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Identification of suitable qPCR reference genes in leaves of Brassica oleracea under abiotic stresses

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Abstract

Real-time quantitative PCR is nowadays a standard method to study gene expression variations in various samples and experimental conditions. However, to interpret results accurately, data normalization with appropriate reference genes appears to be crucial. The present study describes the identification and the validation of suitable reference genes in Brassica oleracea leaves. Expression stability of eight candidates was tested following drought and cold abiotic stresses by using three different softwares (BestKeeper, NormFinder and geNorm). Four genes (BolC.TUB6, BolC.SAND1, BolC.UBQ2 and BolC.TBP1) emerged as the most stable across the tested conditions. Further gene expression analysis of a drought- and a cold-responsive gene (BolC.DREB2A and BolC.ELIP, respectively), confirmed the stability and the reliability of the identified reference genes when used for normalization in the leaves of B. oleracea. These four genes were finally tested upon a benzene exposure and all appeared to be useful reference genes along this toxicological condition. These results provide a good starting point for future studies involving gene expression measurement on leaves of B. oleracea exposed to environmental modifications.

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Acknowledgments

This study was supported by ADEME (Agence De l’Environnement et de la Maîtrise de l’Energie) and the Région Nord Pas-de-Calais through a European Regional Development Fund (No. 38505).

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The authors declare that they have no conflict of interest.

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  1. Laboratoire des Sciences Végétales et Fongiques, Faculté des Sciences Pharmaceutiques et Biologiques, Université Lille Nord de France, EA 4483, Lille 2, B.P. 83, 59006, Lille Cedex, France

    Franck Brulle, Fabien Bernard, Damien Cuny & Sylvain Dumez

  2. LGCgE-Lille 1, Ecologie Numérique et Ecotoxicologie, Université Lille Nord de France, 59650, Villeneuve d’Ascq, France

    Fabien Bernard & Franck Vandenbulcke

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  1. Franck Brulle
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Brulle, F., Bernard, F., Vandenbulcke, F. et al. Identification of suitable qPCR reference genes in leaves of Brassica oleracea under abiotic stresses. Ecotoxicology 23, 459–471 (2014). https://doi.org/10.1007/s10646-014-1209-7

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