Essential oils nano-emulsion confers resistance against Penicillium digitatum in 'Newhall' navel orange by promoting phenylpropanoid metabolism

Inducing natural resistance against pathogen infection in postharvest citrus by exogenous elicitor is a promising alternative to reduce postharvest losses. Here, the potential induced resistance mechanism of blended cinnamaldehyde, carvacrol and eugenol nano-emulsion against P. digitatum in `Newhall' navel orange (Citrus sinensis (L.) Osbeck) was evaluated through combined transcriptomic and metabolomics analysis. Application of nanoemulsion maintained fruit firmness after P. digitatum inoculation, nano-emulsion reduced H2O2 and MDA accumulation of citrus fruit, increased PAL, C4H, 4CL and CAD activity, and higher contents of flavonoids, lignin and total phenolic compounds. Furthermore, higher activities of SOD, CAT, POD and APX in nano-emulsion treated fruit possibly benefited reducing ROS accumulation and lipid peroxidation. The activities of the defense enzymes CHI, GLU were higher in the nano-emulsion treated fruits. RNA-seq identified 653 differentially expressed genes (DEGs) between the control and the nano-emulsion-treated fruit at 48 hpi, including 444 upregulated and 209 downregulated genes. Genes encoding shikimate O-hydroxycinnamoyltransferase (HCT), caffeoyl-CoA O-methyltransferase (CCoAOMT), caffeic acid 3-O-methyltransferase / acetylserotonin O-methyltransferase (COMT), cinnamyl-alcohol dehydrogenase (CAD) and peroxidase (POD) were differentially expressed by nano-emulsion treatment. In addition, 175 differential accumulated metabolites between the control and the nano-emulsion-treated fruit at 48 hpi also were identified, the different accumulated metabolites from phenylpropanoid biosynthesis pathway related with disease resistance including L-phenylalanine, caffeic acid, caffeoyl quinic acid, coniferyl alcohol, coniferaldehyde. The integrated transcriptomic and metabolomic profiling indicated that the differentially expressed genes (DEGs) and the differentially accumulated metabolites (DAMs) were mainly involved in phenylpropanoid biosynthesis pathway. Besides, the nano-emulsion significantly induced accumulation of primary metabolites including amino acids, soluble sugars, organic acids, lipids, sugar and alcohols. All these results indicated that blended essential oil nano-emulsion as an antifungal delivery system to induce resistance against P. digitatum infection of `Newhall' navel orange by promoting phenylpropanoid metabolism.