Selection of internal control genes for real-time quantitative RT-PCR assays in the oomycete plant pathogen Phytophthora parasitica

Fungal Genetics and Biology : FG & B
Hao-Zhi Yan, Ruey-Fen Liou

Abstract

Real-time quantitative reverse transcription-PCR (qRT-PCR) has become one of the most commonly used methods for RNA quantification in recent years. To obtain reliable results with biological significance, it is important that qRT-PCR data are normalized with a proper internal control. In this study, 18 housekeeping genes were selected and evaluated for their potential as a suitable internal control for study of gene expression in the oomycete plant pathogen Phytophthora parasitica. Analysis of qRT-PCR data using the geNorm software indicated that, although commonly used as internal controls, beta-actin (ACT) and translation elongation factor 1alpha (eEF1A) might not be the best choice due to variable expression across different life stages of P. parasitica. Instead, other genes would serve as better controls, including ubiquitin-conjugating enzyme (Ubc), WS21, and beta-tubulin (Tub-b) for 'asexual stage,' Ubc and Tub-b for 'sexual reproduction,' while Ubc and WS21 for the stage of pathogenesis, because of their constant expression levels in each given subset of RNA samples. Although normalization with more than one gene would generate more reliable results, use of a single stably expressed gene as an internal control would suff...Continue Reading

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Citations

Feb 18, 2009·Molecular Genetics and Genomics : MGG·Pamela Hui Peng GanAdrienne R Hardham
Mar 7, 2008·Molecular Plant-microbe Interactions : MPMI·Chih-Hang WuRuey-Fen Liou
Feb 4, 2009·BMC Bioinformatics·Vlad PopoviciPratyaksha Wirapati
Dec 22, 2011·BMC Evolutionary Biology·Anne DiévartChristophe Périn
Jun 3, 2014·BMC Plant Biology·Bao-Zhen FengXiu-Guo Zhang
Oct 1, 2010·Molecular Genetics and Genomics : MGG·Michaela KingGiles E St J Hardy
May 11, 2010·Fungal Genetics and Biology : FG & B·Reena D NarayanAdrienne R Hardham
Oct 24, 2009·Molecular Plant Pathology·Howard S JudelsonReena D Narayan
Jan 6, 2009·Journal of Evolutionary Biology·M S Pankey, J P Wares
Mar 12, 2015·BMC Genomics·Agustina LlanosJean-Luc Parrou

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