RNA extraction from whole fruits comprising the exocarp and the mesocarp (excluding the stones), purity/integrity measurement, cDNA synthesis and RT-qPCR were performed as previously described42. A melt curve analysis was performed at the end of the PCR cycles to check the specificity of the primers. The primers’ amplification efficiencies were determined using a calibration curve consisting of a serial dilution of 6 points (10–2–0.4–0.08–0.016–0.0032 ng/μL).

The expression values were calculated with qBasePLUS (version 3.2, Biogazelle, Ghent, Belgium)102 by using PavACT7 and PaveTIF4E as reference genes, which were sufficient for normalization according to geNORM103.

The log10-transformed NRQ (Normalized Relative Quantities) results were analyzed with IBM SPSS Statistics v20 (IBM SPSS, Chicago, IL, USA). Normal distribution of the data was checked with a Shapiro-Wilk test and graphically with a Q-Q plot. Homogeneity was checked with the Homogeneity of Variance Test. For data following normal distribution and homogeneous, a one-way ANOVA with Tukey’s post-hoc text was performed. For data not following normal distribution and/or not homogeneous, a Kruskal-Wallis test was performed with Dunn’s post-hoc test.

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