Validation of reference genes as internal control for studying viral infections in cereals by quantitative real-time RT-PCR.

BMC Plant Biology
Jana Jarosová, Jiban K Kundu

Abstract

Reference genes are commonly used as the endogenous normalisation measure for the relative quantification of target genes. The appropriate application of quantitative real-time PCR (RT-qPCR), however, requires the use of reference genes whose level of expression is not affected by the test, by general physiological conditions or by inter-individual variability. For this purpose, seven reference genes were investigated in tissues of the most important cereals (wheat, barley and oats). Titre of Barley yellow dwarf virus (BYDV) was determined in oats using relative quantification with different reference genes and absolute quantification, and the results were compared. The expression of seven potential reference genes was evaluated in tissues of 180 healthy, physiologically stressed and virus-infected cereal plants. These genes were tested by RT-qPCR and ranked according to the stability of their expression using three different methods (two-way ANOVA, GeNorm and NormFinder tools). In most cases, the expression of all genes did not depend on abiotic stress conditions or virus infections. All the genes showed significant differences in expression among plant species. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-tubulin (T...Continue Reading

References

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Citations

Jun 5, 2013·Indian Journal of Virology : an Official Organ of Indian Virological Society·S Gadiou, J K Kundu
Oct 1, 2013·Journal of Applied Genetics·Bartłomiej Kozera, Marcin Rapacz
Apr 12, 2011·Journal of Agricultural and Food Chemistry·Angeles ObreroClara I González-Verdejo
May 16, 2012·BMC Plant Biology·Ann J SladeVic C Knauf
Jan 12, 2012·BMC Research Notes·Cécile PetitChristophe Délye
Oct 2, 2012·Molecular Breeding : New Strategies in Plant Improvement·Nancy PodevinSerge Remy
Feb 7, 2015·Plant Biotechnology Journal·Guillermina M MendiondoMichael J Holdsworth
May 4, 2015·Applied Biochemistry and Biotechnology·Filiz GürelDeepak M Kalaskar
Jan 1, 2014·Virus Research·Narelle NancarrowJo E Luck
Jun 11, 2014·Journal of Experimental Botany·Guillermina M MendiondoMichael J Holdsworth
Mar 19, 2016·Molecular Plant Pathology·Damien J LightfootAmanda J Able
Oct 13, 2017·Environmental Toxicology and Chemistry·Zayneb ChaâbeneFranck Vandenbulcke
Apr 21, 2020·The Journal of Pharmacy and Pharmacology·Siva Prasad PandaBikash Ranjan Jena

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Methods Mentioned

BETA
PCR
PCRs
in vitro transcription

Software Mentioned

GeNorm
7300 Real - Time PCR system
NormFinder
BestKeeper

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