Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida)

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dc.identifier.uri http://dx.doi.org/10.15488/503
dc.identifier.uri http://www.repo.uni-hannover.de/handle/123456789/527
dc.contributor.author Klie, Maik
dc.contributor.author Debener, Thomas
dc.date.accessioned 2016-09-02T08:00:21Z
dc.date.available 2016-09-02T08:00:21Z
dc.date.issued 2011
dc.identifier.citation Klie, Maik; Debener, Thomas: Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida). In: BMC Research Notes 4 (2011), 518. DOI: http://dx.doi.org/10.1186/1756-0500-4-518
dc.description.abstract Background: Gene expression studies are a prerequisite for understanding the biological function of genes. Because of its high sensitivity and easy use, quantitative PCR (qPCR) has become the gold standard for gene expression quantification. To normalise qPCR measurements between samples, the most prominent technique is the use of stably expressed endogenous control genes, the so called reference genes. However, recent studies show there is no universal reference gene for all biological questions. Roses are important ornamental plants for which there has been no evaluation of useful reference genes for gene expression studies. Results: We used three different algorithms (BestKeeper, geNorm and NormFinder) to validate the expression stability of nine candidate reference genes in different rose tissues from three different genotypes of Rosa hybrida and in leaves treated with various stress factors. The candidate genes comprised the classical "housekeeping genes" (Actin, EF-1α, GAPDH, Tubulin and Ubiquitin), and genes showing stable expression in studies in Arabidopsis (PP2A, SAND, TIP and UBC). The programs identified no single gene that showed stable expression under all of the conditions tested, and the individual rankings of the genes differed between the algorithms. Nevertheless the new candidate genes, specifically, PP2A and UBC, were ranked higher as compared to the other traditional reference genes. In general, Tubulin showed the most variable expression and should be avoided as a reference gene. Conclusions: Reference genes evaluated as suitable in experiments with Arabidopsis thaliana were stably expressed in roses under various experimental conditions. In most cases, these genes outperformed conventional reference genes, such as EF1-α and Tubulin. We identified PP2A, SAND and UBC as suitable reference genes, which in different combinations may be used for normalisation in expression analyses via qPCR for different rose tissues and stress treatments. However, the vast genetic variation found within the genus Rosa, including differences in ploidy levels, might also influence expression stability of reference genes, so that future research should also consider different genotypes and ploidy levels. eng
dc.language.iso eng
dc.publisher London : Biomed Central Ltd
dc.relation.ispartofseries BMC Research Notes 4 (2011)
dc.rights CC BY 2.0
dc.rights.uri https://creativecommons.org/licenses/by/2.0/
dc.subject Rosa hybrida eng
dc.subject qPCR eng
dc.subject PCR eng
dc.subject.ddc 500 | Naturwissenschaften ger
dc.subject.ddc 580 | Pflanzen (Botanik) ger
dc.title Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida)
dc.type article
dc.type Text
dc.relation.issn 1756-0500
dc.relation.doi http://dx.doi.org/10.1186/1756-0500-4-518
dc.bibliographicCitation.volume 4
dc.bibliographicCitation.firstPage 518
dc.description.version publishedVersion
tib.accessRights frei zug�nglich


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