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

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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, Aegiceras corniculatum, 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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Acknowledgments

This research was supported by the National Natural Science Foundation of China (Nos. 41430966 and 41176101), the Projects of Guangzhou Science and Technology (No. 15020024), the key projects in the National Science and Technology Pillar Program in the Eleventh Five-year Plan Period (No. 2012BAC07B0402), and the projects of the Knowledge Innovation Program of the Chinese Academy of Sciences (No. KSCX2-SW-132).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China

    Ya-Lan Peng & You-Shao Wang

  2. Daya Bay Marine Biology Research Station, Chinese Academy of Sciences, Shenzhen, 518121, China

    You-Shao Wang

  3. Laboratory of Environmental Toxicology, Department of Ecology & Biodiversity, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, China

    Ji-Dong Gu

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  1. Ya-Lan Peng
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  2. You-Shao Wang
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  3. Ji-Dong Gu
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Correspondence to You-Shao Wang.

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Peng, YL., Wang, YS. & Gu, JD. Identification of suitable reference genes in mangrove Aegiceras corniculatum under abiotic stresses. Ecotoxicology 24, 1714–1721 (2015). https://doi.org/10.1007/s10646-015-1487-8

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  • DOI: https://doi.org/10.1007/s10646-015-1487-8

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