Watermelon (Citrullus lanatus) is an economically important horticultural crop known for its sweet red flesh and is a popular summer snack. The southeastern US is an important production region for watermelon, but the hot and humid weather is conducive for the development of fungal diseases. Gummy stem blight (GSB), caused by three species of Stagonosporopsis; S. citrulli, S. caricae and S. cucurbitacearum, is an important disease that can cause severe yield losses worldwide under these favorable conditions. Currently, no resistant cultivars are available for this disease and management depends on preventative fungicide spay programs. Host resistance has been previously identified in C. amarus, an inedible crop wild relative of watermelon. However, different Stagonosporopsis spp. isolates elicit different responses in different resistant host genotypes which complicated resistance breeding. To address these issues, we (i) developed a point-of-care assay that can differentiate S. citrulli from the other two species and (ii) introgressed resistance QTL from wild C. amarus into cultivated watermelon. A dipstick-based DNA extraction method was coupled with an S. citrulli specific loop-mediated isothermal amplification (LAMP) assay to detect as little as 1 pg of DNA with real time fluorescence quantification and endpoint colorimetric detection formats. To address host resistance, we developed high throughput KASP markers spanning the QTL regions, and used marker assisted backcrossing to introgress QTL into the cultivar Crimson Sweet. Eight BC2F3 intogression lines were evaluated in the field for resistance to GSB. Two of the lines showed high levels of resistance to GSB under field conditions. KASP assays were also developed for background selection for known domestication alleles to accelerate selection for fruit quality traits. The developed detection assays, KASP markers and introgression lines can contribute to accelerated breeding for host resistance and general breeding efforts for GSB in watermelon.
