A major QTL region associated with powdery mildew resistance in leaves and fruits of the reconstructed garden strawberry.

Multiple QTLs for powdery mildew resistance were identified in a pre-breeding population derived from the octoploid progenitor species of garden strawberry, including a stable major novel factor on chromosome 3B. Powdery mildew (PM), caused by the biotrophic fungal pathogen Podosphaera aphanis, poses an increasing threat to garden strawberry (Fragaria × ananassa) production worldwide. While a few commercial cultivars exhibit partial resistance, fungicide application remains essential for managing PM outbreaks. However, breeding offers a more sustainable approach for controlling PM. A better understanding of the genetics of resistance is required for informed breeding strategies, e.g. through identifying novel resistance factors derived from the progenitor species of garden strawberry, F. chiloensis and F. virginiana. We conducted genome-wide association (GWA) and multivariate analyses in a reconstructed (ReC) strawberry population to investigate PM resistance under natural infection. Leveraging multi-year field trial data and 20,779 single-nucleotide polymorphism markers, we identified a novel major quantitative trait locus (QTL) on chromosome 3B, designated as q.LPM.Rec-3B.2, that was consistently associated with high PM resistance in both leaves and fruits. Greenhouse validation with a subset of the ReC population confirmed that this QTL region was stable across field and greenhouse environments. Promising candidate genes for resistance, including two for MLO and one for EXO70, were identified within this major QTL. In addition, multi-locus GWA models and non-additive GWA revealed additional resistance QTLs on multiple chromosomes. Despite previous challenges in breeding for robust PM resistance due to its quantitative nature and complex genetic control, our results provide valuable insights into resistance-contributing QTL regions already existing in strawberry, novel wild-derived resistance QTLs not previously known, candidate genes, and pre-breeding germplasm carrying resistance traits as resources for future genome-informed breeding efforts.