Evaluation and selection of candidate reference genes for normalization of quantitative RT-PCR in Withania somnifera (L.) Dunal

PloS One
Varinder SinghPratap Kumar Pati

Abstract

Quantitative real-time PCR (qRT-PCR) is now globally used for accurate analysis of transcripts levels in plants. For reliable quantification of transcripts, identification of the best reference genes is a prerequisite in qRT-PCR analysis. Recently, Withania somnifera has attracted lot of attention due to its immense therapeutic potential. At present, biotechnological intervention for the improvement of this plant is being seriously pursued. In this background, it is important to have comprehensive studies on finding suitable reference genes for this high valued medicinal plant. In the present study, 11 candidate genes were evaluated for their expression stability under biotic (fungal disease), abiotic (wounding, salt, drought, heat and cold) stresses, in different plant tissues and in response to various plant growth regulators (methyl jasmonate, salicylic acid, abscisic acid). The data as analyzed by various software packages (geNorm, NormFinder, Bestkeeper and ΔCt method) suggested that cyclophilin (CYP) is a most stable gene under wounding, heat, methyl jasmonate, different tissues and all stress conditions. T-SAND was found to be a best reference gene for salt and salicylic acid (SA) treated samples, while 26S ribosomal RNA...Continue Reading

References

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Dec 1, 2008·Indian Journal of Microbiology·Pratap Kumar PatiB Singh

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Datasets Mentioned

BETA
AJ236016.1
X55749.1

Methods Mentioned

BETA
reverse-transcription PCR
electrophoresis
PCR

Software Mentioned

MS Excel
StepOne Plus
Excel
RefFinder
BestKeeper
NormFinder
GeNorm
Primer Quest

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