initials: R
authors.lastname: Paczyński
authors.affiliation: University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Physiology, Podłużna 3, 30-239 Kraków, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Rapacz
mesh_terms: D060406:Brassinosteroids / Q000378:metabolism*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D008129:Lolium / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013260:Steroids, Heterocyclic / Q000378:metabolism*; D014908:Triticum / Q000235:genetics* / Q000378:metabolism
chemical_list: D060406:Brassinosteroids; D013260:Steroids, Heterocyclic; C023623:brassinolide
keywords: Brassinosteroids; Gene expression; Real time PCR; Temperature