Exploring the genetic mechanisms underlying superficial scald development and prevention in apple fruit during postharvest storage.

Apple is one of the most valuable fruit consumed worldwide due to its excellent quality features and high degree of acceptation by consumers. However, apples are susceptible to various postharvest disorders, leading to significant economic losses. To slow down food decay and provide constant fresh food to the market, postharvest strategies including decreasing the temperature can slows down the metabolism of fruits, but in parallel can also trigger the occurrence of typical chilling injury related disorders. Among all the physiological disorders, superficial scald is one of the most challenging, causing discoloration and skin browning triggering therefore consumers unacceptance. The development of superficial scald in apple fruit is a complex process influenced by various factors, including genetic background, storage conditions and preharvest management practices. Recent research partially disclosed the molecular mechanisms underlying the development of superficial scald in apple fruit. It has been shown that this disorder is associated with the accumulation of reactive oxygen species (ROS) and the oxidation of alfa-farnesene, a sesquiterpene largely considered as a major actor in the onset of this disorder. Various postharvest treatments have been used to control the development of superficial scald in apple fruit, such as including the application of the ethylene competitor 1-methylcyclopropene (1-MCP) or controlled atmosphere storage. This thesis aimed to identify and characterize the genetic factors involved in symptoms development and to disclose the regulation of this phenomenon. To this end, a comparative multidisciplinary study integrating transcriptomics and metabolomics analysis have been performed in a wide range of storage conditions and across different genetic background. The mechanism of action and effectiveness of two preventing strategies, such as the exogenous application of 1-MCP and storage at low oxygen concentration were assessed in two apple cultivars, ‘Granny Smith’ and ‘Ladina’. The metabolite and transcriptomic profiles revealed that 1-MCP treatment actively stimulated the production of unsaturated fatty acids such as oleic acid (C18:1), linoleic acid (C18:2) and linolenic acid (C18:3), thereby successfully preventing the development of superficial scald in ‘Granny Smith’. ‘Ladina’, on the other hand, was less responsive to these strategies and was characterized by a higher accumulation of very long chain fatty acids (VLCFAs), together with an increased level of fatty acid saturation. Additionally, the integration of the transcriptomic survey and VOCs assessment underlined that storage at low oxygen concentration stimulated the expression of genes involved in anaerobic respiration, such as malate, alcohol dehydrogenase, and pyruvate decarboxylase, leading to the accumulation of ethanol and acetaldehyde in both cultivars. The expression of three members of the VII subgroups of ERF genes was also observed in ‘Granny Smith’, which coordinate the acclimation process to hypoxia in plants. The effect of controlled atmosphere in the prevention of superficial scald was furthermore investigated in fruit of ‘Granny Smith’ stored for different time and employing both a static and dynamic controlled atmosphere approach. The transcriptome analysis found that the duration of storage had an important effect on the coordination of gene expression, and the expression patterns of DEGs identified a distinct networks and types of transcriptomic hubs. Samples that developed superficial scald were characterized by higher concentration of chlorogenic acid and a time-specific increase in the expression of PAL and PPO genes. Prevention of superficial scald was related to a peculiar realigning episodes that involved the accumulation of specific antioxidant metabolites, very long chain fatty acids, and the expression of genes that coordinate the hypoxia acclimation process, such as RAP2-like and PCO. A complementary research was conducted to investigate the connection between the application of 1-MCP, low oxygen, and superficial scald. The study highlighted the possible application of genes involved in key fruit ripening pathways, including ethylene-related pathways and sugar/fermentation metabolism, as potential biomarkers for molecular characterization of the major changes that occur during fruit storage. The specific expression of polyphenol oxidase, along with sorbitol-6-phosphate dehydrogenase, were also used to investigate the onset of superficial scald in apples and correlated with the accumulation of the sugar alcohol sorbitol, which plays a protective role against chilling injuries. Assessing the transcriptional signature of these biomarkers can contribute to the development of new tools for a more informed understanding of the physiological progression of postharvest ripening in apples. Overall, these studies provide insights into the genetic and metabolic mechanisms underlying the development and prevention of superficial scald in apple cultivars during storage. The results obtained here paved the way for the identification of a core set of genes to be used as functional marker suitable to better comprehend this phenomenon and to select new accession characterized by a superior fruit quality and postharvest performance.