Selection of reference genes for reverse transcription-qPCR analysis in the biomonitor macrophyte Bidens laevis L

Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology
Germán LukaszewiczMirta Luján Menone

Abstract

The RT-qPCR has been the method used to analyze gene expression in plants but its benefits have not been completely exploited in the field of plants ecotoxicology when used as molecular biomarkers. The correct use of RT-qPCR demands to establish a certain number of reference genes (RG) which are expected to be invariable in their expression although it does not always happen. The main goals of this work were to: (1) analyze the stability of six potential RG, (2) establish the optimum number of RG, (3) select the most suitable RG to be applied in Bidens laevis under different test conditions and tissues and (4) confirm its convenience by normalizing the expression of one gene of interest under three different challenges. When all data were pooled together, the geNorm algorithm pointed out beta-actin and beta-tubulin (TUB) as the optimal RG pair while NormFinder algorithm selected nicotinamide adenine dinucleotide dehydrogenase (NADHD) and histone 3 (H3) as possessing the most invariable levels of expression. On the other hand, when data were grouped by tissues, ANOVA test selected H3 and TUB, while data grouped by conditions indicated that H3 and NADHD were the most stable RG under this analysis. Therefore, for a general-purpose...Continue Reading

References

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Jun 18, 2011·Virology Journal·Guillermo A MaronicheMariana del Vas

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Citations

Feb 3, 2021·Plant Physiology and Biochemistry : PPB·María Belén FernándezRaúl Cassia

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Datasets Mentioned

BETA
JX843817

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

NormFinder
geNorm
Genex package ( Multid Analyses AB
Basic Local Alignment Search Tool ( BLAST
Genex

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