Sorbitol Mediates NAC17-bZIP1 to Synergistically Regulate Disease Resistance Mechanism in Malus micromalus via Flavonoids

To investigate the effect of sorbitol feeding on immune responses and its underlying mechanism, this study employed crabapple (Malus micromalus), a species native to China, as the experimental material. Detached leaves were subjected to sorbitol feeding experiments, with water feeding as the control. The dynamic changes in lesion area and cell membrane permeability were measured across four groups of leaves following feeding and fungal inoculation, and transcriptome sequencing was performed on these four groups. The results revealed that compared to the control group, leaves fed with sorbitol prior to inoculation exhibited significantly reduced lesion areas and decreased cell membrane permeability. Transcriptome and KEGG pathway analyses indicated that differentially expressed genes were significantly enriched in the phenylpropanoid biosynthesis pathway. HPLC results demonstrated that quercetin and isorhamnetin contents increased significantly after sorbitol feeding, and both compounds were shown to inhibit the mycelial growth of Alternaria alternata, thereby enhancing the disease resistance of crabapple leaves. Among the associated genes, MmF3H1, which regulates the synthesis of quercetin and isorhamnetin, showed the highest level of enrichment. Co-expression trend analysis and qPCR results revealed that MmNAC17 and MmbZIP1 exhibited strong co-expression relationships with MmF3H1 and responded to sorbitol regulation. Functional validation was conducted by constructing overexpression and silencing vectors for MmNAC17/MmbZIP1 and performing transient transformation, thereby elucidating the molecular and biochemical mechanisms underlying the response of M. micromalus to A. alternata. This study provides a theoretical reference for breeding crabapple varieties with enhanced resistance to leaf spot disease.