In vivo and ex vivo study on cell wall components as part of the network in tomato fruit during the ripening process.

Ripening is a process involving various morphological, physiological, and biochemical changes in fruits. This process is affected by modifications in the cell wall structure, particularly in the composition of polysaccharides and proteins. The cell wall assembly is a network of polysaccharides and proteoglycans named the arabinoxylan pectin arabinogalactan protein1 (APAP1). The complex consists of the arabinogalactan protein (AGP) core with the pectin domain including arabinogalactan (AG) type II, homogalacturonan (HG), and rhamnogalacturonan I (RG-I). The present paper aims to determine the impact of a disturbance in the synthesis of one constituent on the integrity of the cell wall. Therefore, in the current work, we have tested the impact of modified expression of the SlP4H3 gene connected with proline hydroxylase (P4H) activity on AGP presence in the fruit matrix. Using an immunolabelling technique (CLSM), an immunogold method (TEM), molecular tools, and calcium mapping (SEM-EDS), we have demonstrated that disturbances in AGP synthesis affect the entire cell wall structure. Changes in the spatio-temporal AGP distribution may be related to the formation of a network between AGPs with other cell wall components. Moreover, the modified structure of the cell wall assembly induces morphological changes visible at the cellular level during the progression of the ripening process. These results support the hypothesis that AGPs and pectins are required for the proper progression of the physiological processes occurring in fruits.

Suppression of a Prolyl 4 Hydroxylase Results in Delayed Abscission of Overripe Tomato Fruits.

The tomato pedicel abscission zone (AZ) is considered a model system for flower and fruit abscission development, activation, and progression. O-glycosylated proteins such as the Arabidopsis IDA (INFLORESCENCE DEFICIENT IN ABSCISSION) peptide and Arabinogalactan proteins (AGPs) which undergo proline hydroxylation were demonstrated to participate in abscission regulation. Considering that the frequency of occurrence of proline hydroxylation might determine the structure as well the function of such proteins, the expression of a tomato prolyl 4 hydroxylase, SlP4H3 (Solanum lycopersicum Prolyl 4 Hydroxylase 3) was suppressed in order to investigate the physiological significance of this post-translational modification in tomato abscission. Silencing of SlP4H3 resulted in the delay of abscission progression in overripe tomato fruits 90 days after the breaker stage. The cause of this delay was attributed to the downregulation of the expression of cell wall hydrolases such as SlTAPGs (tomato abscission polygalacturonases) and cellulases as well as expansins. In addition, minor changes were observed in the mRNA levels of two SlAGPs and one extensin. Moreover, structural changes were observed in the silenced SlP4H3AZs. The fracture plane of the AZ was curved and not along a line as in wild type and there was a lack of lignin deposition in the AZs of overripe fruits 30 days after breaker. These results suggest that proline hydroxylation might play a role in the regulation of tomato pedicel abscission.