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Abstract:
Central metabolism produces carbohydrates and amino acids that are tightly correlated to plant growth and thereby crop productivity. Malate is reported to link mitochondrial respiratory metabolism with cytosolic biosynthetic pathways. Although the function of malate metabolism-related enzymes in providing carbon has been characterized in some plants, evidence conferring this role in the fleshy fruit of cucumber is lacking. Here, radiolabeled bicarbonate fed into the xylem stream from the cucumber roots was incorporated into amino acids, soluble sugars, and organic acids in the exocarp and vasculature of fruits. The activities of decarboxylases, especially the decarboxylation from NADP-dependent malic enzyme (NADP-ME) were higher in cucumber fruit than in the leaf lamina. Histochemical localization revealed that CsNADP-ME2 was mainly located in the exocarp and vascular bundle system of fruit. Radiotracer and gas-exchange analysis indicated that overexpression (OE) of CsNADP-ME2 could promote the carbon flux into soluble sugars and starch in fruits. Further studies combined with metabolic profiling revealed that the down-regulation of CsNADP-ME2 in RNA interference (RNAi) lines caused the accumulation of its substrate malate in the exocarp. In addition to the inhibition of the glycolysis-related genes' expression and the reduction of the activities of the corresponding enzymes, increased amino acid synthesis and decreased sugar abundance were also observed in these lines. The opposite effect was found in CsNADP-ME2-OE lines, suggesting that there may be a continuous bottom-up feedback regulation of glycolysis in cucumber fruits. Overall, our studies indicate that CsNADP-ME2 may play potential roles both in central carbon reactions and amino acid metabolism in cucumber fruits.
