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

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• 作者：Ya-Lan Peng ; You-Shao Wang ; Ji-Dong Gu
• 关键词：Mangrove plants ; Aegiceras corniculatum ; Quantitative real ; time PCR ; Reference gene ; Normalization
• 刊名：Ecotoxicology
• 出版年：2015
• 出版时间：October 2015
• 年：2015
• 卷：24
• 期：7-8
• 页码：1714-1721
• 全文大小：898 KB
• 参考文献：Andersen CL, Jensen JL, ?rntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245-250CrossRef
  Barber RD, Harmer DW, Coleman RA, Clark BJ (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics 21:389CrossRef
  Barsalobres-Cavallari CF, Severino FE, Maluf MP, Maia IG (2009) Identification of suitable internal control genes for expression studies in Coffea arabica under different experimental conditions. BMC Mol Biol 10:1CrossRef
  Basyuni M, Kinjo Y, Baba S, Shinzato N, Iwasaki H, Siregar EBM, Oku H (2010) Isolation of salt stress tolerance genes from roots of mangrove plant, Rhizophora stylosa Griff., using PCR-based suppression subtractive hybridization. Plant Mol Bio Rep 29:533-43CrossRef
  Burchett M, Clarke C, Field C, Pulkownik A (2006) Growth and respiration in two mangrove species at a range of salinities. Physiol Plant 75:299-03CrossRef
  Carvalho K, de Campos MKF, Pereira LFP, Vieira LGE (2010) Reference gene selection for real-time quantitative polymerase chain reaction normalization in “Swingle-citrumelo under drought stress. Anal Biochem 402:197-99CrossRef
  Czechowski T (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139:5-7CrossRef
  Fu X, Huang Y, Deng S, Zhou R, Yang G, Ni X, Li W, Shi S (2005) Construction of a SSH library of Aegiceras corniculatum under salt stress and expression analysis of four transcripts. Plant Sci 169:147-54CrossRef
  Gilman EL, Ellison J, Duke NC, Field C (2008) Threats to mangroves from climate change and adaptation options: a review. Aquat Bot 89:237-50CrossRef
  Huang G-Y, Wang Y-S (2009) Expression analysis of type 2 metallothionein gene in mangrove species (Bruguiera gymnorrhiza) under heavy metal stress. Chemoshpere 77:1026-029CrossRef
  Huang GY, Wang YS (2010) Expression and characterization analysis of type 2 metallothionein from grey mangrove species (Avicennia marina) in response to metal stress. Aquat Toxicol 99:86-2CrossRef
  Jithesh MN, Prashanth SR, Sivaprakash KR, Parida A (2006) Monitoring expression profiles of antioxidant genes to salinity, iron, oxidative, light and hyperosmotic stresses in the highly salt tolerant grey mangrove, Avicennia marina (Forsk.) Vierh. by mRNA analysis. Plant Cell Rep 25:865-76CrossRef
  Marchand C, Lallier-Vergès E, Baltzer F, Albéric P, Cossa D, Baillif P (2006) Heavy metals distribution in mangrove sediments along the mobile coastline of French Guiana. Mar Chem 98:1-7CrossRef
  Migocka M, Papierniak A (2010) Identification of suitable reference genes for studying gene expression in cucumber plants subjected to abiotic stress and growth regulators. Mol Breed 3:343-57
  Nicot N, Hausman JF, Hoffman L, Evers D (2005) Housekeeping gene selection for real-time RT-PCR normalization in potato during biotic and abiotic stress. J Exp Bot 56:2907-914CrossRef
  Peng YL, Wang YS, Cheng H, Sun CC, Wu P, Wang LY, Fei J (2013) Characterization and expression analysis of three CBF/DREB 1 transcriptional factor genes from mangrove Avicennia marina. Aquat Toxicol 140-41:68-6CrossRef
  Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2004-007CrossRef
  Qi J, Yu S, Zhang F, Shen X, Zhao X, Yu Y, Zhang D (2010) Reference gene selection for real-time quantitative polymerase chain reaction of mRNA transcript levels in Chinese cabbage (Brassica rapa L. ssp. pekinensis). Plant Mol Biol Rep 28:597-04CrossRef
  Remans T, Smeets K, Opdenakker K, Mathijsen D, Vangronsveld J, Cuypers A (2008) Normalization of real-time RT-PCR gene expression measurements in Arabidopsis thaliana exposed to increased metal concentrations. Planta 227:1343-349CrossRef
  Selvey S, Thompson E, Matthaei K, Lea RA, Irving MG, Griffiths L (2001) Beta-actin: an unsuitable internal control for RT-PCR. Mol Cell Probe 15:307CrossRef
  Tam N, Wong Y, Wong M (2009) Novel technology in pollutant removal at source and bioremediation. Ocean Coast Manag 52:368-73CrossRef
  Tomlinson PB (1994) The botany of mangroves. Cambridge University Press, Cambridge
  Udvardi MK, Czechowski T, Scheible WR (2008) Eleven golden rules of quantitative RT-PCR. Plant Cell Online 20:1736CrossRef
  Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:research0034CrossRef
  Xu M, Zhang B, Su X, Zhang S, Huang M (2011) Reference gene selection for quantitative real-time polymerase chain reaction in Populus. Anal Biochem 408:337-39CrossRef
  Zhang F-Q, Wang Y-S, Lou Z-P, Dong J-D
• 作者单位：Ya-Lan Peng (1)
  You-Shao Wang (1) (2)
  Ji-Dong Gu (3)
  
  1. State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China
  2. Daya Bay Marine Biology Research Station, Chinese Academy of Sciences, Shenzhen, 518121, China
  3. Laboratory of Environmental Toxicology, Department of Ecology & Biodiversity, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, China
  
• 刊物类别：Earth and Environmental Science
• 刊物主题：Environment
  Environment
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  Ecology
  Environmental Management
  
• 出版者：Springer Netherlands
• ISSN：1573-3017

文摘

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