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Normalization of data, by choosing the appropriate reference genes (RGs), is fundamental for obtaining reliable results in reverse transcription-quantitative PCR (RT-qPCR). In this study, we assessed  Actinidia deliciosa  leaves inoculated with two doses of  Pseudomonas syringae  pv.  actinidiae  during a period of 13 days for the expression profile of nine candidate RGs. Their expression stability was calculated using four algorithms: geNorm, NormFinder, BestKeeper and the deltaCt method. Glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) and protein phosphatase 2A ( PP2A ) were the most stable genes, while β-tubulin and 7s-globulin were the less stable. Expression analysis of three target genes, chosen for RGs validation, encoding the reactive oxygen species scavenging enzymes ascorbate peroxidase (APX), superoxide dismutase (SOD) and catalase (CAT) indicated that a combination of stable RGs, such as  GAPDH  and  PP2A , can lead to an accurate quantification of the expression levels of such target genes. The  APX  level varied during the experiment time course and according to the inoculum doses, whereas both  SOD  and  CAT  resulted down-regulated during the first four days, and up-regulated afterwards, irrespective of inoculum dose. These results can be useful for better elucidating the molecular interaction in the  A. deliciosa / P. s . pv.  actinidiae  pathosystem and for RGs selection in bacteria-plant pathosystems.

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Reference gene selection for normalization of RT-qPCR gene expression data from Actinidia deliciosa leaves infected with Pseudomonas syringae pv. actinidiae