Enrichment of two important metabolites d-galacturonic acid and d-glucuronic acid inhibits MdHb1-mediated fruit softening in apple

Abstract

In apples, fruit firmness is a crucial quality trait influencing fruit storability, transportability, shelf life and consumer preference. However, the genetic network underlying this trait remains unclear. Therefore, the present study investigated the changes in apple fruit at different stages of postharvest storage using a combination of transcriptomic and metabolomic analyses. With prolonged storage, we detected a significant increase in two metabolites, d-galacturonic acid (d-GalUA) and d-glucuronic acid (d-GlcA), which are associated with a key class 1 non-symbiotic haemoglobin (MdHb1). We innovatively found that MdHb1 regulates fruit softening by catalysing the conversion from protopectin to water-soluble pectin. Biochemical analysis demonstrated that MdMYB2/MdNAC14/MdNTL9 transcription factors directly bind to the MdHb1 promoter to activate its transcriptional expression and promote fruit softening. Further injection experiments in apple fruit and histological as well as transmission electron microscopy analyses of the fruit samples revealed that d-GalUA and d-GlcA reduce the transcription of MdHb1, or through the MdMYB2/MdNAC14/MdNTL9-MdHb1 regulatory module, thereby delaying fruit softening. Our study provides novel insights into the role of two important metabolites, d-GalUA and d-GlcA, in the regulation of MdHb1-mediated fruit softening in apples.