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Monilinia blight disease caused by  Monilinia vaccinii-corymbosi  (Reade) Honey ( M.vc ) causes severe damage and economic losses in wild blueberry growing regions. Molecular mechanisms regulating defence responses of wild blueberry phenotypes towards this causal fungus are not yet fully known. A reliable quantification of gene expression using quantitative real time PCR (qPCR) is fundamental for measuring changes in target gene expression. A crucial aspect of accurate normalisation is the choice of appropriate reference genes. This study evaluated the expression stability of seven candidate reference genes ( GAPDH ,  UBC9 ,  UBC28 ,  TIP41 ,  CaCSa ,  PPR  and  RH8 ) in floral tissues of diploid and tetraploid wild blueberry phenotypes challenged with  M.vc.  The expression stability was calculated using five algorithms: geNorm, NormFinder, BestKeeper, deltaCt and RefFinder. The results indicated that  UBC9  and  GAPDH  were the most stable reference genes, while  RH8  and  PPR  were the least stable ones. To further validate the suitability of the analyzed reference genes, the expression level of a pathogenesis related protein gene (i.e.,  PR3 ) was analysed for both phenotypes at four time points of infection. Our results may be beneficial for future studies involving the quantification of relative gene expression levels in wild blueberry species.

Selection and validation of reliable reference genes for gene expression studies from Monilinia vaccinii-corymbosi infected wild blueberry phenotypes