Identification of suitable reference genes in mangrove Aegiceras corniculatum under abiotic stresses

Ya Lan Peng; You Shao Wang; Ji Dong Gu

doi:10.1007/s10646-015-1487-8

Identification of suitable reference genes in mangrove Aegiceras corniculatum under abiotic stresses

Ya Lan Peng, You Shao Wang^*, Ji Dong Gu

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

Gene expression studies could provide insight into the physiological mechanisms and strategies used by plants under stress conditions. Selection of suitable internal control gene(s) is essential to accurately assess gene expression levels. For the mangrove plant, Aegicerascorniculatum, reliable reference genes to normalize real-time quantitative PCR data have not been previously investigated. In this study, the expression stabilities of five candidate reference genes [glyceraldehydes-3-phosphate dehydrogenase (GAPDH), 18SrRNA, β-Actin, 60S ribosomal protein L2, and elongation factor-1-A] were determined in leaves of A. corniculatum treated by cold, drought, salt, heavy metals, and pyrene and in different tissues of A. corniculatum under normal condition. Two software programs (geNorm and NormFinder) were employed to analyze and rank the tested genes. Results showed that GAPDH was the most suitable reference gene in A. corniculatum and the combination of two or three genes was recommended for greater accuracy. To assess the value of these tested genes as internal controls, the relative quantifications of CuZnSOD gene were also conducted. Results showed that the relative expression levels of CuZnSOD gene varied depending on the internal reference genes used, which highlights the importance of the choice of suitable internal controls in gene expression studies. Furthermore, the results also confirmed that GAPDH was a suitable reference gene for qPCR normalization in A. corniculatum under abiotic stresses. Identification of A. corniculatum reference gens in a wide range of experimental samples will provide a useful reference in future gene expression studies in this species, particularly involving similar stresses.

Original language	English
Pages (from-to)	1714-1721
Number of pages	8
Journal	Ecotoxicology
Volume	24
Issue number	7-8
DOIs	https://doi.org/10.1007/s10646-015-1487-8
State	Published - 1 Oct 2015
Externally published	Yes

Keywords

Aegiceras corniculatum
Mangrove plants
Normalization
Quantitative real-time PCR
Reference gene

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10.1007/s10646-015-1487-8

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title = "Identification of suitable reference genes in mangrove Aegiceras corniculatum under abiotic stresses",

abstract = "Gene expression studies could provide insight into the physiological mechanisms and strategies used by plants under stress conditions. Selection of suitable internal control gene(s) is essential to accurately assess gene expression levels. For the mangrove plant, Aegicerascorniculatum, reliable reference genes to normalize real-time quantitative PCR data have not been previously investigated. In this study, the expression stabilities of five candidate reference genes [glyceraldehydes-3-phosphate dehydrogenase (GAPDH), 18SrRNA, β-Actin, 60S ribosomal protein L2, and elongation factor-1-A] were determined in leaves of A. corniculatum treated by cold, drought, salt, heavy metals, and pyrene and in different tissues of A. corniculatum under normal condition. Two software programs (geNorm and NormFinder) were employed to analyze and rank the tested genes. Results showed that GAPDH was the most suitable reference gene in A. corniculatum and the combination of two or three genes was recommended for greater accuracy. To assess the value of these tested genes as internal controls, the relative quantifications of CuZnSOD gene were also conducted. Results showed that the relative expression levels of CuZnSOD gene varied depending on the internal reference genes used, which highlights the importance of the choice of suitable internal controls in gene expression studies. Furthermore, the results also confirmed that GAPDH was a suitable reference gene for qPCR normalization in A. corniculatum under abiotic stresses. Identification of A. corniculatum reference gens in a wide range of experimental samples will provide a useful reference in future gene expression studies in this species, particularly involving similar stresses.",

keywords = "Aegiceras corniculatum, Mangrove plants, Normalization, Quantitative real-time PCR, Reference gene",

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N2 - Gene expression studies could provide insight into the physiological mechanisms and strategies used by plants under stress conditions. Selection of suitable internal control gene(s) is essential to accurately assess gene expression levels. For the mangrove plant, Aegicerascorniculatum, reliable reference genes to normalize real-time quantitative PCR data have not been previously investigated. In this study, the expression stabilities of five candidate reference genes [glyceraldehydes-3-phosphate dehydrogenase (GAPDH), 18SrRNA, β-Actin, 60S ribosomal protein L2, and elongation factor-1-A] were determined in leaves of A. corniculatum treated by cold, drought, salt, heavy metals, and pyrene and in different tissues of A. corniculatum under normal condition. Two software programs (geNorm and NormFinder) were employed to analyze and rank the tested genes. Results showed that GAPDH was the most suitable reference gene in A. corniculatum and the combination of two or three genes was recommended for greater accuracy. To assess the value of these tested genes as internal controls, the relative quantifications of CuZnSOD gene were also conducted. Results showed that the relative expression levels of CuZnSOD gene varied depending on the internal reference genes used, which highlights the importance of the choice of suitable internal controls in gene expression studies. Furthermore, the results also confirmed that GAPDH was a suitable reference gene for qPCR normalization in A. corniculatum under abiotic stresses. Identification of A. corniculatum reference gens in a wide range of experimental samples will provide a useful reference in future gene expression studies in this species, particularly involving similar stresses.

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KW - Aegiceras corniculatum

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Identification of suitable reference genes in mangrove Aegiceras corniculatum under abiotic stresses

Abstract

Keywords

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