Selection and validation of reference genes for quantitative real-time PCR analysis of gene expression in Cichorium intybus

Frontiers in Plant Science
Marianne DelporteDavid Gagneul

Abstract

Plant polyphenols represent a huge reservoir of bioactive compounds. Industrial chicory, an important crop from northwestern Europe, accumulates an original combination of such compounds, i.e., chlorogenic, isochlorogenic, caftaric, and chicoric acids arising from the phenylpropanoid pathway. For a complete understanding of these biochemical pathways, analyses of gene expression using quantitative real-time PCR (qRT-PCR) should be considered. Because cell cultures are a model of choice for specialized metabolism investigations, this study described for the first time the validation of reference genes for this system in chicory. Eighteen potential reference genes were obtained by mining expressed sequence tag databases of chicory for orthologs of Arabidopsis thaliana genes currently used as reference genes. Twelve genes passed the qRT-PCR standard requirements and their expression stability across different samples was tested using three distinct softwares: geNorm, NormFinder, and BestKeeper. In cell cultures grown under various conditions, TIP41 (TIP41 like protein) was shown to be the most stable gene. Further validation of the proposed reference genes was done by normalization of expression levels of a group of genes of inter...Continue Reading

References

Jul 29, 2005·Critical Reviews in Food Science and Nutrition·Augustin ScalbertLiliana Jiménez
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Jan 1, 2014·Evidence-based Complementary and Alternative Medicine : ECAM·Renée A StreetGerhard Prinsloo

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Citations

Mar 24, 2018·Archives of Insect Biochemistry and Physiology·Genhong WangQingyou Xia
Nov 19, 2019·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Wenkai DuChun Liu
Aug 14, 2019·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Yingling WanYan Liu
Oct 30, 2021·Frontiers in Genome Editing·Charlotte De BruynKatrijn Van Laere

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Datasets Mentioned

BETA
DT210806.1
FL679405.1
EH681285.1
EH709287.1
EH675923.1

Methods Mentioned

BETA
PCR
Electrophoresis
454 sequencing

Software Mentioned

GeNorm
R
NormFinder
BestKeeper
R Core
BLASTX
Clath

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