Revisiting cell wall remodeling in Fragaria chiloensis fruit softening: evidence of strong modifications in HG, RG-I...

Fragaria chiloensis is a native, non-climacteric fruit that softens rapidly and intensively during ripening. Previous studies have indicated a reduction in cell wall (CW) material during fruit development, including a decrease in pectin content and changes in pectin solubility. Over time, several cell wall-modifying enzymes have been identified as participants in its softening process, such as FchXTH1, FchRGL, FchEXP2, FchPL and FchPG. By combining various biochemical techniques with the use of specific antibodies targeting different CW domains, an additional perspective on CW modifications was achieved. In the pectin-enriched fraction extracted from F. chiloensis fruit, homogalacturonans with both low and high degrees of methylation were detected, which decreased in abundance throughout development. Rhamnogalacturonan-I (RG-I) domains substituted with arabinose and galactose side chains were also observed, with arabinose chains appearing to degrade in parallel with fruit softening. The presence of dimeric Rhamnogalacturonan-II (RG-II) was detected, revealing the contribution of this pectic domain to maintaining cell wall stiffness during fruit ripening. Additionally, the presence of arabinogalactan proteins (AGPs) in the CW was confirmed. Regarding hemicellulosic polysaccharides, domains corresponding to xyloglucans and arabinoxylans were detected, with xylans present to a lesser extent. Changes in CW domains were tracked throughout the softening of F. chiloensis, confirming strong structural modifications in the CW that can be attributed to the activity of a diverse set of cell wall enzymes.