Selection of reference genes for quantitative real-time PCR analysis in halophytic plant Rhizophora apiculata

PeerJ
Ankush Ashok SaddheKundan Kumar

Abstract

Rhizophora apiculata is a halophytic, small mangrove tree distributed along the coastal regions of the tropical and subtropical areas of the world. They are natural genetic reservoirs of salt adaptation genes and offer a unique system to explore adaptive mechanisms under salinity stress. However, there are no reliable studies available on selection and validation of reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) in R. apiculata physiological tissues and in salt stress conditions. The selection of appropriate candidate reference gene for normalization of qRT-PCR data is a crucial step towards relative analysis of gene expression. In the current study, seven genes such as elongation factor 1α (EF1α), Ubiquitin (UBQ), β-tubulin (β-TUB), Actin (ACT), Ribulose1,5-bisphosphate carboxylase/oxygenase (rbcL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and 18S rRNA (18S) were selected and analyzed for their expression stability. Physiological tissues such as leaf, root, stem, and flower along with salt stress leaf samples were used for selection of candidate reference genes. The high-quality expression data was obtained from biological replicates and further analyzed using five different programs su...Continue Reading

References

Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Feb 1, 2012·Plant Molecular Biology·Fuliang XieBaohong Zhang
Oct 1, 2013·Journal of Applied Genetics·Bartłomiej Kozera, Marcin Rapacz
Mar 27, 2018·Plant Science : an International Journal of Experimental Plant Biology·Rakesh ManukaKundan Kumar

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Citations

Apr 18, 2021·Journal of Biotechnology·Rakesh ManukaSuprasanna Penna

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Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

Bestkeeper
BLASTN
MS Excel
PrimerQuest
NormFinder
Excel
geNorm
SPSS
RefFinder
BLAST

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