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Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
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Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
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Journal Article
Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
2021
Overview
Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression levels in luffa plants exposed to various simulated abiotic stresses [i.e., cold, drought, heat, salt, H
2
O
2
, and abscisic acid (ABA) treatments] were analyzed by qRT-PCR. The stability of the reference gene expression levels was validated using the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. The results indicated that
EF-1α
was the most stably expressed and suitable reference gene overall and for the heat, cold, and ABA treatments. Additionally,
UBQ
expression was stable following the salt treatment, whereas
TUB
was identified as a suitable reference gene for H
2
O
2
and drought treatments. The reliability of the selected reference genes was verified by analyzing the expression of copper/zinc superoxide dismutase (
Cu/Zn-SOD
) gene in luffa. When the most unstable reference genes were used for data normalizations, the resulting expression patterns had obvious biases when compared with the expression patterns for the most ideal reference genes used alone or combined. These results will be conducive to more accurate quantification of gene expression levels in luffa.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 631/449
/ Abscisic Acid - pharmacology
/ Adaptation, Physiological - genetics
/ Drought
/ Droughts
/ Gene Expression Regulation, Plant
/ Humanities and Social Sciences
/ Hydrogen Peroxide - pharmacology
/ Peptide Elongation Factor 1 - genetics
/ Peptide Elongation Factor 1 - metabolism
/ Real-Time Polymerase Chain Reaction - standards
/ Salinity
/ Science
/ Stress, Physiological - genetics
