The Influence of Copper Homeostasis Genes copA and copB on Xylella fastidiosa Virulence Is Affected by Sap Copper Concentration.

Xylella fastidiosa is a xylem-limited plant pathogenic bacterium that causes diseases worldwide in crops such as grape, citrus, and olive. Although copper (Cu)-containing compounds are not used for management of X. fastidiosa-caused diseases, they are widely used in X. fastidiosa hosts in vineyards and orchards. The accumulation of Cu in soils and, therefore, plant saps, could be a challenge for X. fastidiosa survival. Here, the molecular basis of Cu homeostasis was studied in relation to virulence. Although homologous Cu-related genes copA (X. fastidiosa loci PD0100) and copB (PD0101) have been characterized in other bacteria, their functions differ among bacterial species. In vitro, both copA and copB mutants were more sensitive to Cu than the wild-type (WT) strain. Interestingly, the copA mutant was more sensitive to Cu shock, while the copB mutant was more sensitive to chronic Cu treatments. In tobacco greenhouse experiments with normal watering, both mutants reduced virulence compared with WT. But when Cu was added as a drench treatment, both copA and copB mutants had increased disease severity approximately 20 and 50% compared with mutants without Cu added, respectively, which were significantly higher than the approximately 5% observed for WT under the same conditions. These results indicate that the pathogen's Cu homeostasis affects virulence and is influenced by Cu concentration in the environment. Understanding Cu homeostasis in X. fastidiosa will help discern the outcome of Cu treatments and the adaptation of this pathogen to the xylem of plants that have been exposed to high Cu concentrations because of agricultural practices.