Sclerotinia stem rot caused by Sclerotinia sclerotiorum is one of the most destructive diseases in Brassica rapa. Verticillium dahliae Aspf2-like protein (VDAL) is a secretory protein of V. dahliae which has been shown to enhance the resistance against fungal infections in several plants. Nonetheless, the molecular mechanisms of VDAL-primed disease resistance are still poorly understood. In this study, we performed physiological, biochemical, and transcriptomic analyses of Brassica rapa in order to understand how VDAL confers resistance to S. sclerotiorumn infections in plants. The results showed that foliar application of VDAL significantly reduced the plaque area on leaves inoculated with S. sclerotiorum. It also enhanced antioxidant capacity by increasing activities of superoxide dismutase (SOD), peroxidase (POD), peroxidase (APX), glutathione reductase (GR), protoporphyrinogen oxidase (PPO), and defense-related enzymes β-1,3-glucanase and chitinase during the infection periods. This occurred in parallel with significantly reduced relative conductivity at different periods and lower malondialdehyde (MDA) content as compared to sole S. sclerotiorum inoculation. Transcriptomic analysis showed a total of 146 (81 up-regulated and 65 down-regulated) differentially expressed genes (DEGs) in VDAL-treated leaves compared to the control. The most enriched three Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were the mitogen–activated protein kinase (MAPK) signaling pathway, plant hormone signal transduction, and plant-pathogen interaction, all of which were associated with plant immunity. DEGs associated with MAPK and hormone signal transduction pathways were ethylene response sensor ERS2, EIN3 (Ethylene Insensitive3)-binding F-box protein 2 (EBF2), ethylene-responsive transcription factor ERF94, MAPK 9 (MKK9), protein phosphatase 2C (PP2C37), auxin-responsive proteins (AUX/IAA1 and 19), serine/threonine-protein kinase CTR1, and abscisic acid receptors (PLY 4 and 5). Among the DEGs linked with the plant–pathogen interaction pathway were calmodulin-like proteins (CML5, 24, 27), PTI1-like tyrosine protein kinase 3 (Pti13) and transcription factor MYB30, all of which are known to play key roles in pathogen-associated molecular pattern (PAMP)-triggered immunity and effector-triggered immunity (ETI) for hypersensitive response (HR), cell wall reinforcement, and stomatal closure in plants. Overall, VDLA treatment triggered repression of the auxin and ABA signaling pathways and de-repression of the ethylene signaling pathways in young B. rapa seedlings to increase plant innate immunity. Our results showed that VDAL holds great potential to enhance fungal disease resistance in B. rapa crop.

Lobed leaves play a critical role in enhancing the productivity of sprawling crops like zucchini by improving light capture and boosting photosynthesis. However, the genetic basis in zucchini remains largely unknown. Here, we developed an F₂ population from a cross between the entire-leaf cultivar ‘LR’ and the deeply lobed cultivar ‘Xi’. Genetic analysis showed that the non-lobed trait is dominant, with the F₂ segregation ratios (~9 entire:6 shallowly lobed:1 deeply lobed) indicating digenic inheritance. Using bulked segregant analysis sequencing (BSA-seq) and kompetitive allele-specific PCR (KASP) marker analysis, we identified a major effect locus at a 79.8 kb interval on chromosome 10. Within in this interval, gene expression profiling and annotation indicated CpARF6, encoding an auxin response factor, to be the prime candidate gene. Sequencing analysis revealed five nonsynonymous mutations in this gene, including a critical serine-to-leucine substitution at position 335 within the auxin response domain, which is likely a loss function mutation. Our findings establish CpARF6 as a critical regulator of lobed leaf formation in zucchini, providing valuable insights for both leaf development studies and zucchini breeding.