To study the changes in sugar metabolism caused by fungal infection in post-harvest peaches, fruit from two cultivars (‘Baifeng’ and ‘Yulu’) was inoculated with Monilinia fructicola and stored at 10 °C. During disease development, soluble sugar content was monitored, as well as the activities and expression of selected enzymes. Disease progression was accompanied by a decrease in sucrose content and increases in reducing sugars and soluble solids, consistent with higher enzyme activities for acid invertase, neutral invertase and sucrose synthase-cleavage, and lower activities for sucrose synthase-synthesis and sucrose phosphate synthase. Activities of phosphofructokinase, hexokinase, and pyruvate kinase, which are related to hexose metabolism, also increased. These changes stimulate the Embden–Meyerhof–Parnas (EMP) pathway. We conclude that the fungal disease in peach fruit accelerates the decomposition of sucrose, thereby providing more glucose as a substrate to the EMP pathway.

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