Ralstonia solanacearum is a bacterial vascular pathogen causing devastating bacterial wilt. In the field, resistance against this pathogen is quantitative and is available for breeders only in tomato and eggplant. To understand the basis of resistance to R. solanacearum in tomato, we investigated the spatio-temporal dynamics of bacterial colonization using non-invasive live monitoring techniques coupled to grafting of susceptible and resistant varieties. We found four ‘bottlenecks’ that limit the bacterium in resistant tomato: root colonization, vertical movement from roots to shoots, circular vascular bundle invasion, and radial apoplastic spread in the cortex. Radial invasion of cortical extracellular spaces occurred mostly at late disease stages but was observed throughout plant infection. This study shows that resistance is expressed in both root and shoot tissues, and highlights the importance of structural constraints to bacterial spread as a resistance mechanism. It also shows that R. solanacearum is not only a vascular pathogen but spreads out of the xylem, occupying the plant apoplast niche. Our work will help elucidate the complex genetic determinants of resistance, setting the foundations to decipher the molecular mechanisms that limit pathogen colonization, which may provide new precision tools to fight bacterial wilt in the field.
Bacterial, fungal, and viral diseases of tomato (Solanum lycopersicum) are responsible for widespread yield losses, especially in humid growing environments. Chromosome 11 of tomato contains genes that modulate resistance to several prominent tomato pathogens, including bacterial spot caused by Xanthomonas spp., gray leaf spot caused by Stemphylium spp., Fusarium wilt caused by race 2 of Fusarium oxysporum f. sp. lycopersici, and tomato yellow leaf curl virus (TYLCV) caused by begomoviruses. Major resistance loci are quantitative trait locus 11 (QTL-11) and Xv3/Rx4 for bacterial spot, Sm for gray leaf spot, I2 for Fusarium wilt, and Ty-2 for TYLCV. Marker-assisted selection was used to select for rare recombination events that combined these resistance loci into a linked cassette that can be inherited together in future crosses. A pedigree breeding strategy was used with marker-assisted selection and used to identify a novel coupling of Xv3/Rx4 and Ty-2. Recombination between the two genes was estimated as 0.056 cM, demonstrating that effective combinations of resistance can be established using publicly available germplasm. Progeny from the recombinant plants were screened using inoculated seedling trials to confirm resistance. The recombinants identified maintained resistance levels similar to the resistant controls. Trial results suggest that the trait markers on chromosome 11 are tightly linked to the respective resistance loci and are effective for selecting plants with resistance to the target diseases.
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