Brian Michael Ingel; Advisor: Caroline Roper.
Xylella fastidiosa is a xylem-limited bacterium that causes Pierce’s Disease (PD) in grapevine. PD symptom progression is correlated with systemic colonization by X. fastidiosa, which occurs via pit membrane degradation. Previous studies determined that a polygalacturonase, one of several cell wall degrading enzymes (CWDEs) secreted by X. fastidiosa, is necessary for pit membrane degradation. Here, it is demonstrated that the loss of two X. fastidiosa β-1,4-endoglucanases (EGases) in tandem causes significantly slower PD symptom development and progression, and reduces X. fastidiosa virulence and population size. Furthermore, differences in the PD symptom progression rate between two cultivars appear to be caused by variations in pit membrane carbohydrate composition, which influence the rate of enzymatic degradation, and subsequently, the rate of systemic colonization.
How these CWDEs and other virulence factors are secreted has not been fully elucidated. However, several are predicted to be secreted via the Type II secretion system (T2SS). T2SS-deficient X. fastidiosa mutants did not induce PD symptoms, and the mutant bacteria were not recovered from infected vines, indicating that the T2SS and its secreted substrates are necessary for virulence and survival. Analysis of wild type X. fastidiosa and T2SS-deficient mutant secretomes found six potential Type II secreted proteins for further characterization, including three lipases, a β-1,4-cellobiohydrolase, a protease, and a hypothetical protein. However, none of the CWDEs were present in the secretomes, but their high transcript abundance indicates that these enzymes could be post-transcriptionally regulated.
Several PD symptoms appear similar to drought stress symptoms. Under drought conditions, decreased photosynthesis, cessation of plant growth, and carbon starvation occur. Tylose formation in response to X. fastidiosa infection occludes vessels to prevent the spread of pathogens and air embolisms. Interestingly, genes involved in photosynthesis and plant growth are down-regulated during PD symptom development, and tylose formation increases significantly. This increased vessel occlusion in the absence of new vessel formation over time could cause prolonged water stress. Furthermore, starch in the ray parenchyma depletes as PD symptoms progress, indicating that carbon starvation occurs. These results indicate that, along with X. fastidiosa-mediated virulence factors, host responses also contribute to PD.