7 N to 6.9 N. This decline was expected and widely documented due to the action of cell wall degrading enzymes ( Civello et al., 1999, Ferreyra et al., 2007, Salentijn et al., 2003 and Trainotti et al., 2001). The highest transcript accumulation of Exp2 and Exp5 occurred in stages 1 and

2 ( Fig. 1B and C), while the fruit Topoisomerase inhibitor was immature and flesh firmness was high ( Fig. 1A). Expansins, non-enzymatic proteins, are known to act during early stages of fruit development in the process of cell wall polysaccharide solubilisation ( Civello et al., 1999 and Harrison et al., 2001). Therefore, the high transcript accumulation prior to fruit softening confirmed Exp2 and Exp5 as precursors in the softening process in strawberries ( Civello et al., 1999 and Harrison et al., 2001). Transcript accumulation of PLa and PLb was also high at early stages of fruit development (1 and 2) and followed a down-regulation when fruit were turning red ( Selleckchem GSK 3 inhibitor Fig. 1D and E). On the other hand, PLc transcripts accumulated at higher levels during strawberry maturation (stages 4 and 5) ( Fig. 1F). Probably, PLa and PLb are associated with cell division processes while PLc is involved in cell wall disassembly during fruit maturation. PME ( Fig, 1G) and PG ( Fig.

1H), known to be involved in fruit softening ( Castillejo et al., 2004 and Redondo-Nevado et al., 2001), had high transcript accumulation after stage 3, which means that their transcript level during stage 2, although lower than the following

stages, was enough to induce softening, or that the dramatic decline in firmness was not entirely dependent on these two genes. β-Gal transcript accumulation was relatively low, apart from stage 5 (fully ripe stage) ( Fig. 1I). Trainotti et al. (2001) characterised three β-Gal genes; Faßgal1, expressed Carnitine dehydrogenase during maturation, and Faßgal2 and Faßgal3 expressed in green fruit and other tissues. In the current work, β-Gal primers corresponded to Faßgal1, which encode an enzyme acting on galactose, generated from pectin hydrolysis. This way, higher transcript accumulation of β-Gal was expected in an advanced maturation stage, when higher concentration of pectin hydrolysis products is present. Generated from PME and PG action, pectin hydrolysis products serve as substrate for β-galactosidases demonstrating a coordinated process, in which, peaks of transcription occur in order: first Exp2, Exp5, PLa and PLb, then PLc, PME and PG, then β-Gal. Total anthocyanin content increased significantly (P ⩽ 0.05) with fruit development reaching 23.4 mg 100 g−1 during stage 5 ( Table 2). The onset of red colour was correlated with an increase in total anthocyanin content, as expected. The highest total phenolic content was observed during stage 1 (965.5 mg GAE100 g−1) and its levels dropped at stage 2 (628.2 mg 100 g−1), then increased over time reaching 752 mg GAE100 g−1 during stage 5.